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New Legume Hosts of Phakopsora Pachyrhizi Based on Greenhouse Evaluations

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New Legume Hosts of Phakopsora Pachyrhizi Based on Greenhouse Evaluations e-Xtra* New Legume Hosts of Phakopsora pachyrhizi Based on Greenhouse Evaluations T. L. Slaminko, Department of Crop Sciences, University of Illinois, Urbana 61801; M. R. Miles, United States Department of Agriculture-Agricultural Research Service (USDA-ARS), Urbana, IL 61801; R. D. Frederick and M. R. Bonde, USDA-ARS Foreign Disease-Weed Science Research Unit, Ft. Detrick, MD 21702; and G. L. Hartman, USDA-ARS and Department of Crop Sciences, University of Illinois, Urbana 61801 MATERIALS AND METHODS ABSTRACT Species selection. All species tested be- Slaminko, T. L., Miles, M. R., Frederick, R. D., Bonde, M. R., and Hartman, G. L. 2008. New long to the legume subfamily Papilion- legume hosts of Phakopsora pachyrhizi based on greenhouse evaluations. Plant Dis. 92:767-771. oideae and all occur in the southern United States or in major soybean-producing Phakopsora pachyrhizi, the causal organism of soybean rust, was first found in the continental states (25) with the exception of previously United States in 2004 and has been found on soybean, kudzu, Florida beggarweed, and three reported hosts and four unscreened species Phaseolus species in the field. The pathogen has been reported to occur on more than 90 legume in Teramnus, the genus most closely re- species worldwide and it is likely to infect native and introduced legume species in the United lated to Glycine (11). Seed was ordered States. The objective of this study was to determine if 176 species representing 57 genera of legumes, the majority of which are either native or naturalized to soybean-growing areas of the from several repositories in the USDA- United States, could be hosts of P. pachyrhizi. Between one and three accessions of each species, ARS National Plant Germplasm System a total of 264 accessions, were inoculated with a mixture of four isolates of P. pachyrhizi. Sever- (Table 1). ity and sporulation were rated on a 1-to-5 scale at 14 and 28 days after inoculation. P. pachyrhizi Experimental procedures. Between 2 was confirmed by the presence of sporulating uredinia and/or immunological assay on 65 new and 20 seeds per entry per replication were species in 25 genera; 12 of these genera have not been reported previously as hosts. Many of the planted into each 4-cm2 cell in 6 × 12 flats newly identified hosts grow in the southern United States, and like kudzu, could serve as over- containing soilless medium (Sunshine wintering hosts for P. pachyrhizi. Mix, LC1; Sun Gro Horticulture Inc., Bellevue, WA) in a rust-free greenhouse. Additional keywords: host range, Papilionoideae Seed number varied because of seed size, availability, germination, and plant size. Seeds of some species were scarified or pregerminated on water agar to improve Soybean rust is caused by the obligate containing over 650 genera and 18,000 the likelihood of survival. The experimen- fungus Phakopsora pachyrhizi Syd. & P. species (18). The host range of P. pachyr- tal design was a randomized complete Syd. and was first reported in Asia (10). It hizi is restricted to the subfamily Papilion- block with three replications. Two soybean is a serious foliar disease on soybean in oideae, a monophyletic clade within Fa- cultivars, GC00138-29 and UG-5, were Africa, Asia, Australia, and South America baceae (26). The subfamily contains an included in each replication as susceptible (15), but its impact on soybean production estimated 476 genera and 13,860 species checks. The experimental unit was a cell in North America has been limited since its (5). P. pachyrhizi is known to infect 42 containing 1 to 20 plants of a single entry, arrival in 2004 (22) to the southern United genera of papilionoid legumes, but the which represented a unique plant introduc- States (24). limits of its host range are not known. tion (PI). There are 93 hosts of P. pachyrhizi re- Legume systematics might provide useful Inoculation. Four-week-old plants were ported from inoculated and noninoculated insight on the potential host range of P. inoculated with a mixture of four isolates plants (12,17,21,23). All hosts belong to pachyrhizi and its relationship with host of P. pachyrhizi at the USDA-ARS For- the legume family Fabaceae, which is one plants. eign Disease-Weed Science Research Unit of the largest families of flowering plants In the United States, six species have (FDWSRU) Biosafety Level 3 Plant Patho- been reported as hosts of