Evidence for Systemic Infection by Puccinia horiana, Causal Agent of Chrysanthemum White Rust, in Chrysanthemum.

Puccinia horiana, causal agent of the disease commonly known as chrysanthemum white rust (CWR), is a quarantine-significant fungal pathogen of chrysanthemum in the United States and indigenous to Asia. The pathogen was believed to have been eradicated in the United States but recently reappeared on several occasions in northeastern United States. The objective of the study presented here was to determine whether P. horiana could systemically infect chrysanthemum plants, thus providing a means of survival through winters. Scanning and transmission electron microscopy revealed the development of P. horiana on the surface and within leaves, stems, or crowns of inoculated chrysanthemum plants artificially exposed to northeastern U.S. winter temperatures. P. horiana penetrated leaves directly through the cuticle and then colonized the mesophyll tissue both inter- and intracellularly. An electron-dense material formed at the interface between fungal and host mesophyll cells, suggesting that the pathogen adhered to the plant cells. P. horiana appeared to penetrate mesophyll cell walls by enzymatic digestion, as indicated by the absence of deformation lines in host cell walls at penetration sites. The fungus was common in vascular tissue within the infected crown, often nearly replacing the entire contents of tracheid cell walls. P. horiana frequently passed from one tracheid cell to an adjacent tracheid cell by penetration either through pit pairs or nonpitted areas of the cell walls. Individual, presumed, fungal cells in mature tracheid cells of the crown and stems arising from infected crowns suggested that the pathogen might have been moving at least partially by means of the transpiration stream. The demonstration that chrysanthemum plants can be systemically infected by P. horiana suggests that additional disease control measures are required to effectively control CWR.

Effects of frequency of "extreme" temperature highs on development of soybean rust.

Previously, we hypothesized that summer "extreme" diurnal temperature highs in the southeastern United States were responsible for the yearly absence or delay of soybean rust development until fall. Utilizing temperature-controlled growth chambers, a diurnal temperature pattern of 33°C high and 20°C low reduced urediniospore production by 81%. However, that study did not consider the influence of frequency of extreme temperatures on soybean rust. We now report that a temperature high of 35°C for 1 h on three consecutive days, initiated 15 days after inoculation, when lesions had formed, reduced urediniospore production by 50% and required 9 to 12 days for sporulation to resume once the extreme temperature highs ceased. Furthermore, three consecutive days in which the temperature high was 37°C, beginning immediately after inoculation and subsequent dew period, reduced lesion numbers by 60%. The combined effects of reduced numbers of lesions and urediniospores per lesion caused by extreme temperature highs can account for observed absence or delay of soybean rust development in the southeastern United States until fall. A comparison of frequency of extreme temperature highs with numbers of counties reporting presence of soybean rust from 2005 to 2012 verified that extreme temperature highs may be largely responsible for absence or delay of soybean rust development. This is the first report showing the effect of frequency of extreme temperature highs on development of soybean rust. Because the south-to-north progression of soybean rust is required for the disease to occur in the major soybean-production regions of the United States, temperatures in the southeastern United States have a major effect on the entire U.S. soybean industry.

Effects of daily temperature highs on development of Phakopsora pachyrhizi on soybean.

Although considerable information exists regarding the importance of moisture in the development of soybean rust, little is known about the influence of temperature. The purpose of our study was to determine whether temperature might be a significant limiting factor in the development of soybean rust in the southeastern United States. Soybean plants infected with Phakopsora pachyrhizi were incubated in temperature-controlled growth chambers simulating day and night diurnal temperature patterns representative of the southeastern United States during the growing season. At 3-day intervals beginning 12 days after inoculation, urediniospores were collected from each plant and counted. The highest numbers of urediniospores were produced when day temperatures peaked at 21 or 25°C and night temperatures dipped to 8 or 12°C. When day temperatures peaked at 29, 33, or 37°C for a minimum of 1 h/day, urediniospore production was reduced to 36, 19, and 0%, respectively, compared with urediniospore production at the optimum diurnal temperature conditions. Essentially, no lesions developed when the daily temperature high was 37°C or above. Temperature data obtained from the National Climatic Data Center showed that temperature highs during July and August in several southeastern states were too high for significant urediniospore production on 55 to 77% of days. The inhibition of temperature highs on soybean rust development in southeastern states not only limits disease locally but also has implications pertaining to spread of soybean rust into and development of disease in the major soybean-producing regions of the Midwestern and northern states. We concluded from our results that temperature highs common to southeastern states are a factor in the delay or absence of soybean rust in much of the United States.

First Report of Anthracnose of Mile-a-Minute (Persicaria perfoliata) Caused by Colletotrichum cf. gloeosporioides in Turkey.

Mile-a-minute (Persicaria perfoliata (L.) H. Gross; family: Polygonaceae) is an exotic annual barbed vine that has invaded the northeastern USA and Oregon (2). In July of 2010, in a search for potential biological control pathogens (3), diseased P. perfoliata plants were found along the Firtina River near Ardesen, Turkey. Symptoms were irregular dark necrotic lesions along leaf margins and smaller irregular reddish lesions on the lamellae of leaves. Symptomatic leaves were sent to the quarantine facility of FDWSRU, USDA, ARS in Ft. Detrick, MD, for pathogen isolation and testing. Symptomatic leaves were excised, surface disinfested in 0.615% NaOCl, and then incubated for 2 to 3 days in sterile moist chambers at 20 to 25°C. Numerous waxy sub-epidermal acervuli with 84-µm-long (mean) black setae were observed in all of the lesions after 2 to 3 days of incubation. Conidiophores within acervuli were simple, short, and erect. Conidia were one-celled, hyaline, guttulate, subcylindrical, straight, 12.3 to 18.9 × 3.0 to 4.6 µm (mean 14.3 × 3.7 µm). Pure cultures were obtained by transferring conidia onto 20% V-8 juice agar. Appressoria, formed 24 h after placing conidia on dialysis membrane over V-8 juice agar, were smooth, clavate, aseptate, regular in outline, and 6.4 to 10.0 × 5.1 to 7.2 µm (mean 7.5 × 6.6 µm). These characters conformed to the description of Colletotrichum gloeosporioides (Penz.) Penz. & Sacc. (1). A voucher specimen was deposited in the U.S. National Fungus Collections (BPI 882461). Nucleotide sequences for the internal transcribed spacers (ITS 1 and 2), directly sequenced from ITS 1 and ITS 4 standard primers (4), were deposited in GenBank (JN887693). A comparison of these sequences with ITS 1 and 2 sequences of the C. gloeosporioides epitype IMI 356878 (GenBank EU 371022) (1) using BLAST found 479 of 482 identities with no gaps. Conidia from 14-day-old cultures, in an aqueous suspension of 1.0 × 106 conidia ml-1, were spray-inoculated onto healthy stems and leaves of twenty 30-day-old P. perfoliata plants. Another 10 plants were not inoculated. All plants were placed in a dew chamber at 25°C for 16 h with no lighting. They were then placed in a 20 to 25°C greenhouse with a 14-h photoperiod. Light was generated using 400W sodium vapor lights. Lesions developed on leaves and stems of all inoculated plants after 7 days, and symptoms were the same as observed in the field. Each plant was rated weekly for disease severity on a 0 to 10 rating scale where 0 = no disease symptoms and 10 = 100% symptomatic tissue. After 28 days, the average disease rating of inoculated plants was 3.95 ± 0.94. No disease developed on noninoculated plants. C. gloeosporioides was reisolated from all inoculated plants. Host range tests will determine the potential of this isolate as a biological control agent for P. perfoliata. To our knowledge, this is the first report of anthracnose caused by C. gloeosporioides on P. perfoliata. References: (1) P. F. Cannon et al. Mycotaxon 104:189, 2008. (2) J. T. Kartesz and C. A. Meacham. Synthesis of the North American Flora, Version 1.0., North Carolina Botanical Garden, Chapel Hill, N.C. 1999. (3) D. L. Price et al. Environ. Entomol. 32:229, 2003. (4) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. Academic Press, Inc., San Diego, CA, 1990.

