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In addition to causing large losses of nursery stock. W. citricola. in other states and countries. citricola (35). cactorum (3). medicaginis (26). Balci et al.50) as well as forests (4–6. Moreover.. Paul. isolate. The most common species encountered were P. Direct sequencing of the internal transcribed spacer regions (ITS-1 and ITS2) of conserved ribosomal DNA (rDNA) allows investigators to compare sequences and has been employed successfully for . J. Smith. Plant Dis. P. Detecting P. P. In addition.43. Recent international and national surveys for the detection of P. the detection of new hybrids. Species associated with diseased ornamental plants include P. Paul. and R. Nonhosts of P. several new Phytophthora species have been described (10. Phytophthora isolates obtained were identified to species in this present study. Twenty-one nurseries in the fall of 2002 and 24 nurseries in the spring of 2003 were surveyed by Minnesota Department of Agriculture personnel. W. Plants that were previously confirmed as hosts or species associated with P. and the previously identified but undescribed taxon P.21) as well as an additional species. were sampled occaPlant Disease / January 2007 97 ABSTRACT Schwingle. infestans (22). megasperma isolated from alfalfa (Medicago) fields in Minnesota was likely P. several species have been isolated from studies on other crops. citrophthora. and the other appeared to be a new species and is referred to as P. species are reported for the first time from several hosts. however. Fifteen ornamental nurseries in or near Minneapolis and St.14. Most recently. ramorum. P. it has nationwide importance since Minnesota nurseries ship ornamental plants to nearly every state (40). such as P. including the nuclear beta-tubulin (β-tub) gene and mitochondrial encoded cytochrome c oxidase (cox) I and II genes. are discovered in nurseries and in the landscape. europaea. A. The largest nurseries were visited several times each year. A. Two additional isolates obtained did not match known species. alni (14).19). Species identification was carried out using direct sequencing of ITS rDNA.1 billion/year nursery and landscape industry (40). R. Phytophthora species negatively impact ornamental nurseries throughout the world (8.44.16. P.31. A. cryptogea (20). MATERIALS AND METHODS Study sites. Morphological characteristics have been primarily used in the past (8. it is very important to contain their spread as quickly as possible. where they have caused catastrophic damage (10. and mitochondrial coxI genes. the University of Minnesota. alni. They include P. ITS sequences do not clearly resolve all Phytophthora species (15.. no surveys of Phytophthora species had been carried out in Corresponding author: B. P. nicotianae.51). erythroseptica (48). DOI: 10.45). ramorum was not detected.. This work was conducted by the Minnesota Department of Agriculture.6.37). Department of Plant Pathology. University of Minnesota. The P. and the USDA Forest Service. whereas the smaller nurseries were sampled twice in the course of the study. since its host range includes several native and important plants in Minnesota (2. were sampled between April and September in 2004 and 2005. MN1. citricola. Phytophthora isolates were obtained only from symptomatic rhododendrons. P.49). possibly in nursery locations where related but geographically isolated species come in contact. (4) isolated P. When non-native Phytophthora spp. cambivora. Although this work was done in Minnesota. Minnesota ornamental nurseries were surveyed in 2002 and 2003 solely to detect P. Pgchlamydo. A. ramorum (2) were the focus of this study. megasperma.36.25. 91:97-102. sojae (54).20. University of Minnesota. β-tub. 2007. One was similar to P. Schwingle E-mail: schwi035@umn. St. P. and one nursery in Duluth. Samples collected during the surveys were identified to genus by staff in the Plant Disease Clinic. P. Smith. exotic Phytophthora species also threaten natural ecosystems. Blanchette. Since no previous Phytophthora surveys have been carried out in Minnesota. β-tub. hedraiandra. a pathogen which has destroyed entire tracts of California oak forests (45) and threatens forests of the eastern United States (2). J. It is equally important to define the natural geographic ranges of established Phytophthora species so quarantines are not unnecessarily enforced on native pathogens or widespread exotics. cactorum. to differentiate between them (33. sp. alni subsp. Prior to this report. ramorum was of key concern. and two unknown Phytophthora species from forest soils in Minnesota and Wisconsin. and identify indigenous and exotic Phytophthora species present in Minnesota ornamental nurseries.36). 495 Borlaug Hall. The same species have been found in Wisconsin (17. and undescribed species were causing disease in Minnesota ornamental nurseries. P.24. W. alni subsp. and coxI genes and also by studying morphological characteristics.1094 / PD-91-0097 © 2007 The American Phytopathological Society Minnesota despite the problems caused to the state’s $2. P. ramorum. and Blanchette. However. 