NORTH AMERICAN NATIVE ORCHID JOURNAL

Volume 4 September Number 3 1998 a quarterly devoted to the orchids of North America published by the

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NORTH AMERICAN NATIVE ORCHID ALLIANCE
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PROCEEDINGS OF THE 3RD ANNUAL NORTH AMERICAN NATIVE ORCHID CONFERENCE, 8-11 JULY, 1998, LAKE ITASCA, MINNESOTA - Part 1 WHAT MAKES A GOOD ORCHIDIST?……..and more!

NORTH AMERICAN NATIVE ORCHID JOURNAL
(ISSN 1084-7332) published quarterly in March June September December by the

NORTH AMERICAN NATIVE ORCHID ALLIANCE, Inc.
a group dedicated to the conservation and promotion of our native orchids Editor: Paul Martin Brown Assistant Editor: Nathaniel E. Conard Editorial Consultants: Philip E. Keenan Stan Folsom Production Assistant: Nancy A. Webb The Journal welcomes articles, of any length, of both a scientific and general interest nature relating to the orchids of North America. Scientific articles should conform to guidelines such as those in Lindleyana or Rhodora. General interest articles and notes may be more informal. Authors may include line drawings, and/or black and white photographs. Color inserts may be arranged. Please send all inquiries or material for publication to the Editor at PO Box 772121, Ocala, FL 34477-2121 (mid June August: PO Box 759, Acton, ME 04001-0759). 1999 Membership in the North American Native Orchid Alliance, which includes a subscription to the Journal, is $26 per year for United States addresses, $29US in Canada and $32US other foreign countries. Payment should be sent to Nancy A. Webb, 84 Etna St. Brighton, MA 02135-2830 USA. Claims for lost issues or cancelled memberships should be made within 30 days.

NORTH AMERICAN NATIVE ORCHID JOURNAL
CONTENTS NOTES FROM THE EDITOR 191 WHAT MAKES A GOOD ORCHIDIST? The Slow Empiricist 193 PROCEEDINGS OF THE 3RD ANNUAL NORTH AMERICAN NATIVE ORCHID CONFERENCE 8-11 JULY, 1998 LAKE ITASCA, MINNESOTA Part 1 199 PROPAGATING NORTH AMERICAN CYPRIPEDIUM SPECIES FROM SEED: SUCCESSES AND PROBLEMS Bill Steele 200 MANITOBA NATIVE ORCHIDS Lorne Heshka 219 ALLOZYME VARIABILITY IN THE PLATANTHERA CILIARIS COMPLEX Nancy Cowden 241 ORCHID CONSERVATION IN THE 21ST CENTURY: THE VALUE OF INCLUDING MYCORRHIZAL FUNGI TO PRESERVE ENDANGERED SPECIES Lawrence W. Zettler 261 PHOTOGRAPHING NATIVE ORCHIDS Dianne Plunkett Latham Volume 4 Number 3 September 1998

273 3RD ANNUAL NORTH AMERICAN NATIVE ORCHID CONFERENCE FIELD TRIPS, JULY 10 & 11, 1998 Philip E. Keenan 285 LOOKING FORWARD: December 1998 289 th 4 ANNUAL NORTH AMERICAN NATIVE ORCHID CONFERENCE 290 Book Reviews The Genus Cypripedium: Natural History & Cultivation Dr. Guido Braem, Charles & Margaret Baker Wild Orchids Across North America Philip E. Keenan 292
Unless otherwise credited, all drawings in this issue are by Stan Folsom

1. 2. 3. 4. 5. 6. 7.

Color Plates: p. 217 Cypripedium acaule, C. arietinum p. 218 Cypripedium parviflorum var. pubescens flat-petalled type, C. candidum p. 259 Platanthera ciliaris, P. blephariglottis, P. xbicolor p. 260 Spiranthes cernua seedlings p. 271 Encyclia tampensis p. 272 Corallorhiza maculata var. occidentalis forma immaculata, Cypripedium reginae p. 284 Platanthera praeclara, Malaxis paludosa, M. brachypoda, M. unifolia

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 4, number 3, pages 191- 296; issued September 20, 1998. Copyright 1998 by the North American Native Orchid Alliance, Inc. Cover: Cypripedium fasciculatum by Stan Folsom

NOTES FROM THE EDITOR
This has certainly been quite a summer! With raging fires in Florida, torrential rains in the Northeast, extended heat in the Southwest and finally an August when the weather has seemed to settle down and proceed, but through it all there have been many glorious native orchids. For 86 members of the Alliance the 3rd Annual North American Native Orchid Conference in northern Minnesota was certainly a highlight of the summer. Many thanks are in order, but especially to Lorne Heshka for arranging the trip to southern Manitoba; Rob Freeman, Mark Larocque, and Dianne Plunkett for scouting at Itasca; Char Menzel, Nancy Webb, Nancy Cowden and many others who pitched in and helped during the registration and the meeting portion of the conference. Our extended thanks go to all of the staff of Itasca State Park for their help and thoughtfulness during our stay. Two recent deaths in the orchid world should be brought to note: Roberta Case, wife of Fred Case of Saginaw, Michigan passed away this spring. She will be remembered most recently as the co-author, with Fred, of a great new book on Trilliums. Marion Ruff Sheehan of Gainesville, Florida, one of the premier orchid

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illustrators in the world, died in late July. Her work, with that of her husband, Tom Sheehan, has been seen monthly in Orchids (American Orchid Society Bulletin) for many years as Orchid Genera Illustrated and has been assembled recently in a book of the same name. That was only a small portion of the work Marion did, not all of it confined to orchids. Plans are well underway for the 4th North American Native Orchid Conference to be held next April in Florida. Be sure to see the notice in this issue, and register and make appropriate reservations early, as April is a busy time in Florida. Renewals will be sent out in mid September for 1999, with no increase in the membership/subscription rate. Your early renewal helps us plan for the following year. Please note that renewals will be sent to the office in Ocala, Florida and all new memberships, claims and orders for back issues to Nancy Webb in Brighton, Massachusetts. Paul Martin Brown, editor PO Box 772121 Ocala, Florida 34477-2121 tel/fax: 352/861-2565 email: naorchid@aol.com

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WHAT MAKES A GOOD ORCHIDIST?
The Slow Empiricist At the recent 1998 NANOA Conference in Minnesota at Lake Itasca, Paul Catling received the 1998 NANOA Education Award for his contribution to the advancement of knowledge about native orchids. Charles Sheviak and Carlyle Luer were similarly honored at past conferences. These men have been shining lights for the North American native orchid world. These awards got me to thinking. Orchid enthusiasts come in a variety of shapes, sizes and abilities. What is important to one may not be of the slightest interest to another. I contend that there is room for each and every well-intended enthusiast. You will notice that I qualified the enthusiast as being well intended. I did this because there are those people who are self-serving and will employ less than desirable means to achieve their selfish goals. I am thinking of those people who think nothing of digging up wild populations to enhance their personal gardens or enrich their herbaria. There are those who refuse to share information out of professional jealousy. On a lower level, there are those who usurp their field mates' time with choice specimens by taking endless time with their

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photography. There are other poorly-intentioned behaviors but I would like to dwell on the positive contributions the truly well-intentioned enthusiast brings to the mix. I have been impressed with many people I have come in contact with since I started looking at orchids in a serious way. I wanted to profile them in this column but they have been for the most part very self-effacing and shy about having themselves spotlighted. So instead of causing them to be embarrassed by naming them specifically, I have decided to highlight some of their more desirable traits, traits that many good orchidists share. I hope the models of these behaviors recognize themselves and feel a sense of pride in that recognition. As I first stated, enthusiasts come in many forms and have many different motivations. There are the serious students who are advancing our knowledge about orchids with their studies. They work intensely with them and have been exploring such avenues as gene pools, chromosome counts and other very technical aspects to determine new species and validate existing ones. Their enthusiasm has increased our knowledge extensively in the last few years. They have the persistence and the patience to spend long hours analyzing data that they have painstakingly assembled in an effort to unravel some of the origins of certain species. I salute you and applaud your efforts. Then there are the people who are interested in propagating the orchids and getting them to be

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successfully grown in cultivation. They also have to have perseverance, like the analysts of orchid structure, as they struggle to understand the intricacies of the growing medium, the needs of the emerging plants and all the hairy problems that arise between the inception and the fruition of their projects. You, too, are doing remarkable things to advance our understanding. Since my column usually champions the amateur orchidist, I must discuss some of the outstanding beginners I have encountered. Early in my own excursions in the field, I came in contact with a woman who was working to become more knowledgeable about the plants she was seeing. Her method of learning was to take a small notebook with her and perch beside the plants and make a sketch of each new discovery. She did not profess to be an artist but her skill was evident and her detailed drawing gave her an intimate acquaintanceship with her subject. What a wonderful way to become more thoroughly cognizant of an orchid, or, for that matter, any plant's uniqueness. When I was a rank novice, I was also exposed to much kindness on the part of those who had more knowledge of the subject than I had at that time. One woman, a botany professor at a Connecticut college, approached her subject with a common-sense attitude and down-to-earth explanations. She peppered her narration with folksy good humor and was very flexible in her acceptance of others. She had an appetite for anything, and showed an interest in all things that would

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come up. She is truly a light in my advance in understanding. Another woman who showed great sense gave me the ability to accept what comes along with an up-beat attitude. She had some mobility problems but she was content to find things of interest in those areas that she could reach, rather than bemoaning the fact that others were farther afield. I have spent several excursions with her when we elected to remain back in easier terrain. We may have missed some specialties but we also had a wonderful time exploring our own domain and often found some things that the others missed. Lest you think that all amateurs who have anything positive to contribute are of the female gender, I have had male field mates who have taken the time to make sure that I saw a particular plant, or have slowed their pace through the forest to accommodate my speed. The more advanced males in knowledge about orchids were often willing to take the time to explain an obscure point that I might have missed. Some have shared their loupes, their insect repellant, and their water when I was in need. To go back to the professional who contributes to the enthusiasm about orchids, I would point out the great teachers who are out there toiling to spread their knowledge to others. One, in particular, was able to make things clear in his explanations. His classroom teaching style encompassed all his students' levels of understanding and he made sure things were

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comprehended before moving to a new area of explanation. His field trips were always happy events where each new discovery was enthusiastically examined and enjoyed. He made everyone feel important and no one was ever made to feel inadequate because of his level of understanding. Then there are the places of learning that contribute to the empowerment of the enthusiast. Most of the places I have encountered are interested in me and try to provide me with the help I might need. Whether it is as simple as a field station at a national park dispensing information or a university with its myriad tools like libraries, herbaria, etc., the people who run these places usually try to give good, sound advice and help to me in my quests. Their helpful attitudes are sincerely appreciated when one is new to an area or needs help of some kind. Lastly, there are those who write about orchids. They give of their knowledge and expertise to all of us. This is a wonderful sharing of information. Whether you are just an average enthusiast or a college-educated, serious writer, you, through your words, have piqued curiosity, enlarged comprehension, and added to the enjoyment that shared experience brings to those who are similarly engaged. You are a most important link because you keep the flame alive and get the information out to us. To sum up, I would describe the good orchidist as being one who is willing to share, who is deeply

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committed to the field, and who wants to see the world of information about orchids promulgated and proliferated to all who are interested. I thank those who have been positive influences for us. You know who you are. The Slow Empiricist