Characterizing Resistance to Phakopsora pachyrhizi in Soybean.

Resistance in soybean to Phakopsora pachyrhizi, the cause of soybean rust, is characterized by either reddish-brown (RB) lesions or an immune response. The RB type of resistance can be incomplete, as evidenced by the presence of sporulating uredinia within lesions. Susceptibility, on the other hand, is exemplified by tan-colored (TAN) lesions, and can be expressed in gradations of susceptibility or partial resistance that are less well defined. This study evaluated traits associated with incomplete or partial resistance to P. pachyrhizi in soybean by comparing 34 soybean accessions inoculated with four P. pachyrhizi isolates. Six accessions produced RB lesions to all four isolates, while 19 accessions produced TAN lesions, including plant introduction (PI) 200492 (Rpp1) and the susceptible check 'Williams'. Williams had among the largest area under the disease progress curve (AUDPC) values and area under the sporulating uredinia progress curve (AUSUPC) values, while eight accessions had lower AUSUPC values. Of the known sources of single-gene resistance, only PI 230970 (Rpp2), PI 459025B (Rpp4), and PI 594538A (Rpp1b) had lower AUDPC and AUSUPC values than Williams. PI 594538A and PI 561356 had RB lesions and had the lowest AUDPC and AUSUPC values. Of the known sources of single-gene resistance, only PI 230970 (Rpp2) and PI 594538A (Rpp1b) produced fewer and smaller-diameter uredinia than Williams. This study characterized reactions to P. pachyrhizi in 34 accessions based on lesion type and sporulation, and defined incomplete resistance and partial resistance in the soybean-P. pachyrhizi interaction.

First Report of Leaf Spot on Horseweed (Conyza canadensis) Caused by Septoria erigerontis in Turkey.

Horseweed (Conyza canadensis (L).Cronq., Asteraceae) is an invasive exotic weed in Turkey and a problematic native weed in the United States where glyphosate-resistant populations of the weed have developed (2). These characteristics make horseweed a target for biological control efforts. In September 2009, small, brown leaf spots were observed on leaves of C. canadensis in Taflan, Turkey (41°25.398'N, 36°08.352'E). Globose, dark-walled pycnidia were also observed in brown spots on leaves. Diseased tissue was surface disinfested and placed on moist filter paper in petri plates. A fungus designated 09-Y-TR1 was isolated from the diseased leaves. Single-spore isolations were grown on potato dextrose agar (PDA). Cultures on PDA formed dark green-to-black colonies. Pycnidia matured after 3 to 4 weeks when plates were incubated at 23°C with a 12-h photoperiod (black light and cool white fluorescent light). Pycnidia were separate, immersed, and dark brown with a single apical ostiole. Matured conidia were one to three septate, filiform, straight to slightly curved, rounded at the apex, smooth walled, hyaline, and 22 to 40 × 1.4 to 2.5 µm. Morphology was consistent with Septoria erigerontis Peck (3). Comparison of the internal transcribed spacer (ITS) 1 and 2 sequence with available sequences of vouchered S. erigerontis specimens (GenBank EF535638.1, AY489273.1; KACC 42355, CBS 109094) showed 447 of 450 and 446 of 450 identities, respectively. Nucleotide sequences for the ribosomal ITS regions (ITS 1 and 2, including 5.8S rDNA) were deposited in GenBank (GU952666). For pathogenicity tests conidia were harvested from 3-week-old cultures grown on PDA, by brushing the surface of the colonies with a small paint brush, suspended in sterile distilled water, and filtered through cheese cloth. Conidia were then diluted in sterile distilled water plus 0.1% polysorbate 20 to a concentration of 5 × 106 conidia/ml. Stems and leaves of seven 5-month-old seedlings were spray inoculated with 10 ml of this aqueous suspension per plant. Inoculated plants and three noninoculated plants were placed in a dew chamber at 23°C in darkness and continuous dew, and after 48 h, plants were moved to a greenhouse bench. Symptoms were observed 2 days after inoculation. Disease severity was evaluated 2 weeks after inoculation by a rating system with a scale of 0 to 6 based on percentage of plant tissue necrosis, in which 0 = no symptoms, 1 = 1 to 5%, 2 = 6 to 25%, 3 = 26 to 75%, 4 = 76 to 95%, 5 = >95%, and 6 = dead plant. The average disease rating on inoculated plants was 3.55. No disease was observed on noninoculated plants. S. erigerontis was reisolated from all inoculated plants. To our knowledge, this is the first report of leaf spot on horseweed caused by S. erigerontis in Turkey where the fungus may have potential as a classical biological control agent. S. erigerontis has also been reported on C. canadensis in Korea and Portugal (1). In the United States, S. erigerontis has been reported on horseweed in several states (1) and these isolates may have potential as biological control agents of horseweed, particularly glyphosate-resistant horseweed, in the United States. References: (1) D. F. Farr et al. Fungal Databases. Systematic Mycology and Microbiology Laboratory, Online publication. ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ , March 2010. (2) I. Heap. www.weedscience.org , 2006. (3) M. J. Priest. Fungi of Australia: Septoria. ABRS/CSIRO Publishing. Melbourne, 2006.

First Report of Leaf Spot Caused by Cercospora bizzozeriana on Lepidium draba in the United States.