1991 Upper Buford Circle.19. megasperma (21). Phytophthora species are responsible for causing extensive losses of ornamental plants worldwide. Results indicated that several Phytophthora species were more widespread in the nursery industry than previously thought.25). MN.20. Schwingle. ramorum. so other investigators have used additional genome regions. a P.52). citrophthora. indicates that new Phytophthora species are evolving.49. ramorum have led to the finding of previously undescribed Phytophthora species. MN 55108 species identification in several recent studies (5. Phytophthora species associated with diseased woody ornamentals in Minnesota nurseries. and P. P. Although no Phytophthora surveys have been carried out in Minnesota nurseries.52). The objective of the study presented here was to survey.37). Accepted for publication 5 September 2006. Several techniques have been used to identify isolates obtained from Phytophthora surveys. numerous disease surveys have established the Phytophthora species that frequent nurseries and plantations (8. P. but this form of species identification is not always reliable due to the morphological variability and overlap of species’ characteristics within the genus (9. indicating that our taxonomic knowledge of this genus is incomplete and that there are likely many undiscovered species that remain to be identified. Recently.Phytophthora Species Associated with Diseased Woody Ornamentals in Minnesota Nurseries B. such as P. P. quercina–like organism. surveys of ornamental nurseries were performed over 4 years to isolate and identify the Phytophthora species causing diseases of woody plants in Minnesota. Species were identified by direct sequencing of internal transcribed spacer (ITS) rDNA.32.43. cactorum. and P. However. B.

Sporangia were produced by placing a colonized V8 juice agar section from a 1week-old colony in a glass petri dish.0. nicotianae were isolated only in 2003. PCR products were prepared for DNA sequencing by mixing 1 µl EXOSAP-IT (USB Corp. cactorum.9 M boric acid. ramorum were sampled from 12 nursery sites. Several Phytophthora species isolated in 2004 and 2005 were derived from plants which had not been previously reported as hosts for that particular Phytophthora species (Table 2). Small sections from the margins of leaf. citrophthora were the most commonly isolated Phytophthora species during the 2004 and 2005 surveys. P. Cultures were stored at room temperature (20 to 25°C) in the dark and were monitored for hyphal growth for up to 10 days following the initial isolation. Processing of the container mixes and water occurred within 48 and 24 h. ramorum were sampled from 10 nursery sites (nurseries were sampled up to seven times throughout the growing season). respectively. Consensus sequences were matched to those in GenBank for identification to species level using BLASTn searches (1). Two Phytophthora species were isolated from the same plant sample several times in 2004 and 2005. PCR was performed using the following parameters: 5 min at 94°C for the initial denaturation. and a portion of the 28S rRNA gene were amplified by PCR in a PTC-200 Peltier Thermal Cycler or a PTCTable 1. Isolations were made within 48 h of collection.9 M Tris-HCl. P.0× TBE buffer (10× TBE = 0. Ltd. cactorum. They were also isolated in 2003. Diseased tissues from host plants. ITS nucleotide identities from the presumed P. All BLAST search results presented were accompanied by an E value of 0. Of those samples. P. Successful reactions were verified by visualizing PCR products on 1. hedraiandra P. taxon Pgchlamydo.8S. Cultures were then transferred to P10ARP-CMA or PARP-V8. In 2005.6). hedraiandra. DNA extraction. canker. Phytophthora species isolated in 2002 and 2003 from diseased leaves and stems of Rhododendron No. P. During the 2004 growing season. of sampling. citrophthora were isolated from the same plant sample in six separate instances. dieback. wilt) were removed. cactorum and P. stem. Diseased plant survey and isolation methods. were isolated from the same sample once. It was not possible to align the sequences of some isolates due to ambiguous sequence data derived from one sequencing direction (usually the forward direction). nicotianae isolates ranged from 88 to 97%. nicotianae. cactorum and Unknown 1. All isolates were transferred to V8 juice agar (19) slants and kept at 12°C for longterm storage.47). (33) in order to clarify their species identities. PCRs of the coxI and β-tub genes were performed for some isolates using the parameters specified in Kroon et al. citricola and P. and species identification. They were baited as described in Balci and Halmschlager (5. citricola came from the same sample on five separate occasions. which made up 47% of all identified species. respectively. In contrast.sionally if disease symptoms typical of Phytophthora were observed. cactorum and P. DNA was extracted from them with a Qiagen DNeasy Plant Mini Kit using the manufacturer’s instructions. sampling in June yielded significantly higher proportions of Phytophthora isolates (32%) relative to sampling in other months. 