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Part 1 PROPAGATING NORTH AMERICAN CYPRIPEDIUM SPECIES FROM SEED: SUCCESSES AND PROBLEMS Bill Steele MANITOBA NATIVE ORCHIDS Lorne Heshka ALLOZYME VARIABILITY IN THE PLATANTHERA CILIARIS COMPLEX Nancy Cowden ORCHID CONSERVATION IN THE 21ST CENTURY: THE VALUE OF INCLUDING MYCORRHIZAL FUNGI TO PRESERVE ENDANGERED SPECIES Lawrence W. Zettler PHOTOGRAPHING NATIVE ORCHIDS Dianne Plunkett Latham

PROCEEDINGS OF THE 3RD ANNUAL NORTH AMERICAN NATIVE ORCHID CONFERENCE 8-11 JULY, 1998 LAKE ITASCA, MINNESOTA

Part 2 in the December issue is scheduled to include:
Platanthera praeclara, a threatened prairie orchid
Margaret From Recent Research on Minnesota Orchids Welby Smith Color, Form and Variation Paul Martin Brown Cypripedium hybrids of Mahnomen County, Minnesota Rob Freeman Piperias Unraveled Mark Larocque & Paul Martin Brown Recent Advances in the Systematics and Ecology of North American Orchids Dr. Paul M. Catling

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PROPAGATING NORTH AMERICAN CYPRIPEDIUM SPECIES FROM SEED: SUCCESSES AND PROBLEMS
Bill Steele ABSTRACT Eight North American species of Cypripedium have now been grown to flowering from seedlings produced in vitro by asymbiotic culture: pink lady's-slipper, C. acaule, ram's-head lady's-slipper, C. arietinum, California lady's-slipper, C. californicum, small white lady's-slipper, C. candidum, spotted lady's-slipper, C. guttatum, ivory-lipped lady's-slipper, C. kentuckiense, yellow lady's-slipper, C. parviflorum, and showy lady'sslipper, C. reginae. Seedlings of clustered lady'sslipper, C. fasciculatum, sparrow's egg lady's-slipper, C. passerinum, and yellow spotted lady's-slipper, C. yatabeanum, have been produced by asymbiotic methods and have been established ex vitro, but have, to my knowledge, not yet been grown to flowering size. Seedlings of the Central American species Mexican yellow lady's-slipper, C. irapeanum, and downy yellow

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lady's-slipper, C. molle, have been grown in vitro, but have not been established outside of the flask. Only very minor germination of mountain lady's-slipper, C. montanum, seed has been achieved in the lab, and seedlings planted out have survived for but one or two seasons. INTRODUCTION Because of their showy appearance, orchids of the genus Cypripedium have been prized in horticulture. All too frequently, plants in cultivation have been obtained by collecting from wild populations, sometimes to the extent that such populations are jeopardized. Artificial propagation of large numbers of these plants from seed would thus serve simultaneously as a source for horticultural material and as a disincentive for digging wild plants. For this reason, researchers in North America and also Europe and Japan have experimented with artificial propagation of these orchids. Details of the laboratory methods currently used for Cypripedium and other terrestrial orchid propagation may be found in Allen (1996). Production of seedlings in the laboratory does not fully constitute propagation, however. The seedlings must then be grown on to the flowering stage for completion of the orchid life cycle, and for some species the most difficult part of propagation is establishing the plants out of the flask. An excellent summary of present knowledge of horticultural

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requirements can be found in Cribb (1997). In the present report, I summarize the present status of in vitro propagation of the North American Cypripedium species. METHODS Orchid seeds may be germinated in vitro either symbiotically or asymbiotically. In symbiotic germination, the seeds are inoculated with a compatible fungus that provides nutrients to the orchid embryo, just as occurs in nature. The researcher attempts to adjust the cultural medium and conditions such that the fungus grows sufficiently to nurture its orchid partner but not sufficiently that the fungus overwhelms and kills the orchid. In asymbiotic germination, the orchid seeds are distributed on a sterile culture medium that provides the nutrients that a fungus would provide in nature or in symbiotic culture. To date, most of the success in Cypripedium propagation has been obtained using asymbiotic methods. All of my own work has been in asymbiotic culture. One of the main difficulties in germination of Cypripedium seeds is that those of many species are dormant at maturity. The purpose of the dormancy in nature is to prevent germination of the seed at an inopportune time such as just before the onset of winter. Thus to germinate Cypripedium seeds, the dormancy must somehow be circumvented. There are two main ways to deal with the dormancy problem: 1) sow the seeds in culture before

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they are dormant, and 2) remove or neutralize the biochemical agents in the seeds causing the dormancy. Sowing the seeds before they are dormant, sometimes loosely called ―green-podding‖ by orchidists, requires that the seed be harvested in the short time window during which the embryos are growing vigorously before the pod parent introduces dormancy factors into the seeds. A major difficulty in using immature seeds is the determination of when to harvest a seed capsule, for the window for successful embryo culture is only a few days long. In using fully mature seeds, the seed capsule is harvested as close as possible to the time it would split or ―dehisce‖ naturally. Mature seed of most Cypripedium species apparently contains substances that prevent the germination of the seed even when presented with all the mineral nutrients and sugar necessary for growth of the embryo. There are three methods used in an attempt to overcome the dormancy: 1) stratification, 2) oxidation by chemical bleaching, and 3) including a germinationstimulating plant growth regulator or ―hormone‖ in the culture medium. In stratification, the orchid seed is maintained at a temperature near freezing for several months. At this temperature, the seed seems to break down germination-inhibiting substances metabolically. Stratification seems more effective in some species than in others.

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The oxidation of germination-inhibitors is carried out by immersing the seed in a bleaching solution prior to sowing. The bleaching has the dual effect of destroying microorganisms or spores on the seed that would contaminate the culture as well as promoting germination. In practice, two or even all three of these methods are applied to promote germination of a given batch of seed. In one species, C. montanum, no combination of these methods has yet produced successful germination of the mature seed. PRESENT STATUS OF PROPAGATION EFFORTS Table 1 summarizes the current status of artificial propagation of the North American species. In this table, the species are grouped into three classes according to the relative difficulty of propagation on the basis of not only the ease or difficulty of germination and in vitro culture but of establishing out of the flask as well. In the table, asterisks indicate species that have been raised to the flowering stage from seed germinated in vitro. COMMENTS ON INDIVIDUAL SPECIES All the species in the group I consider ―easy‖ to propagate have been grown to flowering size from seedlings produced in the laboratory. The species in this category are both relatively easy to germinate and easy to grow outside the flask. To my knowledge, the first lab-

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grown seedlings of Cypripedium californicum to flower were those raised by Michael Fong (personal communication) of San Jose, California. His four-year-old seedlings produced several flowers. Cypripedium candidum has been grown to flowering by Allan Anderson (personal communication) of the University of Guelph and required nine years to flower. Carson Whitlow (personal communication) of Adel, Iowa has told me that C. kentuckiense typically requires five years to flower after planting-out, and consequently I feel lucky to have had two seedlings flower at four years from the flask. I have had C. parviflorum var. parviflorum flower after two growing seasons in pots, and I had one C. parviflorum var. pubescens flower the second spring after removal from the flask. In my experience, the latter subspecies more typically requires three, four, or more years. As far as I know the first publication of a Cypripedium to be flowered from seed germinated in vitro was by Werner Frosch (1985) of Germany, who raised C. reginae to flowering the third year after sowing. Although I also have had a seedling flower two years after planting-out, three or four years seems more typical for C. reginae.

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TABLE 1. Species

Status of Propagation from Seed to Propagate:

Relatively Easy C. californicum* C. candidum* C. kentuckiense* C. parviflorum* C. reginae*

Species Difficult to Propagate: C. acaule* C. arietinum* C. fasciculatum C. guttatum* C. passerinum C. yatabeanum Species Presently Impossible to Propagate: [C. dickinsonianum] [C. irapeanum] [C. molle] C. montanum

*species that have been raised to flowering from seed germinated in asymbiotic culture. [ ] = Central American species.

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pink lady‘s-slipper Cypripedium acaule

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pink lady‘s-slipper Cypripedium acaule

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ivory-lipped lady’s-slipper Cypripedium kentuckiense

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The group of species that I consider ―difficult‖ to propagate is so primarily because of problems in growing the seedlings once they leave the flask. All of these species germinate fairly easily, and the major difficulty lies in keeping the seedlings growing, or even alive, after planting-out. I was delighted this May to have a Cypripedium acaule seedling flower for the first time, six years after planting-out. I may have been the first person to do this. Approximately a week later, however, Scott Durkee of New Haven, Vermont phoned to say that he had a C. acaule flowering from seed only two years after planting-out. My joy in my achievement was somewhat deflated by Scott‘s superior accomplishment! I have found that C. arietinum seed germinates very readily under the appropriate conditions, but seedlings of this species seem very sensitive to reflasking shock and very often die from being moved from one medium to another. If they survive reflasking, C. arietinum seedlings may grow very large in consideration of the relatively small size of the adult plant and may be even larger than C. reginae seedlings at the time of removal from the flask. The greatest difficulties in raising C. arietinum seedlings is that they require cool growing conditions, a very well-drained medium, and relatively dry growing conditions. If all these demands are met, I have found the seedlings will flower the very next season after planting-out although an additional year or two may be necessary. Cypripedium guttatum also insists on cool growing conditions after planting-out, but Dr. Svante Malmgren (personal communication) in Sweden has carried this species through to flowering. The other species in the

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―difficult‖ group, viz., C. fasciculatum, C. passerinum, and C. yatabeanum have not been flowered from seed to my knowledge, but they germinate readily and can be grown in pots, although with difficulty, and I have little doubt that each will eventually be raised to flowering. Cypripedium passerinum is probably the most difficult of these species to grow outside the flask and seems to require cool temperatures, or the light cycle typical of the high latitudes from which the plant comes, or both in order to survive. Among the species that I consider ―impossible‖ at present, Cypripedium montanum presents extreme difficulty in the germination of its seeds. Despite many experiments by a number of people, only a very few seedlings have been produced by germination of mature C. montanum seed. Peter Wilson (personal communication) in England has perhaps had the greatest success, but he was only able to obtain a few tens of seedlings in many experiments. His limited success involved no unusual treatments or media, and he feels that success depends on the population or the clone from which the seed comes. Immature seed of C. montanum is also difficult to germinate, and I have only ever obtained a handful of seedlings in this manner. Karen May (personal communication), of Bigfork, Montana has worked extensively at germinating immature seed of this species. She has had good success but with seed capsules from only a few of the many plants from which she collected capsules, and she feels

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that ease of germination varies greatly from clone to clone. The other presently ―impossible‖ species are from Central America. Seed of Cypripedium irapeanum and C. molle has been germinated and seedlings grown to the stage for planting-out, but so far no one has been able to grow these seedlings out of the flask. To my knowledge, no significant efforts have been made to grow C. dickinsonianum in vitro. THE FUTURE As growers gain more experience with labpropagated material, the ―easy‖ species should become quite routine, and even now more or less casual gardeners are successfully growing some of these species from lab-propagated seedlings. Before long, someone should succeed in blooming the ―difficult‖ species that have not yet flowered. As more experience is gained with ―difficult‖ species outside the flask, some of these will also become routine, particularly in climates not much different from those in which the plants are native. Transplanted specimens of the Central American species are being grown in cultivation in Mexico, but all of the successful seedling production of which I am aware has been outside that country. What would help greatly with these species is either for seedlings to be shipped to orchidists working near the natural range of these plants or for Central American orchidists to grow the seedlings in vitro.

