RESUMEN
Kluyveromyces marxianus var. marxianus, a causal agent of onion (Allium cepa L.) soft rot, was originally isolated from Walla Walla sweet onion bulbs grown in Oregon and subsequently isolated from onion throughout the southeast Washington onion-growing area, but was not found to be infecting dry bulb storage onions grown in central Washington (1). During September of 2001, a yeast was isolated from dry storage onion bulbs (cv. Teton) grown under sprinkler irrigation in central Washington, exhibiting soft rot symptoms and identified to be K. marxianus var. marxianus (2). Koch's postulate was completed using cv. Teton bulbs surface disinfested with 0.5% NaOCl for 2 min. This isolate and four isolates of Kluyveromyces marxianus var. marxianus (1) were cultured on potato dextrose agar and resuspended to an OD600 = 0.3 (approximately 105 CFU/ml). One-half of a milliliter of each isolate was inoculated to onion using the cut bulb method with three replicates and incubated in a moist chamber at 25°C for 7 days. Onion slices inoculated with the new isolate exhibited soft rot symptoms similar to those caused by the known isolates while no symptoms were observed for the water control. The yeast reisolated from symptomatic tissue was confirmed to be K. marxianus var. marxianus (2). The identification of K. marxianus var. marxianus infecting dry bulb storage onions grown in the Columbia Basin is of interest because the disease can be confused with bacterial soft rot and could become a serious problem in this important storage onion-growing region. References: (1) D. A. Johnson et al. Plant Dis. 72:359, 1988. (2) N. J. W. Kreger-van Rig, ed. The Yeasts: A Taxonomic Study. Elsevier, Amsterdam, 1984.
RESUMEN
The geographic distribution of Iris yellow spot virus (IYSV, Genus Tospovirus, Family Bunyaviridae) in onion (Allium cepa L.) crops in the western United States has increased with the most recent report in Colorado (1,4). Furthermore, the incidence of IYSV has increased significantly in onion crops in the Treasure Valley of southern Idaho and eastern Oregon, where the disease was first detected in the United States (1,2). Surveys of onion seed crops in Washington during the past 2 years showed the presence of plants with symptoms characteristic of IYSV infection, including distinct diamond-shaped chlorotic or necrotic lesions, as well as indistinct circular to irregular, chlorotic or necrotic lesions of various sizes on the scapes of flowering plants. To date, symptomatic plants have been observed in five seed crops in Washington, at incidences ranging from <1% to approximately 20% in individual seed crops. Enzyme-linked immunosorbent assays carried out directly on symptomatic onion samples collected in July 2002, and on Nicotiana benthamiana plants mechanically inoculated with sap from these symptomatic plants, did not detect the presence of IYSV. In late July 2003, symptomatic plants were collected from an onion seed crop in Grant County and tested for IYSV infection by reverse transcription-polymerase chain reaction (RT-PCR). Total nucleic acid was extracted from symptomatic areas of the scapes with the procedure described by Presting et al. (3). Primers specific to the nucleocapsid (NP) gene of IYSV were designed based on sequences in GenBank: 5'-TCA GAA ATC GAG AAA CTT-3' and 5'-TAA TTA TAT CTA TCT TTC TTG G-3' (sense and antisense polarity, respectively). The RT-PCR assay produced an amplicon of the expected size (approximately 700 bp) that was cloned and sequenced. Comparison with the GenBank IYSV gene sequences showed 98% sequence identity of the NP gene. In August 2003, symptoms of IYSV infection were observed in two onion bulb crops, each located within 2 miles of the symptomatic onion seed crop in Grant County. The presence of IYSV in these crops was confirmed by RT-PCR with cloning and sequencing of the amplicon, as described for the seed crop samples. To our knowledge, this is the first confirmation of IYSV in onion bulb and seed crops in Washington, where 16,000 to 18,000 acres of onion bulb crops and 700 to 900 acres of onion seed crops are grown annually (USDA National Agricultural Statistics Service). The increase in prevalence of IYSV in the Pacific Northwest highlights the need for additional research to clarify the epidemiology of this potentially significant pathogen and to develop a regional management program for iris yellow spot. References: (1) J. M. Hall et al. Plant Dis. 77:952, 1993. (2) J. W. Moyer et al. (Abstr.) Phytopathology 93(suppl.):S115, 2003. (3) G. G. Presting et al. Phytopathology 85:436, 1995. (4) H. F. Schwartz et al. Plant Dis. 86:560, 2002.
