NORTH AMERICAN NATIVE ORCHID JOURNAL

Volume 16(1) 2010

IN THIS ISSUE:
GROWING CYPRIPEDIUMS IN CONTAINERS TRANSPLANT METHODS FOR THE ENDANGERED ORCHID SPIRANTHES PARKSII PROPAGATION AND CONSERVATION STATUS OF THE NATIVE ORCHIDS OF THE U.S. ……

and more………….

The North American Native Orchid Journal (ISSN 1084-7332) is a publication devoted to promoting interest and knowledge of the native orchids of North America. A limited number of the print version of each issue of the Journal are available upon request and electronic versions are available to all interested persons or institutions free of charge. The Journal welcomes articles of any nature that deal with native or introduced orchids that are found growing wild in North America, primarily north of Mexico, although articles of general interest concerning Mexican species will always be welcome.

NORTH AMERICAN NATIVE ORCHID JOURNAL
Volume 16 (1) 2010

CONTENTS NOTES FROM THE EDITOR
1 3

LOOKING FORWARD GROWING CYPRIPEDIUMS IN CONTAINERS
Doug Martin, Ph.D. 4

PRELIMINARY RESULTS FOR FIELD ESTABLISHMENT TECHNIQUES OF CALOPOGON TUBEROSUS
Philip J. Kauth, Michael E. Kane & Timothy R. Johnson 12

A PRACTICAL AND INTEGRATED APPROACH TO NATIVE ORCHID CONSERVATION AND PROPAGATION AT THE ATLANTA BOTANICAL GARDEN
Matt Richards & Jenny Cruse Sanders, Ph.D. 25

AN UNDERGRADUATE’S FIRST ADVENTURE INTO FIELD RESEARCH:
AN EPIPHYTIC ORCHID SURVEY IN SOUTHERN FLORIDA

Emily Massey 31

TRANSPLANT METHODS FOR THE ENDANGERED ORCHID SPIRANTHES PARKSII CORRELL
J. Ryan Hammons, Fred E. Smeins & William E. Rogers 38

PROPAGATION AND CONSERVATION STATUS OF THE NATIVE ORCHIDS OF THE UNITED STATES (INCLUDING SELECTED POSSESIONS), CANADA, ST. PIERRE ET MIQUELON, AND GREENLAND
Scott Stewart, Ph.D. & Aaron Hicks 47

SHOULD WE OR SHOULDN'T WE? ETHICS AND ORCHIDS The Slow Empiricist
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RECENT ORCHID LITERATURE OF INTEREST
69 72 ASYMBIOTIC TECHNIQUE OF ORCHID SEED GERMINATION Second revised edition MICROPROPAGATION OF ORCHIDS, Volumes 1 & 2 (2nd Edition)

BOOK REVIEWS

Unless otherwise credited, all graphics were prepared by the authors of the respective papers. The opinions expressed in the Journal are those of the authors. Scientific articles may be subject to peer review and popular articles will be examined for both accuracy and scientific content. Volume 16(1): 1-81 issued January 15, 2010. Copyright 2010 by the North American Native Orchid Journal Cover: Calopogon tuberosus var. tuberosus by Stan Folsom

NOTES FROM THE EDITORS
The first issue of the North American Native Orchid Journal for 2010 is a special issue that is focused on native orchid propagation, cultivation, and reintroduction. The idea for this special issue was borne from conversations among Lawrence Zettler, Aaron Hicks, and the Associate Editor about the need to offer Journal readers an in-depth review of current trends in native orchid propagation and cultivation work. Recognizing the integration of orchid reintroduction with propagation and cultivation work, the Associate Editor has included all three topics in this issue. This issue presents articles by academic and popular authors about native orchid propagation, cultivation, and reintroduction. Doug Martin begins the special issue with detailed advice about cultivating Cypripedium species in containers, followed by Philip Kauth et al. presenting preliminary research results from a Calopogon tuberosus reintroduction study. The special issue continues with Matt Richards and Jenny Cruise Sanders of the Atlanta Botanical Garden discussing practical and integrated approaches to native orchid propagation at the Garden. Next, Emily Massey presents her personal story of orchid field research and reintroduction in southwestern Florida, followed by Ryan Hammons et al.‖s presentation of translocation work with the Federally endangered Spiranthes parksii. The issue concludes with a comprehensive update by Scott Stewart and Aaron Hicks on the propagation and conservation status of orchid species native to the United States, Canada, Greenland, Puerto Rico, and selected possessions. This collection of authors and articles represents a diverse cross section of the cutting edge of orchid propagation, cultivation, and reintroduction. The editors wish to thank all the authors and reviewers who made this special issue possible.

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The electronic format continues to be well received and we now reach more than 1800 readers. Back issues from volume 3 (1997) to present are now available online and you may read the current and back issues at: http://wiki.terrorchid.org/tow:journals The current update of the North American Personal Checklist is also available at that website. The checklist will be updated as needed with new taxa noted.
Paul Martin Brown, Editor naorchid@aol.com 10896 SW 90th Terrace, Ocala, FL 34481 36 Avenue F, Acton, Maine 04001 (June- early October) Scott L. Stewart, Ph.D. Associate Editor slstewar@gmail.com Kankakee Community College Horticulture & Agriculture Programs 100 College Drive Kankakee, Illinois 60901

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LOOKING FORWARD
Future issues scheduled for 2010 of the North American Native Orchid Journal will feature such topics as new taxa in Mesadenus and Corallorhiza, a very special paper on evolutionary classification by Richard Bateman from Kew a new Series HERE AND THERE species found in North America and elsewhere Cypripedium cultivation and hybrids and much more!

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Martin: GROWING CYPRIPEDIUMS IN CONTAINERS

GROWING CYPRIPEDIUMS IN CONTAINERS
Doug Martin, Ph.D. Cypripediums are generally considered hard to grow. However, like all plants, they are adapted to grow under a particular set of conditions in their natural environment. If the grower can provide those conditions, the plants will practically grow themselves. Over the years I‖ve developed methods, most adapted from other growers, which allow me to provide the conditions needed by many cypripediums. While I still have a lot to learn, particularly about growing seedlings and the more demanding species, my system seems to work well for the easier growing species and for hybrids. Not counting seedlings, I now have about two dozen plants of six species and seven hybrids. I‖ve only lost two mature plants in the last four years, both Cypripedium candidum, one of the more demanding species. In this article I‖ll describe cypripediums‖ basic cultural requirements and how I meet them. GROWING CONDITIONS While cypripediums can be grown in garden beds, I prefer to grow mine in containers. This gives me the ability to experiment with growing conditions. I can move them to different locations in the yard with different light levels, and I can easily change the growing medium. Containers: For most orchid growers, growing in containers means using regular flower pots. However, in the wild cypripedium roots spread out in a circle from the crown of the plant, as much as two to three feet in all directions. They also stay shallow, growing in only the top one to two inches of moist, well aerated soil (Stoutamire, 1991). To accommodate this growth habit as much as possible, I use plastic storage containers that are about 16 in. × 10 in. and 7 in. deep (41 × 25 × 18 cm), with one blooming-sized plant per container ( Fig. 1). I‖d like to use larger containers, but they quickly become too heavy and hard to move.
Fig. 1. One of my standard growing containers for cypripediums with a single plant of C. parviflorum var. makasin that I have grown from seed. 4

Martin: GROWING CYPRIPEDIUMS IN CONTAINERS

Although cypripediums can be grown and flowered in regular flower pots, the ones I‖ve seen don‖t seem to grow as large, or have flowers as large, as plants in the wild or in garden beds. I suspect that the roots simply don‖t have room to spread. It‖s sort of a cypripedium bonsai. I think that giving the roots extra room should result in a larger plant. My largest plant of C. reginae seems to support this theory, with three inch flowers on top of two foot tall stalks (Fig. 2). Cypripediums need consistent moisture at their roots. To help accommodate this requirement, my containers mimic the artificial bogs used to grow other North American native orchids. I leave the bottoms of the containers solid and drill holes in the sides about four to five inches (10 to 16 cm) below the surface of the medium. The area below the holes acts as a reservoir and helps to maintain the consistent moisture level that the roots need. It is not necessary to grow them this way, but it gives me a margin of error in watering. Medium: Because cypripediums need air as well as consistent moisture at the roots, I use water retentive growing media that are open, airy, and free draining. For most cypripediums the medium should be neutral to slightly acidic. Most of the mixes I use are 75-80% inert material: Fig. 2. C. reginae flowering in one of my standard gravel, sand, perlite, Turface, or pumice. The containers. The flowers are three inches across and the sand and gravel should be quartz, because too stems are two feet tall. much limestone in the mix will make it too alkaline. The remainder of my growing media is organic material. Some cypripedium growers use peat or chopped tree leaves, but I prefer coir–finely chopped coconut husk fibers. It is moisture retentive like peat, but isn‖t acidic. I add a handful of ground oyster shells to each container to buffer the pH of the medium. Two mixes that I‖ve used with good results are: Medium 1: three parts perlite; one part gravel; one part coir. Medium 2: four parts perlite; one part coir. Water: One of the most important considerations when growing cypripediums is that they require consistent moisture at their roots. The medium should never be allowed to dry out. Cypripediums are sensitive to water quality, so they should only receive water low in dissolved solids such as rain, distilled or RO water.

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Fertilizer: Cypripediums are adapted to grow in nutrient-poor soils. They are light feeders and do not require much fertilizer. In a mostly inorganic medium like I use, they do better with frequent applications of dilute fertilizer. Any good quality fertilizer will do. I use the Michigan State fertilizer at about 25 parts per million of nitrogen. I water with it every other week and use it as a foliar spray on alternate weeks. I fertilize my cypripediums from the time the leaves unfold in the spring until early September. Light: Most cypripediums grow in light shade in open forests or under shrubs. They like morning sun, but must be protected from direct sun during mid-day. I grow mine on the south sides of deciduous trees where they are in shade after about nine or ten o‖clock in the morning. They get about 1,800 foot-candles in mid-day. Some like a bit more light, and I move these away from the tree trunk to where the shade is thinner. Cypripediums can also be grown indoors under lights. As a light source, I prefer the new T-5 fluorescent bulbs. Regular fluorescent bulbs only produce enough light if they are positioned very close to the plants. They are only adequate for seedlings and small plants. High pressure sodium and metal halide lights produce plenty of light but also a lot of heat; the T-5s fall in the middle, with plenty of light but not too much heat. Temperature: While some cypripediums are native to northern areas and require cool temperatures year round, I find that several species and most of the hybrids grow well in my yard in the Kansas City area. Our summer temperatures are often in the 90°s F (mid 30°s C) and even over 100F (38° C). In winter, cypripediums require a cold rest period at near freezing temperatures. The Kansas City area has these conditions from November through March. I grow all of my seedlings, as well as the cooler growing Asian species, Cypripedium macranthos, in a basement lightroom where the temperature stays between 72° F (22° C) to 81° F (27°C). I grow tropical orchids in the lightroom during the winter. When the weather gets warm in May, the tropicals go outside and the cypripediums go inside. The C. macranthos plants were decreasing in size every year when I tried to grow them outside with my other cypripediums. Since I started keeping them in the basement, they are making a comeback. Humidity: Like all orchids, cypripediums prefer humidity above 50%. However, there does seem to be some flexibility. My plants often experience humidity levels of about 25% or less outside during the summer, without any noticeable negative effects. Potting: Cypripediums should be potted with the roots spread out and the growth buds at or just above the surface of the medium (Fig. 3). To accomplish this, I fill the container with medium to about three inches below the top. Then I mound the medium in the center so that it slopes gently down toward the edges. I place the plant on the mound, spread the roots out evenly and add medium until the growth buds are just covered and the surface is level throughout. Then I water the plant and add more medium where it washes down.

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Martin: GROWING CYPRIPEDIUMS IN CONTAINERS

Pests: Deer and rabbits like to eat cypripediums, and squirrels like to dig in the medium. Since we have a lot of all three, I keep my cypripediums in cages made from 1 × 2 in. (2.5 × 5.0 cm) boards covered with chicken wire on the sides and top. Four of the cypripedium containers fit nicely in a 48 × 18 in. (120 × 45 cm) cage. Most of my cages are two feet (60 cm) tall, but the taller C. reginae need one three feet (90 cm) tall.

Fig. 3. Growth buds of C. parviflorum var. makasin showing the proper depth for potting.

Slugs and snails can be a problem, especially if the container is in contact with the ground overnight. These pests can devour an entire seedling or eat through the stem of a blooming sized plant overnight. Because my plants are kept on raised platforms during the growing season, I only worry about slugs and snails in the spring when I uncover containers that have been buried for the winter. I treat with slug pellets containing metaldehyde as part of the move to the growing area. Caterpillars also can reduce a small plant to a leafless stalk seemingly overnight. I check my plants regularly for signs of caterpillar damage. I treat with a dust designed for caterpillars on roses and flowering plants. GROWTH CYCLE Cypripediums are hardy perennial plants with four distinct seasons in their growth cycle. They are dormant during the winter and require a cold rest period. In the spring they break dormancy and grow rapidly, reaching full size and flowering in three to eight weeks, depending on the species. In the late spring and the summer they grow roots, store food and produce the growth buds for the following year. In the fall, the stems and leaves wither and die back to the surface of the medium in preparation for winter dormancy. I treat my plants differently during each of these seasons and will discuss each in turn. Winter Care: Cypripediums are dormant in winter so their needs are simple. They need to be kept cold at temperatures near, but not below, freezing for three to four months. They must not dry out, and they must be protected from temperature extremes; either very low temperatures that could freeze the plant solid, or mid-winter warm spells. If the plants warm up, they may break dormancy and start to grow. The new growths are very tender and will likely be killed when cold temperatures return. The plants should be prepared for winter after the stems have withered and temperatures
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have gotten cold, but before any hard freezes. Here in Kansas City I prepare my cypripediums for winter at the end of November. I prefer to store my plants underground for the winter. I put the containers in a hole deep enough that the surface of the medium is three to four inches below grade level, then cover them with five or six inches of mulch (Fig. 4). Nature maintains the temperature and moisture for me. In colder areas, or where the risk of mid-winter warm spells is greater than here in the Kansas City area, it may be necessary to dig deeper and/or use a thicker layer of mulch. It is important that the hole drains well. I use inexpensive cypress mulch from the local hardware store, but some growers use chopped leaves. I‖ve found that whole leaves pack down and cause the plants to rot. Alternatively, cypripediums can be cover the containers with four to five inches of mulch. stored in an unheated garage, basement or enclosed porch where the temperature stays near freezing. Mulching the surface of the medium will help maintain the moisture level and protect the growth buds from light freezes. I‖ve stored other hardy orchids this way and will have to start doing it with my cypripediums because I‖ve run out of places to dig holes in my yard. Some growers put their plants, pots and all, in plastic bags and store them in a refrigerator, not the freezer, through the winter. The medium should be just barely moist. I‖ve discovered the hard way that if the medium is too wet, the plants will rot. Spring Care: I move my plants from their winter storage to the growing area when the danger of frost is low, but when the daytime temperatures are still cool, preferably in the 50°s F (10° to 15° C). The new shoots are very sensitive to frosts and even a light one will damage the developing leaves and flower buds. A freeze can kill the entire shoot. However, if the daytime temperatures are too high, the new shoots expand too rapidly and produce a weak plant. Based on Kansas City weather, about to first of April I remove the winter mulch, treat for slugs and snails, and move my cypripediums to their growing area. Once the containers are in their growing area, the medium warms up and the plants break dormancy. The growth buds formed the previous year expand and grow to their full size quite quickly. I cover my plants or move them into the garage if a frost is expected. I also cover them or move them into the garage if strong thunder storms or tornadoes are forecast. The only other care necessary at this time is to fertilize and to water if it doesn‖t rain. If the plant is mature, it will bloom now. This all happens in three to eight weeks, depending on the species. Now is the time when I find out how well I grew my plants last year!
Fig. 4. Cypripedium containers being buried for the winter. I

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Martin: GROWING CYPRIPEDIUMS IN CONTAINERS

Summer Care: Summer is when cypripediums grow into larger and stronger plants, but you won‖t see the results until the following year. Cypripediums initiate new roots at the base of the new shoot shortly after flowering. During the summer and into the fall these new roots and the roots from the previous two years elongate, and the plants store energy in the rhizomes and produce the growth buds for next year. The entire above-ground portion of the plant for the next growing season shoots, leaves and flowers - develops within these growth buds (Fig. 5). The care a cypripedium receives during summer will determine how strong a plant and how many flowers, if any, it will produce the next year. During summer, I continue watering and fertilizing as in the spring. The one cultural challenge in the summer is that cypripediums prefer to have their roots cool. I mulch the surface of the medium to protect it and the roots from being heated by the sun. Of course, this also helps to keep the medium from drying out. I use the same mulch used in the winter, and I am careful not to let the mulch touch the stem of the plant as this can cause it to rot. The container can also be set into a shallow, well draining hole in the ground to keep the roots cooler. I watch for caterpillars, slugs and snails, and use an Fig. 5. C. parviflorum var. makasin growth buds are well appropriate treatment immediately if I see developed by early August. They will continue to grow thicker signs of these pests. over the next one to two months as the roots lengthen and the Fall Care: In fall, the cypripediums stop growing and all above-ground growth dies back. I stop fertilizing about the beginning of September. I keep the medium moist and wait for the plants to go dormant, signaled by the leaves turning brown. I do any necessary repotting and dividing in late fall before I put the plants in the ground for the winter. I move my cypripediums to their winter quarters sometime between the first frost and the first hard freeze. New Plants: Cypripedium suppliers ship bare-root dormant plants in either the fall or spring. I prefer to receive them in the fall because spring shipments arrive late in my spring growing season. Most suppliers are located further north than I am, and by the time they can safely ship plants, the weather in my area has already warmed up. When I receive plants in the fall, I seal the bare-root plants in plastic food storage containers with a few drops of water. I then seal the containers in a plastic bag and put them in the refrigerator until spring. Of course, another option is to pot the plants and put them with your other cypripediums for the winter.
plant stores energy in the tubers.

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RECOMMENDED SPECIES AND HYBRIDS The genus Cypripedium has 45 species and is quite diverse. Not surprisingly, some are less demanding in their requirements. Below is a list of some of those species that are considered among the easiest to grow and that I‖ve had success with. Except as noted, I grow these as described above. C. parviflorum: The three varieties of the North American yellow lady‖s-slipper are considered to be among the easiest of the species to grow. The two varieties I have, makasin and pubescens have both grown well for me. C. kentuckiense: The largest species in the genus, it grows naturally in sandy stream banks. Consequently, I use a medium of 50% sand, 30% gravel and 20% peat. However, I would expect it to grow well in either of my standard mixes. C. reginae: This species likes a little more light than others, so I grow it where it gets more morning sun. It grows naturally in more bog-like conditions than other species. I grow mine in a mix of three parts Perlite, one part gravel and two parts ―black peat‖, which is partially composted peat moss. C. formosanum: This is often considered the easiest species to grow. It has rather long rhizomes and the new growths can be several inches away from the previous year‖s, so it requires a large container. Also, it is prone to breaking dormancy during a mid-winter thaw, so extra protection may be needed in winter. C. macranthos: This species grows well if protected from temperatures above 80°F (27°C). I grow these in my basement lightroom during the hot part of the summer. Hybrids: In recent years a number of cypripedium hybrids have become available (Fig. 6). Like most orchids, cypripedium hybrids are easier to grow than the species and are a good choice for the beginning grower.
Fig. 6. C. Hilda (C. ×ventricosum × macranthos), one of the many cypripedium hybrids that are available from commercial growers.

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SOURCES OF CYPRIPEDIUMS Like their tropical cousins, many cypripedium populations in the wild have been decimated by over-collection. Please purchase only artificially propagated cypripediums to help reduce the demand for wild-collected plants. Wild cypripediums have delicate roots that are usually damaged when plants are collected. Most die within two to three years after being removed from the wild. Cypripediums require several years to grow to blooming size and reputable growers price them accordingly. Mature plants sold at bargain prices are almost certainly wild-collected. Many artificially propagated cypripediums are available from reputable nurseries and commercial orchid growers. Some companies that I‖ve found to have consistently high quality plants are: Cyp Haven: http://www.c-we.com/cyp.haven/ Hillside Nursery: http://hillsidenursery.biz/ Itasca Lady Slipper Farm: http://www.ladyslipperfarm.com/ Vermont Lady Slipper Company: http://www.vtladyslipper.com/ Wild Orchid Company: http://www.wildorchidcompany.com/ Spangle Creek Labs: http://www.spanglecreeklabs.com/ SOURCES OF MORE INFORMATION ON GROWING CYPRIPEDIUMS Most of the suppliers listed above provide cultural information on their websites. Holger Perner has an excellent chapter on cultivation in The Genus Cypripedium, by Phillip Cribb. Timber Press, Oregon. 1997. The cypripedium forum: http://www.cypripedium.de/forum/ Doug Martin, 15523 Johnson Dr., Shawnee, KS 66217 bethdougm@kc.rr.com LITERATURE CITED
Stoutamire, W.P. 1991. Central growth cycle of Cypripedium candidum Muhl. root systems in an Ohio Prairie. Lindleyana 6(4): 235-40.