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Among North American species, Cypripedium montanum seems to present the greatest challenge. The plant is not that rare in the wild, so the seed must germinate well in nature. Over a period of several years, I found new seedlings under plants from which I collected seed that I was unable to germinate. Clearly there is a trick to germinating this seed, but all efforts to find the secret have so far been futile.
REFERENCES Allen, C., ed. 1996. North American Native Terrestrial Orchid Propagation and Production. North American Native Terrestrial Orchid Conference, Germantown, Maryland. Cribb, P. 1997. The Genus Cypripedium. Timber Press, Inc., Portland, Oregon. Frosch, W. 1985. Asymbiotische Vermehrung von Cypripedium reginae mit Blüten drei Jahre nach der Aussat. Die Orchidee. 36: 30-32. Bill Steele, Ph.D. Spangle Creek Labs 2295 County Rd. 445, Bovey, MN 55709 scl@uslink.net

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left:

First C. arietinum to flower 1 year after planting out. April 1966. below: My first C. acaule to bloom - - 6 years after planting out. 17 May 1998. Photos by Bill Steele

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Top: Cypripedium parviflorum var. pubescens flat-petalled variant Botton: Cypripedium candidum Lorne Heshka Manitoba 218

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MANITOBA NATIVE ORCHIDS
Lorne Heshka A total of 37 indigenous orchid species are listed for the Province of Manitoba by Manitoba Environment. Four of these species are not discussed in this paper as I have not observed or photographed them in Manitoba. These are: Listera borealis – northern twayblade Listera auriculata – auricled twayblade Malaxis brachypoda – white adder’s mouth Malaxis paludosa – bog adder’s mouth One other orchid that I have not observed is Cypripedium passerinum – sparrow’s egg or Franklin’s lady’s-slipper. Jim Roy has lent me his slides of this species and thus it will be covered in this presentation. The province of Manitoba is located in a transition zone between the Prairies to the west and the Canadian Shield to the east. The province extends from the US border at the 49th parallel in the south to the North West Territories and the Arctic Ocean at Hudson Bay in the north. The vegetation distribution map of the province shows grasslands occupying the south central and southwestern portion of the province. This area is primarily

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agricultural, and except for protected areas, most of the natural orchid habitat has been disturbed. From the south east, in a diagonal band across the province, is the parkland region, primarily a deciduous or mixed deciduous and coniferous forest. Beyond are the lakes and forest of the northern coniferous forest, occupying the largest area of Manitoba. The most northerly portion is the northern transitional forest including the Hudson Bay lowlands and a thin border of tundra along Hudson‘s Bay. The largest number of species of orchids is found in the southeast, while the largest numbers of orchids can be observed further north in the northern coniferous forest. An increasingly popular recreational activity is orchid viewing and orchid photography. This has put some stress on the orchid habitat; however, the benefit is an increased awareness of a need for conservation of habitat for our native orchids. eastern fairy slipper - Calypso bulbosa var. americana. Our earliest-blooming orchid, and arguably the most beautiful, is the eastern fairy slipper. This species, although not common, seems to appreciate a coniferous forest habitat growing over a limestone base. The northern Interlake area provides this habitat and in late May or early June you would be able to find a number of blooming plants.

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ram’s-head lady’s-slipper, Cypripedium arietinum This diminutive species can be found in two distinctly different habitats. In black spruce and tamarack bogs, this plant is normally found either as individuals or occasionally in small clumps, while in drier, often sandy locations, the plants can form significant clumps. The small size of the flower as well as the colour combine to make this a rather inconspicuous plant in the dappled shade of its environment. One of my favourite spots for this orchid is located in Manitoba‘s Interlake region. Here the plants form nice specimen clumps in open areas of a mature white spruce grove. Several orchids make their home in a Jackpine Forest. These forests may be found on a sand base or on the granite outcrops of the Canadian Shield. pink lady’s-slipper - Cypripedium acaule A common orchid in this habitat is the pink lady’sslipper, also known as the moccasin flower. This widespread Cypripedium ranges from the southeastern portion of the province to the Northwest Territories. Most often found in dry pine forests or on the duff covering granite of the Canadian Shield, it is occasionally found in a black spruce and tamarack bog growing in sphagnum moss. The plants in the bogs have always been much reduced in size in comparison to those in drier locations.

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ram’s-head lady’s-slipper, Cypripedium arietinum

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dragon’s mouth Arethusa bulbosa

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Approximately 15 years ago while canoeing with a friend in the Nopiming Provincial Forest in eastern Manitoba we came across a population of Cypripedium acaule that included a scattering of the albiflorum or white-flowered form. Fortunately I insisted we stop and record this on film. Hooker’s orchis - Platanthera hookeri Blooming at the same time as Cypripedium acaule is Platanthera hookeri. Although the green flowers of this species are difficult to see, the two large leaves flat on the ground may help. This plant seems to be equally at home in a black spruce and tamarack forest where the non-blooming plants may be confused with the pad leaved orchis, Platanthera orbiculata. northern slender ladies’-tresses - Spiranthes lacera var. lacera In July, after the Cypripedium acaule have long finished blooming, the delicate spikes of Spiranthes lacera var. lacera make their appearance. A good way to locate these flowers is to crouch down and scan the forest floor for the slender white spikes. checkered rattlesnake orchis - Goodyera tesselata At the same time and in the same areas you may find Goodyera tesselata. Abundant when found in suitable habitat, this, our largest Goodyera, colonises jackpine forests on sand. In moist shady locations, plants up to a foot high may be found.

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small round-leaved orchis Amerorchis rotundifolia

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A common habitat through most of the North is the black spruce and tamarack forest with underlying base of sphagnum. This is a suitable habitat for a number of orchid species. lesser rattlesnake orchis - Goodyera repens In a moist, often dark part of this forest, grows the lesser rattlesnake orchis. This is the smaller of the two Goodyera species we have and often occurs in small colonies. The leaves of this plant, although rather small, are the most attractive of any of our native orchid species. heart-leaved twayblade - Listera cordata var. cordata Normally you could expect to find this species in clean, mossy hummocks under a fairly dense black spruce forest canopy. At my favorite site for this plant it has as its companions ram’s-head lady’s-slipper, the small round-leaved orchis and the blunt-leaved orchid. blunt-leaved rein orchis - Platanthera obtusata The range of this small orchid in Manitoba extends at least to Churchill on Hudson‘s Bay of the Arctic Ocean. It is commonly found in black spruce and tamarack forests throughout the province. I have observed the foliosa form in at least three locations with a colony of several plants in one particular location in southern Manitoba. pad-leaved orchis - Platanthera orbiculata

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The white flowers of this species stand out sharply in the dimly lit shadows of the black spruce and tamarack bogs where it is commonly found. I have photographed this plant on Hecla Island on Lake Winnipeg in mid July, and its range extends much further north. green adder’s-mouth - Malaxis unifolia This is the most common of the three species of Malaxis in the province. Although common, this plant is usually found widely scattered in an area. The flower is best appreciated from the top allowing you to observe each of the freshest flowers. The flowering stem on this orchid elongates as it blooms with seedpods forming at the bottom while there are still unopened buds at the top. small round-leaved orchis - Amerorchis rotundifolia One of the earliest orchid species to bloom in this habitat is small round-leaved orchis. Forming dense colonies in favourable, usually quite moist, locations within a black spruce forest, this dainty orchid must be examined carefully to appreciate its beauty. My wife tells me that when a magnifying glass is used the individual flowers remind her of angels adorned in white gowns spotted with pink. This orchid is commonly found from the south through to the extreme northern reaches of our province. Several of our most showy orchids live in a black spruce, tamarack and sphagnum forest that has an

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open canopy allowing good light to reach the understory. One of the earliest showy orchids that bloom in this habitat is the dragon’s mouth Arethusa bulbosa The vibrant color of this orchid makes it easy to find. Never abundant in Manitoba, finding a group of three or more is a rare treat. This orchid often competes for attention with the grass-pink. The range within Manitoba extends only as far north and west as the northern points of our two largest lakes. grass-pink - Calopogon tuberosus var. tuberosus Years ago, before I caught the native orchid ―virus,‖ I found some pink petals on the ground in an open sphagnum bog while photographing wildflowers. I marked the spot, recorded the date, and made a point of going back to this area the next year to check this out. That was my introduction to the grass-pink and as you can well imagine this new find fascinated me. Since that time I have found colonies of this beautiful orchid in a number of locations, the furthest north being at Hecla Island although I understand their range extends even further north than that. This year has been an especially good year for grass-pink in our area and large colonies exist where previously only single plants were found. The Arctic tundra near the Port of Churchill on the Hudson‘s Bay has only a few orchid species. The most sought after species is the

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sparrow’s egg or Franklin’s lady’s-slipper Cypripedium passerinum This is our northern Cypripedium and although it is probably found in locations south of Churchill, much of its habitat is difficult to access. This orchid blooms in early- to mid-July along with Platanthera obtusata and Amerorchis rotundifolia. One of my favorite orchid haunts is a fen approximately 70 kilometers north of Winnipeg. Springs of slightly alkaline water feed this fen and the open meadows and surrounding forest provide habitat for about twenty different species of orchids along with other interesting plants such as butterworts, horned bladderwort and pitcher plants. large yellow lady’s-slipper - Cypripedium parviflorum var. pubescens Although numerous in this fen, it seldom colonises an area in the same manner in which the small yellow lady’s-slipper can. What it lacks in numbers, however, it makes up in size. This was most dramatically illustrated for me in June of 1996. My wife and I were introducing friends of ours to the wonders of native orchids and had stopped along a roadside where both the large and northern small yellow lady’s-slippers were blooming. We noted a particularly outstanding specimen on the other side of a ditch filled with water. Once a route was discovered across the ditch we were truly astounded by the size of the blooms. The flowers on this plant

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measured 7.5" wide across the petal tips, 4" from top of dorsal sepal to bottom of the pouch, and the pouch itself was 2.25" long. All 5 flowers on this plant were of equal size and the pouches were shaped more like a shoe than a slipper having a flat bottom as opposed to the normal rounded slipper shape. To provide a sharp contrast, a northern small yellow lady’s-slipper was growing immediately adjacent to this giant. Unfortunately when I went to check on this plant this spring (1998) all I found was a hole in the ground. In particular environmental conditions, the petals on this species are found to be flat, as opposed to having the normal curl. My experience with the flat-petalled type is limited to this fen where a mix of the normal flowers were interspersed with a few of the flat-petalled types. showy lady’s-slipper - Cypripedium reginae This regal plant is the floral emblem of the province of Prince Edward Island as well as for the state of Minnesota. The showy lady’s-slipper appreciates the sun for at least a portion of the day and in open locations the plant is often quite short, while in heavily shaded locations the plant will grow to nearly a metre. This species requires an abundance of moisture and in an optimum environment will develop significant colonies. In 1996 I saw my first true albolabium, or whiteflowered form, of this species. These are truly impressive plants and it is important that this colony be protected. The colony is scattered over an area of approx. ½ km. by

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1 ½ km. including a black spruce bog as well as the fen. Some stems have been observed with three blossoms. hooded ladies’-tresses - Spiranthes romanzoffiana This species has a wide range of habitat in Manitoba. It is found growing on small islands in the ponds of the fen as well as throughout the meadows of the fen. It can be found in the sphagnum of the black spruce and tamarack forest where the flower spike is rather lax. In contrast, it grows in the tall grass prairie where the flower spike is a compact spiral. tall white northern bog orchis - Platanthera dilatata var. dilatata This tall white orchid is common in the open grassy meadows of the fen. Companion orchids with a similar blooming date in early July include Liparis loeselii and Platanthera hyperborea as well as Platanthera huronensis. northern green bog orchis - Platanthera hyperborea var. hyperborea This is a common to sometimes abundant orchid in Manitoba in a variety of habitats with ample moisture.
Note: Paul Martin Brown advised that a photograph of a tall, stout plant of the open fen was most likely Platanthera huronensis, the pale green bog orchid.