RESUMEN
Fusarium oxysporum f. sp. cepae and an unidentified Fusarium species have been reported to cause bulb rot of onion (Allium cepa L.) in Washington (1). In August and September 2002, a salmon-pink discoloration was observed on the outer three to four layers of dry scales of approximately 20% of white onion bulbs of cv. Cometa F1, in each of two 20-acre fields in the Columbia Basin of central Washington. Isolations from the discolored areas of the dry scales onto water agar and potato dextrose agar (PDA) yielded fungal colonies characteristic of F. proliferatum (3). The isolates formed long, V-shaped chains of microconidia on polyphialides. Pathogenicity of the isolates of F. proliferatum was tested on white onion bulbs purchased at a local grocery store. The outermost dry scales of each bulb were removed, and the bulb was inoculated by one of three methods: (i) a 5-mm3 section of the fleshy scales was removed using a scalpel, the wound was filled with a 3-mm2 plug of PDA colonized by F. proliferatum, the plug was covered with the section of scale that had been removed, and the inoculation site was covered with Parafilm; (ii) the basal plate of the bulb was dipped into a suspension of 106 microconidia per ml; or (iii) the basal plate was dipped into the spore suspension after wounding by inserting a dissecting needle into the bulb to a depth of 1 cm. A noninoculated bulb provided a control treatment. Bulbs were incubated in a moist chamber at 13°C and examined for discoloration of the outer scales and development of bulb rot. After 2 weeks, salmon-pink discoloration of the outer scales was observed at the inoculation site for both methods of dip inoculation, but not for the plug inoculation method. After 3 weeks, water-soaked, tan to golden, shrunken, soft tissue was observed on the remainder of each dip inoculated bulb, but symptoms of basal rot did not develop. Symptoms were similar to those reported in Idaho for a bulb rot of stored onions caused by F. proliferatum (2). One of the nonwounded inoculated bulbs did not develop a bulb rot, but pinkish discoloration was observed beneath the outer scales and in the neck. F. proliferatum was reisolated from the inoculated bulb tissues. The discoloration observed on the white onions raised concern about the potential for infection to develop into bulb rot in storage. However, thorough curing of the bulbs immediately upon storage restricted infection to the outer dry scales. Similar symptoms were observed at harvest on the bulbs of other white onions in a cultivar trial located near Quincy, WA, although symptoms were not observed on yellow or red cultivars in the trial. The same symptoms were later observed on approximately 70% of bulbs harvested from a 32-acre fresh-market crop of the cv. Sterling in the Columbia Basin. These symptomatic bulbs were rejected for the fresh market. To our knowledge, this is the first report of infection of onion bulbs by F. proliferatum in Washington, which in 2001, had the third largest acreage of onions in the United States after California and Oregon (USDA National Agricultural Statistics Service). References: (1) D. F. Farr et al. Fungi on Plants and Plant Products in the United States. The American Phytopathological Society, St. Paul, MN, 1989. (2) S. K. Mohan et al. (Abstr.) Phytopathology 87:S67, 1997. (3) P. E. Nelson et al. Fusarium species: An Illustrated Manual for Identification. The Pennsylvania State University Press, University Park, 1983.