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Kauth et al.: PRELIMINARY RESULTS FOR FIELD ESTABLISHMENT TECHNIQUES OF CALOPOGON TUBEROSUS

Calopogon tuberosus var. tuberosus common grass-pink

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Kauth et al.: PRELIMINARY RESULTS FOR FIELD ESTABLISHMENT TECHNIQUES OF CALOPOGON TUBEROSUS

PRELIMINARY RESULTS FOR FIELD ESTABLISHMENT TECHNIQUES OF CALOPOGON TUBEROSUS
Philip J. Kauth*, Michael E. Kane & Timothy R. Johnson Abstract
Worldwide habitat loss has led to interest in propagation and reintroduction of orchids. However, scientific investigation regarding successful field establishment remains poorly understood. Previous research has indicated that using dormant storage organs and planting seedlings in areas of reduced competition increased survival of several orchid species. Here we describe methods for establishing Calopogon tuberosus, a North American terrestrial orchid, on the Florida Panther National Wildlife Refuge. Comparative effects of planting seedlings and corms on survival and shoot growth were studied. In addition, seedling survival in burned and unburned plots was studied. While propagule type did not influence survival, date of planting did. A higher percentage of propagules survived when planted in February 2009 during the early growing season. While more seedlings were actively growing in the burned plot during April 2009, seedlings in the unburned plot produced more shoots. The data from this study are being used to develop management plans not only for C. tuberosus, but also other terrestrial orchids.

Introduction The worldwide loss of orchid taxa has led to an abundance of research focused on their conservation, ecology, and reintroduction (Ramsay and Dixon, 2003). Unfortunately, few reports exist that detail management methods for both orchid populations and their habitat (Stewart, 2007). Successful establishment of plants into current or former habitats is often the culmination and goal of orchid conservation research (Batty et al. 2006a). Establishing orchids in the field is challenging because complex ecological requirements of individual taxa are not well-understood (Scade et al., 2006). Successful field establishment of terrestrial orchids has been previously attempted, but only for a few species (McKendrick, 1995; Ramsay and Stewart, 1998; Stewart et al., 2003; Batty et al., 2006b; Scade et al., 2006; Yamato and Iwase, 2008), and few studies have documented field establishment of North American species (Stewart, 2007). Long-term survival of field-transplanted orchids is often very low, in part because efficient methods for establishing orchids are lacking (Batty et al., 2006a). The influence of abiotic and biotic factors on successful field establishment of orchids has not been studied in detail (Scade et al., 2006). However, field establishment of orchids could be an important tool for both conserving orchids and furthering our knowledge of orchid ecology (McKendrick, 1995).
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Kauth et al.: PRELIMINARY RESULTS FOR FIELD ESTABLISHMENT TECHNIQUES OF CALOPOGON TUBEROSUS

A major obstacle to field establishment is initial survival of propagules. Only a few articles highlight techniques for increasing survival of orchid seedlings under in situ conditions (Batty et al., 2006b; Scade et al., 2006; Smith et al., 2009). Batty et al. (2006b) reported higher survival of several Australian orchid species when dormant tubers were reintroduced rather than seedlings. Observations that Thelymitra manginiorum seedlings established more readily than tubers indicate that field performance of different propagules is species-specific (Smith et al., 2009). Competition may also be an important factor to consider for successful establishment (McKendrick, 1995). Dense coverage by native species increased survival of field transplanted orchid species (McKendrick, 1995; Scade et al., 2006; Yamato and Iwase, 2008), but areas of greatest vegetation coverage, including weedy species, impeded total survival (McKendrick, 1995). Calopogon tuberosus var. tuberosus (Linnaeus) Britton, Sterns & Poggenberg, common grass-pink, is a corm-forming terrestrial orchid species found throughout eastern North America from south Florida to Newfoundland, Canada. Calopogon tuberosus is a fairly common native orchid and is an excellent candidate to examine field establishment methods because: 1) seeds germinate readily in vitro, 2) seedlings can be produced in several months (Whitlow, 1996; Kauth et al., 2006; Kauth et al., 2008), and 3) it is a corm-forming species. Because this species produces corms, the role of propagule type on successful field establishment can be studied. The objectives of this study were to: 1) Establish Calopogon tuberosus seedlings at the FPNWR; 2) Compare survival of seedlings and corms of C. tuberosus; 3) Compare survival and growth of seedlings in burned and unburned areas; and 4) Recommend management practices for establishing terrestrial orchids at the FPNWR. The data generated will be used to recommend management practices for the successful conservation of this species and other terrestrial orchids worldwide. Materials and Methods Field Site The Florida Panther National Wildlife Refuge (FPNWR) is located in Collier Co., Florida and consists of 26,400 acres within the Big Cypress Basin (U.S. Fish and Wildlife Service 2009). The FPNWR was established in 1989 to protect the Florida panther and the mosaic of habitats located throughout. As a national wildlife refuge, the FPNWR actively manages the area with invasive plant removal, prescribed burning, native plant propagation, and restoration activities. The FPNWR is divided into 50 fire-management units. Calopogon tuberosus is currently found in units containing wet prairies (Fig. 1). These grass and sedge dominated communities at the FPNWR are found between pine flatwoods dominated by Pinus elliotii (Davis, 1943; Duever et al., 1986). The largest population of C. tuberosus is found in a wet prairie where
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Kauth et al.: PRELIMINARY RESULTS FOR FIELD ESTABLISHMENT TECHNIQUES OF CALOPOGON TUBEROSUS

several hundred plants flower from March through May with peak flowering in mid April. All field plots were established in this area.
B

Fig. 1. Field translocation study at the Florida Panther National Wildlife Refuge. A) Burned (left) and unburned (right) areas in February 2009. B) Burned (background) and unburned (foreground) areas. Yellow flags mark one of the transects. C) Transects and quadrats in the burned area. D) Transect and quadrats in the unburned area in April 2008. E) Close-up of a quadrat.

Seed Source and Preparation Seeds from the FPNWR were collected from mature yellowing capsules in June 2006 and 2007 before capsule dehiscence. Two capsules were collected from at least three plants. Seed capsules were stored over desiccant in laboratory conditions at 23° C for 2 weeks. Seeds were subsequently removed from the capsules and placed in glass scintillation vials over silicagel desiccant, and stored in darkness at -11° C until use. In all experiments, seeds were surface sterilized in sterile scintillation vials for 3 min in a solution of 5 mL absolute ethanol, 5 mL 6% NaOCl, and 90 mL sterile dd water. Seeds were rinsed with sterile dd water after surface sterilization. Solutions were removed with sterile Pasteur pipettes. Seeds were transferred onto the germination medium with a 10µL sterile inoculating loop.
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Kauth et al.: PRELIMINARY RESULTS FOR FIELD ESTABLISHMENT TECHNIQUES OF CALOPOGON TUBEROSUS

Fig. 2. Monthly temperatures recorded at the field transplant site in the Florida Panther National Wildlife Refuge from April 2008-March 2009. Average temperatures represent the mean daily high or low over the entire month. Data were collected with a HOBO H8 Pro series weather station.

Establishment Planting occurred in successive years in April 2008 and February 2009. For the 2008 planting, differences in survival of field transplanted seedlings and corms were examined. For the 2009 planting, the response of planting seedlings in a burned versus unburned area was studied (Fig. 1A, B, C). For all experiments square quadrats 30 cm × 30 cm were constructed from PVC piping (1.5 cm diameter). Each quadrat (Fig. 1E) was divided into 16 sections ca. 7.5 cm × 7.5 cm by using 14 gauge coated electrical copper wire. A HOBO H8 Pro weather station (www.microdaq.com, Ltd., Contoocook, NH) was placed at the site to record daily temperatures and relative humidity (Fig. 2). Comparison of propagule type on field survival Seeds were germinated asymbiotically in vitro beginning March 2007 on P723 medium supplemented with 1% activated charcoal (PhytoTechnology Laboratories, Shawnee Mission, KS). 40 mL of medium was dispensed into square 100 × 15 mm Petri plates (Integrid™ Petri Dish, Becton Dickinson and Company, Franklin Lakes, NJ). Cultures were wrapped in a single layer of Nescofilm (Karlan Research Products, Santa Rosa, CA, USA) and placed under
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Kauth et al.: PRELIMINARY RESULTS FOR FIELD ESTABLISHMENT TECHNIQUES OF CALOPOGON TUBEROSUS

a 12 h photoperiod at 25° C. After 8 weeks culture (May 2007), seedlings were transferred to PhytoTech Culture Boxes (PhytoTechnology Laboratories, Shawnee Mission, KS) containing 100 mL P723 medium. After an additional 30 weeks culture, corms were chilled at 10°C in darkness from October 2007 to January 2008. This was accomplished by removing the shoots and roots from the seedlings, and transferring corms to fresh P723 medium in PhytoTech Culture Boxes. After the chilling period, corms were again transferred to fresh P723 medium in PhytoTech Culture Boxes for an additional 12 weeks under a 12 h photoperiod. Seedlings were subsequently moved to greenhouse conditions April 2008. Seedlings were planted in 9cell pack trays (Model #IKN0809, Hummert International, Earth City, MO) containing Fafard 2 soilless potting mix (Conrad Fafard, Inc., Agawam, MA). Seedlings were covered with clear vinyl humidity domes to prevent desiccation, and placed under 50% shade cloth and a natural photoperiod. Average light levels were 300 µmol m-2 s-1 measured at 12 noon, and average temperatures ranged from 21.6 ± 2° C to 29.3 ± 3° C. After one week humidity domes were removed and seedlings were watered as needed. Three 10 m transects (Fig. 1D) were establish April 23, 2008. Each transect contained four quadrats 2.5 m apart. A randomized block design was used to plant propagules. Corms and seedlings were assigned randomly to a quadrat and quadrat section. Sixteen propagules were used in each quadrat (8 seedlings and 8 corms per quadrat). A total of 192 propagules were planted. Propagules were irrigated with distilled water upon initial planting. Data were collected on 20 May 2008, 9 July 2008, 27 February 2009, and 23 April 2009. Seedling survival in a burned and unburned field plot Seeds were germinated in vitro starting January 2008 on BM-1 Terrestrial Orchid Medium (PhytoTechnology Laboratories, Shawnee Mission, KS) supplemented with 1% activated charcoal. 40 mL of medium was dispensed into square 100 × 15 mm Petri plates. Cultures were placed under a 12 h photoperiod at 25°C. After 8 weeks culture (March 2008), seedlings were transferred to PhytoTech Culture Boxes containing 100 mL BM-1 medium. After an additional 30 weeks culture, corms were transferred to new PhytoTech Culture Boxes containing 100 mL BM-1 medium and chilled at 10°C from October 2008 to December 2008. Seedlings were moved to greenhouse conditions December 2008 until ready for field establishment February 2009. Greenhouse transfer procedures were similar to those previously described. Average light levels were 253 µmol m-2 s-1 measured at 12 noon, and average temperatures ranged from 20.8 ± 2.3°C to 28.8 ± 2.8°C. In January 2009, the wet prairie was burned except the area where Calopogon tuberosus field plots were previously established in 2008. This presented a unique opportunity to compare the effects of planting seedlings in the burned and unburned areas. Two 10 m transects were established in both the burned area and unburned area. Three quadrats were allocated to each transect. Sixteen seedlings were planted in each quadrat for a total of 48 seedlings per transect and 192 seedlings for the experiment.
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Kauth et al.: PRELIMINARY RESULTS FOR FIELD ESTABLISHMENT TECHNIQUES OF CALOPOGON TUBEROSUS

Data Collection and Statistical Analysis For both experiments, survival of all seedlings was recorded. Two different categories were classified in determining propagule survival. Percentage of actively growing green shoots was recorded. Percentage of emergent shoots was recorded when shoots were present, but not necessarily actively growing (i.e. shoots were yellow and brown due to senescence). Seedling leaf measurements were recorded before the February 2009 experiment, and again in April 2009. Shoot emergence data were analyzed using proc glimmix, logistic regression, and leastsquare means in SAS v9.1. Results Comparison of Propagule Type on Field Survival Propagule type (F = 0.50, p = 0.48) did not influence survival, but date (time of data recorded) was significant (F = 20.4, p < 0.0001). At the initial data collection in May 2008, a higher proportion of seedlings (43.8%) had actively growing shoots compared to corms (32.3%) (Fig. 3). After 1 month of field establishment, less than 50% of all propagules had actively growing shoots regardless of treatment. In July 2008, natural leaf senescence had occurred so that no shoots were actively growing. A higher proportion of emergent shoots were observed on seedlings (22.9%) compared to corms (12.5%).

Fig. 3. Survival of Calopogon tuberosus propagules at the Florida Panther National Wildlife Refuge. Histobars represent the mean response of three separate transects each with four quadrats containing 16 propagules. A total of 96 propagules were planted per treatment for a total of 192 propagules. 18

Kauth et al.: PRELIMINARY RESULTS FOR FIELD ESTABLISHMENT TECHNIQUES OF CALOPOGON TUBEROSUS

Fig. 4. Survival of Calopogon tuberosus seedlings in a burned and unburned plot at the Florida Panther National Wildlife Refuge. A) Percentage of plants with actively growing shoots marked by the presence of a growing green shoot. B) Percentage of plants with either have actively growing shoots or previously emerged shoots that senesced. Histobars represent the mean of two transects with three quadrats containing 16 seedlings. Ninety-six seedlings were planted in each treatment for a total of 192 total seedlings. 19

Kauth et al.: PRELIMINARY RESULTS FOR FIELD ESTABLISHMENT TECHNIQUES OF CALOPOGON TUBEROSUS

Data collected during February 2009 occurred during the early growing season in south Florida. The number of green shoots was higher on corms (12.5%) compared to seedlings (10.4%), but this difference was not significant (Fig. 3). In April 2009, no significant difference was observed between the survival of corms (6.25%) and seedlings (8.33%), and the presence of shoots further declined. At this time, one seedling in the early flowering stage established from a corm propagule was observed No shoots were observed in June 2009. Percent of total survivorship of all combined propagules were as follows: 38.0% (May 2008), 18.9% (July 2008), 11.4% (February 2009), and 7.3% (April 2009). Seedling Survival in a Burned and Unburned Field Plot Burning significantly influenced percent of emerged shoots (F = 48.7, p < 0.0001), while the unburned plot influenced the percentage of actively growing shoots (F = 4.32, p = 0.04). Two months after field establishment, the number of actively growing shoots declined in both plots. Three percent and 11% of actively growing shoots were observed in the burned and unburned areas, respectively (Fig. 4B). However, senesced shoots were visible on seedlings in the burned plot, but none in the unburned plot (Fig. 4A). Total survivorship was 7.3% when combining all data. Of the actively growing shoots, shoot lengths were recorded in April 2009 (Table 1). Shoot lengths on all seedlings with actively growing shoots in the burned plot increased, while three of the eleven recorded leaf measurements in the unburned plot decreased (Table 1).
Treatment Unburned Transect 1 1 1 1 1 2 2 2 2 2 2 1 1 2 Quadrat # 1 2 3 3 3 1 2 2 2 3 3 1 2 1 Seedling # 14 3 3 6 13 14 4 9 10 5 12 13 14 3 Height (Feb 2009) 85 25 100 60 90 70 62 76 63 85 26 52 10 95 Height (April 2009) 28 66 75 90 108 92 220 35 72 125 102 91 165 140

Burned

Table 1. Shoot lengths recorded for actively growing Calopogon tuberosus seedlings in February and April 2009. All measurements are in mm. Seedlings were measured in February under greenhouse conditions prior to transplant, and the April data collection was on seedlings after field transplant on the Florida Panther National Wildlife Refuge.

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Kauth et al.: PRELIMINARY RESULTS FOR FIELD ESTABLISHMENT TECHNIQUES OF CALOPOGON TUBEROSUS

Discussion This is the first study of the field establishment of Calopogon tuberosus, and one of the only scientifically documented orchid field establishment studies in North America (Stewart et al., 2003; Zettler et al., 2007). However, conclusive results were not obtained due to the short-term nature of the study, and more results are likely after several years of monitoring. Absence of an actively growing shoot did not indicate propagule death since corms may have been present beneath the soil surface. Their presence beneath the soil was not confirmed in order to minimize soil disturbance. In addition, shoots on field-transplanted seedlings may have senesced naturally because senescence naturally occurs in late May through early June. Field establishment of orchids may depend on propagule type such as seedlings or storage organs. Dormant storage organs, depending on species, were found to successfully survive initial field establishment compared to seedlings (Debeljak et al. 2002; Batty et al. 2006b). Dormant storage organs may be able to survive drought conditions better than seedlings (Batty et al. 2006b); however, results are species specific. Caladenia arenicola and Diuris magnifica established more readily in the field when dormant tubers were planted rather than seedlings, but Thelymitra manginiorum established more readily from seedlings (Batty et al., 2006a). However, no C. arenicola propagules and only 10% of D. magnifica tubers survived into the third growing season. 70% and 35% of T. manginiorum seedlings and tubers, respectively, survived into the third growing season (Batty et al., 2006a). Likewise, Smith et al. (2009) found that 2-3 year old plants (35%) established more readily in the field compared to tubers (11%) after 4 years. In this study, no statistical differences in shoot emergence were observed between corms (6.25%) and seedlings (8.33%) after the first year. The low rates of shoot emergence may have been caused by propagule death or dormancy of corms. Terrestrial orchids can remain dormant for several years (Kery and Gregg 2004) thus long-term monitoring is necessary to observe propagule survival. Throughout 2008-2009, south Florida experienced drought conditions that may have contributed to propagule death. The juvenile state of the propagules (1 year old) may have resulted in poor field establishment as well. Tuber size influenced the survival of several Australian orchids with larger tubers increasing survival compared to smaller tubers (Batty et al., 2006a; Smith et al., 2009). Likewise, larger Calopogon tuberosus corms or more mature plants may have increased survival in the present study due to greater storage reserves that tubers can utilize to sustain drought conditions and initiate growth (Batty et al., 2006a). The influence of competition, shading, and weed coverage influences the establishment of orchids in the field (McKendrick, 1995; Scade et al., 2006). This is the first report comparing field establishment of an orchid in a burned and unburned are in North America. The effects of establishing orchids in burned plots have apparently not been studied, but smoke was shown to be effective at promoting germination of several Australian plant species
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Kauth et al.: PRELIMINARY RESULTS FOR FIELD ESTABLISHMENT TECHNIQUES OF CALOPOGON TUBEROSUS

(Flematti et al., 2004). Although this study investigated seed germination, the study could explain a higher percentage of actively growing shoots in the burned plot. The effects of smoke on emergence of Calopogon tuberosus shoots may warrant investigation. The influence of competition on plant establishment has also been examined. In the present study, more actively growing shoots of Calopogon tuberosus were observed on seedlings in the unburned area during April 2009. Shoots on the seedlings in the burned area were brown and senesced with the exception of three plants. The surrounding native grasses in the unburned area likely shaded the seedlings providing increased survivorship and soil moisture. Seedlings in the unburned area did not receive any level of shading and likely caused seedling desiccation. Shading led to increased survival of several other terrestrial orchids (McKendrick, 1995, 1996; Scade et al., 2006; Yamato and Iwase, 2008), but areas of dense shade and competition can lead decreased seedling survivorship (McKendrick, 1995; Yamato and Iwase, 2008). Fire is a necessary natural disturbance in many ecosystems (Duncan et al., 2008) including wet-prairies in south Florida. Competition with weeds and invasive species during field establishment often reduces the successful field establishment of seedlings (Moyes et al. 2005). Native perennials were established readily in a burned grassland and dolomite glade areas. In addition, reduced weedy species, and prevented forest succession (Moyes et al., 2005; Duncan et al., 2008). While the results of this study are preliminary due to the short-term monitoring of the plots, the techniques employed can be applied to other orchid species worldwide. More definitive results may be observed after another growing season when seedlings in the burned area may re-emerge. Due to the drought conditions the past 2 years in south Florida, additional irrigation may have improved propagule survival. In addition, using symbiotically grown seedlings or inoculating soil with mycorrhizal fungi may have improved seedling survival as well (Batty et al., 2006a; Scade et al., 2006; Smith et al., 2009). Conclusions Based on the success of research with other terrestrial orchids, the following should also be considered to successfully establish Calopogon tuberosus seedlings in the field: 1) Using more mature seedlings to younger seedlings may provide sufficient carbohydrate reserves to survive initial planting. 2) When planting dormant corms, larger corms should be planted. 3) Propagules should be planted at the beginning of the growing season. 4) Due to frequent drought conditions, the effects of supplemental irrigation could be studied. 5) Plots should be monitored for several years to observe successful field establishment. Philip J. Kauth*, Michael E. Kane, Timothy R. Johnson
Plant Restoration, Conservation, and Propagation Biotechnology Program, Environmental Horticulture Department, University of Florida , PO Box 110675, Gainesville, FL 32611, USA. *Corresponding author: email pkauth@ufl.edu 22