The herbaceous perennial Lepidium draba L. is an invasive weed of rangelands and riparian areas in North America and Australia. As of 2002, it had infested 40,500 ha of rangeland in Oregon and large areas in Wyoming and Utah. Little is known of plant pathogens occurring on L. draba, especially in the United States, that could be useful for biological control of the weed. Leaf spots were first noted on a stand of L. draba near Shepherd, MT in 1997. The spots were mostly circular but sometimes irregularly shaped and whitish to pale yellow. The pathogen was erroneously assumed to be Cercospora beticola since its morphological traits closely resembled that species and the area had large fields of sugar beet with heavy Cercospora leaf spot incidence. Diseased leaves of L. draba were collected in 1997 and 2007. Conidia, borne singly on dark gray, unbranched conidiophores produced on dark stromata late in the season, were elongate, hyaline, multiseptate, 38 to 120 × 2 to 6 µm (mostly 38 to 50 × 2 to 5 µm) and had bluntly rounded tips and wider, truncate bases. These characteristics were consistent with the description of C. bizzozeriana Saccardo & Berlese (2). To isolate the fungus, spores were picked from fascicles of conidiophores with a fine-tipped glass rod, suspended in sterile water, and spread on plates of water agar. Germinated spores were transferred to potato dextrose agar (PDA). The ITS1, 5.8S, and ITS2 sequences of this fungus (GenBank Accession No. EU887131) were identical to sequences of an isolate of C. bizzozeriana from Tunisia (GenBank Accession No. DQ370428). However, these sequences were also identical to those of a number of Cercospora spp. in GenBank, including C. beticola. We also compared the actin gene sequences of the Montana isolate of C. bizzozeriana (GenBank Accession No. FJ205397) and an isolate of C. beticola from Montana (GenBank Accession No. AF443281); the sequences were 94.6% similar, an appreciable difference. For pathogenicity tests, cultures were grown on carrot leaf decoction agar. Aqueous suspensions of 104 spores per ml from cultures were sprayed on 6-week-old L. draba plants. Plants were covered with plastic bags and placed on the greenhouse bench at 20 to 25°C for 96 h. Koch's postulates were completed by reisolating the fungus from the circular leaf spots that appeared within 10 days, usually on lower leaves. Spores of C. bizzozeriana were also sprayed on seedlings of sugar beet, collard, mustard, radish, cabbage, and kale under conditions identical to those above. No symptoms occurred. After the discovery of the disease in 1997, plants of L. draba in eastern Montana, Wyoming, and Utah were surveyed from 1998 to 2003 for similar symptoms and signs, but none were found. This, to our knowledge, is the first report of C. bizzozeriana in the United States. The initial report of the fungus in North America was from Manitoba in 1938 (1). It has recently been reported as occurring on L. draba in Tunisia (4) and Russia (3) and is reported as common in Europe (2). A voucher specimen has been deposited with the U.S. National Fungus Collections (BPI No. 878750A). References: (1) G. R. Bisby. The Fungi of Manitoba and Saskatchewan. Natl. Res. Council of Canada, Ottawa, 1938. (2) C. Chupp. A Monograph of the Fungus Genus Cercospora. C. Chupp, Ithaca, NY, 1953. (3) Z. Mukhina et al. Plant Dis. 92:316, 2008. (4) T. Souissi et al. Plant Dis. 89:206, 2005.

First Report of Stem Canker of Salsola tragus Caused by Diaporthe eres in Russia.

Salsola tragus L. (Russian thistle) is a problematic invasive weed in the western United States and a target of biological control efforts. In September of 2007, dying S. tragus plants were found along the Azov Sea at Chushka, Russia. Dying plants had irregular, necrotic, canker-like lesions near the base of the stems and most stems showed girdling and cracking. Stem lesions were dark brown and contained brown pycnidia within and extending along lesion-free sections of the stems and basal portions of leaves. Diseased stems were cut into 3- to 5-mm pieces and disinfested in 70% ethyl alcohol. After drying, stem pieces were placed into petri dishes on the surface of potato glucose agar. Numerous, dark, immersed erumpent pycnidia with a single ostiole were observed in all lesions after 2 to 3 days. Axenic cultures were sent to the Foreign Disease-Weed Science Research Unit, USDA, ARS, Ft. Detrick, MD for testing in quarantine. Conidiophores were simple, cylindrical, and 5 to 25 × 2 µm (mean 12 × 2 µm). Alpha conidia were biguttulate, one-celled, hyaline, nonseptate, ovoid, and 6.3 to 11.5 × 1.3 to 2.9 µm (mean 8.8 × 2.0 µm). Beta conidia were one-celled, filiform, hamate, hyaline, and 11.1 to 24.9 × 0.3 to 2.5 µm (mean 17.7 × 1.2 µm). The isolate was morphologically identified as a species of Phomopsis, the conidial state of Diaporthe (1). The teleomorph was not observed. A comparison with available sequences in GenBank using BLAST found 528 of 529 identities with the internal transcribed spacer (ITS) sequence of an authentic and vouchered Diaporthe eres Nitschke (GenBank DQ491514; BPI 748435; CBS 109767). Morphology is consistent with that of Phomopsis oblonga (Desm.) Traverso, the anamorph of D. eres (2). Healthy stems and leaves of 10 30-day-old plants of S. tragus were spray inoculated with an aqueous suspension of conidia (1.0 × 106 alpha conidia/ml plus 0.1% v/v polysorbate 20) harvested from 14-day-old cultures grown on 20% V8 juice agar. Another 10 control plants were sprayed with water and surfactant without conidia. Plants were placed in an environmental chamber at 100% humidity (rh) for 16 h with no lighting at 25°C. After approximately 24 h, plants were transferred to a greenhouse at 20 to 25°C, 30 to 50% rh, and natural light. Stem lesions developed on three inoculated plants after 14 days and another three plants after 21 days. After 70 days, all inoculated plants were diseased, four were dead, and three had more than 75% diseased tissue. No symptoms occurred on control plants. The Phomopsis state was recovered from all diseased plants. This isolate of D. eres is a potential biological control agent of S. tragus in the United States. A voucher specimen has been deposited with the U.S. National Fungus Collections (BPI 878717). Nucleotide sequences for the ribosomal ITS regions (ITS 1 and 2) were deposited in GenBank (Accession No. EU805539). To our knowledge, this is the first report of stem canker on S. tragus caused by D. eres. References: (1) B. C. Sutton. Page 569 in: The Coelomycetes. CMI, Kew, Surrey, UK, 1980. (2) L. E. Wehmeyer. The Genus Diaporthe Nitschke and its Segregates. University of Michigan Press, Ann Arbor, 1933.

First Report of Leaf Anthracnose Caused by Phomopsis convolvuli on Field Bindweed in Turkey.