155 samples were collected from 21 nurs- eries. polymerase chain reaction (PCR). 18% were associated with Phytophthora.0 or nucleotide identities below 99% were further investigated by sequencing coxI or β-tub genes as well as by studying morphological traits. Complimentary forward and reverse DNA strands were aligned with the software program ChromasPro (version 1. and P. Isolations yielded Phytophthora from 11% of the samples.) and illuminated by a Dark Reader transilluminator (Clare Chemical Research. 1 150 Minicycler using the universal primers ITS1 and ITS4 (53). potting mixes of host plants and surface water from irrigation water holding ponds were sampled and stored in plastic bags and sterile plastic containers. ITS-1. Technelysium Pty. P. at 4°C until they were processed. and root lesions were removed and submerged in P10ARP-CMA. and species identification was not corroborated with morphological data because cultures did not survive in storage for a sufficient period. cactorum and P. citricola P.) and optimized by manual alignment. or PARP-V8 selective medium (20). and 1 min at 72°C. P. and incubating on a lab bench for 24 h. ITS-2. and stored at 4°C until isolations were made in the laboratory.8S. A final extension was run for 5 min at 72°C. CO). and of this total. citrophthora P. 186 diseased plants known to be hosts or plants associated with P.5 µl PCR product and performing the following thermocycler program: 15 min at 37°C and 15 min at 80°C. placed in plastic bags. a previously identified but undescribed taxon (12). 392 diseased plants known to be hosts or plants associated with P. and P. which displayed symptoms indicative of Phytophthora infection (i. citricola and P. 1 min at 50°C. A portion of the 18S rRNA gene. P. The BLAST results for these isolates were also corroborated with published morphological data (19. 10 mM EDTA). 15% yielded Phytophthora isolates. cactorum P. 18% yielded Phytophthora. Isolates sharing a nucleotide identity with a voucher specimen below 99% were verified morphologically. leaf necrosis.. When morphological characteristics were necessary to corroborate BLAST results. 98 . Isolates with BLAST search results accompanied with E values >0. Of those samples. citricola and P. Inc. P. 5. a Phytophthora-selective medium (30). Phytophthora species associated with diseased Rhododendron in 2002 and 2003 are presented in Table 1. gametangia and oospores were measured on V8 juice agar media after 1 week of growth. but not as frequently as P. Their length:breadth ratios and shapes were compared with published accounts (19.0. citricola. All sequencing was carried out in an ABI 377 automated DNA fragment analyzer at the University of Minnesota BioMedical Genomics Center. a recently described leaf pathogen of a Viburnum species (16) and a newly introduced organism to the United States (46). P.22. and ITS-2 rDNA between the various isolates of the respective Phytophthora species and GenBank accessions. PCR products were stained with SYBR green I stain (Cambrex Corp. citrophthora were isolated commonly in 2002 and totaled 91% of the identified Phytophthora species. OH) and 2. Dolores. RESULTS In the 2002 survey of Rhododendron. The ends of the consensus ITS sequences were trimmed to remove 18S and 28S sequences according to sequence boundaries described previously (34). In the 2003 survey. 302 diseased samples from 24 nurseries were collected. flooding with 40 ml of 1.5% soil extract (29). then 34 cycles of 1 min at 94°C. The production of chlamydospores and hyphal swellings by isolates were two additional morphological traits that aided in corroborating species identity. citrophthora were also baited from potting mixes containing plants that harbored these respective Phytophthora species on diseased aboveground tissue. making up 16.47). Like the 2004 survey. After subcultures grew on selective media for 1 to 4 weeks. taxon Pgchlamydo. During 2004. 91 No. respectively (Table 2). pH 8. Sampling in June yielded significantly higher proportions of Phytophthora isolates (23%) than did sampling in July through September.0% agarose gels in 1.e.. Cleveland. of isolates 24 10 15 8 3 BLAST search identitiesa 99–100% 99–100% 98–100% 99–100% 99% Phytophthora species P. Diseased roots were washed in sterile tap water for 20 s and then blotted dry with sterile paper before submerging in selective media. 0. P. As in the 2002 and 2003 surveys. Plant tissue was the primary material used for isolation. taxon Pgchlamydo a These values represent the range of nucleotide identities in the ITS-1. but potting mix and irrigation water were sampled occasionally.. and 17% of the total number of species identified. Plant Disease / Vol. 55. respectively. was found in both years. and P. 5.