Kauth et al.: PRELIMINARY RESULTS FOR FIELD ESTABLISHMENT TECHNIQUES OF CALOPOGON TUBEROSUS

Literature Cited
Batty, A.L., M.C. Brundrett, K.W. Dixon, and K. Sivasithamparam. 2006a. New methods to improve symbiotic propagation of temperate terrestrial orchid seedlings from axenic culture to soil. Australian Journal of Botany 54: 367-74. Batty, A.L., M.C. Brundrett, K.W. Dixon, and K. Sivasithamparam. 2006b. In situ seed germination and propagation of terrestrial orchid seedlings for establishment at field sites. Australian Journal of Botany 54: 375-81. Davis, J.H. 1943. The Natural Features of Southern Florida. The Florida Geological Survey, Tallahassee, FL.311. Debeljak, N., M. Regvar, K.W. Dixon, and K. Sivasithamparam. 2002. Induction of tuberisation in vitro with jasmonic acid and sucrose in an Australian terrestrial orchid, Pterostylis sanguinea. Plant Growth Regulation 36: 253-60. Duever, M.J., J.E. Carlson, J.F. Meeder, L.C. Duever, L.H. Gunderson, L.A. Riopelle, T.R. Alexander, R.L. Myers, and D.P. Spangler. 1986. The Big Cypress National Preserve. National Audubon Society, New York. Duncan, R.S., C.B. Anderson, H.N. Sellers, and E.E. Robbins. 2008. The effect of fire reintroduction on endemic and rare plants of a southeastern glade ecosystem. Restoration Ecology 16: 39-49. Flematti, G.R., E.L. Ghisalberti, K.W. Dixon, and R.D. Trengove. 2004. A compound from smoke that promotes seed germination. Science 305: 977 Kauth, P. J., W.A. Vendrame, and M.E. Kane. 2006. In vitro seed culture and seedlings development of Calopogon tuberosus. Plant Cell, Tissue and Organ Culture 85: 91-102. Kauth, P. J., M.E. Kane, W.A. Vendrame, and C. Reinhardt-Adams. 2008. Asymbiotic germination response to photoperiod and nutritional media in six populations of Calopogon tuberosus var. tuberosus (Orchidaceae): evidence for ecotypic differentiation. Annals of Botany 102: 783-93. Kery, M., and K. Gregg. 2004. Demographic analysis of dormancy and survival in the terrestrial orchid Cypripedium reginae. Journal of Ecology 92: 686-95. McKendrick, S.L. 1995. The effects of herbivory and vegetation on laboratory-raised Dactylorhiza praetermissa (Orchidaceae) planted into grassland in Southern England. Biological Conservation 73: 215-20. McKendrick, S.L. 1996. The effects of shade on seedlings of Orchis morio and Dactylorhiza fuchsii in chalk and clay soil. New Phytologist 134: 343-52. Moyes, A.B., M.S. Witter, and J.A. Gamon. 2005. Restoration of native perennials in a California annual grassland after prescribed spring burning and solarization. Restoration Ecology 13: 659-66. Ramsay, M.M. and J. Stewart. 1998. Re-establishment of the lady's slipper orchid (Cypripedium calceolus L.) in Britain. Botanical Journal of the Linnaean Society 126: 173-81. Ramsay, M.M. and K.W. Dixon. 2003. Propagation science, recovery, and translocation or terrestrial orchids. In K. W. Dixon, S. P. Kell, R. L. Barrett, and P. J. Cribb [eds.], Orchid Conservation. Natural History Publications (Borneo), Kota Kinabalu, Sabah, Malaysia. Scade, A., M.C. Brundrett, A.L. Batty, K.W. Dixon, and K. Sivasithamparam. 2006. Survival of transplanted terrestrial orchid seedlings in urban bushland habitats with high or low weed cover. Australian Journal of Botany 54: 383-89. Smith, Z. F., E.A. James, M.J. McDonnell, and C.B. McLean. 2009. Planting conditions improve translocation success of the endangered terrestrial orchid Diuris fragrantissima (Orchidaceae). Australian Journal of Botany 57: 200-209. Stewart, S.L. 2007. Integrated conservation of Florida Orchidaceae in the genera Habenaria and Spiranthes: model orchid conservation systems for the Americas. PhD Dissertation, University of Florida, Gainesville, FL, USA. Stewart, S.L., L.W. Zettler, J. Minso, and P.M. Brown. 2003. Symbiotic germination and reintroduction of Spiranthes brevilabris Lindley, an endangered orchid native to Florida. Selbyana 24: 64-70. U.S. Fish and Wildlife Service. 2009. Florida panther national wildlife refuge fact sheet. http://www.fws.gov/southeast/pubs/facts/flpcon.pdf. Last accessed 7 October, 2009. 23

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Whitlow, C.E. 1996. Mass production of Calopogon tuberosus, pp.5-10. In C. Allen [ed.], North American Native Terrestrial Orchids: propagation and production. North American Native Terrestrial Orchid Conference, Germantown, Maryland. Yamato, M. and K. Iwase. 2008. Introduction of asymbiotically propagated seedlings of Cephalanthera falcata (Orchidaceae) into natural habitat and investigation of colonized mycorrhizal fungi. Ecological Research 23: 329-37. Zettler L.W., S.B. Poulter, K.I. McDonald, and S.L. Stewart. 2007. Conservation-driven propagation of an epiphytic orchid (Epidendrum nocturnum) with a mycorrhizal fungus. HortScience 42: 135-39.

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Richards & Sanders: A PRACTICAL AND INTEGRATED APPROACH TO NATIVE ORCHID CONSERVATION AND PROPAGATION AT THE ATLANTA BOTANICAL GARDEN

A PRACTICAL AND INTEGRATED APPROACH TO NATIVE ORCHID CONSERVATION AND PROPAGATION AT THE ATLANTA BOTANICAL GARDEN
Matt Richards & Jenny Cruse Sanders Ph.D. In recent years interest has grown in the propagation of rare orchids native to North America. For the Atlanta Botanical Garden it stems originally from the need to reproduce valuable collections held in public trust for display and education interests in the Fuqua Orchid Center (Fig. 1). Now the focus has grown to include work on propagating natives specifically for conservation purposes. These effective methods of propagation and production have not always been a result of scientific research backed by statistical analysis, but still have contributed greatly to our ongoing commitment to sustain and improve in situ orchid populations in the Southeastern United States (Fig. 2). As the program continues to grow, the Garden maintains its project driven approach to assist in the protection of habitat as well as to produce plants suitable for conservation work while developing a balance between scientific research, practical propagation and ensuing horticultural practices. For decades, the Garden‖s conservation program has been forged by following through project driven goals and objectives. Work has been focused on forming relationships between private-landowners, federal, state, and local agencies. Needs are defined to outline the roadmap towards reaching each particular goal. The Georgia Plant Conservation Alliance was formed in 2005 and is inclusive of many levels of contributors in the native plant conservation community. As a founding member, the Garden has united with other botanical gardens, institutions, universities, individuals, governmental and non-governmental agencies to provide a service to the broad scheme of plant conservation (Fig. 3). Through this alliance, many of the collaborators have been able to facilitate rare plant conservation without the obstacles presented by having a formal organization. All parties involved with each specific project have identified their particular talents, resources, and capabilities they can contribute to the overall project. These services may include diplomatic relations, propagation and production, on the ground labor, or motivating an army of dedicated volunteers to see the project through. The Atlanta Botanical Garden has found its niche in rare plant propagation and horticultural excellence (Fig. 4). All participants contribute in some way or another, and all of the work is
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Richards & Sanders: A PRACTICAL AND INTEGRATED APPROACH TO NATIVE ORCHID CONSERVATION AND PROPAGATION AT THE ATLANTA BOTANICAL GARDEN

done in close collaboration in large part due to the coordination efforts carried out by the GPCA Conservation Coordinator, Jennifer Ceska at the State Botanical Garden of Georgia. Orchids in Georgia continue to require field surveys to assess the current status of state element occurrence records. We also attempt to identify suitable habitat and additional populations of native species. The Georgia Orchid Initiative through the Atlanta Botanical Garden ultimately aims to resurvey all orchid species known to occur in state and catalog localities with GPS and GIS technologies. Through tissue culture, field surveys, horticultural experiments, and ex situ propagation, the orchid research team is taking an integrated and collaborative approach to better understand the geography, natural history and reproductive biology of native orchids. This initiative hopes to provide an increased understanding of the biology and geographic distribution of native terrestrial orchids in Georgia (Fig. 5). The Garden often works with the GADNR (Georgia Department of Natural Resources) to identify the needs for orchid surveys and species recovery in the state. We have decided it would make most sense to concentrate our efforts using the state ranks, surveying the element occurrence records first of S1, S2 and S3 species, and then move through the list to S5 respectively. In addition, peripheral habitat is identified using satellite imagery, soil maps, GIS technologies, and ground reports to better assess the status of the species. Once populations have been located, we begin work on assessing the habitat threats, and potential management strategies (if any) (Fig. 6). For some of these orchid populations there is a need to safeguard populations through propagation and ex situ collections, especially those included in the S1/S2 G1/G2 rarity status. This propagated material is grown in an organized and indexed manner so that it may be used responsibly to augment populations or for future reintroduction into conservation lands (if deemed necessary and appropriate). One example would be that of Platanthera chapmanii (Chapman’s fringed orchid) (Fig. 7). Known historically from parts of south Georgia, the species had not been documented in the state for nearly a century. In September of 2009 a GPCA member, Dr. Richard Carter of Valdosta State University confirmed extant populations of the species he had seen earlier in 2006. Soon after the discovery was documented, seed was collected from two populations for in vitro propagation. At the time of this writing, stage-3 germination had been achieved (Fig. 8). With some good horticulture and a bit of luck, the future of this population of rare orchids is better protected. The need to propagate the species was prioritized by assessment of habitat. The habitat was surveyed by members of the GPCA and considered to be in extreme danger of alteration, subject to road improvements, herbicide applications, and timber activities. Measures will soon be taken to develop further conservation strategies. Additionally, Platanthera integrilabia (monkey-face orchid) occurs in a handful of counties in the State of Georgia. We have worked together with the GADNR to survey extant populations, additional habitat on conservation lands, and any new 'undiscovered
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Richards & Sanders: A PRACTICAL AND INTEGRATED APPROACH TO NATIVE ORCHID CONSERVATION AND PROPAGATION AT THE ATLANTA BOTANICAL GARDEN

populations' of the species. Since 2007, surveys have been active for the species. Meanwhile, propagation has been ongoing, safeguarding populations on private land in ex situ collections produced from seed with the hope of augmenting these populations one day (Fig. 9). Although one of the rarest orchids in North America, it is oddly very easily propagated from seed. The seed is sterilized using 90 ml RO water, 10 ml Clorox, and .01 ml Tween 20 shaking vigorously for 10 minutes. The seed is poured into a sterile filter and rinsed several times using sterile RO water. The seed is allowed to air dry within a laminar flow hood, and then sprinkled onto sterile media. This species is easily germinated on ½-strength P668 from PhytoTech Labs. After approximately 5 months, the seedlings are transferred to ¾-strength P668. Although it will germinate and grow on various other media, the cost and simplicity of this procedure effectively produces reproductive plant material in a short amount of time (Fig. 10). The process is typically adapted to the natural bio-rhythm of the species. The seed is harvested and sown at the time it would naturally fall to the ground. It is placed in dark cabinets after sowing, and then into cold storage during the winter months. During this time, the seedlings continue to develop. The ensuing spring allows the seedlings to emerge into photosynthetic growth under artificial lighting while still in sterile culture. Soon after the emergence of the first shoot, the plants are transferred onto fresh media and allowed more space to grow. When the plants are large enough to be transferred into pots and soilless media in our greenhouses, they are typically overwintered in vitro and potted up immediately following 120 days at ca. 38 F. By following the natural biorhythm of the plant, it becomes easier to transition them between growing stages and eventually back into their native habitat. This method would typically allow for outplanting at the time the species goes dormant. The propagation of our native orchids has been well documented in the past and it is common to find available scientific and popular articles regarding propagation of many of our natives. There are also many books that have been published on the subject of orchid propagation. The Garden approaches each species independently, researches what has been done by others in the past with the species and then develops a plan to tackle the first obstacle of germination. We sow our orchids using various methods, some are green capsule, some are dry seed, some are sterilized in different fashions, and many different media are used. Since 2002, records have been kept defining the work done with each orchid species. Although much of this remains unpublished, the database is a critical tool used daily in the Tissue Culture Lab. We can refer back many years to varying treatments and procedures used at propagating hundreds of species. With this information available, our staff can better develop an experiment that could eventually lead to successful propagation. Generally speaking, we sow most seed on several different media, and develop a plan to simulate natural bio-rhythm of the plant to what extent is possible. Once germination has been achieved, we will select several replate media for trial. Once the best media for replating is determined through observation and analysis of growth, the plants are transferred to quickly produce healthy plants suitable for transfer to ex vitro culture. Again, each species is treated independently and
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Richards & Sanders: A PRACTICAL AND INTEGRATED APPROACH TO NATIVE ORCHID CONSERVATION AND PROPAGATION AT THE ATLANTA BOTANICAL GARDEN

our talented and experienced staff develops the appropriate soil amendments to achieve desired growth and continues to refine the protocol for each species during greenhouse culture. Orchids are one of the most charismatic groups of plants, and yet few people are aware that so many species of orchids are native to Georgia. At the Atlanta Botanical Garden we reach a large audience with approximately 350,000 visitors per year, and we can inform this audience about native orchid species (Fig. 11). In 2007 we educated more than 14,500 students through educational programs (tours, outreach, afterschool, camps) developed for kindergarten through eighth grade, and 4971 adults through tours, lectures, training programs and family programming. Information learned through the Georgia Orchid Initiative will be made available to teachers as a resource for teaching about native plant species. Display plantings and interpretive signage will also be developed in the conservation garden, the children‖s garden, and the garden discovery carts will inform visitors about our research on native orchids in Georgia and North America (Fig. 12). Matt Richards & Jenny Cruse Sanders Ph.D., Atlanta Botanical Garden, 1345 Piedmont Ave NE,
Atlanta, Georgia 30309 mrichards@atlantabotanicalgarden.org; jsanders@altantabotanicalgarden.org www.atlantabotanicalgarden.org www.atlantabotanicalgarden.org/site/conservation/native_plants www.uga.edu/gpca/

The Atlanta Botanical Garden (Garden) has served the Southeastern region as both a horticultural resource and a place of enjoyment since 1976. The Garden has two facilities under its stewardship – 30 acres in the heart of Midtown Atlanta and 185 acres in Gainesville, Georgia (Smithgall Woodland Garden). Ranked as one of the top ten botanical gardens in the United States, the Garden develops and maintains plant collections for display, education, research, conservation, and enjoyment. The Garden offers stunning garden displays and exceptional education programs for people of all ages. Many of its collections of rare and endangered plant species cannot be seen anywhere else in the world, and its conservation work, both nationally and internationally, is critical to preserving our natural heritage. The Fuqua Orchid Center opened to the public in 2002 providing an exciting opportunity to further develop and display its already distinguished orchid collection. The display glasshouses maximize and augment the existing tropical lowland orchid collections and provide specialized facilities for new collections of orchids that grow at high elevations. Back-up greenhouse facilities for orchid care and a Tissue Culture Lab for plant propagation are also included in this center as are greenhouse facilities to propagate, and safeguard rare indexed plant populations of the southeastern United States for conservation purposes. Outdoors, and adjacent to the Fuqua Orchid Center is the Conservation Garden that highlights native bog habitats of the southeastern United States including coastal plain, cataract, and mountain bogs. For more information on programs, hours of operation, events, and classes, please visit www.atlantabotanicalgarden.org.

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Richards & Sanders: A PRACTICAL AND INTEGRATED APPROACH TO NATIVE ORCHID CONSERVATION AND PROPAGATION AT THE ATLANTA BOTANICAL GARDEN

Fig. 1 (left) Atlanta Botanical Garden Conservation Garden D. Lentz Fig. 2 (right) Cypripedium kentuckiense (ivory-lipped lady‖s-slipper) out-planting S. Larson

Fig. 3 (left) GPCA members at 2009 fall meeting Okefenokee Swamp. J. Ceska Fig. 4 (right) ABG Conservation Greenhouse facility. M. Richards

Fig. 5 (left) New population of Listera smallii (Small‖s twayblade) discovered in 2008.
B. Wilson

Fig. 6 (right) Survey of Platanthera spp. emerging after late winter burn in a coastal plain bog. L. Kruse 29

Richards & Sanders: A PRACTICAL AND INTEGRATED APPROACH TO NATIVE ORCHID CONSERVATION AND PROPAGATION AT THE ATLANTA BOTANICAL GARDEN

Fig. 7 (left) Platanthera chapmanii
R. Carter.

Fig. 8 (right) Stage 3 germination of Platanthera chapmanii
M. Richards

Fig. 9 (left) Platanthera integrilabia growing in vitro
R. Gagliardo

Fig. 10 (right) Platanthera integrilabia produced from seed flowering in the conservation greenhouse.
M. Wenzel

Fig. 11 (left) Visitors reading an interpretive display of Epipactis gigantea (stream orchid), a Flagship Species for the North American Region Orchid Specialist Group. M. Richards Fig. 12 (right) Hybrid swarm of Platanthera grown from seed at ABG on display in the Fuqua Orchid Center. M. Richards

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Massey: AN UNDERGRADUATE‖S FIRST ADVENTURE INTO FIELD RESEARCH: AN EPIPHYTIC ORCHID SURVEY IN SOUTHERN FLORIDA

AN UNDERGRADUATE’S FIRST ADVENTURE INTO FIELD RESEARCH:
AN EPIPHYTIC ORCHID SURVEY IN SOUTHERN FLORIDA

Emily Massey My interest in orchids began when I was an undergraduate student at Illinois College (IC), a small liberal arts institution located in Jacksonville, Illinois. There, I worked with Dr. Lawrence Zettler in the Orchid Recovery Program. This program focuses on the propagation, study, and reintroduction of threatened and endangered orchid species. While at IC, I participated in a number of studies with several different orchids. These projects included two studies involving symbiotic seed germination. In the first study, we examined crossing effects on seed viability, germination, and protocorm growth in Platanthera leucophaea (Nuttall) Lindley, the eastern prairie fringe orchid. Seed germination, propagation, and reintroduction of Epidendrum nocturnum Jacquin, the night-fragrant epidendrum was examined in our second study (Massey et al., 2007). I also participated in a study to asymbiotically propagate several epiphytic south Florida orchids such as E. amphistomum A. Richard, the dingyflowered star orchid; E. rigidum Jacquin, the rigid epidendrum; Polystachya concreta (Jaquin) Garay & Sweet, the yellow helmet orchid; Prosthechea cochleata (Linnaeus) W.E. Higgins var. triandra (Ames), the Florida clamshell orchid; and Vanilla phaeantha Reichenbach f., the oblong-leaved vanilla orchid. All of these studies were conducted in the laboratory, except for the reintroduction of Epidendrum nocturnum, which is an endangered Florida epiphyte with night fragrant flowers that are believed to be pollinated by a species of hawkmoth. Although I liked lab work, it was this study that introduced me to field research and it was one of the best experiences I had while working with Dr. Zettler. The project took place in the fall of 2005 at the Florida Panther National Wildlife Refuge (FPNWR). The FPNWR is located 20 miles east of Naples in Collier County, Florida and was established as a safe haven for the diminishing Florida panther population and other threatened and endangered animal and plant species.
31

Fig. 1 Image of me and the two other students (William Kutosky, and Kris McDonald) reintroducing E. nocturnum at the Florida Panther National Wildlife Refuge in 2005
S.L. Stewart

Massey: AN UNDERGRADUATE‖S FIRST ADVENTURE INTO FIELD RESEARCH: AN EPIPHYTIC ORCHID SURVEY IN SOUTHERN FLORIDA

For about a week, two of my lab mates and I visited the refuge to reintroduce Epidendrum nocturnum seedlings propagated in our lab. At the time of the reintroduction, the cool, murky water at most sites was waist and chest deep for my 5‖2‖‖ stature (Fig. 1). You definitely had to be careful where you were walking, or I should say feel around where you were stepping, because you sure could not see through the water beneath your feet. This was due to the presence of tannins that darken the water into a coffee-like brew. This was a very

Fig. 2. One of the many swamp buggies in the fleet.

intimidating project and I was a little apprehensive at first. We were venturing out into the wilderness, with the possibility of running into an alligator or worse, and we were on foot. However, the longer we worked, the less this seemed to matter. I may have had to wring out my clothes every night of the trip, but this field research experience was one of the most memorable moments of my life. I had many other new experiences as well. I got to take my first spin in a swamp buggy, which is basically a very large, open-air vehicle resembling a monster truck without a top (Fig. 2). Another new experience and probably one of the more amusing moments of this excursion took place when a local news reporter and cameraman came out to the refuge to capture our efforts. The cameraman must have known what he was getting into because he showed up wearing boots and worn clothing. Perhaps the reporter should have consulted with him before he dressed that morning because he wore shiny dress shoes, khaki pants, a very neat button down shirt, and a tie. Needless to say the reporter was a bit out of his element, but he was a good sport about it. With a smile, he waded out into the swampy water after a few minor wardrobe adjustments (i.e., rolled up his pant legs a good three or four inches and donned a pair of borrowed boots) to film a portion of the piece.
32

Massey: AN UNDERGRADUATE‖S FIRST ADVENTURE INTO FIELD RESEARCH: AN EPIPHYTIC ORCHID SURVEY IN SOUTHERN FLORIDA

We eventually reintroduced 43 Epidendrum nocturnum seedlings back into the wild. Unfortunately, this part of Florida had just been damaged by Hurricane Wilma. This stripped many of the trees‖ upper canopy, exposing the seedlings to higher light levels and fewer than 10% of our reintroduced seedlings remained one year later. Another issue was that we had little idea of what trees to affix these seedlings and what microhabitat conditions they needed (i.e., epiphytic associates, location on the tree, and the light levels required). The locations we selected for these plants were based on observations made by the staff at the refuge and students performing research on site. The FPNWR is home to about 27 orchid species in 17 genera with many of these species being threatened or endangered. It is possible that the survival of many of Fig. 3. Image of one transect at the study site. these species hinges on the habitat (i.e., tree species in the area), the microhabitat (i.e., substrate of establishment and epiphytic associates), and other factors (i.e., light level to which they are exposed). However, little to no data has been collected on the orchid microhabitats of these species at the refuge. This leads us to the study at hand. My project surveyed an area of the FPNWR for epiphytic orchids and the mircohabitats associated with them. The site was classified as a slough transitioning to a floodplain swamp and was believed to consist mainly of pop ash (Fraxinus caroliniana), pond apple (Annona glabra), and baldcypress (Taxodium distichum) for epiphytic orchids and catalogued the microhabitats associated with them. Some of the species I surveyed were orchids that I had worked with in the Orchid Recovery Program back in Illinois. Again, I was working in some of the same sites I had visited two years ago, but the terrain was slightly different. For one, the atmosphere was very different. The cooler fall weather had been replaced by the hot and very humid summer months. The site was no longer flooded and I could see where I was stepping most of the time. Despite this, I still encountered some obstacles. About once a week, I experienced tiny paper cuts on my exposed arms and legs, cuts that were the direct result of the very tall and sharp saw-grass (Cladium jamaicense), which in some spots was taller than me. The saw-grass was also an area of concern because alligators often find this habitat to be conducive for nest building. I encountered 3
33

Massey: AN UNDERGRADUATE‖S FIRST ADVENTURE INTO FIELD RESEARCH: AN EPIPHYTIC ORCHID SURVEY IN SOUTHERN FLORIDA

many more mosquitoes than I had in the fall and began each day by spraying myself with bug spray. Fortunately the only animals I came across were deer and a couple of harmless snakes. The data were collected in June and July of 2007 by another student, Cabrina Hamilton, and me. Data were collected for this project along 30 transects, 140 m long and 10 m apart for a total area sampled of 42,000 m2 (Fig. 3). An orchid was counted in the survey if it was within 1 meter from the ground (Fig. 4). A midday light measurement was also collected for each plant using a Sper Scientific, Broad Range LUX/FC meter (840022) and recorded in Lux. Some other data collected consisted of the phorophyte (i.e., a plant on which epiphytes grow) for each orchid, the substrate on which the orchid was established (i.e., moss, bark of host tree, lichens, or a combination of any two), the diameter of the part of the tree closest to the orchid was measured in centimeters, and the orientation of the orchid in regards to substrate tilt (i.e., located on the trunk, an angled or a horizontal limb, or on a fallen tree) along with the directionality of the orchid (i.e., facing N, E, W, S, NE, NW, SE, and SW). The orchid‖s epiphytic associates were measured (i.e., vascular plants like bromeliads and ferns and non-vascular organisms like lichens and mosses). We also subjectively determined the percentage of the area in the microhabitat they comprised and estimated the number of species present.