P. pachyrhizi: gen Containment Facility at Ft. Detrick, soybean (22); kudzu, Pueraria lobata MD (13). The inoculum consisted of an (Willd.) Ohwi (8); Florida beggarweed, equal mixture of isolates from Brazil (BZ Corresponding author: G. L. Hartman E-mail: [email protected] Desmodium tortuosum (Sw) DC. (23); dry 01-1), Paraguay (PG 01-2), Thailand (TH bean, Phaseolus vulgaris L.; lima bean, P. 01-1), and Zimbabwe (ZM 01-1) (14). Trade and manufacturers’ names are necessary to lunatus L.; and scarlet runner bean, P. coc- Spores were removed from liquid nitrogen report factually on available data; however, the cineus L. (12). storage, heat shocked at 40°C for 5 min, USDA neither guarantees nor warrants the standard With the appearance of soybean rust in and allowed to rehydrate by incubating of the product, and the use of the name by USDA over water in an enclosed petri plate for 12 implies no approval of the product to the exclusion the continental United States in 2004 (22), of others that may also be suitable. there are many new potential legume hosts h (14). Distilled water with 0.01% (v/v) of P. pachyrhizi. Many legume species in Tween 20 (Sigma, St. Louis, MO) was *The e-Xtra logo stands for “electronic extra” the United States were either not previ- added to the dry spores and the mixture and indicates that a supplementary figure and ously tested or were not in the same geo- was stirred with a glass rod. The spore table not included in the print edition are graphic region as the fungus. These legu- suspension was then filtered through a 53- available online. minous species may aid in the over- µm nylon screen into a beaker to remove Accepted for publication 8 January 2008 wintering of the fungus and provide a debris and clumps of spores, and the spore source of inoculum at the beginning of the concentration was determined with a soybean-growing season. The objective of hemacytometer. Distilled water was added doi:10.1094/ PDIS-92-5-0767 this study is to identify leguminous hosts to adjust the final spore concentration to This article is in the public domain and not copy- that are either native or naturalized to the 25,000 spores per ml. Each flat was inocu- rightable. It may be freely reprinted with custom- ary crediting of the source. The American Phyto- southern United States or other major soy- lated with 25 ml of the spore suspension pathological Society, 2008. bean-growing areas. with an atomizer at 138 kPa. Flats were Plant Disease / May 2008 767 placed in a dew chamber at 20 to 22°C for (Sylvania, Danvers, MA). The plants were mist chamber in the greenhouse at 20 to 16 h and then transferred to a greenhouse rated 14 days after inoculation (DAI) and 25°C with natural light. Mist was provided at 20 to 25°C with a 16-h photoperiod. reinoculated with the same procedure as for 1 min at 20 min intervals for the dura- Supplemental lighting was provided with described above. After incubation in the tion of the experiment. The plants were 1,000-watt Metalarc high-intensity lamps dew chamber, the plants were placed in a rated as before at 14 DAI. Table 1. List of legume hosts that were susceptible when inoculated with Phakopsora pachyrhizi in the greenhouse and the lesion type and severity ratings for each species Severitya Seed New New Lesion Standard Scientific name Accessionb sourcec genusd speciesd typee No.f Mean Range error Alysicarpus rugosus PI 286530 S9 x RB 9 1.9 1–4 0.39 Astragalus canadensis PI 232539 W6 x RB 8 1.8 1–3 0.25 A. cicer PI 362119a, PI 452451a, W6 BL, RB 16 3.2 2–4 0.16 PI 576968a A. crassicarpus DLEG 900280 DLEG RB 4 1.3 1–2 0.25 A. glycyphyllos PI 206882, PI 420665a W6 RB 10 1.6 1–2 0.16 Baptisia alba var. Ames 27423a, PI 636376 NC7 x x RB 10 2.0 1–3 0.26 macrophylla B. australis Ames 24958, PI 443123 NC7 x BL, RB 11 1.5 1–3 0.21 B. bracteata var. Ames 3095a NC7 x BL, RB 6 2.5 1–3 0.34 laevicaulis Cajanus cajan NSL 73128, PI 520598a NPGS BL, RB 9 3.6 2–5 0.38 Calopogonium caeruleum PI 362125a S9 RB, TAN 5 4.0 3–5 0.45 Calopogonium PI 204364a, PI 279595a, S9 x RB, TAN 24 3.1 2–5 0.15 mucunoides PI 286288a, PI 322302a Caragana arborescens PI 310390a, PI 369217a, NC7 x x RB 27 1.9 1–3 0.14 PI 371524a, PI 633648a, PI 636378a Centrosema virginianum PI 322350a, PI 386281a S9 x BL, RB 8 1.9 1–4 0.35 Cologania angustifolia DLEG 900669a, DLEG DLEG x x RB, RED 12 3.7 2–5 0.26 var. angustifolia 990127a Cologania angustifolia DLEG 890367Da, DLEG x RB, TAN 11 3.1 2–4 0.21 var. stricta DLEG 990130a Cologania lemmonii DLEG 880053Da DLEG x RB 1 4.0 4 . Crotalaria incana PI 263427a, PI 336996a S9 x RB, TAN 14 1.8 1–3 0.19 C.
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