f The ITS sequence of Unknown 2 matched that of a P.c Malus ‘Snowdrift’c Malus ‘Brauzam’ Acer platanoides Symptoms Leaf spot Leaf spot Leaf lesion Dieback. Ribes alpinum Viburnum prunifoliumc Quercus macrocarpa Quercus macrocarpa Acer tatricum Acer spp. In addition. Its coxI sequence data matched P. Viburnum trilobum Hibiscus sp. leaf margin necrosis Leaf tip necrosis Discolored cambium Leaf lesion Leaf lesion Leaf margin necrosis Dieback. wilt Root rot Leaf spot Dieback Dieback. e Unknown 1 produced poor ITS sequence data that matched P. sp. All BLAST search results presented were accompanied by an E value of 0. leaf lesion Dieback Leaf lesion Dieback Leaf lesion Leaf Spot Dieback. leaf tip necrosis No. citricola DQ486659 DQ486660 DQ486661 DQ486662 DQ486663 DQ486664 DQ486665 DQ486666 P.0. Forsythia ‘Northern Gold’c Hamamelis virginianac Lonicera spp. dieback. c These host genera are not proven hosts of the respective Phytophthora species. of isolates 1 1 1 2 1 2 1 1 1 1 1 3 1 1 1 1 1 2 1 2 3 1 1 1 1 1 1 37 1 1 5 1 1 1 1 1 2 1 1 1 1 1 7 2 1 1 1 1 2 1 1 1 1 1 1 BLAST search identitiesa 100% 100% 99% 99–100% 99% 99–100% 100% 100% 100% 100% 100% 100% 99% 100% 100% 98% 100% 100% 100% 99–100% 98–100% 100% 99% 100% 100% 99% 100% 99–100% 100% 100% 99–100% 100% 99% 99% 99% 99% 99% 100% 99% 99% 100% 100% 99–100% 99–100% 100% 99% 100% 100% 100% 100% 100% 99% n/ad n/ae n/af GenBank accessionb DQ486654 DQ486655 DQ486656 DQ486657 P. leaf lesion Dieback. Froebelc Syringa vulgaris Tilia americana Viburnum prunifoliumc P. stems.8S. Morphologically. resulting in high nucleotide identity (99%) and an E value of 0. and ITS-2 rDNA between the various isolates of the respective Phytophthora species and GenBank accessions. wilt Leaf lesion Leaf lesion Dieback. Myrica pensylvanicac Prunus maackii Pyrus calleryana Rhododendron ‘Henry’s Red’ Rhododendron ‘Pahjola’s Daughter’ Rhododendron cvs. rubi GenBank strain with a high nucleotide identity (99%). leaf lesion Dieback Leaf tip necrosis Stem rot.c Magnolia stellatac Pyrus calleryanac Quercus rubra Rhododendron ‘Mikkeli’ Rhododendron ‘Rosy Lights’ Rhododendron sp. gonapodyides. gonapodyides. The same isolate had high nucleotide identity (99%) with coxI of a P. dieback. and roots of ornamental plants Phytophthora species P.c Taxus cvs. hedraiandra P. 5. Phytophthora species isolated in 2004 and 2005 from diseased leaves. b Accessions to GenBank were made for a representative number of isolates for each species. Magnolia stellata Rhododendron ‘Mikkeli’ Rhus aromaticac Rhododendron ‘Henry’s Red’c Taxus sp.0. Morphologically. leaf lesion Leaf lesion Leaf and petiole necrosis. cambivora GenBank specimen. MN1 did not match any species in GenBank well despite nonambiguous sequence data. Rhus aromaticac Spiraea × bum. taxon Pgchlamydo P. drechsleri more so than P. Plant Disease / January 2007 99 . d ITS and β-tub sequences of P. root rot Stem rot Leaf margin necrosis Canker.Table 2. leaf lesion Canker. it represented P. leaf lesion Dieback. cultures representing these accessions will be deposited in the American Type Culture Collection (ATCC). fragariae var.c Rosa sp. cambivora P. drechsleri and resulted in an E value of 2 × 10-82. nicotianae DQ486667 DQ486668 DQ486669 DQ486670 DQ486671 DQ486672 DQ486658 P. citrophthora Dictamnus alba-purpureusc Hamamelis virginianac Myrica pensylvanicac Rhododendron ‘Haaga’ Rhus aromaticac Rhus spp. the isolate matched both species. root rot. stunting Wilt Canker. sp. root rot Leaf margin necrosis Leaf lesion Leaf tip necrosis Leaf lesion Canker Leaf lesion Canker. MN1 Unknown 1 Unknown 2 a These values represent the range of nucleotide identities in the ITS-1.c Euphorbia sp. megasperma P. Isolates sharing a nucleotide identity with a voucher specimen below 99% were verified morphologically. cactorum Host Acer rubrum Acer tatricum Calycanthus floridusc Lonicera cvs. leaf lesion. leaf lesion Leaf lesion Leaf margin necrosis Crown and root rot Leaf spot Dieback Dieback.c Malus ‘Beacon’ Malus cvs. leaf spot Leaf spot Bleeding canker Leaf lesion Canker Leaf lesion Dieback. Syringa ‘Miss Ellen Willmott’c Syringa vulgaris ‘Common Purple’c Syringa cvs. Rhododendron cvs. petiole necrosis.