4

5

6

7

Figs. 4-7. Mature orchids sampled at the survey site. Campylocentrum pachyrrhizum is an example of a leafless orchid [5], Prosthechea cochleata var. triandra an orchid with leaves and visible pseudobulbs [6], and Epidendrum amphistomum an orchid with leaves and no visible pseudobulbs [7]. 34

Massey: AN UNDERGRADUATE‖S FIRST ADVENTURE INTO FIELD RESEARCH: AN EPIPHYTIC ORCHID SURVEY IN SOUTHERN FLORIDA

8

9

Figs. 8, 9. Mature orchids in flower during the study: Epidendrum amphistomum [8] and Polystachya concreta [9].

The orchids themselves were divided into three categories based on their morphological differences: leafless (Fig. 5) (i.e., Campylocentrum pachyrrhizum (Reichenbach f.) Rolfe, crooked-spur orchid; ribbon orchid and Harrisella porrecta (Reichenbach f.) Fawcett & Rendle, the leafless harrisella), orchids with leaves and visible pseudobulbs (Fig. 6) (i.e., Encyclia tampensis (Lindley) Small, the Florida butterfly orchid; P. concreta; and P. cochleata var. triandra), and orchids with leaves and no visible pseudobulbs (Fig. 7) (i.e., 7 8 6 Epidendrum amphistomum, E. nocturnum, and E. rigidum). They were further subdivided 9 into their stages of development. The plants without leaves were separated by the number of green roots: seedlings (>3 green roots), juveniles (3-5 green roots), and mature plants (with flowering or fruiting bodies or >5 green roots). The plants with leaves were separated into seedling (plant ≤0.5 cm), juvenile (plant ≥0.5 cm and ≤10 cm), and mature (flowering or fruiting bodies or plant ≥10 cm) plants. The number of green roots (leafless orchids) and the number5of green leaves were counted (orchids with leaves). If a plant was in flower or fruiting, 7 then we also counted the number of flowers and capsules. During the study the only orchids in flower were E. amphistomum (Fig. 8) and P. concreta (Fig. 9) and the only orchid seen in fruit was E. amphistomum. We sampled 419 orchids in total with a majority of the orchids surveyed being juveniles with fewer mature plants and seedlings. Of the mature plants, E. amphistomum were fruiting (2) and flowering (7). Polystachya concreta was also in flower (1). Most of the orchids surveyed were found on pop ash (Fraxinus caroliniana) (371) with 100%, 89%, and 88% of them being leafless, leaves with pseudobulbs, and leaves without pseudobulbs respectively. Leafless orchids were observed on trunks or branches <51 cm in diameter whereas orchids with leaves and visible pseudobulbs, as well as orchids with leaves and no visible pseudobulbs were noted on trunks and branches between 11-110 cm. All of the seedlings sampled occurred on moss, and it appeared that the juvenile and mature plants were either on moss or a combination of moss and bark. A majority of the epiphytic orchids without leaves were found on horizontal substrates, whereas orchids with leaves were affixed to branches/trunks at a 45 degree angle or a vertical position. Moreover, the majority of the orchids were oriented 4 on 8 substrates that received little direct sunlight (N, NE position). Many of the epiphytic 9
35

Massey: AN UNDERGRADUATE‖S FIRST ADVENTURE INTO FIELD RESEARCH: AN EPIPHYTIC ORCHID SURVEY IN SOUTHERN FLORIDA

associates consisted of mosses and ferns (e.g., resurrection fern, Pleopeltis spp.), as well as bromeliads, vines and occasionally lichens (Massey et. al., 2008). Taken together, it appears the orchids at this site are established on moss or a combination of moss and bark of the phorophyte, which is largely pop ash (F. caroliniana) with all of the seedlings being established on moss. These orchids were often facing in a northerly or northeasterly direction and either on branches or trunks tilted at an angle or vertical for the orchids with leaves and horizontal for the leafless orchids. Currently, the Orchid Recovery Program at Illinois College is propagating many of the orchid species surveyed in this study with the hopes of reintroducing them. Despite the thoroughness of this study, more research is needed before we can give orchids reintroduced in this area a fighting chance. For instance, the data in this study seem to indicate that the epiphytic orchids grow prominently on pop ash (F. caroliniana). We are unsure if this result is due to there being more trees of this species in the area or if there is a physical (i.e., bark texture, moisture capabilities, or the arrangement of the canopy cover) or chemical association between the orchids surveyed and this species of tree. A future study could survey the trees in this area or analyze the bark of all of the species of trees indicated in this study to better understand this relationship. Also it seemed that despite the orchids growing in close proximity to lichens few of them were established on or very close to lichens. Perhaps there is a reason for this dissociation. I suggest further studying of the orientation and tilt for these and other orchid species and more data collection of the phorophytes and branch diameter, especially for the orchids with leaves. In addition to these factors, studies regarding the stage of the orchid best suited for reintroduction should be assessed. Our data indicate that juveniles (i.e., plant ≥0.5 cm and ≤10 cm) were highly abundant at the site so perhaps plants should only be reintroduced if they are at a juvenile or mature plant stage. Further study of these threatened and endangered orchids is needed. Many of these species are in danger of being poached, having their habitats destroyed by humans and hurricanes, and having their territory encroached upon by exotic species. Hopefully, this study will promote future research aimed at improving the survival of both Florida orchids and other threatened and endangered species through reintroduction or better protection and management of their habitats. Acknowledgements
Many people were influential from the conception of this study to completion of this manuscript. Cabrina Hamilton for her aid in data collection, Dr. Lawrence Zettler for asking me to join his lab and fostering my love of research, Larry Richardson and the U.S. Fish and Wildlife Service for allowing me to come and work on the Florida Panther Refuge, Dr. Scott Stewart for helping with the formation of the study, Illinois College and the Charles and Dorothy Frank Scholarship for funding my study, Dr. Elizabeth Rellinger for her all of her patience in helping me with statistical analysis, the refuge staff for their assistance during the survey, and for everyone who spent their precious time reviewing this article. I kindly thank all of them for their support.

36

Massey: AN UNDERGRADUATE‖S FIRST ADVENTURE INTO FIELD RESEARCH: AN EPIPHYTIC ORCHID SURVEY IN SOUTHERN FLORIDA

Emily Massey, Department of Environmental Horticulture, University of Florida, PO Box 110675,
Gainesville, FL, 32611. emassey@ufl.edu As for my future plans, this study was a great experience and while I still enjoy laboratory work, it really sparked my interest in field research too. Currently, I am a graduate student enrolled at the University of Florida earning my Master‖s degree in Environmental Horticulture. My proposed project examines water relationships, specifically the affects of stress, and growth in two tree species. Although this is not based in ecological restoration, this study will provide a good basis for future research. Eventually I plan on returning to more ecologically based projects and securing a research position.

Literature Cited:
Massey, E.E., K. Hamilton, S.L. Stewart, L.W. Richardson, and L.W. Zettler. 2008. Substrate preferences of epiphytic orchids (seedlings, juveniles, mature plants) within the Florida Panther National Wildlife Refuge. Illinois State Academy of Science 101: 62-63. Massey, E.E. and L.W. Zettler. 2007. An expanded role for in vitro symbiotic seed germination as a conservation tool: Two case studies in North America (Platanthera leucophaea and Epidendrum nocturnum). Lankesteriana 7(1-2): 303-08.

37

Hammons, Smeins & Rogers: TRANSPLANT METHODS FOR SPIRANTHES PARKSII

Spiranthes parksii Navasota Ladies’-tresses Grimes County, Texas
38

Hammons, Smeins & Rogers: TRANSPLANT METHODS FOR SPIRANTHES PARKSII

TRANSPLANT METHODS FOR THE ENDANGERED ORCHID SPIRANTHES PARKSII CORRELL
J Ryan Hammons, Fred E. Smeins & William E. Rogers ABSTRACT
Spiranthes parksii (Navasota ladies‖ tresses) is an endangered terrestrial orchid endemic to the Post Oak Savanna ecosystem in central-east Texas. Methods of whole plant transplantation are needed to conserve individuals that will be destroyed by development activities. A soil-intact and a bare-root method were evaluated. Spiranthes parksii and its congener, S. cernua can be distinguished when in flower, but are indistinguishable from one another based on morphology of their leaf rosettes. Unknown leaf rosettes of S. parksii or S. cernua were transplanted into areas where S. parksii and S. cernua were known to co-occur. Compared to percent production of leaf rosette and flower production of undisturbed individuals on-site, transplanted individuals by both methods have been successful.

INTRODUCTION Spiranthes parksii, Navasota ladies’-tresses, is an endangered orchid endemic to centraleast Texas within the Post Oak Savanna Ecoregion where it co-occurs with its congener S. cernua which has a broad distribution across eastern North America (Pelchat, 2005; Brown, 2008). Spiranthes parksii has also been found further east in the Pineywoods Ecoregion, however, vegetation documented at these occurrences was similar to the Post Oak Savanna, and not typical of the Pineywoods (Bridges & Orzell, 1989). The Post Oak Savanna Ecoregion is dominated by native bunchgrasses and forbs with scattered clumps of trees and shrubs, primarily post oak (Quercus stellata) (TPWD, 2009). Other common woody species are blackjack oak (Quercus marilandica), black hickory (Carya texana), American beautyberry (Callicarpa americana), yaupon (Ilex vomitoria), farkleberry (Vaccinium arboreum), winged elm (Ulmus alata), eastern redcedar (Juniperus virginiana), and water oak (Quercus nigra) (Brezanson, 2009). Common grass species are little bluestem (Schizachyrium scoparium), other bluestems (Andropogon spp.), Indiangrass (Sorghastrum nutans), purpletop (Tridens flavus), curly threeawn (Aristida desmantha), and longleaf spikegrass (Chasmanthium sessilifloraum). This system was originally maintained as a savanna by frequent fires and grazing by bison, and with their absence, tree/shrub species increase and grasses/forbs decrease (TPWD, 2009). Within the Post Oak Savanna, Spiranthes parksii typically occurs on sparsely vegetated areas along the upper reaches of ephemeral and intermittent drainages. Individuals are also found away from drainages along game/livestock trails and/or in small herbaceous openings at a tree/shrub dripline where a herbaceous patch meets a tree/shrub community (Hammons, 2008; USFWS, 2009). Spiranthes cernua, nodding ladies’-tresses, also occurs in these habitats.

39

Hammons, Smeins & Rogers: TRANSPLANT METHODS FOR SPIRANTHES PARKSII

A solid waste landfill is needed for Bryan/College Station, Texas and surrounding areas. During construction, an estimated 379 Spiranthes parksii plants will be destroyed. In order to meet mitigation requirements, the United States Fish and Wildlife Service (USFWS) Biological Opinion required 57 hectares of deed restricted areas be purchased around the landfill footprint to protect and conserve S. parksii plants that occurred in those areas and to serve as recipient sites for transplanted individuals. As well, the Biological Opinion permits research to develop procedures for successful transplantation of at-risk plants to protected areas. It is our goal to explore soil-intact and bare-root methods of transplantation. METHODS Both Spiranthes parksii and S. cernua are perennial and produce a leafless inflorescence during mid-fall (Oct.-Nov.). A basal rosette of leaves is produced between November and April, which is followed by a dormant underground stage until the next flowering season. Identification of the two species is apparent during flowering; however, they cannot be differentiated during the leaf rosette stage of growth. All transplantations occurred at the end of leaf rosette growth to minimize disturbance during the growing period. Additionally, transplantation occurred when soil moisture was at field capacity. All were placed in deed restricted areas where other Spiranthes parksii/S. cernua flowering individuals were previously documented. Plant locations were marked in the field with survey flags and GPS positions so they could be re-visited to monitor survival. Additionally, several hundred undisturbed S. parksii/S. cernua leaf rosettes were marked in the same area to monitor survival compared with transplants. All transplanted individuals and between 22 and 540 undisturbed leaf rosettes were monitored for flowering and leaf rosette production each year after transplantation. Root Tuber Distribution and Bare-Root Transplantation Based on size and length of rosette leaves, six small and four large individuals were excavated in spring 2007. Length of each leaf and root tuber was measured, and each were summed to give total leaf length and total root tuber length to 1) determine if leaf size and root tuber size are correlated, and 2) determine the size and extent of root tubers so that root tubers would not be damaged during transplantation. For this study, a root tuber constitutes any underground structure growing from the bud zone. Soil was removed from the root tubers, individuals were wrapped in a wet paper towel and transplanted to deed restricted areas within two hours following excavation (Fig. 1). In spring 2008, an additional cohort of 57 Spiranthes cernua/S. parksii leaf rosettes and two known S. parksii were re-located and transplanted. In spring 2009, 14 known S. parksii individuals were transplanted. Of these, six had <5 cm of one root tuber taken for examination of mycorrhizal fungi infection and isolation in the laboratory.

40

Hammons, Smeins & Rogers: TRANSPLANT METHODS FOR SPIRANTHES PARKSII

Soil-intact Transplantation In spring 2007, a 20 cm diameter PVC pipe was used to excavate individuals while keeping the soil intact around root tubers. The PVC pipe was cut into 15 cm lengths and beveled at the bottom so it could be hammered into the soil around a leaf rosette. A shovel was then placed underneath the PVC pipe so that soil within the PVC pipe could be excavated. After excavation, plants were transplanted to deed restricted areas within approximately two hours. A hole was carefully dug in the deed restricted areas to fit the diameter and depth of the transplant inside the PVC pipe. After placing the transplant and PVC pipe in the pre dug hole, the PVC pipe was removed and soil was fed into the cracks around the transplant to fill any large air spaces (Fig. 2).

Fig. 1. Methodology for bare-root transplantation. a) shovel buried deep beneath plant and soil slightly raised, b) individual carefully taken out of soil with most soil removed so measurements could be taken, c) root tubers wrapped in a wet paper towel, and d) stored for transport to deed restricted areas.

RESULTS Root Tuber Demographics and Bare-Root Transplantation For the 10 bare-root transplants in spring 2007, total leaf length for the small individuals ranged from 5 to 11 cm, while total leaf length for the large individuals ranged from 22 to 32 cm. The number of root tubers per individual ranged from 2 to 8. Total leaf length and total root tuber length were positively correlated (R2 = 0.84; p= .000). The maximum depth of a root tuber from the base of the stem was 9 cm, while the maximum lateral distance was 8 cm. Root tubers were found to be both exhausted and not exhausted in S. parksii/S. cernua individuals, as noted by Wells et al. (1991; Fig. 3).
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Hammons, Smeins & Rogers: TRANSPLANT METHODS FOR SPIRANTHES PARKSII

Fig. 2. Methodology for soil-intact transplantation. a) PVC section centered around plant and hammered into ground, b) shovel slid underneath PVC section to be lifted out, c) transplants placed for transportation, and d) hole dug to fit PVC, transplant placed in pre-dug hole, PVC removed, and soil fed into cracks where PVC was to rid of any air spaces.

Fig. 3. Spiranthes rosette individual that does not have remnants of an exhausted root tuber (left) and one with two exhausted root tubers (right).

With the exception of leaf rosette production in 2008, subsequent production of the 10 bare-root transplants have had a higher percent production than undisturbed Spiranthes cernua/S. parksii individuals also originally found in spring 2007 (Fig 4). Individual plants
42

Hammons, Smeins & Rogers: TRANSPLANT METHODS FOR SPIRANTHES PARKSII

show no consistent pattern. One individual remained dormant for 2 flowering and 2 leaf rosette stages, but emerged as a leaf rosette in fall 2009. Another has formed a flowering stalk and leaf rosette for all stages of growth monitored thus far. However, none flowered as S. parksii. Soil-Intact Transplants – Spring 2007 Flower and leaf rosette production of soil-intact transplants has been similar to undisturbed Spiranthes parksii/S. cernua leaf rosettes on site, and has surpassed percent production of undisturbed individuals (Fig. 4). These plants have also exhibited considerable variability. Some have remained dormant for as many as four growing seasons before emerging as a flowering stalk or leaf rosette. Five flowered as S. parksii, of which two have flowered all three consecutive years. Other individuals have flowered as S. cernua or remain unknown as to the species due to herbivory before identification could be confirmed.

Fig. 4. Percent production of S. parksii/S. cernua transplanted and undisturbed leaf rosettes (spring 2007) each growing season post-transplantation. Numbers in bars represent the number of individuals observed each growing season.

Bare-Root Transplants – Spring 2008 Percent leaf rosette and flower production of these transplants have been consistently lower than undisturbed leaf rosettes on site (Fig. 5). However, one individual flowered as Spiranthes parksii in 2008, and other individuals are still producing vegetatively including one of the known S. parksii. Sixteen appeared to be destroyed by feral hogs during winter of 2008 after transplantation. Despite this disturbance, three individuals transplanted the area emerged as leaf rosettes in 2009.
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Hammons, Smeins & Rogers: TRANSPLANT METHODS FOR SPIRANTHES PARKSII

Fig. 5. Percent production of S. parksii/S. cernua transplants and undisturbed leaf rosettes each growing season post-transplantation. Numbers in or above bars represent the number of individuals observed each growing season.

Spiranthes parksii Bare-Root Transplants – Spring 2009 Four of the 14 (28%) flowered and four (28%) produced a rosette of leaves in fall 2009 following transplantation. One of the six which had <5 cm of a root tuber taken flowered, while three produced a rosette of leaves. DISCUSSION Transplanted individuals for both methods appeared in subsequent years. Bare-root transplants from spring 2007 have had a higher percentage of post-production than those in spring 2008 and soil-intact in spring 2007. However, this could be due to a low sample size. While percentages are consistently lower for bare-root transplants in spring 2008, one of 59 has flowered as Spiranthes parksii and some are still persisting vegetatively. Soil-intact transplants have produced the best results for S. parksii since five individuals have flowered as S. parksii, of which two have flowered all three flowering seasons monitored. Additionally, percent production of these has been higher than undisturbed plants in the last three growing seasons. Bare-root transplantation of 14 known S. parksii from spring 2009 which flowered at least once in the previous two years have produced inflorescences and leaf rosettes after transplantation, including those that had <5 cm of a root tuber removed.