RESUMEN
Nine fields direct-seeded with onion (Allium cepa L.) were surveyed in central Washington in the spring and summer of 2001 for Botrytis species associated with onion seed crops produced in this semiarid region. Forty plants were sampled from each field in a 'W' pattern in April, and 20 plants were similarly sampled from each field in June and July. Each plant was placed in a separate plastic bag, stored at 4 ± 2°C for 3 to 5 weeks, sliced lengthwise using a knife sterilized with 70% ethyl alcohol, incubated in a moist chamber for 5 days, and examined under a dissecting microscope. Fungal growth resembling Botrytis spp. was transferred to acidified potato dextrose agar (PDA) for species identification based on colony morphology, rate of growth, and spore and sclerotium characteristics (3). Cultures were incubated on a laboratory bench at 24 ± 4°C with 8 to 16 h of daylight. A species resembling B. porri (3) was detected in 3 fields in April at an incidence ranging from 3 to 28%, and in 2 of the same 3 fields in each of June and July at incidences ranging from 5 to 10%. Infected plants were asymptomatic at the time of sampling. The isolates formed brown, cerebriform sclerotia and sporulated sparsely. Subsamples of seed harvested from each field were assayed for Botrytis spp. To detect internal infection, 400 seeds from each of the nine fields were soaked in 0.525% NaOCl for 60 s, triple-rinsed in sterile deionized water, air dried, placed on a selective agar medium (2) with 20 seed per 9-cm-diameter petri plate, and incubated at 24°C (12 h day/night) for 14 days. Seeds were examined 5, 10, and 14 days after plating, and fungi resembling Botrytis spp. were transferred to acidified PDA for species determination. Isolates resembling B. porri were detected in 0.75% of seed from two of the three fields in which this species was isolated from plant samples. The internal transcribed spacer 1 region of ribosomal DNA of four isolates of the putative B. porri (two from plant samples and two from seed) were sequenced, and all four sequences matched that of B. porri registered in GenBank (Accession No. Z99666) most closely. Botrytis porri is a pathogen of garlic (A. sativum L.), leek (A. porrum L.), and wild garlic (A. vineale L.), but can infect onion and shallot (A. ascalonicum L.) when inoculated on these hosts (1). To our knowledge, this is the first report of natural infection of onion by B. porri, and the first report of seedborne B. porri on onion. References: (1) W. R. Jarvis. Pathology. Page 62 in: Botryotinia and Botrytis Species: Taxonomy, Physiology, and Pathogenicity. Canada Department of Agriculture, Monograph No. 15, 1977. (2) G. Kritzman and D. Netzer. Phytoparasitica 6:3, 1978. (3) A. H. Presly. Plant Pathol. 34:422, 1985.
RESUMEN
The cost of managing late blight in potatoes during a severe epidemic caused by new, aggressive strains of Phytophthora infestans in the Columbia Basin of Washington and Oregon in 1995 was documented. The mean number of fungicide applications per field varied from 5.1 to 6.3 for early- and midseason potatoes, and from 8.2 to 12.3 for late-season potatoes in the northern and southern Columbia Basin, respectively. In 1994, a year when late blight was not severe, the mean number of fungicide applications per field made to early- and midseason potatoes was 2.0; whereas late-season potatoes received a mean of 2.5 applications. The mean per acre cost of individual fungicides applied varied from $4.90 for copper hydroxide to $36.00 for propamocarb + chlorothalonil. Total per acre expenses (application costs plus fungicide material) for protecting the crop from late blight during 1995 ranged from $106.77 to $110.08 for early and midseason potatoes in different regions of the Columbia Basin and from $149.30 to $226.75 for lateseason potatoes in the northern and southern Columbia Basin, respectively. Approximately 28% of the crop was chemically desiccated before harvest as a disease management practice for the first time in 1995, resulting in an additional mean cost of $34.48/acre or $1.3 million for the region. Harvested yields were 4 to 6% less than in 1994. The total cost of managing late blight in the Columbia Basin in 1995 is estimated to have approached $30 million.