Loesel‘s twayblade - Liparis loeselii Although this is a common orchid of the fen, because of its size and color it is often difficult to find among the grasses of the open meadows. It was quite by accident that I first discovered this diminutive species. Exploring a side drive off a gravel road in a boggy area,

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three tiny plants caught my attention and we were able to add another species to the list of 10 we had already found in bloom that day. A careful check of the surrounding area revealed a colony of some 40 plants. In a wet floating bog in the southeastern region of the province we find two of our less common orchids. rose pogonia - Pogonia ophioglossoides At the extreme north-western extent of its range the rose pogonia is not a common orchid in our province. However, in appropriate habitat it is abundant and in this particular bog which extends several kilometres, it is impossible to estimate the number of plants except to say that there must be many thousands. In this same bog we find another very uncommon orchid in Manitoba, the green fringed orchis or ragged orchis - Platanthera lacera. Out in the open bog the flowers are a greenish creamy white with deep fringes while the flowers on plants found on the trail to this bog some 100 meters away are green in color with shallow fringes.
Note: Paul Martin Brown suggested that some of the latter may be Platanthera xandrewsii although the nearest known population of Platanthera psycodes is approximately 150 kilometers away.

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A small colony of the plants along the trail were associated with a larger colony of approximately 50 plants of Platanthera hyperborea. This species has been recorded and positively identified as far north as Hecla Island on Lake Winnipeg, approximately 300 km north of the US border. The specific directions to this small colony on Hecla were recorded. However, no one has been able to locate these plants since they were first observed some 20 years ago. small purple fringed orchis - Platanthera pyscodes Only one small colony of this species is known to exist in Manitoba. This colony is located at the extreme south east corner of the province and to reach this site you are within 50 meters of the US-Canada border when you leave the paved highway. Several years ago a highway was built through the centre of the colony. Fortunately a few plants survived in a moist deciduous forest on either side of the highway. northern bracted green orchis - Coeloglossum viride var. viride This orchid appears to prefer a dry habitat in a deciduous or mixed deciduous/coniferous forest. However this spring I found a couple of plants growing within 20 centimeters of the edge of a small stream. early coralroot - Corallorhiza trifida This is the earliest of the saprophytic orchids to bloom in Manitoba and is the most common. It also has the most extensive range, from the US border through

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to the northern border with the Northwest Territories. The plants I have found have always been associated with moist locations and I have even found them growing in shallow water in a cedar bog. Whether they actually grew in the water or were flooded after they appeared could not be determined. spotted coralroot - Corallorhiza maculata var. maculata The bright red stems of this saprophytic orchid first caught my attention in a small clearing in a mature white spruce stand. All of the plants I have seen since have been associated with white spruce. Commonly found in cottage country, most cottage owners do not realize that the flowers they are mowing are orchids. striped coralroot - Corallorhiza striata var. striata This member of the coralroot genus seems to be much more tolerant of a variety of habitats than the other Manitoba coralroots. I have found it in a pine forest along with Cypripedium acaule, in mixed forests of white spruce and aspen, as well as moist rich soils adjacent to a small stream. The striped coralroot is reasonably common throughout its range in Manitoba and its striking appearance makes it easy to find. northern small yellow lady’s-slipper – Cypripedium parviflorum var. makasin Readily forming large clumps, this orchid will quickly colonise habitat it finds to its liking. In 1996 I stumbled across just such a location near Lake Winnipeg, approximately 70 km north of Winnipeg. While checking

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out a location where I had, in previous years, found a scattering of plants, to my astonishment, an east-facing slope of a gravel road to a moist ditch contained literally hundreds of plants. It appeared as though someone had taken a pepper shaker and spread the seeds over a patch approximately 5 metres wide by 12 metres long. A follow-up visit to this site in 1997 found that little had changed. This is one species of Cypripedium that can be successfully rescued from a threatened area. A single plant I rescued from a roadside before a road grader‘s blade and transplanted to my yard, has, within the space of a few years multiplied to over 90 blooming stems, many of which contain two flowers. The fragrance emanating from this clump is overwhelming. The northern small yellow lady’s-slipper normally appreciates relatively open sites. However, I have found plants growing in fairly heavily wooded locations. The plants in this type of habitat are often single stems of considerable height, often growing in excess of 20 inches. small white lady’s-slipper – Cypripedium candidum The flowers of this orchid tend to open early in June while the leaves are still tightly curled about the stem. An endangered species, this orchid unfortunately is found in habitat that is most suitable for agriculture. It is still found in several protected locations in Manitoba.

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The Tall Grass Prairie is home to several colonies. A companion orchid in the Tall Grass Prairie sites is Great Plains ladies’-tresses – Spiranthes magnicamporum Blooming in mid-September, this plant is well hidden among the Blue Stem Grass of the prairie. Its wonderfully fragrant flowers do make it an easy orchid to locate, as one can smell this orchid long before you actually see it. The final species is one of the orchids featured at this conference, the western prairie fringed orchis – Platanthera praeclara. Although this species is found in only one area in Manitoba, the numbers are incredible. In 1996 over 20,000 flowering plants were counted. The actual number of plants in this area, including seedlings and non-blooming plants, is difficult to comprehend. Although the land where these orchids are found is owned by a number of different organizations, the overall management is overseen by the Nature Conservancy of Canada. Observation of Malaxis paludosa- the bog adder’smouth in Manitoba
During the Friday, July 10, 1998 field trip to observe and photograph the Malaxis paludosa near Itasca State Park, I remarked that an area I frequently search for orchids, approximately 50 kilometres north of Winnipeg was remarkably similar in many respects to the Itasca site. On July

Note of Interest:

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13, 1998, Jim Roy of Winnipeg and I decided that we would check this area for the bog adder’s-mouth. After 2 hours of unproductive searching we decided to look in an ecological reserve on the opposite side of the highway. We immediately found some nice specimens of Spiranthes romanzoffiana and when I set up my tripod to photograph a plant, I noted three plants of Malaxis paludosa directly below the camera. The “search image” from the Itasca observations assisted in focusing on these plants. The companion orchids within a metre of the Malaxis paludosa were; Cypripedium arietinum – ram’s-head lady’s-slipper, Platanthera hyperborea – northern green bog orchis and Spiranthes romanzoffiana – hooded ladies’-tresses. In discussion with Dr. Karen Johnson, the Curator of Botany at the Museum of Man and Nature in Winnipeg, I learned that this species is considered rare in Manitoba and there have been only 3 “confirmed” observations in Manitoba, the last in 1985. There have been 7 other “unconfirmed” observations. None of the previous observations have been in this particular ecological reserve and none as near to the city of Winnipeg. Lorne Heshka, 1204 De Graff Place, Winnepeg, Manitoba R2g 1y8 Canada e-mail: lheshka@escape.ca

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above:

Platanthera blephariglottis white fringed orchid
right:

Platanthera ciliaris orange fringed orchid

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ALLOZYME VARIABILITY IN THE PLATANTHERA CILIARIS COMPLEX
Nancy Cowden The genus Platanthera consists of approximately 200 species distributed throughout the north temperate regions of the world (Luer, 1975). North American Platanthera taxa are assembled into five sections, some more easily circumscribed than others, as Luer (1975) so eloquently stated the matter. Among these intrageneric groups, section Blephariglottis may arguably be the most easily defined, with members having a distinctly fringed lip, more or less. Examples of this assemblage include: white fringed orchid, Platanthera blephariglottis, for which the section was named, with its white flowers and fringed labellum; orange fringed orchid, P. ciliaris, similar to P. blephariglottis but characteristically orange in color; and monkey-face orchid, P. integrilabia, white like P. blephariglottis but bearing an only slightly serrated lip; and purple flowered taxa such as large purple fringed orchid, P. grandiflora. Within section Blephariglottis, the P. ciliaris complex displays a particularly intriguing group of variations on a common theme: floral color white,

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orange, or some intermediate shade, lips highly fringed to barely serrate, individual flowers small (approximately 1.0 cm across the lateral sepals) or comparatively large (2.0 cm or more across the lateral sepals), all typically bearing 40 or more flowers per inflorescence. In his study James Folsom delimited the Platanthera ciliaris complex as P. ciliaris, P. blephariglottis, P. cristata, and P. integrilabia as well as Chapman's hybrid fringed orchid, P. xchapmanii, Canby's hybrid fringed orchid, P. xcanbyi, and bicolor hybrid fringed orchid, P. xbicolor, all hybrid taxa (Folsom, 1979, 1984). Folsom‘s primary investigation considered the appropriate taxonomic status of P. chapmanii, using floral morphology, pollination biology, and cytology to evaluate whether Chapman’s orchid should be recognized at the specific level. Because P. chapmanii is a purported hybrid between P. ciliaris and P. cristata, Folsom focused his attention on these taxa, excluding the remaining members of the complex (Folsom, 1979). Morphological comparisons among the taxa that he examined led Folsom to propose that P. chapmanii functions as an independent species; however, morphology could not provide conclusive evidence of parentage (Folsom, 1984). Taken as a whole, the floral morphological variation and gradation among and within the taxa comprising the Platanthera ciliaris complex would, I suspect, make a thorough morphometric survey a true

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exercise in frustration. I found while working with P. ciliaris and P. blephariglottis, as well as their presumed hybrid P. xbicolor, that the only floral characteristic capable of separating them from one another in a principle components analysis was floral color (Cowden, 1993). Contrasting somewhat with my results, Judith Robertson‘s study of disjunct populations clearly demonstrated ecotypic differences in the floral morphology and pollinator complements of coastal plain and montane populations of P. ciliaris (Robertson, 1987; Robertson and Wyatt, 1990a, 1990b). Although allied by visible similarities, the P. ciliaris complex‘s evolutionary relationship might better be evaluated by nonmorphological methods. Generally speaking, Platanthera taxa for which we have chromosome counts prove to have a somatic complement of 42 chromosomes. There are exceptions, as Sheviak and Bracht (1998) have determined for some members of the P. hyperborea complex. For members of the P. ciliaris complex, diploid chromosome counts prove to be 42 for P. ciliaris, P. cristata, P. blephariglottis, and P. chapmanii (Love and Simon, 1968; Folsom, 1979; Dressler, 1981; Tanaka and Kamemoto, 1984). I am unaware of reported chromosome counts for P. integrilabia, P. xbicolor, or P. xcanbyi, but it certainly seems reasonable to suspect that these taxa would possess similar chromosome counts. While morphology and cytology seem to confirm what we suspect about relationships within the