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Hammons, Smeins & Rogers: TRANSPLANT METHODS FOR SPIRANTHES PARKSII

Pileri (1998) noted that after excavating five Spiranthes cernua plants to analyze the root tubers for mycorrhizal infection, all but one plant that was destroyed by a small mammal survived transplantation by reappearing the next year. She also noted that they were better able to survive when transplanted during the vegetative or early reproductive phases. However, others believe, or have found, that bare-root transplanting of terrestrial orchids is unsuccessful (Ferry, 2008; Steinauer, 2008). In this study, S. cernua (spring 2007) and S. parksii (spring 2008 and spring 2009) responded positively to bare-root transplantation. The three S. cernua that flowered after bare-root transplantation in spring 2007 were of the larger leaf rosettes. The success of these could be due to large underground root tubers which could be used to offset the effects of disturbance caused by transplantation. As well, mature S. parksii transplanted in spring 2009 could also be using underground reserves to offset the effects of transplantation. Previous efforts of soil-intact transplantation of Spiranthes parksii using 15 cm diameter irrigation pipe at the TMPA Gibbons Creek Lignite Mine conservation areas yielded positive results (Parker 2006). However, quantitative data and long-term observations were not made. Efforts of soil-intact transplanting in other terrestrial orchids have been unsuccessful, as with Isotria medeoloides (Brumbeck, 1996). In this study, both Spiranthes parksii and S. cernua responded well to this method. In fact, compared to undisturbed leaf rosettes at the study site, percent production of soil-intact transplants have been greater in the last three growing seasons. This might be due to placement of transplants since they were placed in areas of ideal habitat of S. parksii/S. cernua. Undisturbed leaf rosettes may be persisting in areas which have become unfavorable for flowering due to woody encroachment. However, quantitative data would need to be collected to verify this. While all transplants were placed in areas where Spiranthes parksii/S. cernua occurred, placement could possibly be influencing post-production since microhabitats vary greatly within a savanna patchwork. Additionally, initial size of leaf rosettes prior to transplantation could affect post-production. However, detailed analysis of microhabitats and plant sizes would need to be conducted to pursue these hypotheses. CONCLUSIONS While both methods of transplantation have yielded positive post-production in individuals, if given the time and labor, the soil-intact method would be preferred. Not only has this method yielded higher survival, but the intact soil may contain tubers of plants other than the target individual. Upon digging up one Spiranthes parksii for bare-root transplanting in spring 2009, another individual was found dormant as a root tuber. This was also seen when taking soil samples around individual plants. Upon returning to the laboratory to sieve soil samples, a Spiranthes spp. root tuber was found. Comparison of transplanted individuals to undisturbed plants of the same species is critical in giving accurate results of success or failure. If given only the results of transplanted individuals in this study, one might conclude individuals are dying due to transplantation.
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Hammons, Smeins & Rogers: TRANSPLANT METHODS FOR SPIRANTHES PARKSII

However, transplanted and undisturbed individuals have both declined and/or fluctuated in subsequent production after transplanting was initiated. Long-term monitoring of these individuals is crucial to clarify life history characteristics and environmental variables that influence the persistence of undisturbed and transplanted individuals.
ACKNOWLEDGEMENTS We would like to thank the Brazos Valley Solid Waste Management Agency (BVSWMA) for funding and HDR, Inc. for assistance with this research. Individuals to thank are Linda Langlitz, Josh Grace, Martha Ariza, and Trey Witcher for their assistance in transplanting and monitoring of individual plants.

J Ryan Hammons, Fred E. Smeins & William E. Rogers
Department of Ecosystem Science and Management, Texas A&M University, College Station, Tex. ryanhammons2000@yahoo.com

LITERATURE CITED
Brown, P.M. & S.N. Folsom. 2008. Field Guide to the Wild Orchids of Texas. Gainesville: University Press of Florida. Bezanson, D. 2000. Natural Vegetation Types of Texas and Their Representation in Conservation Areas. The University of Texas at Austin. http://www.abisw.org/bezanson/ Bridges, E.L. and S.L. Orzell. 1989. Additions of Noteworthy Vascular Plant Collections from Texas and Louisiana, with Historical, Ecological and Geographical Notes. Phytologia 66: 12-69. Brumback, W.E. and C.W. Fyler. 1996. Small Whorled Pogonia (Isotria medeoloides) Transplant Project. In Falk, D.A., C.I. Millar, and M. Olwell. 1996. Restoring Diversity: Strategies for Reintroduction of Endangered Plants. Washington, D.C.: Island Press, Ferry, R.J. 2008. Relocating Terrestrial Orchid Plants. North American Native Orchid Journal 14: 179-82. Hammons, J.R. 2008. Demographic, Life Cycle, Habitat Characterization and Transplant Methods for the endangered orchid, Spiranthes parksii Correll. M.S. Thesis, Department of Rangeland Ecology and Management, Texas A&M University, College Station, Texas. Parker, K.M. 2006. Personal communication. Texas Ecological Services, College Station, Texas. Pelchat, C. 2005. Spiranthes parksii Correll – Navasota Ladies‖ Tresses. McAllen International Orchid Society Journal 6: 9-15. Pileri, V.S. 1998. Root morphology, distribution of mycorrhizae, and nutrient status of the terrestrial orchid Spiranthes cernua. M.S. Thesis, Department of Biology, University of Nebraska at Omaha, Omaha, Nebraska. Steinauer, G. 2008. Transplanting a Rare Orchid. Nebraska Game and Parks Commission Annual Report of the Wildlife Conservation Fund. Texas Parks and Wildlife Department. Accessed 2009. Post Oak Savanna and Blackland Prairie Wildlife Management. http://www.tpwd.state.tx.us/landwater/land/habitats/post_oak/ United States Fish and Wildlife Service. 2009. Navasota Ladies‖-Tresses (Spiranthes parksii) 5-Year Review: Summary and Evaluation. Austin Ecological Services Field Office, Austin, Tex.

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PROPAGATION AND CONSERVATION STATUS OF THE NATIVE ORCHIDS OF THE UNITED STATES (INCLUDING SELECTED POSSESIONS), CANADA, ST. PIERRE ET MIQUELON, AND GREENLAND
Scott Stewart, Ph.D. & Aaron Hicks INTRODUCTION Conservation of biodiversity has become a primary biological, economic, and humanistic concern as the global community faces the sixth great extinction event in the Earth's history (Canadell and Noble, 2001). The implementation of conservation efforts must begin with careful planning, otherwise we risk the Johnny Appleseed effect of biodiversity conservation—we scatter our efforts into the wind and whichever efforts result in fruitful conservation we consider successful and all others we consider unproductive. This naive approach to the conservation of biodiversity runs the risk of missing important biotic components of global biodiversity. This paper is an attempt to gather verifiable propagation, cultivation, and conservation status information on the native orchids of the United States, Canada, and associated foreign lands in one document that may be used to guide future orchid conservation efforts in these regions. We have chosen to include all orchid species and varieties considered native to the United States, Canada, Puerto Rico, the U.S. Virgin Islands, Guam, Saint Pierre et Miquelon islands, and Greenland. Species and varieties considered as introduced, exotic, escaped from cultivation, and waifs have been excluded. Color and growth forms have also been excluded as their taxonomic status and genetic stability are often controversial. All propagation and conservation data has been verified through scientific publications and personal communications with experts in orchid propagation, cultivation, and conservation. In general, the most recent taxonomic checklist proposed by Brown (2009) has been used throughout. The current work represents the first effort to gather such a volume of specific information for such a large number of species and widespread geographic area, and should be considered a working draft. The authors invite comments and additional verifiable data from readers.

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GEOGRAPHIC ANALYSIS Although national boundaries are political rather than biological, a brief analysis may be enlightening with respect to the conservation of orchid species. From a financial standpoint, the geographic areas reported here encompass a significant proportion of global productivity (Table 1).
Table 1—Gross domestic products of major regions in current work. Data summarized from World Bank, World Development Indicators, and CIA World Factbook. Country United States Canada Puerto Rico US Virgin Islands Guam Total Gross Domestic Product (US $ billions) 14200 (2008) 1400 (2008) 67.9 (2001) 2 (1993) 2.5 (2005 est.) 15672

With approximately 410 taxa, this comes out to about $38.3 billion in average domestic productivity per species in the geographic regions surveyed. A conservative figure for the number of species globally is approximately 24,000 taxa, with the regions surveyed here making up only 1.7% of the total. With global productivity calculated at $60.6 trillion in 2008 (World Bank), a figure of $2.52 billion in productivity per species is reached, a substantially smaller figure. To paraphrase the International Union for Conservation of Nature (IUCN), if orchid growers cannot pull a plant back from the brink of extinction, what hope is there for other plant families? To extend this statement, if economic powerhouses with all their resources cannot preserve their own species—which are relatively few in number when compared to the global diversity—what hope is there for other orchids? Hawaii Despite the abunance of hybrids and introduced species, the true native orchid taxa of Hawaii are limited to three species, one of which (Platanthera holochila, puahala-a-kane; Fig. 1) is listed as

Fig. 1. Platanthera holochila (puahala-a-kane) in natural habitat in Hawaii.
L. Zettler

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threatened under the U.S. Endangered Species Act. There has been some propagation work with this species—symbiotic germination efforts using mycobionts isolated from local Hawaiian populations were unsuccessful while symbiotic efforts using non-Hawaiian mycobionts have been successful, and there was no desire to introduce non-native mycobionts from outside the islands. There has been reasonable success in propagating the species using asymbiotic methods (McDonald et al., 2006; L. Zettler, personal communication). Guam Of the species native to Guam, only a small number have been successfully propagated and brought into horticultural cultivation. Little is known of the plants of the island, although the genera represented should be considered generally straightforward in asymbiotic culture systems. It seems likely they would present few difficulties in terms of artificial propagation. Nothing is known of the natural mycobionts or symbiotic culture requirements of the orchids of Guam. Western United States and Canada

Fig. 2. Cypripedium californicum (California lady‖s-slipper) photographed in southern Oregon, U.S.A.
S.L. Stewart

This region presents myriad natural biomes: ranging from desert, to Pacific rain forest, to true Arctic environments in the northernmost portions of Alaska. Several species native to this region are showy (Cypripedium californicum, California lady's-slipper; Fig. 2) and warrant additional propagation effort. Even in the arid desert states orchids are surprisingly well-represented, found in all but three and ten counties of Arizona and New Mexico respectively (Coleman, 2002).
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Federally protected species of interest in this region include Piperia yadonii (Yadon's piperia), which has eluded attempts at cultivation, although there has been some recent interest in the ecology and propagation of the species (George et al., 2009; Sharma et al., 2007; R. Buck, personal communication). A locally endemic species, Spiranthes delitescens (Canelo Hills ladies'-tresses), has proven to be remarkably easy to grow in culture (Hicks, 2007), producing large numbers of plants from friable callus when stressed in sterile tissue culture. Seedlings have flowered in cultivation at University of Arizona. A total of four populations are known, although a fifth has been reported (M. Falk, personal communication). Another species of interest in the region is Spiranthes infernalis (Ash Meadows ladies'tresses), known from a cluster of populations over an area of approximately 28 acres in Nye County, Nevada. Estimates as to its total population numbers vary, but the species‖ global population is estimated in the low one thousands, possibly as low as 1107 (Morefield, 2001). There has reportedly been an effort to propagate the species at Royal Botanic Gardens at Kew. Central United States and Canada With the change of the American prairie, Platanthera leucophaea (eastern prairie fringed orchid; Fig. 3) has dwindled in numbers and is currently listed as Federally threatened. Asymbiotic efforts to propagate the species have met with limited success (Stoutamire, 1996); however, symbiotic propagation efforts have been highly successful (Stewart, 2000; Zettler, 1999; Zettler et al., 2005; Zettler et al., 2001). Also afforded Federally threatened status, P. praeclara has been the subject of extensive asymbiotic and symbiotic propagation efforts. Asymbiotic efforts with P. praeclara have been met with reasonable success after lengthy culture periods and multiple cold treatments of in vitro seed (From and Read, 1998). Sharma et al. (2003) reported the successful symbiotic propagation of the species. Also native to this region is the Federally threatened Texas endemic Spiranthes parksii (Navasota ladies’-tresses). There has been some success in propagating this species from seed by researchers at the Atlanta Botanical Garden. Wilson (web page) reports the species has proven to be remarkably easy to cultivate using asymbiotic techniques. Additional asymbiotic and symbiotic propagation efforts are currently underway (R. Hammons, personal communication).
50

Fig. 3. Platanthera leucophaea (eastern prairie fringed orchid) photographed in southern Wisconsin, U.S.A.
P. M. Brown.

Stewart & Hicks: PROPAGATION AND CONSERVATION STATUS OF NATIVE ORCHIDS

Eastern United States and Canada, excluding Florida The eastern United States are home to a number of Platanthera species that are statelisted as endangered, in many cases. Many of these species have proven to be resistant to traditional propagation techniques, and success has been sporadic to nonexistent. Similar to P. leucophaea, many species have been severely impacted by a variety of anthropogenic factors such as urbanization, agriculture, and fire suppression. In addition to the many Platanthera species of this region, Isotria medeoloides (small whorled pogonia), known from the eastern United States, is listed as Federally threatened, and had been the focus of many ecological and horticultural studies. The species has resisted numerous attempts at artificial propagation; however, new efforts by the Smithsonian Environmental Research Center and the U.S. Park Service are planned (J. O'Neill, personal communication). Florida With more than a hundred species known from Florida alone, it is practical to treat the state as a separate entity for conservation purposes. In addition to the intrinsic diversity present in Florida is the urban development the state has undergone, in conjunction with the influx of invasive species that further threatens the state's native species. Perhaps of greatest interest is the leafless Dendrophylax lindenii (ghost orchid; Fig. 4), which is an endangered species under state law. However, the plant has proven to be remarkably easy to grow in vitro, provided seeds can be reliably produced. Unfortunately, mortality is high amongst deflasked plantlets and growth is slow; it seems likely there is some aspect of the species' cultivation that remains cryptic such that it may eventually be brought into cultivation—although this is a theme that has been repeated for many decades without realization (Correll, 1978). Afforded similar state-level protection is Cyrtopodium punctatum (cigar orchid), which forms large (to 1.5 meter), robust plants whose populations have been decimated by poaching and habitat
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Fig. 4. Dendrophylax lindenii (ghost orchid) photographed in southwestern Florida, U.S.A.
S. L. Stewart

Stewart & Hicks: PROPAGATION AND CONSERVATION STATUS OF NATIVE ORCHIDS

alteration. However, the species forms large capsules producing similarly large quantities of seeds that germinate and grow on a variety of media with no special requirements in asymbiotic culture. From a mechanical standpoint, the roots are not like those of most orchids, forming tangled mats in vitro, resulting in damage when deflasked. From this, it may be best to plant seedlings in individual tubes. Considerable research effort has been focused on the asymbiotic propagation, reproductive biology, and potential for reintroduction of the cigar orchid (Dutra, 2008; Dutra et al., 2009a,b). Preliminary symbiotic germination and reintroduction of the species has been successful (Stewart and Richardson, 2008; S. Stewart, unpublished data). Another state-listed endangered species is Basiphyllaea corallicola (Carter's orchid), which is easy to propagate asymbiotically, forming new pseudobulbs with fresh shoots every few months. Similarly protected, both Epidendrum nocturnum (night-fragrant epidendrum) and Macradenia lutescens (Trinidad macradenia) offer no difficulties in vitro. A symbiotic propagation protocol has even been developed for E. nocturnum (Zettler et al., 2007) and preliminary reintroductions have taken place (Stewart, 2008). Another species whose numbers have declined to the point where it has been listed as endangered by the state is Tolumnia bahamensis (Florida's dancing lady); germination is straightforward, while subsequent culture is made difficult by the usual cultural quirks within the Oncidiinae in that differentiation is slow, resulting in large clusters of protocorms without roots. These problems usually resolve with subsequent replating, resulting in large numbers of plants that eventually form stout seedlings with good roots. Mortality is high when deflasked. Bletia purpurea (pine-pink) is another Floridian species whose populations face decline as urbanization in southern Florida increases. The germination, in vitro culture, and transfer to the greenhouse of the species is quite easy (Dutra et al., 2008); however, the species is rarely seen in the commercial marketplace. Although not afforded protection, Eulophia alta (wild coco) is a strong grower in sterile flask, although others have noted that it germinates and grows quite readily from seed sown directly in rich earth. Johnson et al. (2007) presented a side-by-side comparison of asymbiotic versus symbiotic germination this species, demonstrating preliminary evidence of an advantage during symbiotic germination and subsequent in vitro seedling development. Several other Florida native species are considered endangered, and lack active conservation efforts, such as Beloglottis costaricensis (Costa Rican ladies'-tresses), Bulbophyllum pachyrachis (rat-tail orchid), Cyclopogon cranichoides (speckled ladies'-tresses), Epidendrum amphistomum (dingy-flowered star orchid), Ionopsis utricularioides (delicate ionopsis), Liparis elata (tall twayblade), Mesadenus lucayanus (copper ladies'-tresses), Polystachya concreta (pale-flowered polystachya), Vanilla barbellata (worm-vine), and Vanilla mexicana (thin-leaved vanilla), among others. The State of Florida affords some 54 taxa endangered status, 16 more are protected as threatened, and two more—Encyclia tampensis (Florida butterfly orchid) and Epidendrum magnoliae var. magnoliae (green-fly orchid; syn. Epidendrum conopseum)—are protected as commercially exploited.
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A number of Florida native orchids are also found throughout the Greater and Lesser Antilles—including Cuba, Puerto Rico, and the U.S. Virgin Islands. Cuba is not treated in this paper. While the degree of this wider geographic distribution can be seen in comparing the native orchid flora of Florida to that of Puerto Rico, other islands in the Antilles may also possess some species known from Florida. For example, Dendrophylax lindenii, Cyrtopodium punctatum, Eulophia alta, and Bletia purpurea are all known from Cuba and Florida (Llamacho and Larramendi, 2005). Puerto Rico Much of the primary forest of Puerto Rico was removed early in the 20th century and it is difficult to state with certainty that any species of orchid were lost. However, two endemic species—Lepanthes caritensis (Carite babyroot orchid) and the Federally endangered Lepanthes eltoroensis (Luquillo Mountain babyroot orchid)—are known only from very small populations (Tremblay et al., 1998) and a third Puerto Rican orchid, Cranichis ricartii (Puerto Rico helmet orchid), is Federally endangered and may be extirpated. Relatively little is known about the propagation of most orchids found in Puerto Rico. U.S. Virgin Islands Twenty-six orchid species are known from the U.S. Virgin Islands, all of which are also found in Puerto Rico (Ackerman, 1995). St. Thomas is the most species-rich with 23 species, followed by St. John (13 species), and St. Croix (9 species). Taken as a whole, 29 species are known from both the U.S. and British Virgin Islands. Only Eulophia alta and Erythrodes hirtella (false helmet orchid) are known from the British Virgin Islands and not from the U.S. Virgin Islands. Ackerman (1995) commented that additional species may have been present in both the U.S. And British Virgin Islands at one time, but rapid urbanization on the populated islands may have caused some species to become extirpated from the islands. As in Puerto Rico and Florida, the U.S. Virgin Islands also have a number of exotic orchids that have escaped cultivation and become established on various islands, including Spathoglottis plicata (Philippine ground orchid; St. Thomas), Vanilla mexicana (Mexican vanilla; St. Croix), and V. planifolia (commercial vanilla; St. Croix, St. John, and St. Thomas). HORTICULTURAL NOTES Several of the genera of interest here remain recalcitrant to existing propagation techniques. Genera such as Corallorhiza remain cryptic in their in vitro cultural needs, although Jay O'Neill (personal communication) notes that sporadic germination on commercial media has occurred, while semi-wild field plantings have been met with good success, including one recruit growing to flower. The reliance of mycotropic interactions for the seed germination and plant development of saprophytic terrestrial genera (i.e., Corallorhiza, Hexalectris) may be the root cause for their difficulty in in vitro propagation. The genus Cypripedium is by far the most successful in terms of propagation and commercialization, with a number of species being made available for sale on a regular basis. In fact, the majority of the North American species are available through a number of
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reputable companies as mature plants, and it is expected that this market has considerable potential for gardeners that would like to engage in the horticultural cultivation of these species. Several species have been presented as candidates for commercial propagation; prices and availability have improved over the past several years. The genus is desirable and conspicuous, making it an ideal subject for commercialization in order to potentially reduce the collection of plants from the wild. Cyrtopodium punctatum is perhaps one of the most underrepresented species with respect to propagation. It is so fecund and such a strong grower that it would seem to be an ideal candidate for commercial exploitation in its native state of Florida. The decimation of wild populations of the species implies its desirability could prove useful to nurseries that are willing to produce specimens for local growers. Similarly, Eulophia alta, Epidendrum nocturnum, and Encyclia tampensis are readily propagated; Eulophia alta could probably be introduced into popular culture much in the same way as the other two have. Occasionally available from commercial vendors, Epipactis gigantea Fig. 5. Calypso bulbosa var. occidentalis (western fairy-slipper) photographed in southern Oregon, U.S.A. (stream orchid) is an ideal S.L. Stewart candidate for western gardens. Both Calypso bulbosa var. americana (eastern fairy-slipper) and C. bulbosa var. occidentalis (western fairy-slipper; Fig. 5) are diminutive, but showy, native orchids with tremendous horticultural potential. The species has frustrated many individuals attempting in vitro propagation with sporadic germination and limited subsequent growth. Ashmore (1999) has reported the successful in vitro asymbiotic propagation and ex vitro cultivation of both Calypso varieties, and numerous man-made Calypso hybrids. The scaling-up of such work
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would be required before Calypso would be regularly available to gardeners; however, such propagation, hybridization, and horticultural selection work are important first-steps toward commercialization of these showy native orchids. The genus Goodyera has been popular for many years as a plant for culture in terraria; other allied members of the genus have been available as jewel orchids in the trade for years, with few—if any—from artificial propagation. Propagation of jewel orchids for the purposes of horticulture is most commonly by division. It seems likely most, if not all, of the native Goodyera species in the horticultural trade have been wild-collected. It would be desirable to provide for this genus in artificial propagation as well. The genus Habenaria is almost unknown with respect to commercial asymbiotic seed propagation, as with members of the genus Listera, Malaxis, Hexalectris, Piperia, Platanthera, and Spiranthes. There is little to say about this otherwise unrelated group other than to say that new techniques, media, and mycobionts will have to be developed to master this group of plants. The exception would be the genus Spiranthes, which has proven to be prolific in sterile culture for the most part. The propagation of Habenaria species has received considerable attention in recent years, particularly as a part of habitat or integrated conservation efforts in Florida (Stewart, 2007; Stewart and Kane, 2006a, b; Stewart and Zettler, 2002). Tropical epiphytic genera, including Bulbophyllum, Dendrobium, Encyclia, Epidendrum, Lepanthes, Maxillaria, Oncidium, Pleurothallis, and so forth, have a better outlook than terrestrial species, if for no better reason conventional commercial orchid propagation laboratories are better suited to the propagation of these plants, many of which can be produced in large numbers if the economic incentives are present, and fresh, clean material can be produced. CONCLUDING REMARKS In total, there are six species afforded Federal protection as endangered species, the highest level of protection: Cranichis ricartii, Lepanthes eltoroensis, Piperia yadonii, Platanthera holochila, Spiranthes delitescens, and Spiranthes parksii. Three species—Isotria medeoloides, Platanthera praeclara, and P. leucophaea—are considered Federally threatened. Of these, only four—Spiranthes delitescens, S. parksii, Platanthera praeclara, and P. leucophaea—have been artificially produced in substantial quantities. Several other species are limited to small colonies in single geographic areas (Platanthera pallida, Spiranthes infernalis), exist as exceedingly small populations (Lepanthes caritensis), have cryptic germination requirements (Calypso bulbosa, Isotria medeoloides, Platanthera spp.), are achlorophyllous (Corallorhiza spp.), or survive transfer from sterile tissue culture rarely or not at all (Dendrophylax lindenii, some Cypripedium spp.). Indeed, listing or upgrading some species from threatened to endangered status is probably merited, although from a conservation perspective this makes attempts to propagate these species markedly more difficult. While most species that have been targeted in propagation efforts have been brought into cultivation, a few remain refractory to existing techniques—or the material provided for
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propagation has been so scant that insufficient experiments could be run. A few, such as Platanthera species, have sporadic success that is reminiscent of that with Paphiopedilum species; if this is the case, perhaps we will see enhanced germination with subsequent generations, with each passage through sterile flask (or symbiotic culture) selecting those seeds that are likely to germinate under such conditions. It is our hope that, given the resources of the geographic region discussed here, all species could eventually be propagated and made available for culture in public and private gardens, or reintroduction if the need arises. Asymbiotic and symbiotic propagation status and current conservation status for species found within the geographic region of the United States, Canada, Puerto Rico, the U.S. Virgin Islands, Guam, Greenland, and Saint Pierre et Miquelon is found in Table 2. ACKNOWLEDGEMENTS The authors would like to thank the numerous individuals who contributed personal communications and information for this paper. Paul Martin Brown was instrumental during editing and revision of this work. Scott Stewart, Ph.D., Director, Horticulture & Agriculture Programs, Kankakee Community College,
Kankakee, IL 60901 sstewart@kcc.edu Aaron Hicks, The Orchid Seedbank Project, P.O. Box 7042, Chandler, AZ 85246, ahicks51@cox.net