Field bindweed (Convolvulus arvensis L.; Convolvulaceae) is a troublesome perennial weed found among many important crops in the world (1). In May of 2007, dying field bindweed plants were found along the edge of a wheat (Triticum aestivum L.) field between Bafra and Taflan, Turkey (41°34.395'N, 35°52.215'E). Lesions on leaves were irregular and variable in size and dark black with green margins. Severely diseased leaves were wilted or dead. Fruiting bodies were not evident on field-collected material. Diseased tissue was surface disinfested and placed on moist filter paper in petri plates. Numerous pycnidia with alpha conidia were observed after 2 weeks. A fungus, designated 24-6, was isolated from the diseased leaves. Cultures on potato dextrose agar (PDA) were floccose with white mycelia and small black stromata. Alpha conidia from pycnidia on inoculated plants were biguttulate, one celled, hyaline, oblong to ellipsoid, and 7.0 to 12.8 × 3.0 to 5.5 µm (mean 10.0 × 3.9 µm). Neither beta conidia nor the teleomorph, Diaporthe sp., were observed on diseased tissue or in cultures. Morphology was consistent with that of Phomopsis convolvuli Ormeno-Nunez, Reeleder & A.K. Watson (2). Alpha conidia were harvested from 12-day-old cultures grown on PDA by brushing the surface of the colonies with a small paint brush, suspending the conidia in sterile distilled water, and filtering through cheesecloth. The conidia were then resuspended in sterile distilled water plus 0.1% polysorbate 20 to arrive at a concentration of 107 conidia/ml. Stems and leaves of seven plants at the 3- to 5-leaf stage were spray inoculated with 10 ml per plant of this aqueous suspension. Inoculated plants and two noninoculated plants were placed in a dew chamber at 24°C in darkness and continuous dew. After 48 h, plants from the dew chamber were moved to a greenhouse bench. Disease severity was evaluated 1 week after inoculation with a rating system based on a scale from 0 to 4, in which 0 = no symptoms, 1 = 1 to 25% necrosis, 2 = 26 to 50% necrosis, 3 = 51 to 75% necrosis, and 4 = 76 to 100% necrosis (2). The average disease rating on inoculated plants was 3.75. No disease was observed on noninoculated plants. P. convolvuli was reisolated from all inoculated plants. Comparison of the internal transcribed spacer (ITS) 1 and 2 sequences with available sequences of a vouchered P. convolvuli specimen (GenBank Nos. U11363, U11417; BPI 748009, FAU649) showed 192 of 193 and 176 of 179 identities, respectively, for the two regions. Nucleotide sequences for the ribosomal ITS regions (ITS 1 and 2, including 5.8S rDNA) were deposited in GenBank (Accession No. FJ710810), and a voucher specimen has been deposited with the U.S. National Fungus Collections (BPI 878927). To our knowledge, this is the second report in the world of leaf anthracnose on field bindweed caused by P. convolvuli. The first report was from Canada (3) of an isolate that was later patented for biological control of C. arvensis (4). References: (1) L. Holm et al. The World's Worst Weeds. University Press of Hawaii, Honolulu, 1977. (2) J. Ormeno-Nunez, et al. Can. J. Bot. 66:2228, 1988. (3) J. Ormeno-Nunez et al. Plant Dis. 72:338, 1988. (4) A. K. Watson et al. U.S. Patent 5,212,086, 1993.

First Report of Leaf Spot Caused by Cercospora bizzozeriana on Hoary Cress in Russia.

Hoary cress (Lepidium draba (L.) subsp. draba (synonym = Cardaria draba (L.) Desv.) (1), family Brassicaceae, is a common weed in Russia but it is an aggressive invasive weed in the northwestern United States. In the summer of 2006, dying hoary cress plants were found near Kugoyeyskoye in the Krylovskoy area of the Krasnodar Region of Russia. Plants had grayish white leaf spots on most of the leaves. In some cases, the diseased leaf spots dropped out of the leaves producing shot-holes. In most cases, the leaf spots coalesced and the leaves wilted and died. Diseased leaves were collected, air dried, and sent to the quarantine facility of the Foreign Disease-Weed Science Research Unit (FDWSRU), USDA/ARS, Fort Detrick, MD. The air-dried leaves were observed microscopically, and numerous conidiophores and conidia were observed on both sides of leaves within and around the lesions. The fungus isolated (DB06-018) conformed to the description of Cercospora bizzozeriana Saccardo & Berlese (2). Conidiophores were 1 to 5 geniculate, unbranched, pale olive-brown, and uniform in color and width (4 µm). Conidia were multiseptate, hyaline, cylindric, straight to slightly curved, and measured 57 to 171 µm (average 103) long × 3.8 to 6.7 µm (average 4.6) wide. Leaves of rosettes (10 to 15 cm in diameter) of four hoary cress plants were spray inoculated with an aqueous suspension of conidia (1 × 105/ml) and mycelia harvested from 6- to 8-day-old cultures grown on V8 medium. Inoculated plants and two noninoculated plants were placed in a dew chamber at 20°C in darkness and continuous dew. After 96 h, plants were moved from the dew chamber to a greenhouse bench. All plants were watered twice daily. After 12 days, symptoms were observed on all inoculated plants. Symptoms were identical to those observed in the field in Russia. No symptoms were observed on noninoculated plants. C. bizzozeriana was reisolated from the leaves of all symptomatic plants. Nucleotide sequences were obtained for the internal transcribed spacer regions ITS1 and ITS2 and the 5.8S ribosomal RNA gene (GenBank Accession No. EU031780) and aligned with the same sequences obtained from another C. bizzozeriana isolate (GenBank Accession No. DQ370428) collected in Tunisia. There was 100% alignment of the two sequences with no gaps. Both isolates of C. bizzozeriana are destructive pathogens on hoary cress and locally severe epidemics have been observed in both Russia and Tunisia (4). This fungus has also been reported in North America (3) and has the potential as a biological control agent where the weed is a problem. To our knowledge, this is the first report of C. bizzozeriana on L. draba subsp. draba in Russia. A voucher specimen has been deposited with the U.S. National Fungus Collections (BPI 878175). Live cultures are being maintained at FDWSRU. References: (1) I. A. Al-Shehbaz and K. Mummenhoff. Novon 12:5, 2002. (2) C. Chupp. A Monograph of the Fungus Genus Cercospora. C. Chupp, Ithaca, New York, 1953. (3) I. L. Conners. Res. Bra. Can. Dep. Agric. 1251:1, 1967. (4) T. Souissi et al. Plant Dis. 89:206, 2005.

First Report of Leaf Spot Caused by Colletotrichum cf. linicola on Field Bindweed in Turkey.

