Lindsey J. du Toit

Iris yellow spot virus: An Emerging Threat to Onion Bulb and Seed Production

This paper reviews the history of Iris yellow spot virus in the USA and the world, epidemiology of Iris yellow spot and the onion thrips vector, and management of Iris yellow spot and thrips. Future prospects and outlook are also discussed.

Verticillium Wilt in Spinach Seed Production

There are no previous reports of Verticillium wilt in fresh and processing spinach (Spinacia oleracea) crops in the United States. In 2002, a hybrid spinach seed crop in the Pacific Northwest developed late-season wilt symptoms. Assays of the harvested seed and stock seed of the male and female parents revealed 59.5, 44.0, and 1.5%, respectively, were infected with Verticillium dahliae. Assays of 13 stock or commercial seed lots grown in 2002 and 62 commercial lots harvested in 2003 in Denmark, Holland, New Zealand, and the United States revealed the prevalence of Verticillium spp. in commercial spinach seed. Sixty-eight lots (89%) were infected with Verticillium spp. at incidences ranging from 0.3 to 84.8%. Five spinach seed isolates of V. dahliae were pathogenic on each of three spinach cultivars by root-dip inoculation. V. dahliae was detected on 26.4% of the seed from 7 of 11 inoculated plants but on none of the seed from 6 control plants, demonstrating systemic movement of V. dahliae. Seed-to-seed transmission was also demonstrated by planting naturally infected seed lots. This is the first report of Verticillium wilt of spinach in the primary region of spinach seed production in the United States.

Population analyses of the vascular plant pathogen Verticillium dahliae detect recombination and transcontinental gene flow.

The fungal pathogen Verticillium dahliae has resulted in significant losses in numerous crops in coastal California, but lettuce remained unaffected until the mid-1990s. Since then outbreaks have decimated entire fields, but the causes of this sudden susceptibility of lettuce remain elusive. The population structure of V. dahliae isolated from coastal California (n=123) was investigated with 22 microsatellite markers, and compared with strains from tomato in central California (n=60), spinach seed imported from Washington State and Northern Europe (n=43), and ornamentals from Wisconsin (n=17). No significant differentiation was measured among hosts in coastal California or with the spinach and Wisconsin ornamental sampling groups. In contrast, the tomato sampling group was significantly differentiated. Significant gene flow was measured among the various geographic and host sampling groups, with the exception of tomato. Evidence of recombination in V. dahliae was identified through gametic disequilibrium and an exceedingly high genotypic diversity. The high incidence of V. dahliae in spinach seed and high planting density of the crop are sources of recurrent gene flow into coastal California, and may be associated with the recent outbreaks in lettuce.

Carrot Purple Leaf: A New Spiroplasmal Disease Associated with Carrots in Washington State

During the growing seasons of 2003 and 2004, a disease occurred in several carrot crops in south central Washington with symptoms suggestive of infection by phytopathogenic mollicutes (phytoplasmas and spiroplasmas). In the fall, many affected carrot plants exhibited extensive purple or yellow-purple leaf discoloration, general stunting of shoots and taproots, and formation of bunchy, fibrous secondary roots. For detection of the putative causal agents, polymerase chain reaction (PCR) assays were performed using primers specific to phytoplasmas as well as primers specific to plant-pathogenic spiroplasmas. Restriction fragment length polymorphism (RFLP) analyses of PCR-amplified 16S rDNA sequences revealed that about 81% of affected plants showing dark purple or yellow-purple leaf symptoms tested positive for Spiroplasma citri. Of affected plants showing mild purple discoloration of leaf margins, 18% tested positive for a phytoplasma strain belonging to the clover proliferation group (16SrVI), subgroup 16SrVI-A, and 11% for another phytoplasma strain belonging to the aster yellows group (16SrI), subgroup 16SrI-A. Nucleotide sequence analysis of cloned 16S rDNA confirmed the phytoplasma group affiliations. Some symptomatic plants were co-infected with S. citri and either aster yellows phytoplasma or clover proliferation group phytoplasma. To our knowledge, this is the first documentation of spiroplasma infection of carrot in the United States.

A real-time, quantitative PCR seed assay for Botrytis spp. that cause neck rot of onion

A real-time fluorescent polymerase chain reaction (PCR) assay was developed using SYBR Green chemistry to quantify the Botrytis spp. associated with onion (Allium cepa) seed that are also able to induce neck rot of onion bulbs, i.e., B. aclada, B. allii, and B. byssoidea. The nuclear ribosomal intergenic spacer (IGS) regions of target and nontarget Botrytis spp. were sequenced, aligned, and used to design a primer pair specific to B. aclada, B. allii, and B. byssoidea. Primers and amplification parameters were optimized to avoid amplifying the related species B. cinerea, B. porri, and B. squamosa, as well as Sclerotinia sclerotiorum and isolates of 15 other fungal species commonly found associated with onion seed. The primers reliably detected 10 fg of genomic DNA per PCR reaction extracted from pure cultures of B. aclada and B. allii. Conventional assays of surface-disinfested and nondisinfested seed on an agar medium were used to determine the incidence of neck rot Botrytis spp. associated with each of 23 commercial onion seed lots, and the real-time PCR assay was used to determine the quantity of DNA of neck rot Botrytis spp. in each seed lot. A linear relationship could not be found between the incidence of seed infected with the neck rot Botrytis spp. using the conventional agar seed assays and the quantity of DNA of the neck rot Botrytis spp. detected by the real-time PCR assay. However, the real-time PCR assay appeared to be more sensitive than the conventional agar assay, allowing detection of neck rot Botrytis spp. in 5 of the 23 seed lots that tested negative using the conventional agar seed assay.