REFERENCES FOR TEXT
Ashmore, S. 1999. Calypso bulbosa—hybridization and cultivation. North American Native Orchid Journal 5:27-32. Buck, R. 2006. Personal communication with A. Hicks. Brown, P.M. 2009. Personal checklist of the wild orchids of North American, north of Mexico. North American Native Orchid Journal Special Publication #4. Canadell, J. and I. Noble. 2001. Challenges of a changing earth. Trends in Ecology and Evolution 16:664-66. Coleman, R. 2002. The Wild Orchids of Arizona and New Mexico. Ithaca, N.Y.: Comstock Books. Cornell University Press. Correll, D.S. 1978. Native Orchids of North America North of Mexico. Palo Alto, Calif.: Stanford University Press, Dutra, D. 2008. Reproductive biology and asymbiotic seed germination of Cyrtopodium punctatum: an endangered Florida native orchid. Masters Thesis, University of Florida. Dutra, D., M.E. Kane, and L. Richardson. 2009a. Asymbiotic seed germination and in vitro seedling development of Cyrtopodium punctatum: a propagation protocol for an endangered Florida native orchid. Plant Cell, Tissue and Organ Culture 96:235-43. Dutra, D., M.E. Kane, C. Reinhardt Adams and L. Richardson. 2009b. Reproductive biology of Cyrtopodium punctatum in situ: implications for conservation of an endangered orchid. Plant Species Biology 24:92-103. Dutra, D., T.R. Johnson, P.J. Kauth, S.L. Stewart, M.E. Kane, and L. Richardson. 2008. Asymbiotic seed germination, in vitro development, and greenhouse acclimatization of the threatened terrestrial orchid Bletia purpurea. Plant Cell, Tissue and Organ Culture 94:11-21. Falk, M. 2008. Personal communication with A. Hicks. From, M. and P. Read. 1998. Platanthera praeclara: strategies for conservation and propagation. North American Native Orchid Journal 4:299-312. George, S., J. Sharma, and V.L. Yadon. 2009. Genetic diversity of the endangered and narrow endemic Piperia yadonii (Orchidaceae) assessed with ISSR polymorphisms. American Journal of Botany 96:2022-30. Hammons, R. 2009. Personal communication with S. Stewart. Hicks, A. 2007. On the germination and subsequent culture of Spiranthes delitescens Sheviak in sterile culture. Orchid Digest 71:158-60. 56

Stewart & Hicks: PROPAGATION AND CONSERVATION STATUS OF NATIVE ORCHIDS Johnson, T.R., S.L. Stewart, D. Dutra, M.E. Kane, L. Richardson. 2007. Asymbiotic and symbiotic seed germination of Eulophia alta (Orchidaceae)—preliminary evidence for the symbiotic culture advantage. Plant Cell, Tissue and Organ Culture 90: 313-23. Llamacho, J.A. and J.A. Larramendi. 2005. The Orchids of Cuba. Lleida: Spain.Greta Editores, McDonald, K., S. Hopkins, S. Perlman, and L.W. Zettler. 2006. The status and propagation of the Federally endangered Hawaiian endemic, Platanthera holochila (Orchidaceae). Southeastern Biology 53:209. Morefield, J.D. 2001. Nevada Rare Plant Atlas. Nevada Natural Heritage Program, Carson City, Nevada. O'Neill, J. 2008. Personal communication with A. Hicks. Sharma, J., M.L. Ishida, and V.L. Yadon. 2007. Mycorrhizal diversity of an endemic terrestrial orchid. Lankesteriana 7: 215-18. Sharma, J., L.W. Zettler, J.W. Van Sambeek, M.R. Ellersieck, and C.J. Starbuck. 2003. Symbiotic seed germination and mycorrhizae of the Federally threatened Platanthera praeclara (Orchidaceae). American Midland Naturalist 149: 104-20. Stewart, S.L. 2000. Symbiotic seed germination of the Federally threatened eastern prairie fringed orchid, Platanthera leucophaea (Nuttall) Lindley, and three Habenaria species from Florida. North American Native Orchid Journal 6: 180-92. Stewart, S.L. 2007. Integrated conservation of Florida Orchidaceae in the genera Habenaria and Spiranthes: model orchid conservation systems for the Americas. Ph.D. Dissertation, University of Florida. Stewart, S.L. 2008. Orchid reintroduction in the United States: a mini-review. North American Native Orchid Journal 14: 54-59. Stewart, S.L. and L.W. Richardson. 2008. Orchid flora of the Florida Panther National Wildlife Refuge. North American Native Orchid Journal 14: 70-104. Stewart, S.L. and M.E. Kane. 2006a. Asymbiotic seed germination and in vitro seedling development of Habenaria macroceratitis (Orchidaceae), a rare Florida terrestrial orchid. Plant Cell, Tissue and Organ Culture 86: 147-58. Stewart, S.L. and M.E. Kane. 2006b. Symbiotic seed germination of Habenaria macroceratitis (Orchidaceae), a rare Florida terrestrial orchid. Plant Cell, Tissue and Organ Culture 86: 159-67. Stewart, S.L. and L.W. Zettler. 2002. Symbiotic germination of three semi-aquatic rein orchids (Habenaria repens, H. quinqueseta, H. macroceratitis) from Florida. Aquatic Botany 72: 25-35. Stoutamire, W. 1996. Seeds and seedlings of Platanthera leucophaea (Orchidaceae). In: C. Allen (Ed.), North American Native Terrestrial Orchids, Propagation and Production. North American Native Terrestrial Orchid Conference, Germantown, Maryland, pp. 55-61. Tremblay R.L., J.K. Zimmerman, L. Lebrón, P. Hayman, I. Sastre, F. Axelrod, and J. Alers-Garcia. 1998. Host specificity and low reproductive success in the rare endemic Puerto Rican orchid Lepanthes caritensis. Biological Conservation 85: 297-304. Wilson, H.D. Spiranthes parksii—endangered orchid of the Texas post oak savannah. http://botany.csdl.tamu.edu/FLORA/hdwsp/sp_pro.htm, accessed 6 December 2009. Zettler, L. 2009. Personal communication with S.L. Stewart and A. Hicks. Zettler, LW. 1999. A report on the use of fungi to germinate seeds of Platanthera integra, P. leucophaea, Spiranthes ovalis var. erostellata, and Encyclia tampensis. North American Native Orchid Journal 5: 232-47. Zettler, L.W., S.B. Poulter, K.I. McDonald, and S.L. Stewart. 2007. Conservation-driven propagation of an epiphytic orchid (Epidendrum nocturnum) with a mycorrhizal fungus. HortScience 42: 135-39. Zettler, L.W., K.A. Piskin, S.L. Stewart, J.J. Hartsock, M.L. Bowles, and T.J. Bell. 2005. Protocorm mycobionts of the Federally threatened eastern prairie fringed orchid, Platanthera leucophaea (Nutt.) Lindley, and a technqiue to prompt leaf elongation in seedlings. Studies in Mycology 53: 163-71. Zettler, L.W., S.L. Stewart, M.L. Bowles, and K.A. Jacobs. 2001. Mycorrhizal fungi and cold-assisted symbiotic germination of the Federally threatened eastern prairie fringed orchid, Platanthera leucophaea (Nuttall) Lindley. American Midland Naturalist 145: 168-75.

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Stewart & Hicks: PROPAGATION AND CONSERVATION STATUS OF NATIVE ORCHIDS Table 2—Propagation and conservation statuses for orchids species native to survey regions. Geographic range key: US=United States (including Alaska), HI=Hawaii, CAN=Canada, SPM=St. Pierre et Miquelon, G=Greenland, PR=Puerto Rico, VI=U.S. Virgin Islands, and GU=Guam. Propagation status key: 1=available commercially on a regular basis, 2=has been available commercially in recent past, 3=is routinely maintained in laboratory culture, 4=has been produced experimentally in laboratory culture, 5=experimental culture has had limited success, and 6=experimental culture has been unsuccessful. Conservation status key: F=Federally protected as endangered, f=Federally protected as threatened, S=state protected as endangered, s=state protected as threatened, o=state protected by other designation. Propagation data from botanic gardens have been capped at ―3‖ as their role generally does not involve the production of plants for commercial ventures; nd = no data.
Geographic Range Taxa Amerorchis rotundifolia Anoectochilus sandvicensis Aplectrum hyemale Arethusa bulbosa Basiphyllaea corallicola Beloglottis costaricensis Bletia patula Bletia purpurea Brachionidium ciliolatum Brachionidium sherringii Brassavola cucullata Brassavola nodosa Brassia caudata Broughtonia domingensis Bulbophyllum guamense Bulbophyllum longiflorum Bulbophyllum pachyrhachis Bulbophyllum profusum Calanthe triplicata Calopogon barbatus Calopogon multiflorus Calopogon oklahomensis Calopogon pallidus Calopogon tuberosus var. simpsonii Calopogon tuberosus var. tuberosus Calopogon × fowleri Calopogon × goethensis Calopogon × obscurus Calopogon × simulans Calopogon × vulgaris Calypso bulbosa var. americana Calypso bulbosa var. occidentalis Campylocentrum micranthum Campylocentrum pachyrrhizum Campylocentrum pygmaeum Cephalanthera austiniae Cleistesiopsis bifaria Cleistesiopsis divaricata Cleistesiopsis oricamporum Cleistesiopsis × ochlockoneensis US, CAN,G HI US, CAN US, CAN, SPM US, PR US US, PR US PR PR PR, VI PR US PR GU GU US GU GU US US US US US US, CAN, SPM US US US US US US, CAN US, CAN PR US, PR PR US, CAN US US US US

In vitro Propagation Asymbiotic nd nd 2,5 nd 3, 4 nd 4, 5 2, 4 nd nd 2, 3 1, 2, 3 3 nd 2 3 nd 6 3 3 3, 6 3 3 3, 4 1, 3, 5 nd nd nd nd nd 4 4 6 6 nd nd 5, 6 3 nd nd

In vitro Propagation Symbiotic nd nd 4 nd nd nd nd 6 nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd 5 5 nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd

Conservation Status S, s nd S,s,o S, s, o S S nd S nd nd nd nd S nd nd nd nd nd nd nd S nd nd S, o S, o nd nd nd nd nd S, s, o S, s nd S nd nd nd nd nd nd

Reference(s) 11 22 3,11,14,23,24 11 2, 11, 23 11 11, 14, 23 2, 6, 11, 24 11 11 2, 8, 11, 24 1, 2, 11, 23, 24 11, 24 11 9, 20, 23 9, 20 18 9, 20, 23 9, 20, 24 11, 24 11, 14 11, 24 11, 24 6, 11 1, 2, 6, 11, 13, 14, 24 19 19 19 19 19 11, 23 11, 14 2, 11 11, 23 11 11, 18 11, 14, 19, 24 11, 19, 24 19 19

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Stewart & Hicks: PROPAGATION AND CONSERVATION STATUS OF NATIVE ORCHIDS
Taxa Cochleanthes flabelliformis Coeloglossum viride var. virescens Coeloglossum viride var. viride Coelogyne guamensis Comparettia falcata Corallorhiza bentleyi Corallorhiza maculata var. maculata Corallorhiza maculata var. mexicana Corallorhiza maculata var. occidentalis Corallorhiza maculata var. ozettensis Corallorhiza mertensiana Corallorhiza odontorhiza var. odontorhiza Corallorhiza odontorhiza var. pringlei Corallorhiza striata var. striata Corallorhiza striata var. vreelandii Corallorhiza trifida Corallorhiza wisteriana Corymborkis forcipigera Corymborkis veratrifolia Cranichis muscosa Cranichis ricartii Cranichis tenuis Cyclopogon cranichoides Cyclopogon elatus Cyclopogon miradorense Cypripedium acaule Cypripedium arietinum Cypripedium californicum Cypripedium candidum Cypripedium fasciculatum Cypripedium guttatum Cypripedium kentuckiense Cypripedium montanum Cypripedium parviflorum var. makasin Cypripedium parviflorum var. parviflorum Cypripedium parviflorum var. pubescens Cypripedium passerinum Cypripedium reginae Cypripedium yatebeanum Cypripedium × alaskanum Cypripedium × andrewsii nm. andrewsii Cypripedium × andrewsii nm. landonii Cypripedium × columbianum Cypripedium × herae Cyrtopodium macrobulbon Cyrtopodium punctatum Dactylorhiza aristata var. aristata Dactylorhiza aristata var. kodiakensis Dactylorhiza praetermissa var. praetermissa Deiregyne confusa Dendrobium guamense Dendrobium phillippense Range PR US, CAN US, CAN GU PR US US, CAN, SPM US US, CAN US US, CAN US, CAN US, CAN US, CAN US, CAN US, CAN, SPM, G US PR GU US, PR PR PR US, PR US, PR, VI PR US, CAN, SPM US, CAN US US, CAN US US, CAN US US, CAN US, CAN US US, CAN, SPM US, CAN US, CAN US US US, CAN US, CAN US, CAN US, CAN US US, PR US US CAN US GU GU Asymbiotic 4, 6 nd nd nd 3, 5 nd nd nd nd nd nd 5 nd nd nd nd nd nd nd 3 nd nd 6 6 nd 1, 2, 3 1, 2 1, 5 1, 3, 5 2, 6 1 1, 3, 5 1, 5 1, 4 1, 3, 6 1, 3, 6 1 1, 4, 5 1 1 1, 3 nd nd nd 6 1, 3, 4 5 nd 1 nd nd nd Symbiotic nd nd nd nd nd nd 6 nd nd nd nd 4 nd nd nd nd nd nd nd nd nd nd nd nd nd 4 nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd 4 nd nd nd nd nd nd Status nd nd nd nd nd nd o nd nd nd nd S, s, o nd S, o nd S, s, o S, o nd nd S F, S nd nd S nd S, o S, s, o nd S, s, o o nd S, o nd S, o S, s, o S, s, o nd S, s, o nd nd nd nd nd nd nd S nd nd nd nd nd nd Reference(s) 11, 23, 24 18 18, 19 9, 20 2, 11, 23 11 11, 22 18, 19 11 11 11 3, 11 11 11 11 11 11 11 9 11, 24 11 8 11, 23 8, 11, 23 11 1, 5, 11, 16, 24 5, 11, 16 1, 5, 11, 14 1, 5, 11, 14 5, 11, 14 5, 11, 14 5, 11, 13, 14, 15,16, 17, 24 11, 14, 16 1, 5, 11 11, 14, 15, 17, 24 1, 5, 11, 14, 15, 16, 17, 24 11, 16 1, 5, 11, 14, 15, 16, 17 5, 11 11, 16 1, 11, 15, 16 11 11 19 2, 11, 18, 19 2, 6, 11, 14, 24 11, 14 11 16, 18, 19 11 9, 20 9, 20

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Stewart & Hicks: PROPAGATION AND CONSERVATION STATUS OF NATIVE ORCHIDS
Taxa Dendrobium scopa Dendrophylax lindenii Dendrophylax porrectus Dichaea hystricina Dichaea latifolia Dichaea pendulata Dichromanthus cinnabarinus Dichromanthus michuacanus Didymoplexis fimbriata Dilomilis montana Domingoa haematochila Elleanthus cordidactylus Eltroplectris calcarata Encyclia gravida Encyclia isochila Encyclia pygmaea Encyclia rufa Encyclia tampensis Epidendrum acunae Epidendrum anceps Epidendrum amphistomum Epidendrum antillanum Epidendrum boricuarum Epidendrum ciliare Epidendrum floridense Epidendrum jamaicense Epidendrum magnoliae var. magnoliae Epidendrum magnoliae var. mexicanum Epidendrum miserrimum Epidendrum mutelianum Epidendrum nocturnum Epidendrum ramosum Epidendrum rigidum Epidendrum secundum Epidendrum strobiliferum Epidendrum tridens Epidendrum vincentinum Epipactis gigantea Eria rostriflora Erythrodes hirtella Erythrodes plantaginea Eulophia alta Eulophia macgregorii Eulophia pulchra Eurystyles ananassocomos Galeandra bicarinata Galearis spectabilis Geodorum densiflorum Goodyera oblongifolia Goodyera pubescens Goodyera repens Goodyera tesselata Range GU US US, PR PR PR PR US US GU PR PR PR US, PR PR PR US, PR US US US PR, VI US, PR, VI PR PR PR, VI US PR US US PR PR US, PR PR US, PR PR US PR PR US, CAN GU PR PR US, PR GU GU PR US US, CAN GU US, CAN US, CAN US, CAN, SPM US, CAN Asymbiotic nd 1, 3 nd nd nd 3 nd nd 6 nd nd nd 2, 6 nd nd nd 3, 4 1, 3 nd 2, 3 3 nd nd 2, 3 3, 6 6 3 3 nd nd 1, 3, 4 nd 3, 4 1 nd nd nd 1, 4 nd nd nd 2, 4, 5 nd nd nd nd 5, 6 nd 1 4, 5, 6 6 6 Symbiotic nd 5 nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd 4 nd nd nd nd nd nd nd nd 4 nd nd nd 3, 4 nd nd nd nd nd nd nd nd nd nd 4 nd nd nd nd 6 nd nd 4 nd nd Status nd S nd nd nd nd nd nd nd nd nd nd S nd nd S nd o S nd S nd S nd nd nd o nd nd nd S nd S nd S nd nd o nd nd nd nd nd nd nd S S, s, o nd S, o S, o S, o S, s, o Reference(s) 9, 20 2, 6, 11, 12, 24 18, 19 11 11 1, 8 11 19 9, 20, 23 11 11 11 2, 11, 23 11 11 11 2, 11, 23 2, 11, 22, 24 11 8, 11, 23, 24 11, 18, 22, 24 11 11 8, 11, 23, 24 11, 23, 24 11, 23 11, 22, 24 11, 24 11 11 2, 6, 11, 22, 24 11 11, 22, 24 11, 23 11 11 8 2, 11, 16, 24 9, 20 11 11 1, 2, 6, 11, 14, 24 9, 20 9, 20 11 11 1, 3, 11, 22, 24 9, 20 11, 16 2, 3, 11, 14, 22, 24 11, 14 11, 23