One (Unknown 1. cambivora and a P. Another species (Unknown 2. fragariae var. Indiana. citricola was baited from potting mix that contained plants from which no Phytophthora isolates were recovered. P. rubi GenBank submission (accession AF266761). gonapodyides. Balci and Halmschlager commonly isolated P. and it has an ITS profile most similar to that of P. P. alni subsp. It resembled P. We cultured 18 isolates sharing the same diseased tissue with a different Phytophthora species in our 2005 surveys (i. 2. fragariae var. citrophthora was the most common Phytophthora species isolated from effluent holding ponds in three commercial nurseries in California. citricola was isolated from an irrigation water holding pond in 2004. cambivora in producing bullate oogonia and hyphal swellings. In addition to being baited from potting mix. from ornamental nursery plants in Oregon (43) and Ohio (49). this nucleotide identity was low (92% [434/472]) and the E value was high (10-171). on the two parental species occupying similar areas on the same host plant (11). alni (14). and Unknown 1 was isolated from Malus. P. rubi and P. Morphologically. (38). and the resulting hybrids display morphological characteristics of P. the species whose ITS rDNA most closely resembles that of P. citricola from 88% of all diseased rhododendron tissues tested. in 2003 and 2005 from Minnesota nurseries. 100 Plant Disease / Vol. Several species were identified in the 2004 and 2005 surveys that were not isolated in previous surveys (Table 2).3% of the 2005 samples). taxon Pgchlamydo was isolated from new hosts. sp. cambivora. and it best matched P. and West Virginia (4) and from stream water in the southern Appalachian Mountains (28). insolita. primarily because it produces chlamydospores (12. alni subsp. Phytophthora species identified from surveys in 2004 and 2005 were isolated from many host genera rather than from only Rhododendron species. DISCUSSION Surveys completed during 2002 to 2005 revealed the most frequently isolated Phytophthora species from Minnesota ornamental nurseries were P. In the United States. citrophthora. The ITS sequence derived from Unknown 2 shared 99% (768/770) nucleotide identity with a P. Orlikowski and Szkuta (42) isolated P. the ITS rDNA was sequenced several times. It closely resembled both P. and Puerto Rico (19). This is exceedingly important since Phytophthora hybrids have been found to infect different hosts than parent species (11. P. but P.24). drechsleri (accession L76549). In five nurseries in Poland. citricola (27). fragariae var.0). sharing 99% (869/879) nucleotide identity. drechsleri produced an E value of 2 × 10-82 and a nucleotide identity of 99% (162/164). citricola was also isolated there (36). alni has proven to be a very damaging pathogen of Alnus (11.e. sp. and hybrids can cause widespread damage on these new hosts (24). despite unambiguous sequence data. Since P. E value = 0. Bolivia. named Group K isolates. Ohio. cambivora. However. but they are homothallic. MN1 after P. However. (13) suggested that isolates previously declared P. Results from the current study suggest that the P. However. citricola in surveys of Austrian (5) and Turkish (6) woodlands. 91 No. cambivora occasionally produces oogonia in single culture (23). rubi could not reliably be differentiated from P. and P. In addition. A sequence produced from the β-tub gene of this isolate compared most closely with that of a P. rubi. but no natural hosts are known (19). fragariae var. Nevertheless. Further investigation of Unknown 1 is needed to establish its taxonomic status. cambivora (accession DQ202503). and its host range must be investigated to determine if it is a threat to natural ecosystems as well as to ornamental plants. insolita GenBank submission (accession AY564073) (nucleotide identity = 95% [798/836]. megasperma were two of these species. Another isolate (P. sp. rubi in being homothallic. restriction fragment length polymorphism (RFLP). citrophthora and P. gonapodyides. quininea (accession DQ275189). was discovered in soil from a citrus orchard in Taiwan and found to be pathogenic to