Field bindweed (Convolvulus arvensis L., Convolvulaceae) is one of the most problematic weeds in the world (1) and a target of biological control efforts (2). In the summer of 2006, dying field bindweed plants were found in a wheat field near Bafra, Turkey (41°21.197'N, 36°12.524'E). Plants had water-soaked lesions that developed into necrotic leaf spots on most of the leaves, particularly along the leaf margins, and on some stems. In most cases, the leaf spots coalesced, causing the leaves and later plants to wilt and die. Diseased leaves and stems were taken to the Phytopathology Laboratory of the Faculty of Agriculture, Ondokuz Mayis University, Samsun, Turkey. Diseased tissue was surface disinfested and placed on moist filter paper in petri dishes. Numerous acervuli with setae and conidia typical of a Colletotrichum sp. were observed after 2 to 5 days. A fungus, designated 06-01, was isolated from the diseased leaves. Stems and leaves of seven 12-week-old plants were spray inoculated in the laboratory with an aqueous suspension of conidia (106 spores per ml; 10 ml per plant) harvested from 6- to 8-day-old cultures grown on malt extract agar. The plants and two noninoculated checks were placed in a dew chamber at 22°C in darkness and continuous dew. After 48 h, plants from the dew chamber were moved to a greenhouse bench. All plants were watered twice daily. Symptoms were observed 5 days after inoculation. No symptoms were observed on noninoculated plants. Isolate 06-01 was reisolated from all inoculated plants. In the field, 20 inoculated plants became diseased after 20 days with approximately 36% diseased leaf tissue from which 06-01 was consistently reisolated. Diseased tissue and cultures of the fungus were sent to the Foreign Disease-Weed Science Research Unit, USDA/ARS, Fort Detrick, MD. The fungus conformed to the description of Colletotrichum linicola Pethybr. & Laff., which was noted as distinct from C. lini (3). The original description is also different than the description of C. lini (Westerdijk) Tochinai by Sutton (4). Acervuli were sparse, subepidermal, and erumpent. Conidia were hyaline, oblong or cylindrical or somewhat spindle-shaped with dull-pointed ends, guttulate, and 14 to 19 × 4 to 5 µm (mean 17 × 4 µm). Conidiophores were short, simple, hyaline, and emerged from subepidermal stroma. Setae were simple, erect, 3-septate, and dark with hyaline tips. DNA sequences were obtained for the internal transcribed spacer regions (GenBank Accession No. EU000060) and compared with other sequences in GenBank. Sequences from 06-01 matched 100% with one isolate of C. linicola and 99% with two other isolates of C. linicola. These isolates formed a unique clade. However, 06-01 was also 99% identical to other species of Colletotrichum. Thus, species identification is inconclusive. Isolate 06-01 is a destructive pathogen on field bindweed, and severe disease can be produced by inoculation of foliage with an aqueous suspension of conidia. To our knowledge, this is the first report of Colletotrichum on field bindweed. A voucher specimen has been deposited with the U.S. National Fungus Collections (BPI 878174). References: (1) L. Holm et al. The World's Worst Weeds. University Hawaii Press, Honolulu, Hawaii, 1977. (2) G. Defago et al. BioControl 46:157, 2001. (3) G. H. Pethybridge and H. A. Lafferty. Sci. Proc. R. Dublin Soc. 15:359, 1918. (4) B. C. Sutton. The Coelomycetes. Commonw. Mycol. Inst., Kew, England, 1980.

First Report of Anthracnose of Salsola tragus Caused by Colletotrichum gloeosporioides in Russia.

In October of 2006, dying Salsola tragus L. (Russian thistle, tumbleweed), family Chenopodiaceae, plants were found along the Azov Sea at Chushka, Russia. Approximately 40 plants in the area were diseased and almost 80% of these were dying. Plants were approximately 1 m tall × 0.5 m wide. Dying plants had irregular, necrotic lesions along the length of the stems. Leaves of these plants were also necrotic. Lesions on stems and leaves were dark brown and usually coalesced. Diseased stems were cut into 3- to 5-mm pieces, disinfested in 70% ethyl alcohol, and then placed onto the surface of potato glucose agar (PGA). Numerous, waxy, subepidermal acervuli with 110 µm long (mean) black setae were observed in all of the lesions after 2 to 3 days. Conidiophores were simple, short, and erect. Conidia were one-celled, hyaline, ovoid to oblong, falcate to straight, and measured 12.9 to 18.0 × 2.8 to 5.5 µm (mean 15.6 × 4.2 µm). Appressoria formed 24 h after placing conidia on a dialysis membrane over 20% V8 juice agar. Appressoria measured 4.0 to 13.9 × 2.4 to 8.8 µm (mean 7.0 × 5.2 µm). These characters conformed to the description of Colletotrichum gloeosporioides (Penz.) Penz. & Sacc. in Penz. (1). A voucher specimen was deposited with the U.S. National Fungus Collections, Beltsville, MD (BPI 878389). Nucleotide sequences for the internal transcribed spacers (ITS 1 and 2) were deposited in GenBank (Accession No. EU530697) and aligned with ITS sequences of two other isolates from S. tragus. There was 100% similarity to each isolate, one from Greece (Accession No. DQ344621) and one from Hungary (Accession No. EU805538). Axenic cultures on PGA were sent to the Foreign Disease-Weed Science Research Unit, USDA, ARS, Fort Detrick, MD for testing in quarantine. Conidia were harvested from 14-day-old cultures grown on 20% V8 juice agar, and healthy stems and leaves of 30-day-old plants of S. tragus (13 plants) were spray inoculated with an aqueous conidial suspension of 1.0 × 106 conidia/ml plus 0.1% v/v polysorbate 20. Another 13 control plants were sprayed with water and surfactant without conidia. Plants were placed in an environmental chamber at 100% humidity for 16 h in the dark at 25°C. After approximately 24 h, all plants were transferred to a greenhouse at 20 to 25°C, 30 to 50% relative humidity, and natural light augmented by 12-h light periods with 500 W sodium vapor lights. Lesions developed on stems of all inoculated plants after 7 days. After 14 days, nine plants were dead and all inoculated plants were dead after 3 weeks. No symptoms developed on control plants. C. gloeosporioides was reisolated from stem pieces of all inoculated plants, and the morphology of the reisolated pathogen was the same as that of the initially isolated pathogen. To our knowledge, this is the first report of anthracnose caused by C. gloeosporioides on S. tragus in Russia. Reference: (1) B. C. Sutton. Page 15 in: Colletotrichum Biology, Pathology and Control. J. A. Bailey and M. J. Jeger, eds. CAB International, Wallingford, UK, 1992.

Comparative Susceptibilities of Legume Species to Infection by Phakopsora pachyrhizi.