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Stewart & Hicks: PROPAGATION AND CONSERVATION STATUS OF NATIVE ORCHIDS
Taxa Govenia floridana Govenia utriculata Gymnadeniopsis clavellata var. clavellata G. clavellata var. ophioglossoides Gymnadeniopsis integra Gymnadeniopsis nivea Habenaria alata Habenaria amalfitana Habenaria distans Habenaria eustachya Habenaria macroceratitis Habenaria monorrhiza Habenaria odontopetala Habenaria quinqueseta Habenaria repens Hapalorchis lineatus Helleriella punctulata Heterotaxis sessilis Hexalectris spicata var. arizonica Hexalectris spicata var. spicata Hexalectris grandiflora Hexalectris nitida Hexalectris revoluta var. colemanii Hexalectris revoluta var. revoluta Hexalectris warnockii Ionopsis satyrioides Ionopsis utricularioides Isochilus linearis Isotria medeoloides Isotria verticillata Jacquiniella globosa Jacquiniella teretifolia Koellensteinia graminea Leochilus puertoricensis Lepanthes caritensis Lepanthes dodiana Lepanthes eltoroensis Lepanthes rubipetala Lepanthes rupestris Lepanthes sanguinea Lepanthes selenitepala Lepanthes veleziana var. retusicolumna Lepanthes veleziana var. veleziana Lepanthes woodburyana Lepanthopsis melanantha Liparis elata Liparis guamensis Liparis hawaiensis Liparis liliifolia Liparis loeselii Liparis saundersiana Liparis vexillifera Range US PR US US US US PR, VI PR US, PR PR US PR, VI PR US US, PR PR PR US US US US US US US US PR US, PR, VI PR US, CAN US, CAN PR PR PR PR PR PR PR PR PR PR PR PR PR PR US, PR US, PR, VI GU HI US, CAN US, CAN PR PR Asymbiotic 6 nd 3 nd 5 5 nd nd 5 nd 2, 4 6 2, 3 3 2, 3 nd nd nd nd 3 nd nd nd nd nd nd 4 2, 6 6 6 nd nd 6 nd 6 nd nd nd nd nd nd nd nd nd 5, 6 5 nd nd 3 4 nd nd Symbiotic 6 nd 4, 5 nd 5 nd nd nd 5 nd 4 nd 3 3 4 nd nd nd nd nd nd nd nd nd nd nd nd nd 6 nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd 6 4 nd nd nd Status S S nd nd S, s S, s nd nd S nd nd nd nd nd nd nd nd nd nd S, s, o nd S nd nd o nd S nd f, S, s, o S, s, o nd nd nd nd nd nd F, S nd nd nd nd nd nd nd S S nd nd S, s S, s, o nd nd Reference(s) 6, 11, 24 11 18, 19, 22, 24 18, 19 11, 22, 24 11, 24 8, 11 11 6, 11 11 6, 11, 23 8, 11, 23 6, 11, 23 6, 11, 24 6, 11, 14, 24 11 11 19 18, 19 11, 24 11 11 19 19 11 11 8, 11, 23, 24 2, 11, 23 3, 11, 22 11, 14, 24 11 11 11, 23 11 2, 11 11 11 8 11 11 11 11 11 11 11, 23, 24 8, 11, 24 9, 20 22 3, 11, 24 11, 23 11 11 ×

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Stewart & Hicks: PROPAGATION AND CONSERVATION STATUS OF NATIVE ORCHIDS
Taxa Liparis × jonesii Listera auriculata Listera australis Listera banksiana Listera borealis Listera convallarioides Listera cordata var. cordata Listera cordata var. nephrophylla Listera smallii Listera ×veltmanii Luisia teretifolia Lycaste barringtoniae Macradenia lutescens Malaxis abieticola Malaxis bayardii Malaxis brachypoda Malaxis corymbosa Malaxis diphyllos Malaxis major Malaxis massonii Malaxis paludosa Malaxis porphyrea Malaxis soulei Malaxis spicata Malaxis unifolia Malaxis wendtii Maxillaria acutifolia Maxillaria coccinea Maxillaria parviflora Mesadenus lucayanus Microthelys rubrocallosa NervillIa platychila Nervillia aragoana Nervillia jacksoniae Nidema ottonis Oncidium altissimum Oncidium floridanum Oncidium meirax Pelexia adnata Phreatia minutiflora Phreatia thompsonii Piperia candida Piperia colemanii Piperia cooperi Piperia elegans subsp. decurtata Piperia elegans subsp. elegans Piperia elongata Piperia leptopetala Piperia michaelii Piperia transversa Piperia unalascensis Piperia yadonii Range US US, CAN US, CAN US, CAN US, CAN US, CAN, SPM US, CAN, SPM US, CAN US US, CAN GU PR US US US, CAN US, CAN US US, CAN PR PR US, CAN US US PR, US US, CAN, SPM US PR PR US, PR US, PR, VI US GU GU GU PR PR, VI US PR US, PR GU GU US, CAN US US US US, CAN US, CAN US US US, CAN US, CAN US Asymbiotic nd nd nd nd 6 nd nd nd 6 nd 6 nd 2 nd nd nd nd nd nd nd nd nd nd nd 5 nd nd 3 nd nd nd nd nd nd nd nd 5 nd 5 nd nd nd nd nd nd nd nd nd nd nd nd nd Symbiotic nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd 5 nd nd nd nd 6 nd nd nd 5 5 nd nd 6 5 nd Status nd S, s S, s, o nd o S, s, o S, s, o nd S, s, o nd nd nd S nd S, o S, s, o o nd nd nd S nd o nd S, s, o nd nd nd S S nd nd nd nd nd nd S nd S nd nd nd nd nd nd nd nd nd nd nd nd F Reference(s) 11 11 11 11 11, 23 11 11 11 11, 24 11 9, 20, 23 11 2, 11 21 11 11 11 11 11 11 11 11 11 11 11, 24 11 11 2, 11 11 11 19 9, 20 9, 20 9, 20 11 8, 11 6, 11, 24 11 11, 14 9 9, 20 6, 11 11 11 11 6, 11 6, 11 11 11 6, 11 6, 11 11

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Stewart & Hicks: PROPAGATION AND CONSERVATION STATUS OF NATIVE ORCHIDS
Taxa Platanthera aquilonis Platanthera blephariglottis Platanthera brevifolia Platanthera chapmanii Platanthera chorisiana Platanthera ciliaris Platanthera conspicua Platanthera convallariifolia Platanthera cristata Platanthera dilatata var. albiflora Platanthera dilatata var. dilatata Platanthera dilatata var. leucostachys Platanthera flava var. flava Platanthera flava var. herbiola Platanthera grandiflora Platanthera holochila Platanthera hookeri Platanthera huronensis Platanthera hyperborea Platanthera integrilabia Platanthera lacera Platanthera leucophaea Platanthera limosa Platanthera macrophylla Platanthera obtusata subsp. obtusata Platanthera obtusata subsp. oligantha Platanthera orbiculata Platanthera pallida Platanthera peramoena Platanthera praeclara Platanthera psycodes Platanthera purpurascens Platanthera shriveri Platanthera sparsiflora Platanthera stricta Platanthera tescamnis Platanthera tipuloides var. behringiana Platanthera yosemitensis Platanthera zothecina Platanthera × andrewsii Platanthera × apalachicola Platanthera × beckneri Platanthera × bicolor Platanthera × canbyi Platanthera × channellii Platanthera × correllii Platanthera × enigma Platanthera × estesii Platanthera × evansiana Platanthera × folsomii Platanthera × hollandiae Platanthera × keenanii Range US, CAN US, CAN, SPM US US US, CAN US, CAN US US US US, CAN US, CAN, SPM US, CAN US, CAN US, CAN US, CAN, SPM HI US, CAN US, CAN G US US, CAN, SPM US, CAN US US, CAN US, CAN, SPM US CAN, SPM, US US US US, CAN US, CAN, SPM US US US US, CAN US US US US US, CAN US US US US US US US, CAN US US US CAN US Asymbiotic nd 3, 5 nd 2, 5 nd 2, 3, 5 3 nd 5 nd 5, 6 nd 5 nd 6 4, 5 5 nd nd 3, 5 3, 6 3, 5 nd nd nd nd nd nd 4 3, 6 4, 6 nd nd nd nd nd nd nd nd nd 5 nd 3 nd nd nd nd nd nd nd nd nd Symbiotic nd nd nd nd nd 5 nd nd 4, 5 nd nd nd nd nd nd 6 nd nd nd 4 3 3 nd nd nd nd nd nd nd 3 nd nd nd nd nd nd nd nd nd nd 6 nd nd nd nd nd nd nd nd nd nd nd Status nd S, s, o nd nd s S, s nd nd S, s, o nd S, s, o nd S, s, o S, s, o S, s, o F S, s, o o nd S, s o f, S, s, o o s o nd S, s, o nd S, s f S, s, o nd nd o o nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd 11, 14, 18, 24 11 11, 23, 24 11 1, 11, 14, 22, 23, 24 18, 24 21 11, 22, 24 18 1, 11, 14, 24 11 11, 24 11 11, 24 22 11, 24 11 25 11, 14, 22, 24 11, 24 6, 11, 14, 22, 24 11 11 11 11 11 11 11, 23, 24 1, 11, 14 1, 11, 14, 24 11 19 11, 18 11 19 11 19 11 11 6 19 19, 24 19 19 19 19 11 19 19 19 19 Reference(s)

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Stewart & Hicks: PROPAGATION AND CONSERVATION STATUS OF NATIVE ORCHIDS
Taxa Platanthera × kelleyi Platanthera × lassenii Platanthera × lueri Platanthera × osceola Platanthera × reznicekii Platanthera × smithii ×Platanthopsis vossii Platythelys sagreana Platythelys querceticola Pleurothallis appendiculata Pleurothallis aristata Pleurothallis domingensis Pleurothallis gelida Pleurothallis obovata Pleurothallis pruinosa Pleurothallis pubescens Pleurothallis racemiflora Pleurothallis ruscifolia Pleurothallis wilsonii Pogonia ophioglossoides Polystachya concreta Polystachya foliosa Ponthieva brittoniae Ponthieva racemosa Ponthieva ventricosa Prescottia oligantha Prescottia stachyodes Prescottia pellucid Prosthechea boothiana var. erythronioides Prosthechea cochleata var. triandra Prosthechea pygmaea Pseudorchis straminea Psilochilus macrophyllus Psychilis kraenzlinii Psychilis krugii Psychilis macconnelliae Psychilis monensis Pteroglossaspis ecristata Pteroglossaspis pottsii Rhynchophreatia micrantha Sacoila lanceolata Sacoila paludicola Sacoila squamulosa Scaphyglottis modesta Schiedella arizonica Spiranthes amesiana Spiranthes brevilabris Spiranthes casei var. novaescotiae Spiranthes casei var. casei Spiranthes cernua Spiranthes delitescens Spiranthes diluvialis Range US US US US CAN US US US US PR PR PR US, PR PR PR PR PR PR PR US, CAN, SPM US, PR, VI PR, VI US US, PR, VI PR US, PR, VI PR, VI PR US US, PR, VI US, PR CAN PR PR PR PR, VI PR US US GU US, PR, VI US US PR US US US CAN US, CAN US, CAN US US, CAN Asymbiotic nd nd 3 5 nd nd nd nd nd nd nd nd 4, 6 nd nd nd 5 nd nd 2, 3 2, 3 2, 3 nd 5, 6 nd 5 nd nd 3 1 nd nd nd 4 nd nd 6 3 5 nd 2, 3, 4, 5 nd nd nd nd nd 4 nd nd 2, 3, 5 3 6 Symbiotic nd nd nd 6 nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd 4 nd nd nd nd nd nd nd nd nd nd 6 nd nd nd nd nd 3, 4 nd nd 4 4 nd Status nd nd nd nd nd nd nd nd nd nd nd nd S nd nd nd nd nd nd S, s, o S nd S S nd nd nd nd nd nd S nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd S nd nd nd F nd Reference(s) 19 11 19, 24 19, 24 19 19 19 18 11 11 11 11 11, 23, 24 11 11 11 2, 11 11 11 11, 23, 24 2, 8, 11, 23 2, 8, 11, 23 11 6, 8, 11, 24 11 8, 14 8 8 19, 24 6, 8, 19 11 11 11 11, 23 11 8 8, 23 18, 24 19, 24 9 1, 8, 24 18, 19 18, 19 11 18, 19 11 6, 11 18 18 1, 2, 6, 14, 19, 24 2 18, 23

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Stewart & Hicks: PROPAGATION AND CONSERVATION STATUS OF NATIVE ORCHIDS
Taxa Spiranthes eatonii Spiranthes floridana Spiranthes infernalis Spiranthes lacera var. gracilis Spiranthes lacera var. lacera Spiranthes longilabris Spiranthes lucida Spiranthes magnicamporum Spiranthes ochroleuca Spiranthes odorata Spiranthes ovalis var. erostellata Spiranthes ovalis var. ovalis Spiranthes parksii Spiranthes praecox Spiranthes romanzoffiana Spiranthes stellata Spiranthes sylvatica Spiranthes torta Spiranthes tuberosa Spiranthes vernalis Spiranthes × borealis Spiranthes × eamesii Spiranthes × folsomii Spiranthes × intermedia Spiranthes × itchetuckneensis Spiranthes × meridionalis Spiranthes × simpsonii Stelis perpusilliflora Stelis pygmaea Stenorrhynchos speciosum Taeniphyllum marianense Tetramicra canaliculata Tipularia discolor Tolumnia bahamensis Tolumnia prionochila Tolumnia variegata Trichocentrum carthagenense Trichocentrum maculatum Trichosalpinx dura Triphora amazonica Triphora craigheadii Triphora gentianoides Triphora hassleriana Triphora latifolia Triphora rickettii Triphora surinamensis Triphora trianthophoros var. texensis Triphora trianthophoros var. trianthophoros Tropidia polystachya Vanilla barbellata Vanilla claviculata Vanilla dilloniana Range US US US US, CAN US, CAN US US, CAN US, CAN US, CAN US US, CAN US US US US, CAN, SPM US US US, PR, VI US US US, CAN US US US US US US, CAN PR PR PR GU PR, VI US US PR, VI PR, VI US US PR PR US US PR US US PR US US US, PR US, PR, VI PR PR, US Asymbiotic nd 4 nd 4, 6 nd nd nd 2, 3, 5 nd 1, 3, 4 3 nd 4, 5 5 5 nd 5 nd 6 2, 3 nd nd nd nd nd nd nd nd nd 4 nd 2, 3 3, 5, 6 1, 3 2 1, 3 nd nd nd nd nd 4 nd nd 6 nd nd 3 6 nd nd nd Symbiotic nd 6 nd nd nd 3, 4 nd 4 nd 4 4 nd nd nd nd nd nd nd 5 4 nd nd nd nd nd nd nd nd nd nd nd nd nd 4 nd 4 nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd nd Status nd nd nd nd nd s S, s, o S, s, o S, s, o S, o nd nd F, S nd S, s, o nd nd nd S, s, o S, s, o nd nd nd nd nd nd nd nd nd nd nd nd S, s, o S nd nd nd S nd S S nd nd S nd nd nd nd S S nd S Reference(s) 11 6, 11 11 14, 24 18 11, 22 11 1, 11, 14, 24 11 1, 6, 11, 24 18, 22, 24 18 11, 24 11 2, 11, 14 19 18 8, 18 6, 11, 23 1, 11, 24 11 19 19 11 11 11 11 11 11 11, 24 9, 20 2, 8, 11, 23 3, 11, 14, 24 2, 11, 24 8, 11, 23 8, 11, 24 18 11 11 11 11 11, 24 11 11 11, 23 11 18 18, 24 11, 24 8, 11 11 11

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Stewart & Hicks: PROPAGATION AND CONSERVATION STATUS OF NATIVE ORCHIDS
Taxa Vanilla mexicana Vanilla phaeantha Vanilla poitaei Wullschlaegelia aphylla Zeuxine fritzii Range US, PR, VI US PR PR GU Asymbiotic nd nd nd Nd Nd Symbiotic nd nd nd nd nd Status S S nd nd nd Reference(s) 8, 11 11 11 11 11, 20

References for Table 2: 1. From, M. 2009. Personal communication with A. Hicks. 2. Hicks, A. 2009. Personal communication. 3. O'Neil, J. 2009. Personal communication with A. Hicks. 4. Sheviak, C. 2009. Personal communication with A. Hicks. 5. Steele, W. 2009. Personal communication with A. Hicks. 6. Stewart, S. 2009. Personal communication. 7. Whitten, M. 2009. Personal communication with A. Hicks. 8. Ackerman, J.D. 1995. An orchid flora of Puerto Rico and the Virgin Islands. New York Botanical Garden Press. 9. Raulerson, L. 2006. Checklist of plants of the Mariana Islands. University of Guam Herbarium Contribution 37: 1-69. 10. Stone, B.C. 1970. Flora of Guam. University of Guam Press. 11. USDA PLANTS Database: http://plants.usda.gov/. 12. Hermann, P. 2009. Personal communication with A. Hicks. 13. Whitlow, C. 2009. Personal communication with A. Hicks. 14. Stoutamire, W. 2009. Personal communication with A. Hicks. 15. Vermont Ladyslipper web page: http://www.vtladyslipper.com/. 16. Fraser's Thimble Farms web page: http://www.thimblefarms.com/. 17. Zielinski, R. 2009. Personal communication with A. Hicks. 18. Brown, P.M. & S.N. Folsom. 2003. The Wild Orchids of North America, north of Mexico. University Press of Florida. 19. Brown, P.M. 2009. Personal checklist of the wild orchids of North America, north of Mexico. 20. Raulerson, L. and A.F. Rinehardt. 1992. Ferns and Orchids of the Mariana Islands. American Printing Corporation. 21. Flora of North America web page: http://www.fna.org/. 22. Zettler, L. 2009. Personal communication with S. Stewart. 23. Meyers, T. 2009. Personal communication with A. Hicks. 24. Richards, M. 2009. Personal communication with A. Hicks. 25. Brown, P.M. 2009. Personal communication with S. Stewart.

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Empiricist: SHOULD WE OR SHOULDN'T WE?

SHOULD WE OR SHOULDN'T WE? ETHICS AND ORCHIDS
The Slow Empiricist
I've written about how the orchid enthusiast likes to play with creating a new and better specimen that meets some criteria of esthetics that the orchid world deems desirable. I have opted for leaving the natives as they are and enjoying them for their unique and irreplaceable beauty. Does that stance mean that I am opposed to the idea of reintroducing a native orchid to a habitat that once contained these plants? Not necessarily. If the plants were lost to the area because of man's interference such as clearing the land and building on the site, as Disney World in Florida did, when it obliterated thousands of yellow fringeless orchids to put in a parking lot, then I feel that it is perfectly acceptable to reintroduce the plants to the area if possible. The 'if possible' raises interesting philosophical and ethical questions. How ethical is it to play with these orchids and take the chance that you can introduce them to a spot if you fail? I remember a few years ago an attempt was made to plant seedlings of an orchid along a likely spot in Goethe State Forest in Dunnellon, Florida. Many of the plantlets did not survive. The plants that were reintroduced in areas where there were other plants did well and are still surviving after seven years or so. The ones that failed were put in likely habitat but something was missing and they declined and disappeared. More work was needed to make such an attempt possible. Like all experiments, how ethical is it to let living things, even plants, open to the possibility of failure and their ultimate demise? Then there is the possibility of the reintroduced plants taking over the area. If the yellow fringeless orchids mentioned above were put back in an area adjacent to the Disney lot that had other native species in it and the reintroduced plants really took and flourished like they had before the bulldozers obliterated them would they annihilate the other natives in the area? Would this be acceptable if the annihilated species weren't orchids? Although it might seem farfetched, suppose the reintroduced orchids could hybridize with other native orchids growing nearby and produce stunning new crosses or even a new species. It would be exciting and probably very valuable in the long run and I'm not sure of the consequences such an occurrence would make.

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Empiricist: SHOULD WE OR SHOULDN'T WE?