apple and cucumber fruits in inoculation trials. Not only are P.18). Unknown 2 resembled P. cultured from Malus ‘Brauzam’. Table 2) not found in previous surveys was isolated from Acer platanoides (Norway maple). The coxI gene of Unknown 1 was sequenced. P. P. in part. and isozyme profiles . Table 2). MN1 is heterothallic. drechsleri and not P. sp. fragariae–like species have been shown to hybridize (11).. Both P. cambivora and morphology similar to P. quininea. fragariae using ITS data. sp. Surveys of potting mixes and irrigation water were not conducted in 2005. Since these species are common to many areas of the world. fragariae var. Subsequent investigations showed these two Phytophthora varieties could be differentiated by sequencing coxI and II genes (37). citrophthora found in nurseries. The morphology of Unknown 1 was more typical of P. The A2 mating type of P. and P. The likelihood of Phytophthora hybrids developing in nursery settings depends. Unknown 1 is likely to be P. and the BLAST search match to P. the species whose β-tub gene most closely resembles that of P. Cooke et al. drechsleri than of P. Due to low ITS and β-tub nucleotide identities and different sexual reproduction character between our isolate and that of known species. gonapodyides belonged to a separate taxon because they produced chlamydospores and spherical hyphal swellings that were unlike P. sp. alni suggests it may be hybrid in origin. cambivora and P. 1 fragariae var. fragariae var. P. drechsleri was shown to be pathogenic on Malus roots and stems by Matheron et al. quininea. and a Taxus sp. P.P. sp. gonapodyides (accession AY564181). had an ITS sequence that best matched P. P. rubi in being homothallic and P. (15) demonstrated that P. Rhododendron spp. rubi and P. MN1. given its production of chlamydospores and hyphal swellings. P. quininea and P. citricola. fragariae var. P. This demonstrates that Phytophthora species commonly occupy the same niche in nursery environments. differed from known species in their mitochondrial DNA. A standard hybrid between these two parent species was named P. Unknown 2 resembles P. Unknown 2 was isolated from a variegated Norway maple tree. and from recycled irrigation water in ornamental nurseries in Germany (50). ramorum (accession AY616757) shared the next highest ITS nucleotide identity (E value = 5*10-136) with P. cambivora in producing bullate oogonia and hyphal swellings. The fact that the isolate we collected from a diseased Norway maple has gene sequences similar to both P. cambivora. P. insolita are homothallic species (19).19). the isolation of these species from Minnesota nurseries was not surprising. P. Two additional species not found in previous surveys were isolated from Malus (crabapple) cultivars. MN1. This conclusion was supported by another study that found those same isolates. MN1 appears to represent an undescribed taxon. Though the identity of Unknown 1 remains debatable because of poor ITS sequence data. citricola. Unlike the 2002 and 2003 surveys. and careful investigation of Phytophthora isolates should determine if they are suspected hybrids. alni subsp. the presence of a similar organism in Minnesota is alarming. Brasier et al. The previously identified but yet undescribed species P. MN1) cultured from a Malus species had highest ITS nucleotide identity with P. Morphologically. citricola was collected from oak forest soils in Illinois. resulting in ambiguous base-calling. but they also are encountered in natural areas. cactorum. is a pathogen of Cinchona species in Peru. Irrigation water in six Virginia nurseries contained P. MN1 belongs to an undescribed and unknown taxon. This is the first report of this taxon. fragariae var. Two other unknowns were isolated in the 2004 and 2005 surveys. The coxI sequence of Unknown 2 shared highest nucleotide identity with P.. cactorum. the coxI gene of the isolate best matched that of P. gonapodyides. and P. All three species are common in ornamental nurseries around the world and have been isolated from container mixes from nurseries in the southeastern United States (20). rubi is a pathogen solely of Rubus idaeus (19).