Knowledge of the host range of Phakopsora pachyrhizi is important to agriculture in the United States because of the distinct possibility that economic losses could occur to crops other than soybean. Furthermore, it is possible that alternative hosts could provide a means of overwintering of the pathogen, providing inoculum to initiate epidemics in future years. To clarify the potential importance of soybean rust on nonsoybean legumes and their role in overwintering of the disease, multiple accessions of clover, cowpea, pea, kudzu, lima bean, snap bean, and single accessions of coffee senna, Florida beggarweed, hemp sesbania, hyacinth bean, partridge pea, and showy crotalaria were inoculated under greenhouse conditions with urediniospores of P. pachyrhizi; infected soybean plants served as a control. The four criteria used to assess susceptibility were lesion density, proportion of lesions with sporulating uredinia, average number of uredinia per lesion, and average uredinia diameter, each determined 2 weeks following inoculation. Based on lesion densities, percentage of lesions with sporulation, and average numbers of uredinia per lesion, soybean, kudzu, and pea were the most susceptible species, followed by snap bean. However, because infected pea plants defoliated rapidly, urediniospore production presumably was limited, lessening the potential for epidemics on pea. Cultivars of snap bean produced numerous brown to reddish-brown lesions, many of which sporulated, but numbers of uredinia per lesion were lower than on soybean, kudzu, or pea. The presence of both tan (susceptible) and reddish-brown (resistant) lesions on kudzu demonstrated physiological differentiation on that host. Some kudzu plants appeared to be potentially excellent hosts for overwintering of the disease. The average number of uredinia per lesion appeared to be a valid measurement with which to compare host susceptibilities, and may have epidemiological significance. High susceptibility of a host was characterized by numerous uredinia with a wide range of sizes within individual lesions. In contrast, low susceptibility to rust was characterized by no or a few small uredinia.

Effects of temperature on urediniospore germination, germ tube growth, and initiation of infection in soybean by phakopsora isolates.

ABSTRACT Temperature is a critical factor in plant disease development. As part of a research program to determine how specific environmental variables affect soybean rust, we determined temperature effects on urediniospore germination and germ tube growth of four isolates of Phakopsora pachyrhizi, one each from Brazil, Hawaii, Taiwan, and Zimbabwe, and an isolate of P. meibomiae from Puerto Rico, collected over a 25-year period. Also compared were the effects of temperature during a night dew period on initiation of disease by the P. pachyrhizi isolates. All variables were fit to a nonlinear beta function with temperature as the independent variable. Minimum, maximum, and optimum temperatures, along with shape parameters of the beta function for each variable, were statistically analyzed. All Phakopsora isolates behaved similarly as to how temperature affected urediniospore germination, germ tube growth, and initiation of disease. The results suggest that P. pachyrhizi has changed little in the past few decades with respect to how it responds to temperature and that previously collected research data continues to be valid, simplifying the development of soybean rust disease models.

Slender Wheatgrass is Susceptible to Smut Caused by Ustilago phrygica from Turkey.

Slender wheatgrass (Elymus trachycaulus (Link) Gould ex Shinners subsp. trachycaulus), family Poaceae, tribe Triticeae, is a native North American grass that is used as a livestock forage. Ustilago phrygica, a systemic ovary-smut fungus, is native to Turkey and West Asia and is pathogenic on Aegilops spp. and Taeniatherum caput-medusae (L.) Nevski subsp. asperum (Simonk.) Melderis (medusahead), an invasive weed in the western United States that is targeted for biological control. An isolate of the fungus (U.S. National Fungus Collections, BPI 871725; GenBank Accession No. DQ139961) was collected from medusahead in Turkey and screened for possible use in classical biological control of this weed. Screening was done in quarantine in a BSL-3 facility of the Foreign Disease-Weed Science Research Unit, USDA, ARS, Ft. Detrick, MD. The focus of screening was determination of host range of the fungus among related native and agriculturally important grasses in North America. A procedure was developed to consistently and quickly produce disease on medusahead and other grasses. Without vernalization, plants inoculated with U. phrygica will not produce smutted spikes (seedheads). Teliospores of the fungus were vacuum inoculated (1) onto caryopses (seeds) of medusahead and slender wheatgrass, which were then placed on moist germination paper in a petri dish or on moist vermiculite in plastic boxes. The dishes, sealed with Parafilm, and the boxes, covered with lids, were placed in a dark refrigerator at 3°C. After 8 weeks, all seedlings were transplanted into pots on a greenhouse bench at 22 to 25°C and 14 h light (photosynthetic photon flux density [PPFD] 620 µmol·s-1·m-2). The plants began to flower and produce smutted spikes 40 days later. These tests were repeated once. Fourteen of sixty medusahead plants from inoculated caryopses incubated on germination paper and nine of twenty-four plants from caryopses incubated on vermiculite became smutted and produced numerous smutted spikes per plant. Partial systemic infection was the norm, and all diseased plants had some spikes that were not diseased. One slender wheatgrass plant of nine plants grown from inoculated caryopses incubated on germination paper was also smutted and produced three diseased spikes. Nielsen (2) indicated susceptibility of slender wheatgrass to U. phrygica, but only as a single entry in a table under the synonym Agropyron trachycaulum (Link) Malte ex H. F. Lewis in a report on susceptibility of Aegilops spp. to U. phrygica. Because this is an obscure mention of the susceptibility of slender wheatgrass to U. phrygica, the fungus-host association does not explicitly appear in literature and is absent from relevant databases. Our tests with the fungus confirm that slender wheatgrass is susceptible to U. phrygica and lead us to conclude that the fungus would not be a good candidate for classical biological control of medusahead in North America. This formal report should establish this fungus-host association in literature and ensure reference in plant disease databases. References: (1) C. C. Allison. Univ. Minn. Agric. Exp. Stn. Tech. Bull. August:1, 1936. (2) J. Nielsen. Can. J. Bot. 70:581, 1992.

First Report of Leaf Spot Caused by Cladosporium herbarum on Centaurea solstitialis in Greece.

Centaurea solstitialis L. (yellow starthistle), family Asteraceae, an invasive weed in California and the western United States, is targeted for biological control. In the summer of 2003, an epidemic of unknown etiology on dying C. solstitialis plants was observed near Kozani, Greece (40°22'07″N, 21°52'35″E, elevation, 634 m). Plants had necrotic light brown leaf spots on the lower leaves and the decurrent leaf bases along the stems. Often, necrotic lesions extended along the stems to the capitula. Virtually all plants in a solid stand of C. solstitialis (approximately 0.5 ha) showed disease symptoms. Diseased plants were collected, air dried, and sent to the quarantine facility of the Foreign Disease-Weed Science Research Unit (FDWSRU), USDA/ARS, Fort Detrick, MD. On the basis of culture growth (45-cm diameter after 2 weeks at 25°C on malt extract agar), fungal morphology (1), and comparison with 21 internal transcribed spacer sequences in GenBank, the putative causal organism was identified as Cladosporium herbarum (Pers.:Fr.) Link. (teleomorph = Davidiella tassiana (De Not.) Crous & U. Braun). Sixteen C. solstitialis plants in the rosette stage and 16 plants in the bolted stage were inoculated with an aqueous suspension of spores (106 conidia ml-1) and placed in an environmentally controlled chamber at 25°C with 8 h of dew and 12 h of light daily. Plants in the rosette stage were resistant, but the fungus was very aggressive on bolted plants. Within 4 to 6 days of inoculation, necrosis developed on leaves and stems and then spread up the stems to the capitula, often resulting in plant death. The fungus also infected developing flowers. Cladosporium herbarum was reisolated from each of the 16 bolted C. solstitialis plants in two separate tests at the FDWSRU and from all bolted inoculated plants at the European Biological Control Laboratory (EBCL) in Greece. In the greenhouse at the EBCL, the pathogen readily spread to (and was isolated from) another 10 noninoculated C. solstitialis plants in close vicinity to the inoculated C. solstitialis plants. Results of host range tests will establish if this isolate of Cladosporium herbarum has the potential as a biological control agent of C. solstitialis in the United States and does not pose a threat to other Centaurea spp. used in horticulture. A voucher specimen has been deposited with the U.S. National Fungus Collections (BPI 863446). Live cultures are being maintained at the FDWSRU and EBCL, Greece. To our knowledge, this is the first report of a disease caused by Cladosporium herbarum on C. solstitialis. Reference: (1) M. H. M. Ho et al. Mycotaxon 72:115, 1999.