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Platanthera ciliaris complex, neither of these methods provide the precision necessary to answer the question: to what degree are these taxa related? Put a slightly different way, does the P. ciliaris complex, as defined by Folsom (1979), represents a natural assemblage of closely related plants? In order to quantitatively evaluate these questions, I employed starch gel, protein electrophoresis. As a method, protein electrophoresis has been widely used by population biologists for over thirty years but has seen few applications to orchid taxa. The technique requires that active proteins, specifically enzymes, be liberated from individual organisms, usually by grinding up tissue in an appropriate extraction buffer. The extract is then introduced to a gel matrix, often either potato starch or polyacrylamide, and individual enzymes are separated by electrical charge and/or molecular size by running an electrical current across the gel and its complementary buffer system. Afterward, horizontal slices of the gel can be assayed for particular enzymes by supplying the appropriate enzyme substrates, cofactors, and other necessary components as well as a dye that interacts with the specific enzyme‘s activities (Wendel and Weeden, 1989). Variable forms of the enzyme, known as allozymes if coded for by a single gene or isozymes if coded for by different genes, appear as bands that migrate different distances through the gel matrix. Because distinct enzyme forms are inherited in a Mendelian fashion, much like flower color, these markers may be used to decipher mating patterns within

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populations, as well as gene flow among populations and among closely related species (Hamrick, 1989; Hamrick and Godt, 1990). For my study, leaf tissue was obtained from a total of 21 populations of Platanthera taxa, including P. ciliaris, P. blephariglottis, P. xbicolor, P. cristata, P. chapmanii, and P. integrilabia. I was unable to sample material from P. xcanbyi. When population size permitted a minimum of twenty individuals per population were sampled. No more than one lower leaf was removed from each plant; in many cases one-half of the basal-most leaf provided sufficient tissue for analyses. Enzyme activity in leaf material was maintained by cold storage until assays could be carried out. In all, nine enzymes, including 12 purported loci, were resolved using three buffer systems (Table 1). Banding patterns resulting from selective staining procedures were used to infer allozyme variability. Allelic designations for each individual were determined by comparing Rf values to an internal standard run on each gel. Unambiguous confirmation of allelic comparability within and between taxa was achieved by electrophoresing hypothesized variants side by side on the same gel. Resulting banding patterns conformed to those detected for other diploid plant taxa both in terms of number of loci expressed and quaternary structure (Wendel and Weeden, 1989; Weeden and Wendel, 1989).

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Determinations of population-level allelic frequencies, percent polymorphic loci, and alleles per locus were performed using the computer program BIOSYS-1, release 1.7 (Swofford and Selander, 1989). Species-level allele frequencies were calculated by combining data from all populations of a taxon. Percent polymorphic loci and alleles per locus for species were calculated according to the method of Hamrick and Godt (1990). Species-level phenograms were generated using the unweighted pair-group method with arithmetic averaging (UPGMA) developed by Sneath and Sokal (1973). Nei's unbiased genetic identity, I, (Nei, 1978) was calculated for each taxon and provided values utilized by UPGMA. A total of 446 individuals from six Platanthera taxa were analyzed. Five members of the Platanthera ciliaris complex (P. ciliaris, P. blephariglottis, P. xbicolor, P. chapmanii, and P. cristata) consistently shared at least one allele per locus. The frequency of shared alleles varied both within and among the five taxa, indicating no taxonomically diagnostic variations. Intraspecific comparisons of mean unbiased genetic-identity values ranged from 0.777 in Platanthera integrilabia to 0.999 in P. xbicolor. Mean genetic identities among interspecifics were much more variable, with P. blephariglottis and P. xbicolor sharing a mean value of 0.964 while the majority of pairs exhibited lower genetic identities (Table 2). As is depicted in the UPGMA phenogram of Nei‘s genetic identities, high genetic

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similarity values occurred among P. ciliaris complex members (Fig. 1). First to cluster were P. blephariglottis and P. xbicolor at 0.989, closely followed by P. ciliaris and P. chapmanii which joined together at 0.971. The two pairs of taxa link at the 0.930 identity level, and P. cristata joined the group at 0.797. Also included in the P. ciliaris complex, P. integrilabia clusters with the previous five taxa at a relatively low value of 0.465. Genetic similarity among Platanthera ciliaris complex members is further illustrated by examining the mean frequency of private alleles (Table 3). No unique alleles were detected in either P. ciliaris or P. xbicolor. A review of allelic frequencies on a locus-by-locus basis shows that, with the exception of allele ―c‖ at PGM1(phosphoglucomutase, locus 1), P. blephariglottis populations surveyed resolved no unique alleles. With the aforementioned deviation, all P. blephariglottis allozymes were also common to P. ciliaris, P. xbicolor, and P. chapmanii. Private alleles were more frequent in other members of the Platanthera ciliaris complex, including P. cristata, with a mean value of 0.500 unique alleles per locus and P. integrilabia, at 0.600 (Table 3).

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In practical terms, what do these results suggest? First and foremost, with the exception of Platanthera integrilabia, members of the P. ciliaris complex display a very high level of genetic identity. Surveys such as those reported by Gottlieb (1981) and Crawford (1983) revealed that genetic identity values lower than 0.80 are typical of congeneric taxa. A corollary of this assertion is that conspecific populations regularly have genetic identity values ranging from 0.80 to 1.00 (Gottlieb, 1981; Crawford, 1983). While I would not suggest that P. ciliaris and most of its complex members are conspecifics, allozyme data point even more strongly than do morphological observations to the conclusion that P. ciliaris, P. blephariglottis, P. xbicolor, and P. chapmanii are recent derivatives of a common origin. Platanthera cristata, sharing a genetic identity with the aforementioned taxa of 0.797, is likely also to have a recent, shared evolutionary history. In contrast, P. integrilabia has a comparatively low genetic identity value (0.465) with other members of the P. ciliaris complex, perhaps suggesting a more distant relationship. At present, no other studies have compared allozyme diversities among members of an orchidaceous complex. However, there have been several studies that investigated intrageneric genetic variation in terrestrial orchids. Scacchi, De Angelis, and Lanzara (1990) found that comparisons of genetic identity values among 11 Orchis species ranged from 0.000 to 0.767 with a mean congeneric identity of 0.194. Low identity values were also obtained by the same researchers for comparisons

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among three Epipactis species (Scacchi, Lanzara, and De Angelis, 1987). An average congeneric identity of 0.197 was reported by Case (1994) for comparisons among five North American Cypripedium species; the greatest genetic similarity, 0.794, was found for the paring of yellow lady's-slippers, C. calceolus (C. parviflorum), and small white lady's-slipper, C. candidum, while the lowest value, 0.000, was obtained for showy lady'sslipper, C. reginae and ram's-head lady's-slipper, C. arietinum. For the Platanthera taxa examined in this study, all, except P. integrilabia, share genetic identity values higher than those previously reported for other orchid congeners. Allozyme data do support the grouping of Platanthera ciliaris, P. blephariglottis, P. cristata, P. chapmanii and P. xbicolor into an assemblage conveniently referred to as the P. ciliaris complex. However, P. integrilabia proves to be a genetically more distinct member of the genus. Is P. integrilabia sufficiently similar to the other taxa examined to be grouped in the section Blephariglottis, as defined by Luer (1975)? At this point data are equivocal, needing a more thorough allozyme investigation of other taxa included in this group. It is clear that our concepts of intrageneric relationships in the genus Platanthera will continue to be fine-tuned as further investigations progress. This, undoubtedly, will also prove true of other genera in the Orchidaceae.

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Nancy Cowden, Ph.D., Assistant Professor of Biology, Department of Biology, Randolph-Macon Women's College, 2500 Rivermont Ave., Lynchburg, VA 24503 Ncowden@rmwc.edu

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Literature cited Case, M.A. 1994. Extensive variation in the levels of genetic diversity and degree of relatedness among five species of Cypripedium (Orchidaceae). American Journal of Botany 81(2): 175-184. Cowden, N.E. 1993. A biosystematic study of Platanthera ciliaris, P. blephariglottis, and P. xbicolor, emphasizing the northern portion of their range. Ph.D. thesis. Miami University, Oxford, Ohio. Crawford, D.J. 1983. Phylogenetic and systematic inferences from electrophoretic studies. In S.O. Tanksley and T.J. Orton [eds.], Isozymes in plant genetics and breeding, part A, 257-287. Elsevier Science Publishers, Amsterdam, Holland. Dressler, R.L. 1981. The orchids: Natural history and classification. Harvard University Press, Cambridge, Massachusetts. Folsom, J.P. 1979. Investigations into the nature of the putative hybrid Platanthera xchapmanii (Orchidaceae). M.A. thesis. Vanderbilt University, Nashville, Tennessee. Folsom, J.P. 1984. A reinterpretation of the status and relations of the taxa in the Platanthera ciliaris complex. Orquedia 9: 321-345. Gottlieb, L.D. 1981. Electrophoretic evidence and plant systematics. Annals of the Missouri Botanical Garden 64: 161180.

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Hamrick, J.L. 1989. Isozymes and the analysis of genetic structure in plant populations. In D.E. Soltis and P.S. Soltis [eds.], Isozymes in plant biology, 87-105. Dioscorides Press, Portland, Oregon. Hamrick, J.L. and M.J.W. Godt. 1990. Allozyme diversity in plants. In A.H.D. Brown, M.T. Clegg, A.L. Kahler, and B.S. Weir [eds.], Plant population genetics, breeding, and genetic resources, 43-63. Sinauer, Sunderland, Massachusetts. Love, A. and W. Simon. 1968. Cytotaxonomical notes on some American orchids. Southwestern Naturalist 13: 335-342. Luer, C.A. 1975. Platanthera. In The native orchids of the United States and Canada excluding Florida, 176-242. New York Botanical Garden, Bronx Nei, M. 1978. Estimation of average heterozygosity and genetic distance from a small number of individuals. Genetics 89:583-590. Robertson, J.L. 1987. Pollination and reproductive biology of Platanthera ciliaris in disjunct populations in the southeastern United States. M.S. thesis. University of Georgia, Athens, Georgia. Robertson, J.L. and R. Wyatt. 1990a. Evidence for pollination ecotypes in the yellow-fringed orchid, Platanthera ciliaris. Evolution 44: 121-133. Robertson, J.L. and R. Wyatt. 1990b. Reproductive biology of the yellow-fringed orchid, Platanthera ciliaris. American Journal of Botany 77: 388-398. Scacchi, R., G. De Angelis, and P. Lanzara. 1990. Allozyme variation among and within eleven Orchis species (fam. Orchidaceae), with special reference to hybridizing aptitude. Genetica 81: 143-150.

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Scacchi, R., P. Lanzara, and G. De Angelis. 1987. Study of electrophoretic variability in Epipactis helleborine (L.) Crantz, E. palustris (L.) Crantz and E. microphylla (Ehrh.) Swartz (fam. Orchidaceae). Genetica 72: 217-224. Sheviak, C. J. and M. Bracht. 1998. New chromosome number determinations in Platanthera. North American Native Orchid Journal 4(2): 168-172. Sneath, P.H.A. and R.R. Sokal. 1973. Numerical taxonomy. W.H. Freeman, San Francisco, California. Soltis, D.E., C.H. Haufler, D. Darrow, and G. Gastony. 1983. Starch gel electrophoresis of ferns: a compilation of grinding buffers, gel and electrode buffers, and staining schedules. American Fern Journal 73: 9-27. Swofford, D.L. and R.B. Selander. 1989. BIOSYS-1. A computer program for the analysis of allelic variation in population genetics and biochemical systematics. Release 1.7. Illinois Natural History Survey, Champaign, Illinois. Tanaka, R. and H. Kamemoto. 1984. Chromosomes in orchids: counts and numbers. In J. Arditti [ed.], Orchid biology: reviews and perspectives, III, 323-410. Cornell University Press, Ithaca, New York. Weeden, J.F. and N.F. Wendel. 1989. Genetics of plant isozymes. In D.E. Soltis and P.S. Soltis [eds.], Isozymes in Plant Biology, 46-72. Dioscorides Press, Portland, Oregon. Wendel, N.F. and J.F. Weeden. 1989. Visualization and interpretation of plant isozymes. In D.E. Soltis and P.S. Soltis [eds.], Isozymes in plant biology, 5-45. Dioscorides Press, Portland, Oregon.