Gapped BLAST and PSI-BLAST: A new generation of protein database search programs. 81:1349-1357. 8:595-597. Gilli. 17. C. and P. N.. L. Phytopathology 77:1475-1482. T. and Maxwell. Jeffers. St. 1997.13). hedraiandra closely resembles P. Cooke.. Anonymous. M. isolates of P. LITERATURE CITED 1. 25:661-664. Ph.. P. cambivora and P. 7. and Schoelz.. Cultural characters. Creation of species hybrids of Phytophthora with modified host ranges by zoospore fusion. Surveys in Minnesota and in other regions of the United States must continue. B. Fortunately. L. Mycologia 97:218-228. This. Reemergence of potato and tomato late blight in the United States.41). Kong. Animal and Plant Health Inspection Service (APHIS). Pathol. C. Phytophthora species in oak ecosystems in Turkey and their association with declining oak trees. C. S. 1999. Washington. T.43.. E. J. Winton. 22..) Phytopathology 95:S46. Brasier... taxon Pgchlamydo in Minnesota.. hedraiandra isolates were associated with diseased rhododendrons. Pages 351-364 in: Phytophthora: Its Biology. Hansen.S.. Sandy Gould. J. and Hansen. sp. Gibbs. Brasier. A. E. Sue Cohen. Multiple new phenotypic taxa from trees and riparian ecosystems in Phytophthora gonapodyides-P.. and Goodwin. Denman. E. N. Erwin. nicotianae. and Rose. M. Hamm. they vary in ITS sequence from P. P. hedraiandra is a proven pathogen of Viburnum species in Italy and Spain causing collar and root rot as well as leaf lesions (7. 108:1172-1184. 50:481-487. M. Natl. D. P. and more work must be done to investigate how the host range of P. C.. T. E.D. C. and its variants: Designation of emerging heteroploid hybrid pathogens spreading on Alnus trees. 1983. 8. Mycol. Hwang. 83:1129-1136. 52:694-702. taxon Pgchlamydo were also recently baited from roots of alder trees in three nurseries in Bavaria (31). K. K. L. 2003. and water in France (12. 2000. F. Eggers.. 23. 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This is the first report of P.. C.50). M. Brasier et al. Species such as P. They were isolated from Prunus species in New York and Michigan and Malus pumila in New York (39). cherry (Prunus) roots in Cheltenham. megasperma ITS clade 6. 21. 25. In addition. South America (25). A. The discovery of P. O. H. 1980. and Hansen. C. W. K. H. Our identification of cultures isolated in 2002 and 2003 surveys produced the first report of P. E. M. Ferguson. Plant Pathol. E. M. M. Duncan. Comparison of two media selective for Phy- Plant Disease / January 2007 101 . Species of the Phytophthora megasperma complex. Beales. For. Rep. Balci.. Greslebin. Finally. P.. C. L. J. P. E. St. Madison. Brasier. Brasier. R. S. Z. taxon Pgchlamydo. ACKNOWLEDGMENTS We thank Ann Chaptman for her help in collecting and processing twice as many plant samples in 2005 as in 2004. and Ribeiro. 20.. Cooke. J. L. No. M. A. 28. D. sp. Miller. and Jeffers. and Man In’t Veld.. Plant Dis. Zhang. 27. 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