First Report of Anthracnose of Salsola tragus Caused by Colletotrichum gloeosporioides in Greece.

In early October of 2005, dying Salsola tragus L. (Russian thistle, tumbleweed), family Chenopodiaceae, plants were found along the Aegean Sea at Kryopigi Beach, Greece (40°02'29″N, 23°29'02″E, elevation 0 m). All of the 30 to 40 plants in the area were diseased and approximately 80% were dead or dying. All plants were relatively large (approximately 1 m tall × 0.5 m diameter), and living portions of diseased plants were flowering. Dying plants had irregular, necrotic lesions extending the length of the stems. Leaves of these plants were also necrotic. Lesions on stems and leaves were dark brown and usually coalesced. Diseased stem pieces were taken to the European Biological Control Laboratory, USDA, ARS at the American Farm School in Thessaloniki, Greece. There, diseased stem pieces were surface disinfested for 15 min with 0.5% NaOCl and placed on moist filter paper in petri dishes. Numerous, waxy subepidermal acervuli with black setae were observed in all lesions after 2 to 3 days. Conidiophores were simple, short, and erect. Conidia were one-celled, hyaline, ovoid to oblong, falcate to straight, 12.9 to 18.0 × 2.8 to 5.5 µm (mode 16.1 × 4.5 µm). These characters conformed to the description of Colletotrichum gloeosporioides (Penz.) Penz. & Sacc. in Penz. (2). Conidia were placed on modified potato carrot agar and axenic cultures from these isolations were sent to the quarantine facility of the Foreign Disease-Weed Science Research Unit, USDA, ARS, Fort Detrick, MD for testing. On the basis of DNA sequences, two variants within S. tragus have been described in California and named "Type A" and "Type B" (1). Conidia were harvested from 14-day-old cultures grown on 20% V8 juice agar, and healthy stems and leaves of 18 30-day-old plants of S. tragus Type A and 10 Type B plants were spray inoculated with an aqueous conidial suspension (1.0 × 106 conidia/ml plus 0.1% non-ionic surfactant). Three control plants of each type were sprayed with water and surfactant only. Plants were placed in an environmental chamber (18 h of dew in darkness at 25°C). After 1 day, all plants were transferred to a greenhouse (20 to 25°C, 30 to 50% relative humidity, and natural light augmented with 12-h light periods with 500-W sodium vapor lights). Lesions developed on stems of inoculated Type A plants after 5 days. After 14 days, all inoculated Type A plants were dead. Lesions on Type B plants were small and localized; all plants were diseased but no plants died. No symptoms occurred on control plants. C. gloeosporioides was reisolated 14 to 21 days after inoculation from stem pieces of all inoculated plants of both types of S. tragus. This isolate of C. gloeosporioides is a destructive pathogen on S. tragus Type A and is a potential candidate for biological control of this weed in the United States. To our knowledge, this is the first report of anthracnose caused by C. gloeosporioides on S. tragus in Greece. A voucher specimen has been deposited with the U.S. National Fungus Collections, Beltsville, MD (BPI 871126). Nucleotide sequences for the internal transcribed spacers (ITS 1 and 2) were deposited in GenBank (Accession No. DQ344621) and exactly matched sequences of the teleomorph, Glomerella cingulata. References: (1) F. Ryan and D. Ayres. Can. J. Bot. 78:59, 2000. (2) B. C. Sutton. Page 15 in: Colletotrichum Biology, Pathology and Control. J. A. Bailey and M. J. Jeger, eds. CAB International Mycological Institute, Wallingford, UK, 1992.

First Report of an Ovary Smut of Italian Thistle Caused by a Microbotryum sp. in Greece.

Italian thistle (Carduus pycnocephalus L.), family Asteraceae, is a common weed in Greece. It is also a problematic invasive weed in the western United States and a target of biological control efforts. In May 2005, smutted capitula of Italian thistle were found in an abandoned field in Halkiades, Greece. A total of 38 smutted plants, representing approximately 20% of those plants present, were found in a portion of the field that was lightly infested with Italian thistle. In most cases, capitula of all diseased flowers were smutted. In one or two cases, capitula on some branches of the plants were smutted, whereas capitula on other branches were healthy. Diseased capitula were noticeably more globose than healthy ovoid capitula, and diseased capitula did not open completely. When diseased capitula were split open, the ovaries in all florets within the capitula were filled with powdery masses of smut teliospores. Diseased capitula were collected, air dried, and sent to the quarantine facility of the Foreign Disease-Weed Science Research Unit (FDWSRU), USDA/ARS, Fort Detrick, MD. Teliospores within the capitula were extracted and observed microscopically. Teliospores of isolate DB05-014 were relatively uniform in shape and size, globose, 12.0 to 17.3 × 12.3 to 18.0 µm (mean 14.5 × 15.1 µm), violet tinted pale to medium yellowish-brown; wall reticulate appearing as coarse, radiate wings on the spore margin, 5 to 7 polyangular meshes per spore diameter, muri, 0.7 to 2.0 µm high in optical median view appearing as gradually narrowing blunt spines, 0.5 to 1 µm wide at their basis; in scanning electron microscopy (SEM), the meshes were subpolygonal, wall and interspaces were finely verruculose. Teliospores were more globose and slightly smaller than the description of Microbotryum cardui (A. A. Fischer Waldh.) Vánky (2), but the mean sizes were within the described range. When compared with teliospores of M. cardui on C. acanthoides, the numbers of polyangular meshes per spore diameter were within the range of the description using SEM, but the muri were about one-half of the height of those described. Nucleotide sequences for the internal transcribed spacers (ITS 1 and 2) and 5.8S ribosomal region (GenBank Accession No. AY280460) were aligned with sequences of other smut fungi using the BLAST algorithm of the National Center for Biotechnology Information. The closest alignment of DB05-014 was with M. scorzonerae (590 of 627 bp identities or 94% with 2% gaps). No sequences of M. cardui were available for comparison, but only M. cardui has been reported on Carduus spp. (1,2). Another smut reported on a Carduus sp. is Thecaphora trailii (1). DB05-014 is a likely variant of M. cardui from a previously unknown host. Italian thistle is an annual plant that reproduces solely by seeds (achenes). Because of the lack of seed production on smutted plants and the systemic nature of the disease, this fungus has great potential as a biological control agent for Italian thistle in the United States. A voucher specimen has been deposited with the U.S. National Fungus Collections (BPI 871812). To our knowledge this is the first report of a Microbotryum sp. parasitizing C. pycnocephalus. References: (1) K. Vánky. European Smut Fungi. Gustav Fischer Verlag, Stuttgart, Germany, 1994. (2) K. Vánky and D. Berner. Mycotaxon 85:307, 2003.