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Werth, C.I. 1985. Implementing an isozyme laboratory at a field station. Virginia Journal of Science 36: 53-76.

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Cowden: ALLOZYME VARIABLILITY IN THE PLATANTHERA CILIARIS COMPLEX Table 1. Enzyme systems, enzyme abbreviations, and associated electrophoretic buffer systems used for allozyme resolution. Numbered systems, with the exception of 8H*, appear in Soltis et al. (1983); LiOH is as in Werth (1985).

Enzyme
Glutamate dehydrogenase- (E.C. 1.1.1.118) Glucose-6-phosphate isomerase- (E.C. 5.3.1.9) Isocitrate dehydrogenase (E.C. 1.1.1.42) Leucyl amino peptidase (E.C. 3.4.11.1) Malate dehydrogenase (E.C. 1.1.1.37) Phosphogluconate dehydrogenase (E.C. 1.1.1.44) Phosphoglucomutase (E.C. 5.4.2.2) Shikimic acid dehydrogenase (E.C. 1.1.1.25) Superoxide dismutase (E.C. 1.15.1.1) Triose phosphate isomerase (E.C. .3.1.1)

Enzyme abbreviation GDH GPI IDH LAP MDH PGDH PGM SkDH SOD TPI

Buffer system LiOH,7 9,11 11 7,9 9 9,11 11 8H LiOH 8H

*System 8H is a modification of Soltis et al. (1983) developed by R. James Hickey. Gel and electrode buffers are as follows: Gel buffer, pH 7.6 Electrode buffer, pH 8.0 Tris...........……………………3.996g LiOH............….1.636g Citric acid, monohydrate......1.15 g Boric acid.......16.262g LiOH...........………………….0.168g Dilute to 1 liter Boric acid.....…………………1.855g Dilute to 1 liter

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Table 2. Matrix of mean genetic similarity (Nei's unbiased identity) for intraspecific (along the diagonal) and interspecific (below the diagonal) comparisons among all Platanthera taxa. Range of genetic variation appears in parantheses. N = number of populations for each taxon.
N ciliaris 9 ciliaris 0.908 (0.736-1.000) blephariglottis 5 0.891 (0.712-0.991) xbicolor 2 0.896 (0.793-0.980) chapmanii 2 0.889 (0.723-0.998) cristata 1 0.789 (0.649-0.902) integrilabia 2 0.471 (0.394-0.544) 0.940 (0.888-0.992) 0.964 (0.933-0.998) 0.853 (0.728-0.961) 0.741 (0.663-0.875) 0.466 (0.393-0.550) 0.999 (0.999-0.999) 0.855 (0.815-0.890) 0.771 (0.770-0.772) 0.460 (0.448-0.477) 0.895 (0.895-0.895) 0.819 (0.806-0.829) 0.433 (0.398-0.503) ***** (*****-*****) 0.307 (0.254-0.360) 0.777 (0.777-0.777) blephariglottis xbicolor chapmanii cristata integrilabia

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Figure 1. Phenogram of relationships among Platanthera ciliaris complex members, constructed from Nei’s unbiased identity values (Nei, 1978).

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Table 3. Proportion of polymorphic loci (P) and mean number of alleles per locus (A), for populations (p) and taxa (t) and the average frequency of private alleles per locus (p(1)) for each taxon. Platanthera taxon ciliaris blephariglottis xbicolor chapmanii cristata integrilabia Pp 0.305 0.216 0.250 0.500 0.166 0.291 Pt 0.666 0.500 0.250 0.583 0.166 0.500 Ap 1.324 1.250 1.250 1.541 1.166 1.333 At 1.833 1.583 1.250 1.667 1.166 1.583 p(1) 0.000 0.030 0.000 0.142 0.500 0.600

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ORCHID CONSERVATION IN THE 21ST CENTURY: THE VALUE OF INCLUDING MYCORRHIZAL FUNGI TO PRESERVE ENDANGERED SPECIES
Lawrence W. Zettler Introduction Orchids have evolved a unique mycorrhizal symbiosis in which the fungal partner is consumed as an energy source by the orchid (=mycotrophy). In nature, the orchid life cycle is completely dependent on the availability of specific fungi to initiate seed germination and seedling development. As orchid habitats continue to be destroyed by human-related factors, a pressing need arises to secure mycorrhizal fungi for use in orchid conservation efforts projected for the next century. This paper will briefly describe the orchid-fungal symbiosis, explain how fungi can be used to grow orchids from seed, and emphasize the importance of the symbiosis for conservation during the 21st century. The Orchid-Fungal Symbiosis and its Significance The orchid-fungal symbiosis begins when an orchid seed‘s embryo becomes infected by a Rhizoctonialike fungus (Basidiomycotina). Upon entering the suspensor region of the embryo, the fungus proliferates

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within cortical cells of the host, forming ball-like masses of hyphae, termed pelotons (Fig. 1). Pelotons are distinctive structures unique to an orchid mycorrhiza, and are usually confined to cortical regions of root-like organs. In a stable symbiosis, orchids are able to control the spread of fungal infection by producing natural fungicides (phytoalexins) and by digesting pelotons (Fig. 2). The latter process yields carbohydrates (energy), previously tied up in fungal hyphae, to the host orchid. As such, orchids are mycotrophs or ―fungus feeders.‖ Because there is no evidence that the fungus benefits from this association and it appears to be harmed but rarely killed, most authorities now view the symbiosis as a form of parasitism by the orchid on the fungus. The ability of orchids to obtain energy from fungi, in addition to sunlight (photosynthesis), clearly demonstrates that these plants are heterotrophic, especially as seedlings. Once mature, orchids are believed to exist as both heterotrophs and autotrophs, depending on a given species‘ needs. With this alternative nutritional capability, the Orchidaceae has evolved to become the world‘s most diverse plant family with an estimated 30,000+ species. Orchid seeds, for example, have no need for weighty food reserves (endosperm) making them capable of long-distance wind dispersal into new habitats favoring speciation. As another example, some terrestrials such as pink lady'sslipper, Cypripedium acaule, perform ―disappearing acts,‖ i.e., they remain below ground for several years as mycotrophs, then reappear in an autotrophic capacity perhaps to maximize reproductive success in favorable years.

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While the utilization of fungi as an energy source appears to have benefited orchids as a whole, it has its costs. Without fungi in the natural habitat, seeds would fail to germinate, abruptly ending the orchid life cycle. This absolute requirement for fungi has made growing orchids from seed a complicated and difficult process compared to other plants. For endangered orchids, this dilemma poses a serious challenge to conservation efforts. Artificial attempts to propagate orchids from seed on nutrient media without fungi have been largely successful for the epiphytes (Fig. 3) but unsuccessful for many terrestrials. If orchids are artificially propagated with little regard for fungi and reintroduced into ―restored‖ habitats, they will not be able to spawn seedlings unless suitable fungi are available for germination. As native orchid habitats continue to be destroyed by human-related factors, there arises an urgent need to isolate, screen, and preserve mycorrhizal fungi. One means to propagate orchids and screen fungi for conservation purposes is to employ the technique of symbiotic seed germination.

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Symbiotic Seed Germination: An Overview Symbiotic seed germination seeks to mimic nature by exposing orchid seeds to fungi to initiate the germination process, usually in the laboratory. It is considered a valuable conservation tool because it can be used to rapidly germinate seeds of endangered species that often resist attempts at artificial (asymbiotic) propagation. It also provides a means to screen fungi for their effectiveness at initiating a symbiosis. In addition, it permits fungal-infected seedlings to survive soil transfer better than those grown without fungi. Its primary drawback lies in the investigator, because few are trained in mycology or are willing to work with two organisms (orchids and fungi) instead of one (the orchid). The most widely used symbiotic technique involves sowing surface-sterilized seeds in vitro over a low-nutrient medium such as oatmeal agar within a Petri dish or test tube slant. Upon the introduction of an effective fungus followed by incubation in darkness, orchid seeds often germinate rapidly (ca. 2 weeks) and at high percentages (>50%), depending on the species and seed pretreatment used. Under optimal conditions, orchid embryos undergo rapid and dramatic change in size and appearance, transforming into leaf-bearing seedlings from masses of undifferentiated cells. Most seedlings can be exposed to white light upon leaf expansion until tissues develop green pigmentation and presumably initiate photosynthesis. Eventually, such seedlings are suitable for transfer ex vitro, and usually

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survive provided they are acclimated in a greenhouse on a mixture of moist, pre-inoculated soil and Sphagnum moss (Fig. 4). In cooler climates, seedlings should be transplanted into suitable habitats during spring, after the threat of frost. Presumably, fungal-inoculated seedlings have the ability to digest pelotons as a food source, making their transition to adulthood more likely. It is also assumed that fungal inoculum from the seedlings may permeate into the surrounding substrate and spawn additional seedlings. In 1993, a new symbiotic technique was developed that has the potential to revolutionize our understanding of orchid ecology and greatly benefit conservation efforts. It involves placing terrestrial orchid seeds in packets constructed from nylon plankton netting followed by burial in a natural substrate (Fig. 5). Mycorrhizal fungi in the substrate are permitted to pass through the netting and into the orchid embryos. Packets are then retrieved and inspected for fungalinfected seedlings. For the first time, fungi can be isolated from young seedlings in the natural habitat—a process that would practically be impossible considering the minute size of seeds and seedlings. Clearly, this technique provides a quantitative means to study orchid seed germination in situ. It also represents an instrument Zettler used to extract the most effective fungal isolates responsible for germination. To a mycologist, the technique can be viewed as a way to ―bait‖ fungi from soil. Once extracted from orchid tissues, fungi can be screened for their effectiveness at promoting in vitro germination, followed by permanent deposition into an

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international culture collection such as the University of Alberta Microfungus Collection and Herbarium (UAMH). In light of ongoing habitat destruction, I strongly urge other investigators to consider using both techniques in order to preserve the often-overlooked fungal component. Conclusion The ecology and distribution of orchid mycorrhizal fungi in nature remains poorly understood despite their importance to the Orchidaceae. Recent evidence suggests that orchid fungi may persist as independent saprophytes with a wider distribution than the orchids they support. Some orchid fungi may even form partnerships with members of other plant families. Until it can be demonstrated that habitat destruction does not contribute to the loss of the fungal taxa required by orchids, every attempt should be made to include fungi in conservation efforts. This includes the fungi of epiphytic and terrestrial taxa alike, despite the relative ease by which the former group can be cultivated from seed in the absence of fungi. It is simply too easy to focus on the highly visible orchid plant while ignoring the rarely seen, poorly understood, yet critical fungal component. The use of in situ symbiotic seed germination provides hope for isolating the most effective fungi. In vitro symbiotic techniques provide the method to screen fungal isolates, and cultivate orchid seedlings with a mycotrophic capability.