I have never been unhappy about introduced species such as the Epipactis helleborine, the broad leaved helleborine, coming into the United States and colonizing areas because it doesn't drown out the competition. Or at least so far it hasn't. Another orchid from overseas is Zeuxine, the lawn orchid, that has populated Florida from north to south and isn't invasive. It isn't like the Brazilian pepper that has overgrown south Florida pushing out desirable species in its relentless search for new places to grow. Does this mean we could take a native of say Western Asia and introduce it successfully here in the U.S. and feel justified if it takes off like the Brazilian pepper? Being able to introduce native species into the landscape does raise some concerns for fragile habitats if it becomes common practice for orchids to be propagated by anyone, especially if they proliferate wherever they are introduced. Are amateur gardeners well enough schooled to be able to go to a local nursery and buy orchids that have been proven to survive in an area and put them willy-nilly wherever they want? I am especially wary of those individuals who profess to want to improve a species and start that process of refining the native orchid to produce a 'superior' plant. But, of course, I don't care for all the hybridizing that goes on amongst the orchid hobbyists. I know this is highly unlikely in my time but who really knows what man's meddling might produce? So should we or shouldn't we? I say let's proceed with caution and keep in mind all the possibilities of our actions. We could be creating a monster that will comeback to haunt us or we could be creating a blessing in repopulating an area with species that should never have been obliterated in the first place. Your Slow Empiricist

Spiranthes brevilabris, short-lipped ladies‖-tresses Goethe State Forest, Levy Co., Florida
P.M. Brown

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RECENT ORCHID LITERATURE OF INTEREST

RECENT ORCHID LITERATURE OF INTEREST
From: 2009. American Journal of Botany 96: 2022-30. Genetic diversity of the endangered and narrow endemic Piperia yadonii (Orchidaceae) assessed with ISSR polymorphisms Sheeja George, Jyotsna Sharma, and Vern L. Yadon Abstract: Highly endangered plants that are also narrow endemics are generally found to be genetically depauperate and thus are exceedingly susceptible to ecological and anthropological threats that can lead to their extinction. Piperia yadonii is restricted to a single California county within a biodiversity hotspot. We used nine primers to generate intersimple sequence repeat (ISSR) data to assess its genetic diversity and structure. Within each population, 99% of the loci were polymorphic, expected heterozygosity was low, and a majority of the loci were shared with few other populations. Forty percent of the total variation could be attributed to population differentiation while the rest (60%) resides within populations, and the genetic distances between populations were independent of the corresponding geographical distances. High divergence among populations is likely due to fragmentation and limited gene flow. Each population contains several private loci, and ideally, each should be protected to preserve the overall diversity of the species. Because P. yadonii currently retains a modest amount of genetic variation among individuals within populations, preserving and expanding the habitat at each site to allow natural expansion of populations would be additional strategies for its conservation before populations become too small to persist naturally. From: 2009. Annals of Botany 104(3): 543-56 Terrestrial orchid conservation in the age of extinction Nigel D. Swarts and Kingsley W. Dixon Background: Conservation through reserves alone is now considered unlikely to achieve protection of plant species necessary to mitigate direct losses of habitat and the pervasive impact of global climate change. Assisted translocation/migration represents new challenges in the face of climate change; species, particularly orchids, will need artificial assistance to migrate from hostile environments, across ecological barriers (alienated lands such as farmlands and built infrastructure) to new climatically buffered sites. The technology and science to underpin assisted migration concepts are in their infancy for plants in general, and orchids, with their high degree of rarity, represent a particularly challenging group for which these principles need to be developed. It is likely that orchids, more than any other plant family, will be in the front-line of species to suffer large-scale extinction events as a result of climate change.

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RECENT ORCHID LITERATURE OF INTEREST

Scope: The South West Australian Floristic Region (SWAFR) is the only global biodiversity hotspot in Australia and represents an ideal test-bed for development of orchid conservation principles. Orchids comprise 6 % of all threatened vascular plants in the SWAFR, with 76 out of the 407 species known for the region having a high level of conservation risk. The situation in the SWAFR is a portent of the global crisis in terrestrial orchid conservation, and it is a region where innovative conservation solutions will be required if the impending wave of extinction is to be averted. Major threatening processes are varied, and include land clearance, salinity, burning, weed encroachment, disease and pests. This is compounded by highly specialized pollinators (locally endemic native invertebrates) and, in the most threatened groups such as hammer orchids (Drakaea) and spider orchids (Caladenia), high levels of mycorrhizal specialization. Management and development of effective conservation strategies for SWAFR orchids require a wide range of integrated scientific approaches to mitigate impacts that directly influence ecological traits critical for survival. Conclusions: In response to threats to orchid species, integrated conservation approaches have been adopted (including ex situ and translocation principles) in the SWAFR with the result that a significant, multidisciplinary approach is under development to facilitate conservation of some of the most threatened taxa and build expertise to carry out assisted migration to new sites. Here the past two decades of orchid conservation research in the SWAFR and the role of research-based approaches for managing effective orchid conservation in a global biodiversity hotspot are reviewed. From: 2009. Botanica Helvetica 119: 69-76. Mathematical inference of the underground clonal growth of Epipactis helleborine (L.) Crantz (Orchidaceae, Neottieae) Anna Jakubska-Busse, Malgorzata Dudkiewicz, Pawel Jankowsi, and Radoslaw Sikora Abstract: Epipactis helleborine (L.) Crantz (Orchidaceae, Neottieae) can spread by sexual or vegetative propagation. The choice of strategy likely depends on the environmental conditions. The rhizome is the organ of vegetative reproduction; hence, it is crucial to understand its development. Unfortunately, it is hardly possible to investigate rhizome morphology directly, since E. helleborine is a protected species in most European countries. The goal of our investigation was to infer the growth patterns of underground parts of an orchid population from long-term annual observations of its aboveground shoots. We implemented the Minimum Spanning Tree method to determine a likely set of underground connections between shoots and to simulate the annual growth of new rhizomes. Furthermore, we modeled the spatial distribution of shoots with a density kernel estimator to compare the density gradients with the direction of growth of the rhizomes. Observed shoot numbers fluctuated between 72 and 183 from year to year. Our results suggest that (1) vegetative reproduction prevails in the studied population, (2) the population consists of about a dozen clones with a diameter of up to 6 m, (3) rhizomes produce up to five new shoots at one branch end per year, (4) rhizomes develop in the direction of decreasing
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RECENT ORCHID LITERATURE OF INTEREST

population density, and (5) nodes of rhizomes may produce new offshoots after up to 7 years of dormancy. From: 2008. Floriculture, Ornamental and Plant Biotechnology 5: 375-91. Techniques and applications of in vitro orchid seed germination Philip J. Kauth, Daniela Dutra, Timothy R. Johnson, Scott L. Stewart, Michael E. Kane, and Wagner Vendrame Abstract: In nature orchid seeds germinate only following infection by mycorrhizal fungi that provide the developing embryo with water, carbohydrates, minerals, and vitamins. Orchid seeds were first germinated at the base of wild-collected potted orchids, but germination was unreliable and seedling mortality rates were high. In vitro germination techniques, which were developed in the early 1900s, have resulted in more reliable germination and propagation of many orchid taxa. The earliest in vitro orchid seed germination techniques utilized mycorrhizal fungi found in nature to simulate germination and seedling development. In 1922 Lewis Knudson germinated orchid seeds in vitro by sowing seeds on sterile nutrient medium amended with sucrose. This technique is known as asymbiotic seed germination since no fungal mycobiont is used to promote germination. For both symbiotic and asymbiotic orchid seed germination, many conditions must be address such as photoperiod, temperature, and mineral nutrition. In the case of symbiotic germination, another important factor is fungal compatibility. In recent years, the limitations that seed dormancy poses to the germination of orchid seed have also been examined. In this chapter techniques and applications of asymbiotic and symbiotic seed germination will be discussed in relation to photoperiod, temperature, nutrition, seed dormancy, and fungal mycobionts.

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BOOK REVIEWS ASYMBIOTIC TECHNIQUE OF ORCHID SEED GERMINATION SECOND REVISED EDITION Aaron J. Hicks, Chapter 7 by Scott Stewart, PhD. The Orchid Seedbank Project. 2009. Raven Roost Books. 185 pp. ISBN 0-9673049-3-8 $54.00 http://www.ravenroostbooks.com/?page=shop/flypage& product_id=1464&keyword=Hicks&searchby=author&o ffset=0&fs=1&CLSN_513=1259942277513a4de039774047 3a50e1 Literature on orchid seed germination is abundant but scattered throughout scientific publications not readily available to the general public. Several books have been published that describe basic instructions for orchid seed sowing including Orchids from Seed by P.A. Thompson, Home Orchid Growing by R.T. Northern, and Growing Orchids from Seed by P. Seaton and M. Ramsay. In writing Asymbiotic Technique of Orchid Seed Germination, Mr. Hicks hoped to provide a manual that will “clarify the entire flasking process for all readers.” There is no doubt that Mr. Hicks wanted to create a comprehensive guide involving as much information on orchid seed germination as possible. In doing so, I believe the book actually provided too much unnecessary information that could potentially leave some confused and overwhelmed. Upon receiving the book, I was impressed with its quality. The black and white and glossy cover caught my eye immediately, and the scanning electron microscopy image of the orchid seeds was an excellent choice. The paper used in the book‖s production was good quality. I was also impressed with the figures and diagrams throughout the book. I was especially pleased that many figures were in color, which enhanced the overall quality of the book. Mr. Hicks also did a good job at providing a list of companies that sell tissue culture equipment and supplies. I was pleased with many aspects of the book. The section in Chapter 3 on pollination was well written and useful, and the figures were an excellent addition. Likewise the sections on harvesting and storing orchid seed were welcomed because many hobbyists have questions concerning these topics. Mr. Hicks also does a thorough job describing aseptic sowing areas, and includes information for constructing a glove box. In Chapter 4 the techniques for sowing orchid seed were described quite thoroughly. I was pleased that Mr. Hicks included descriptions of using both loose, mature seed and green capsules. Mr. Hicks gives the readers several options to surface sterilize seed, and provides two excellent figures that helped to clarify the text. Appendix I, which was the most useful section of the book, outlined the actual flasking technique that was easy-to-follow.
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Dr. Stewart‖s contribution on symbiotic germination was a welcomed edition since this technique is popular for conservation purposes. In addition, no other book to my knowledge provides steps outlining symbiotic germination. Dr. Stewart provided thorough background and history on the orchid seed/fungal relationship. The glossary at the end of the chapter is a necessary addition for those not familiar with the scientific terms. Although the highly scientific writing may be daunting for the hobbyist, this chapter would be a necessary edition to any orchid seed scientist‖s library. I found that much of the information presented was unnecessary. Related information, such as that regarding the green capsule technique, was scattered throughout the book making it difficult to read at times. Chapter 6 on advanced techniques could be absorbed into other chapters. Mr. Hicks devotes several pages in Chapter 6 to media modifications, which I believe could be included in the media section in Chapter 3. Having an entire chapter solely devoted to media would have been a welcomed revision. Also, the section in Chapter 6 on osmotic strength and phenolics could have been edited out. I also did not find the section in Chapter 6 on new directions in seed disinfection useful since Mr. Hicks already described several techniques in Chapter 4. That being said, I also felt that there were too many techniques discussed for disinfecting seeds, many of which would not be used. I was very puzzled why Mr. Hicks even mentioned using chlorine gas as a sterilization technique considering this is incredibly dangerous. Presenting two or three useful techniques would make the text less confusing. I also would have recommended combining Chapters 1 and 2 and increasing the information on the unique biology and anatomy of orchid seeds. Overall I thought the book was a good attempt at providing a comprehensive guide to orchid seed germination. However, the book could have been simpler with less information. Mr. Hicks even states in Appendix I that much of the information is of little or no practical value for the beginner. I agree with his statement. I would recommend the book to hobbyists and growers who want a book with enormous amounts of information on the subject. Philip Kauth, Ph.D.
Plant Restoration, Conservation, and Propagation Biotechnology Program, Environmental Horticulture Department, University of Florida , PO Box 110675, Gainesville, FL 32611, USA. pkauth@ufl.edu

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MICROPROPAGATION OF ORCHIDS, VOLUMES 1 & 2 (2ND EDITION) Joseph Arditti. 2008. Wiley-Blackwell Publishing, Hoboken. Hardcover. Vol. I (pages 1–756), Vol. II (pages 757–1523). $475 http://www.wiley.com/WileyCDA/WileyTitle/productCd1405160888.html More than 30 years ago, the first volume of Joseph Arditti, PhD‖s, Orchid Biology, Reviews and Perspectives series was published. In the initial volume, an appendix was added that served as a manual to growing orchids via clonal propagation and tissue culture — a relatively new and exciting branch of orchid biology at that time. Largely because of the appendix, demand for the first volume of Orchid Biology continued, even after going out of print in 1990. Consequently, Arditti and fellow colleague Robert Ernst, PhD, embarked on a mission to expand the appendix and publish it as a separate book, Micropropagation of Orchids. Published in 1993, the initial volume of Micropropagation of Orchids combined the Orchid Biology appendix with more recent procedures and methods leading up to 1990. What resulted was a massive (682 page), informative book that also remained in demand after its printing. Although Arditti officially retired in 2001, he was compelled to write a second edition of Micropropagation of Orchids, largely because of a wealth of new publications on the subject that surfaced between 1990 and 2000. Hence, the 2nd edition of Micropropagation of Orchids was subsequently born. As predicted, the 2nd edition is even more massive than the first, encompassing two volumes. Both are housed by a handsome, durable sleeve that measures 10½ inches (26 cm) in height, 3½ inches (9 cm) in width and 8 inches (20 cm) in depth. Volume I encompasses the set‖s first three chapters. In typical fashion, Chapter 1 is a historical account of orchid micropropagation, complete with informative text, black-andwhite portraits of noteworthy specialists and other select images. The chapter concludes with some predictions and potential breakthroughs that lie ahead. Those who have read the first Micropropagation of Orchids will find that Chapters 2 and 4 have been revised and rewritten. In Chapter 2 (General Outline of Techniques and Procedures), new information was added that should be useful to many readers, ranging from the educated beginner to the seasoned specialist. Many of the chapter‖s 74 pages contain various tables and figures that nicely supplement the text. Among the topics addressed include media components, plant growth regulators, pH, stock solutions, media preparation, sterile technique and culture conditions (e.g., illumination), among others. Given that the author was unaware of published information on the techniques aimed at how to deal with/handle internal culture contamination, the second edition of Micropropagation of Orchids, like the first, does not address the issue. However, Chapter 2 does provide a section on anticontaminants (antibiotics) and their use. The remaining 600-plus pages of Volume I are dedicated to Chapter
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3, Methods for Specific Genera. In this chapter, 59 genera of terrestrial and epiphytic orchids alike are addressed, beginning with Acampe and ending with Lycaste. Well-known genera such as Cattleya, Dendrobium and Encyclia are discussed along with the mixed company of lesserknown examples, as well as hybrid-derived genera. For each, tables outlining various media are provided. Unfortunately but understandably, the font size of each table is extremely small (about like reading the label of a pinned insect specimen in an entomology collection). Those who have bifocals will be well-served. In Volume II, Chapter 3 continues to march through an additional 52 genera, beginning with Malaxis and ending with Zygopetalum. Taken together, a total of 111 genera are outlined by this huge chapter spanning both volumes. In Chapter 4, the author provides a brief summary of some broad generalizations about orchid tissue culture that many readers should appreciate, given the often unpredictable nature of the practice, and the suffocating wealth of new information. Volume II nears completion with a succulent references section that consumes 76 pages. A few of the newer references are accompanied by links to the Internet. The book concludes with a series of eight useful appendices: 1) General Information on Supplies, Equipment, Terms, and Reagents; 2) Sources of Supplies and Equipment; 3) Sites of Interest on the World Wide Web; 4) Light; 5) Formulary; 6) various units and other values used in micropropagation; 7) Additional Information; and 8) Plant Preservative Mixture. A glossary and index aptly follow. Aside from being an excellent resource like its predecessor, I especially liked the various historical insets that were strategically dispersed throughout Chapter 3 (e.g., Origin of the term protocorm, page 254; use of vanilla by Aztec Emperor Montezuma, page 1291). This approach was both informative and captivating, and served as a reminder of why we go to such extremes in our quest to propagate these plants. In closing, Micropropagation of Orchids, 2nd edition, does not cater to those interested in related fields of study (e.g., bioengineering, cytogenetics, molecular biology, seed germination), as the author had intended. Instead, the book lives up to its billing as a useful resource strictly dedicated to micropropagation. As Arditti states in the book‖s preface, new methods will undoubtedly surface in the coming years, but this edition of Micropropagation of Orchids will be his last treatment. Secure a copy for your personal library before it is out of print once and for all. Lawrence W. Zettler, Ph.D.
Illinois College, Jacksonville, Illinois. lwzettle@.ic.edu This review previously appeared in ORCHIDS magazine from the American Orchid Society and is reprinted by permission.

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FROM THE SWAMPS OF SOUTH FLORIDA TO THE WILDS OF NORTHERN ALASKA…. TO WINDSWEPT NEWFOUNDLAND AND THE BIG BEND OF

WEST TEXAS

WILD ORCHIDS….
from the University Press of Florida

by Paul Martin Brown & Stan Folsom

Ordering information from University Press of Florida www.upf.com or 1-800-226-3822 or for signed and inscribed copies from the authors at naorchid@aol.com
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Native orchids are increasingly threatened by pressure from population growth and development but, nonetheless, still present a welcome surprise to observant hikers in every state and province. Compiled and illustrated by long-time orchid specialist Paul Martin Brown, this pocket guide to the woodland and bog rein orchids forms part of a series that will cover all the wild orchids of the continental United States and Canada. Brown provides a description, general distributional information, time of flowering, and habitat requirements for each species as well as a complete list of hybrids and the many different growth and color forms that can make identifying orchids so challenging.

Lady's-slippers in Your Pocket
A Guide to the Native Lady's-slipper Orchids, Cypripedium, of the United States and Canada
34 color photos, 2008 $9.95, 1-58729-655-1, 978-1-58729-655-0

Ladies'-tresses in Your Pocket
A Guide to the Native Ladies'-tresses Orchids, Spiranthes, of the United States and Canada
30 color photos, 2008 $9.95, 1-58729-656-X, 978-1-58729-656-7

Grass-pinks and Companion Orchids in Your Pocket
A Guide to the Native Calopogon, Bletia, Arethusa, Pogonia, Cleistes, Eulophia, Pteroglossaspis, and Gymnadeniopsis Species of the Continental United States and Canada
43 color photos, 3 drawings, 2008 $9.95, 1-58729-700-0, 978-1-58729-700-7

Twayblades and Adder’s-mouth Orchids in Your Pocket
A Guide to the Native Liparis, Listera, and Malaxis Species of the Continental United States and Canada
49 color photos, 3 drawings, 2008 $9.95, 1-58729-702-7, 978-1-58729-702-1

Fringed Orchids in Your Pocket
A Guide to Native Platanthera Species of the Continental United States and Canada
47 color photos, 3 drawings, 2009 $9.95, 1-58729-812-0, 978-1-58729-812-7

Woodland and Bog Rein Orchids in Your Pocket
A Guide to Native Platanthera Species of the Continental United States and Canada
50 color photos, 3 drawings, spring 2010 $9.95, 1-58729-8627, 978-1-58729-862-2

Each laminated guide is 16 3/4 x 16 7/8 inches and folds to 4 1/8 x 9 inches.

OR FROM THE AUTHORS AT NAORCHID@AOL.COM

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Genera Orchidacearum
The six volumes of Genera Orchidacearum will provide a complete, robust classification of the orchids, descriptions of individual species, and cultivational information. The series, superbly illustrated with color photographs and line drawings of all the genera, will be an indispensable reference tool for scientists and for orchid breeders, collectors and enthusiasts. Order from http://www.oup.com/us/catalog/general/subject/GeneralScience/? view=usa&ci=9780198507130

NOW AVAILABLE!

Genera Orchidacearum
Volume 5 Epidendroideae (Part 2) Alec M. Pridgeon, Phillip Cribb, Mark W. Chase, Finn N. Rasmussen $195.00 Hardback, Nov 2009 ISBN13: 9780198507130 ISBN10: 0198507135 The first reference work of the world's orchid genera that reelects their long evolutionary history, this fifth volume treats 186 genera of the largest subfamily, Epidendroideae.

Genera Orchidacearum
Volume 4: Epidendroideae (Part 1) Alec M. Pridgeon, Phillip Cribb, Mark W. Chase, Finn N. Rasmussen $329.00 Hardback, May 2006 ISBN13: 9780198507123I SBN10: 0198507127 This fourth volume in the series treats the first 210 genera of the largest subfamily of Orchidaceae, Epidendroideae

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Genera Orchidacearum
Volume 3: Orchidoideae (Part 2), Vanilloideae Alec M. Pridgeon, Phillip J. Cribb, Mark W. Chase, Finn N. Rasmussen $195.00 Hardback, Jul 2003 ISBN13: 9780198507116 ISBN10: 0198507119 This third volume describes the remaining 105 genera of subfamily Orchidoideae and all 15 genera of subfamily Vanilloideae.

Genera Orchidacearum
Volume 2 Orchidoideae (Part 1) Alec M. Pridgeon, Phillip J. Cribb, Mark W. Chase, Finn N. Rasmussen $300.00 Hardback, Mar 2001 ISBN13: 9780198507109 ISBN10: 0198507100

Genera Orchidacearum
Volume 1: General Introduction, Apostasioideae, Cypripedioideae Alec M. Pridgeon, Phillip J. Cribb, Mark W. Chase, Finn Rasmussen $250.00 Hardback, Nov 1999 ISBN13: 9780198505136 ISBN10: 0198505132

Genera Orchidacearum, Volume 6 in preparation - due 2010

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