First Report of Smut Caused by Microbotryum silybum on Ivory Thistle.

Silybum eburneum Coss. & Durieu. (ivory thistle) and S. marianum (L.) Gaertn. (milk thistle) are dominant, invasive weeds in northern Tunisia (1). S. marianum is also invasive in the United States and targeted for biological control. The smut fungus Microbotryum silybum Vánky & Berner is a naturally occurring pathogen of S. marianum in Greece (2) but not in Tunisia or the United States. To assess the safety of the fungus for biological control in the United States, plants related to S. marianum were evaluated for susceptibility to M. silybum in the quarantine facility of the Foreign Disease-Weed Science Research Unit (FDWSRU), USDA/ARS, Fort Detrick, MD. Because of the close genetic relationship of S. eburneum to S. marianum, both were tested for susceptibility under greenhouse conditions at the FDWSRU. All inoculations were done by placing 5 mg of teliospores of M. silybum in the central whorl of rosettes with three to five true leaves. Individual plants in soil-filled pots were placed in a controlled chamber at 16°C with 10 h of light daily. Photon flux density in the chamber was 34 µmol·m-2·s-1 supplied by three 1.8-m long 115W fluorescent tubes and three 52W incandescent bulbs. The central whorl was misted with distilled water twice daily for 2 weeks and the temperature was then lowered to 8°C for 6 weeks. The plants were transferred to a greenhouse bench at 22 to 25°C with 14 h of light daily. Photon flux density on the bench was 620 µmol·m-2·s-1 provided by two 500W sodium vapor lamps, one 1,000W metal halide lamp, and incidental sunlight. After approximately 7 weeks, plants of each species had fully developed capitula that flowered normally, produced no flowers, or formed abnormal flowers. Abnormal capitula contained powdery masses of teliospores in the ovaries of the florets. In contrast to systemic infections that were observed in the field (2), different branches of bolted plants bore both diseased and normal capitula. In turn, diseased capitula of both species were either completely diseased (all florets filled with teliospores) or partially diseased. Four of ten S. marianum plants and six of nine S. eburneum plants were diseased. Pathogenicity tests were repeated four times with similar results. In Greece, field inoculation of S. marianum with 5 mg of teliospores produced an average of 89% diseased plants with an average of 250 g of teliospores produced per plant. A similar level of disease is possible for S. eburneum under field conditions. Teliospores from smutted ovaries of both plant species conformed to the description for M. silybum (2). Both species are annual plants that reproduce solely by seeds. Since M. silybum prevents seed production, this fungus has great potential as a biological control agent in the United States and Tunisia. A voucher specimen has been deposited with the U.S. National Fungus Collections (BPI 863477). Nucleotide sequences for the internal transcribed spacer region are available in GenBank (Accession No. AY285774). To our knowledge, this is the first report of M. silybum parasitizing S. eburneum. References: (1) G. Pottier-AlaPetite. Flore de la Tunisie: Angiospermes-Dicotylédones, Gamopétales, Tunis, 1981. (2) K. Vánky and D. Berner. Mycotaxon 85:307, 2003.

First Report of a Leaf Spot Caused by Cercospora bizzozeriana on Lepidium draba subsp. draba in Tunisia.

Lepidium draba (L.) subsp. draba (synonym = Cardaria draba (L.) Desv.), commonly known as white-top or hoary-cress (1), family Brassicaceae, is a common weed and emerging problem in wheat in Tunisia. It is also a problematic invasive weed in the northwestern United States and a target of biological control efforts. During the summer of 2002, dying L. draba plants were found around Tunis, Tunisia. Plants had grayish white leaf spots on most of the leaves. In some cases, the leaf spots dropped out of the leaves producing "shot-holes". In most cases, the leaf spots coalesced, and the leaves wilted and died. Diseased leaves were collected, air-dried, and sent to the quarantine facility of the Foreign Disease-Weed Science Research Unit (FDWSRU), USDA/ARS, Fort Detrick, MD. The air-dried leaves were observed microscopically, and numerous conidiophores and conidia were observed on both sides of the leaves within and around the lesions. The fungus isolated (DB03-009) conformed to the description of Cercospora bizzozeriana Saccardo & Berlese (2). Conidiophores were unbranched, pale olive-brown, 1 to 5 geniculate, and uniform in color and width. Conidia were hyaline, straight to slightly curved, multiseptate, and 57 to 171 × 3.8 to 6.7 µm (average 103 to 4.6 µm). Stems and leaves of 12 rosettes (10 to 15 cm in diameter) of 6-week-old L. draba plants were spray inoculated with an aqueous suspension of conidia (1 × 105/ml) harvested from 6- to 8-day-old cultures grown on carrot leaf decoction agar. Six of the plants and two noninoculated plants were placed in a dew chamber at 22°C in darkness and continuous dew. The other half of the plants and two noninoculated plants were placed on a greenhouse bench at approximately 25°C and covered with clear polyethylene bags. After 72 h, plants from the dew chamber were moved to a greenhouse bench, and the bagged plants were uncovered. All plants were watered twice daily. After 9 days, symptoms were observed on the plants that had been bagged but not on the plants from the dew chamber. Symptoms were identical to those observed in the field in Tunisia and included "shot holes". No symptoms were observed on noninoculated plants. C. bizzozeriana was reisolated from the leaves of all symptomatic plants. Completion of Koch's postulates was repeated with an additional five plants. This isolate of C. bizzozeriana is a destructive pathogen on L. draba subsp. draba, and severe disease can be produced by inoculation of foliage with an aqueous suspension of conidia. This isolate is a good candidate for mycoherbicide development in Tunisia where the weed and pathogen are indigenous. However, some commercially grown Brassica species were found susceptible to this isolate, which will preclude its use as a classical biological control agent in the United States. To our knowledge, this is the first report of C. bizzozeriana on L. draba subsp. draba in Tunisia. A voucher specimen has been deposited at the U.S. National Fungus Collections (BPI 843753). Live cultures are being maintained at FDWSRU and the Institut National Agronomique de Tunisie, Tunis, Tunisia. References: (1) I. A. Al-Shehbaz and K. Mummenhoff. Novon 12:5, 2002. (2) C. Chupp. A Monograph of the Fungus Genus Cercospora. C. Chupp, Ithaca, New York, 1953.