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Zettler: ORCHID CONSERVATION Acknowledgments I extend thanks to Marta R. Bowen (Springfield, IL) for critiquing the text, and to Paul Martin Brown for providing the photograph depicted in Fig. 3. FURTHER READING: Arditti, J. 1966. Orchids. Scientific American 214: 70-78. Clements, M. A. 1988. Orchid mycorrhizal associations. Lindleyana 3: 73-86. Gill, D. E. 1996. The natural population ecology of temperate terrestrials: pink lady‘s-slippers, Cypripedium acaule. In: North American native terrestrial orchid propagation and production (conference proceedings), National Arboretum, Washington, D.C. (ed. C. Allen) pp. 91-106. Rasmussen, H. N. 1995. Terrestrial Orchids: From Seed to Mycotrophic Plant. Cambridge University Press, Cambridge. Rasmussen, H. N. and D. F. Whigham. 1993. Seed ecology of dust seeds in situ: A new study technique and its application in terrestrial orchids. Am. J. Botany 80: 1374-1378. Rasmussen, H. N. and D. F. Whigham. 1998. Importance of woody debris in seed germination of Tipularia discolor (Orchidaceae). Am. J. Botany 85: 829-834. Zelmer, C. D. and R. S. Currah. 1995. Evidence for a fungal liaison between Corallorhiza trifida (Orchidaceae) and Pinus contorta (Pinaceae). Can. J. Botany 73: 862-866. Zettler, L. W. 1997. Orchid-fungal symbiosis and its value in conservation. McIlvainea 13: 40-45. Zettler, L. W. 1997. Terrestrial orchid conservation by symbiotic seed germination: Techniques and perspectives. Selbyana 18(2): 188-194. Lawrence W. Zettler, Ph.D., Department of Biology, The Illinois College, 1101 West College Ave., Jacksonville, Illinois 62650 USA. Dr. Zettler teaches organismal biology at The Illinois College and directs undergraduate student research in collaboration with The Morton Arboretum in Lisle, IL where he serves as a Research Associate. E-mail: lwzettle@hilltop.ic.edu

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DON'T FORGET THE 1999 4TH NORTH AMERICAN NATIVE ORCHID CONFERENCE!
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Encyclia tampensis Florida butterfly orchid FL P.M. Brown

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Left: Corallorhiza maculata var. occidentalis forma immaculata western spotted coralroot, yellow spotless from MN

Below: Cypripedium reginae showy lady's-slipper MN Dianne Plunkett Latham

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PHOTOGRAPHING NATIVE ORCHIDS
Dianne Plunkett Latham Photographing wildflowers such as native orchids need not be difficult or involve a large amount of expensive equipment. Whereas an avid nature photographer could use a triple flash/tripod set-up with a diffusion tent, I don‘t recommend it, as you may need to bring along a Sherpa to tote all your equipment into the field! To spare my back (and the environment) I use a minimum of equipment. Depending on the film speed preferred, a tripod may be unnecessary. I recommend basic equipment as follows: a single lens reflex camera (which is NOT set on automatic), a flash unit, and magnification equipment such as a macro lens or stacking close-up lenses. If you have a little extra space in your backpack, I would also add one or two of the commercially available expandable reflectors. The Camera A single lens reflex camera is necessary so that you can preview your photographs before snapping the shutter and confirm that the subject of interest is in focus. Do not use the automatic setting if you happen to have that feature, as camera light meters can be fooled

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in the dim light of the forest floor. It is better to log your shots for a few rolls. That is, keep track of your shutter speeds and F stops for each photograph. Review the photographs with the settings in various light conditions, thereby learning what range of settings work well for the film speed you prefer. You should then bracket your shots from among the range of useful settings. For example, if you are shooting in full sun, and you want maximum depth of field, a standard setting is achieved as follows. Set the shutter speed equal to the ASA of the film you are using (i.e., ASA 64 is 1/60 of a second) and use an F stop of 16. To bracket this you could either vary the shutter speed setting (1/30 or 1/125 for example), or you could vary the depth of field setting (F11 or F22 for example). Be careful if you vary both the shutter speed and the F stop as you may cancel out any change in lighting. A shutter speed of 1/125 at F8 admits the same amount of light as a shutter speed of 1/ 60 at F11, for example. The only difference one would see in these settings is that of a greater depth of field at the F11 setting. The higher the F stop, the deeper the area which is in focus. It is not unusual for me to shoot a 24-exposure roll per blossom trying many different settings. Somewhere on the roll you will indeed get the perfect shot! An automatic camera will not let you have the necessary level of control. Flash You will not need a very powerful flash as most wildflower photographs are taken at close-up range. To determine the correct setting for a flash exposure, you

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could review the flash specifications, dust off your physics formulas on the fall-off of light, and measure your distance to the subject. You can then precisely determine what shutter speed and F stop is correct for your speed film at a specific distance. That would be too much work for me, however. It is far easier to log a range of settings on a roll of shots using your preferred film and flash. From this you can tell what settings work best at your typical working distance for wildflower close-ups. Some photographers set up two auxiliary flashes mounted on tripods at either side of the subject. This is in addition to the main flash on the camera, which is also mounted on a tripod. A triple flash set-up will yield a very balanced light. Auxiliary flashes are activated by a slave unit attached to the auxiliary flash. The slave unit senses the burst of the main flash and trips the auxiliary flash. This necessitates three tripods (and a Sherpa) however, to say nothing of disturbing the environment while setting up all the tripods. I don‘t recommend this type of tripod set-up in fragile areas such as where orchids typically grow. Close-up Equipment To achieve sufficient magnification, a set of stacking close-up lenses, a macro lens, a reversal ring, a bellows, a set of extension tubes, or combination thereof, is necessary for wildflower photography. The least expensive way to achieve magnification is to use

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close-up lenses. A typical set of three close-up lenses can be purchased for less than $35.00. This is also one of the lightest weight methods of achieving magnification. A typical set includes magnification lenses of +1, +2 and +4. Such a set of close-up lenses can be combined for +3, +5, +6, or +7 magnification. The least light is lost with this method of magnification because the focal length is not significantly extended. Most of my orchid photographs were taken with these. I presently use a 90mm Tamron macro lens. With the increased focal length of the macro lens, the working distance is greater from the subject to the camera flash than with close-up lenses. As a consequence, the F-stop must be opened wider when using a macro lens since the camera with flash attachment will be back farther from the subject. A reversal ring is another inexpensive, lightweight method of achieving magnification. With a reversal ring, take the regular lens, in my case an F1.8, 55mm lens, reverse the direction of the lens, and attach the lens by screwing it on to the barrel of the camera using the reversal ring. This will yield about a one-to-one life-size photograph. Reversing a 24mm wide-angle lens will yield about a 2.5 magnification. This is the magnification method used by Lorne Heshka for his excellent program on Manitoba's Native Orchids during the July 1998 NANOA conference. An advantage of reversing the lens is that no additional light is necessary.

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The disadvantage of a reversal ring is that you cannot use the lens focusing system but must focus by moving the camera, thus it is convenient to have a focusing rail. With the reversal ring you lose the bright wide open lens focusing because the automatic diaphragm lever is facing away from the camera. You can use your finger, however, to push this lever to open the lens diaphragm for focusing and release it to stop-down mode when releasing the shutter. Another alternative for magnification is to use a bellows and rack out the lens to the desired length for the intended magnification. Using a bellows has the advantage of allowing magnification on a sliding scale rather than a fixed magnification system such as when reversing a lens. A bellows has the disadvantage of being heavy, bulky, time consuming to set up, and requires more light because of the increased focal length. A final magnification option to consider is to add one or more extension tubes. This could be done in combination with the bellows. This has the disadvantage of increasing the focal length, thereby necessitating greater illumination. Extension tubes are a relatively lightweight, relatively inexpensive way of achieving magnification. Reflectors

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If you do not want to use an auxiliary flash, it is wise to add some reflectors to your backpack to bounce in additional sunlight. Compact, expandable reflectors, which fold into a small area, are commercially available. When released, they form a circle of white, silver or gold material. I use the LITEDISC 12 inch diameter white reflector and the 22 inch diameter white/silver reflector manufactured by Photoflex. They fold to a five-and-onehalf inch diameter and a 9-inch diameter size respectively. Alternatively, one can make his or her own reflector by covering any size or shape piece of cardboard, or stiff material, with a foil such as Reynolds Wrap. If you don‘t have a friend to hold the reflectors for you, be sure to bring a few of your gardening sticks to lean them against. Bamboo sticks take up little room in your pack and can be driven into the ground with minimum impact to the environment . Film I have a strong preference for Kodachrome 25 film because it provides the greatest resolution (detail). Such slow film, however, will generally require the use of a tripod, unless the subject is in quite a bright light. In general, you want to use the slowest film available that you are willing to consider in order to obtain the greatest detail. The slower the film, the greater the detail. If Kodachrome 25 is too slow for your patience, try Ektachrome 64. Higher speed film is useful if there is much wind, or if conditions are very dark, and you do not want to increase light artificially.

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Tripod A tripod is advisable, especially if you are using slow-speed film in low-light conditions. I use a Gitzo tripod with a Bogen head. The advantage of a Gitzo tripod is that the legs can be spread out nearly flat to the ground. The advantage of the Bogen head is that it has a shorter column than the standard Gitzo head, allowing the tripod to be positioned close to the ground. The tripod head can be swiveled and tilted in many directions. Using a strap to sling the tripod over your shoulder will make hiking easier. When working with a tripod, using a cable release is always recommended, to eliminate any movement of the camera. Natural Backgrounds Having a complementary yet contrasting background is the objective of wildflower photography. This can be done by several means. Blurring the natural green background is very pleasing. A pink lady’sslipper, Cypripedium acaule, photographed at St. Croix. State Park in Minnesota with a +6 close-up lens was taken with Kodachrome 25 at a 1 second F16 exposure using a tripod and flash. The one-second exposure brings the light from the green background through. An alternative example showing more of the background in another pink lady’s-slipper photograph taken at Land O‘ Lakes State Forest in Minnesota using Kodachrome 25 with a one second exposure at F16 with a flash. The

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detail is beautiful, but a very still day is required for these exposures! Black Backgrounds An alternative to using a background that shows the color of the environment, is to use settings which produce a black background. Although I find it more pleasing to use the color of the natural background, this is not always possible. In the attached showy lady’sslipper, Cypripedium reginae, which I photographed at Ft. Snelling State Park in Minnesota, I had a wire fence in the background. To eliminate distracting backgrounds, choose settings such that the fall-off of light will result in a black background. Black backgrounds will also give the maximum contrast for those photographs in which you want the subject to stand out with maximum impact. The following sequence shows the variety of equipment and film that can be used to achieve a black background. The C. reginae black background which eliminated the fence was achieved through the use of Ektachrome 64 and an exposure of 1/60 of a second at F22 with a flash. I call this type of a shot a ‗grab‘ shot because the camera can be hand held without the need of a tripod. A +4 close-up filter was used as well. A similar black background effect can be achieved by using Kodachrome 25 with a 1/60 second F16 exposure using close up filters as was done with a western prairie fringed orchid, Platanthera praeclara. A black background was also achieved in the attached western spotted coralroot, yellow spotless form, Corallorhiza maculata var. occidentalis forma immaculata, taken at Scenic

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State Park in Minnesota, using Kodachrome 64 with a one second, F16 flash exposure, including a 90mm macro lens with 3 extension tubes plus a tripod. Another example of a dark background was achieved with a Calypso taken on Flower Pot Island near the Bruce Peninsula in Lake Huron, Ontario. It was shot with Kodachrome 25 at a ½ second, F22 exposure with flash and a tripod using +6 close-up lenses. In summary, when using Kodachrome 25 and close-up lenses, a one second F16 exposure will bring through the background whereas a 1/60 second F16 exposure at a similar magnification and flash will cause the background to black out. Artificial Backgrounds An example is of a Case’s ladies’- tresses, Spiranthes casei, that showed the ditch habitat near Spooner, Wisconsin. It was taken on Kodachrome 25 with a one second, F16 exposure using a tripod and flash. To achieve a more effective close-up and remove distractions in the background, I had a friend hold the blue reverse side of my jacket in the background. The photograph was taken at a one second, F11 exposure with a +6 magnification on a tripod with a flash. Another technique for replacing distracting backgrounds is to move to your setting something natural from elsewhere in the environment. Examples could include a piece of birch bark or a moss covered rock. Be sure to return whatever you have chosen to the spot from which you found it so as to have minimal impact on the environment. When photographing in the field, be

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careful not to trample immature plants and other companion plants. Resist the urge to snip off the flowers of adjacent plants which may be a distraction in your background; a native plant population needs the flower‘s seeds to sustain itself. Instead, use one of the artificial background or black background techniques, which I have described, to remove any distracting elements. Composition There are some important principles to keep in mind when composing your photograph. These include positioning of the center of interest, positioning leading line diagonals from the left side of the photograph, and selecting an odd number of subjects. Resist the urge to place a subject in the center of a photograph, as such a composition is static. It is better to place a subject on one of the ‗golden thirds‘. For this placement, imagine a tic-tack-toe image imposed upon your lens. Place the subject of interest on one of the four intersections. If you have diagonal line, it should originate on the left side of the photograph because the eye, in our culture, reads from left to right. If you have multiple subjects of interest, it is best to have an odd number of subjects, as the placement will appear less static then an even number of subjects. Three large yellow lady’s-slippers, Cypripedium parviflorum var. pubescens were photographed on Dorcas Bay in the Bruce Peninsula, Ontario on Kodachrome 25 film at 1/60 F11 with a flash. Try back lighting or side lighting for more impact. For further tips on composition read the excellent article on Picture Perfect

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Orchids by the Slow Empiricist in the June 1998 issue of the North American Native Orchid Journal at pp. 133 – 147. More Information Photographing wildflowers, especially native orchids, has been an interest of mine for nearly 20 years. An excellent way to learn more about photography is to join a nature photography organization. I am indebted to the Minnesota Nature Photography Club for their many lectures and field trips through which I learned much. Contact your local photography store to obtain the names of your local camera clubs.

Dianne Plunkett Latham, 7013 Comanche Ct., Edina, MN 55439 E-mail: PlunkettDi@msn.com Dianne is the former President of the Minnesota Nature Photography Club and the past chair of the Minnesota Exhibition of Botanical Photography. She is a three-star exhibitor in the Photography Society of America Nature Division

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3RD ANNUAL NORTH AMERICAN NATIVE ORCHID CONFERENCE FIELD TRIPS, JULY 10 & 11, 1998
Philip E. Keenan Lake Itasca State Park, Minnesota, is near the dividing line between the forested and open prairie of northwestern Minnesota, just southwest of Bemidji, a small city of 11,000 people with a convenient air link to Minneapolis. This allowed for a fair diversity of orchids seen on the two days of field trips. In a wooded bog adjacent to the park an unusual juxtaposition of three species of adder's-mouth orchids, Malaxis, occurred in full bloom: bog adder'smouth, M. paludosa, white adder's-mouth, M. brachypoda, and green adder's-mouth, M. unifolia. The very rare M. paludosa produced nearly one hundred stems of perhaps the smallest terrestrial orchid flowers on the North American continent. It was interesting to note that here in the shade of a wooded bog the flowers were noticeably smaller than those in an open, sunlit bog in Anchorage, Alaska a few years ago. Large numbers of blunt-leaved rein orchis, Platanthera obtusata, in prime condition, were scattered among heart-leaved twayblades, Listera cordata, most of which were past

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prime. Many people were attracted to several good specimens of pad-leaved orchis, Platanthera orbiculata. Also noted were the almost ubiquitous northern green bog orchis, Platanthera hyperborea, just another green orchid to many, but not all, participants—who ran the gamut from beginner to expert—and the recently differentiated pale-green bog orchis, P. huronensis. In the ditches, a few remaining showy lady's-slippers, Cypripedium reginae, still in good color, provided a final photographic opportunity, and one clump of long bracted green orchis, Coeloglossum viride var. virescens, did likewise. (As in many areas of the country, this spring was two weeks earlier than usual in Minnesota). The northern small yellow lady's-slipper, Cypripedium parviflorum var. makasin, was one of the few orchids not in bloom. Also seen at this site were large yellow lady's-slipper, Cypripedium parviflorum var. pubescens, lesser rattlesnake orchis, Goodyera repens, ram's-head lady's-slipper, Cypripedium arietinum, small roundleaved orchis, Amerorchis rotundifolia, and hooded ladies'-tresses, Spiranthes romanzoffiana, which were all present either in the woods or along the adjacent road, For an excellent total of sixteen species and varieties. Despite the large number of people, foot traffic was carefully controlled, resulting in minimal damage. The highlight in the field was the second day‘s drive of more than three hours (each way) to extreme southern Manitoba‘s Tall-grass Prairie Preserve near Tolstoi, Manitoba. Why Manitoba and not the Minnesota prairie, you ask? Because this was a poor year

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in Minnesota where apparently it never produces the quantity of prairie fringed orchids that Manitoba consistently does. Perhaps 10,000 flowering, 3-foot, sturdy stems of western prairie fringed orchid, Platanthera praeclara, waved back and forth in moderate breezes under a blistering sun. The size and substance of the intensely white lips is most impressive. For example, the middle segment of the thrice-divided lip measures just under two inches in diameter! This is larger than any other fringed orchid, including the eastern prairie fringed, Platanthera leucophaea, which it clearly surpasses in size and beauty. The one advantage the eastern has over the western is its sensational strong mid-day fragrance. The western offers its fragrance, interestingly, only at dusk. The western prairie fringed orchid is rated second only to Cypripedium reginae as the most spectacular North American native orchid north of Florida.

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Philip E. Keenan, 31 Hillcrest Dr., Dover, NH . 03820 Philip is a regular contributor to the Journal and is the author of a myriad of articles on native orchids as well as a new book, Wild Orchids Across North America, to be published by Timber Press this fall. E-mail: sk2325@server1.rscs.net

Platanthera praeclara western prairie fringed orchis

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LOOKING FORWARD
December 1998 PROCEEDINGS OF THE ANNUAL NORTH AMERICAN NATIVE ORCHID CONFERENCE Part 2 …….and more!

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4th ANNUAL NORTH AMERICAN NATIVE ORCHID CONFERENCE
April 8-13, 1999
University of South Florida Tampa, Florida

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Thursday & Friday - April 8 & 9 Field trips in north-central Florida Ocala/Gainesville area Saturday & Sunday - April 10 & 11 Conference Sessions at USF Monday & Tuesday - April 12 & 13 Field trips in south-central Florida Sebring/Naples area

Registration Fee of $45 per person includes all field trips and conference sessions A schedule of the field trips and suggested list of accommodations will be sent upon registration Space is limited to 75 so register early Send your check for $45 per person to North American Native Orchid Alliance PO Box 772121 Ocala, Florida 34477-2121
The 1998 conference was sold out several months prior to the July dates. Do not be one of the members who will have to be turned away for the 1999 conference. Register and make reservations early! Last year we had several members who made their airline and hotel reservation in December but neglected to register for the conference until the last minute. This year when the conference is full we will not be able to take add-ons even if you have made other paid reservations!

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BOOK REVIEWS:
The Genus Paphiopedilum, Natural History and Cultivation Dr. Guido Braem, Charles O. and Margaret L. Baker 1998 ISBN 0-9665337-0-4 Botanical Publishers, Inc., Kissimmee, FL Distributed by Alpha Orchids, Miami, FL 180pp. Paper. 9x12" $49.95 47 color photographs, 3 color paintings, 71 line drawings, 6 maps The latest addition to the global orchid literature is one of the most beautifully produced books of its type. The large page format, superbly printed color (mostly full page), illustrations and vast amount of both botanical and horticultural information leave little room for criticism. Although not dealing with native North American orchids, this volume is the first of a series that will eventually cover all species of Paphiopedilum, Phragmipedium and related genera, and Cypripedium. The latter will be of special interest to native orchid enthusiasts. The first section of this volume deals with taxonomy—a subject that is very dear to me—and

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critical to anyone who wants to truly understand the subject. Dr. Braem has clearly, accurately, and, at times, in great detail, given a crash course in botanical nomenclature and taxonomy, contrasted it to horticultural nomenclature, and given examples of perpetuated errors and pitfalls. He often cites both historical and contemporary examples of both proper and erroneous taxonomic citations. The section is an absolute must for all native orchid enthusiasts—whether rank amateur or professional! The second section is a detailed history and synopsis of the subfamily Cypripedioideae and the genus Paphiopedilum from both the historical and contemporary perspective. The majority of the section is devoted to a complete listing of all species in their respective sections. The last section is the bulk of the volume which contains detailed accounts of 31 species (of the 89 listed), with dramatic full-page color photographs, historical information, habitat data and just about everything you could possibly want to know about the culture of each individual species. An index is provided to taxa and it will be cumulative in each forthcoming volume. Only very few typographical errors were found: lilly for lily and a footnote when no footnote reference appeared. If I could have suggested anything different it would have been to include more in situ photographs and a Literature Cited for each volume. Although there are copious literature references throughout the text, the Literature

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Cited is scheduled for the concluding volume. The remaining species of Paphiopedilum are scheduled for Volumes 2 and 3 with a projected publication date of late autumn 1998 for Volume 2 and spring 1999 for Volume 3. Although not inexpensive, it is certainly worth every dollar for the quality of both the information and the publication. I thoroughly recommend this to all lovers of slipper orchids everywhere. PMB

Wild Orchids Across North America

A Botanical Travelogue
Philip E. Keenan

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1998. Timber Press, Portland, OR ISBN 0-88192-452-0 $39.95 320 pp., 170 color photographs, hardcover, 6x9" Memoirs and travelogues have long been a popular vehicle for sharing personal experiences on a wide variety of subjects. Most natural history tomes of this type are very general and have little specific information to generate vivid images for the reader. Not so with Philip Keenan's assemblage of more than two decades of traveling throughout most of North America in search of many of the wild orchids of that continent. Keenan‘s descriptive prose is a fitting complement for his brilliant full-color images of wild orchids taken in situ from coast to coast. Having accompanied him on many of these forays I can say that each vignette or anecdote brings me back to the day of discovery! For those who have not been there, his photographs will give you a taste of what can be waiting for you. Both the very rare and the frequent and familiar are treated with the same reverence and respect. For those who have lived in the Northeast the wide-spread pink lady's-slipper is still be to admired as much as the rapidly decreasing fairy-slipper. Keenan takes us to such diverse places as Newfoundland, Alaska (and its breathtaking Kodiak Island), the pinelands of New Jersey, and the open savannas of southeastern North Carolina. A few days in the 'sky islands' of southeastern Arizona yield several sought-after species. Although not

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a rabid 'life-lister' as are so many in various natural history fields, Keenan appreciates each new species and joins many of us in completing the study of all species within a genus. This volume is both an excellent addition to the native orchidist‘s library or a perfect gift for anyone who appreciates nature. I can think of no other book on native orchids, or wildflowers in general, that would be as ideal as a first volume to intrigue and infect someone with an appreciation for the natural world around them. PMB

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