Efficacy of Hot Water and Chlorine for Eradication of Cladosporium variabile, Stemphylium botryosum, and Verticillium dahliae from Spinach Seed

Cladosporium variabile, Stemphylium botryosum, and Verticillium dahliae are seedborne and seed-transmitted pathogens of spinach. Spinach seed treatments in 1.2% NaOCl for 10 to 60 min, or hot water (40, 45, 50, 55, and 60°C) for 10 to 40 min, were evaluated for eradication of these fungi from seed. C. variabile and V. dahliae were largely eradicated by chlorine treatment for ≥ 10 min. Although chlorine treatment reduced the incidence of S. botryosum, this fungus was not eradicated after 60 min in chlorine. Seed germination was not affected adversely by chlorine treatment, even after 60 min. In contrast, germination was reduced significantly by hot water treatment at 50°C for ≥30 min or 55 or 60°C for ≥10 min. C. variabile was eradicated from seed treated in 40°C water for 10 min. V. dahliaewas eradicated from seed treated at 55°C for ≥30 min or 60 °C for ≥10 min. S. botryosum was eradicated from a lightly infected seed lot (5% incidence) by hot water treatment at 55 or 60 °C for ≥10 min, but could not be eradicated from two heavily infected lots (>65% incidence), even at 60°C for 40 min. Using precisely controlled parameters, chlorine or hot water seed treatments can be used to eradicate C. variabile and reduce the incidence of S. botryosum and V. dahliae in spinach seed without damaging germination.

Seedborne Cladosporium variabile and Stemphylium botryosum in spinach.

Assays of 77 spinach (Spinacia oleracea) seed lots produced in the United States, Denmark, the Netherlands, or New Zealand in 2000 to 2003 showed that Stemphylium botryosum, causal agent of Stemphylium leaf spot, was present in every lot, at a mean incidence of 29.1% per lot. Either Cladosporium variabile, causal agent of Cladosporium leaf spot, or the morphologically similar species C. macrocarpum, was present in 37 of the 77 lots, at a mean incidence of 1.8% per lot. Some seed isolates of S. botryosum and C. variabile proved pathogenic on spinach. Nonpathogenic isolates resembling C. variabile were identified as C. macrocarpum by the absence of torulose aerial hyphae. Pathogenic isolates of S. botryosum were also detected in each of 12 seed lots stored for up to 11 years at 4.4°C and 60% relative humidity. C. variabile or C. macrocarpum was detected in only 2 of the 11 lots, which had been stored for 3 and 8 years. Component seed assays demonstrated that S. botryosum and C. variabile (or C. macrocarpum) were internal and external in spinach seed. S. botryosum was detected in 5 to 76% of the embryos of five seed lots, but the two Cladosporium species were detected in only 0 to 1% of the embryos of these lots. This suggests greater potential difficulty at eradicating S. botryosum than C. variabile from infected spinach seed using seed treatments.

Evaluation of an Aeroponics System to Screen Maize Genotypes for Resistance to Fusarium graminearum Seedling Blight

A noncirculating aeroponics system was evaluated as a method for rapid screening of maize genotypes for resistance to Fusarium graminearum seedling blight/root rot. The system allows for nondestructive, repetitive sampling of seedlings for assessing disease progress and seedling growth. Shoot growth and root rot were assessed at 3-day intervals, and final shoot and root dry weight were determined 15 days after inoculation. The nine hybrids screened differed in severity of root rot as early as 6 days after inoculation, indicating differences in resistance to F. graminearum. Inoculation did not always significantly affect shoot growth, root dry weight, or shoot dry weight, but differences in these agronomic traits were observed among hybrids.LH119 × LH51 and Pioneer Brand 3379 showed the greatest resistance to root rot. Area under-disease progress curve and a critical stage of disease assessment (9 days after inoculation) gave similar rankings of hybrids for root rot resistance, indicating that a single disease assessment (versus multiple assessments) may be adequate in screening for resistance with this aeroponics system.

Prevalence of Botrytis spp. in Onion Seed Crops in the Columbia Basin of Washington

Of 12 onion seed lots harvested in the semi-arid Columbia Basin of Washington in 1999 or 2000, 8 were infected and 10 infested with Botrytis aclada at incidences of 1 to 10% and 2 to 26%, respectively. Twenty to forty plants were sampled from each of nine direct-seeded, biennial seed crops in April, June, and July 2001 and assayed for Botrytis spp. Six direct-seeded crops were sampled in October and November 2001 and April, June, and July 2002. One bulb-to-seed crop was sampled in April, June, and July 2002. The incidence of B. aclada increased through each season, reaching 100% in most fields by July. Infections were primarily asymptomatic, with no apparent relationship between plant infection and infection of harvested seed. B, cinerea, B. squamosa, and B. porri were detected in 16, 4, and 4% of the fields, respectively, at lower incidences than B. aclada. Harvested seed from 15 of the fields were infected with B. aclada at <1 to 28%. B, cinerea, B. porri, and B. squamosa were detected in three, three, and none of the harvested lots, respectively.

Reactions of processing sweet corn hybrids to Gibberella ear rot

Gibberella ear rot has increased in prevalence recently on late-season processing sweet corn grown in the Midwestern United States. Little information is available about the reactions of hybrids currently grown for processing. A total of 52 processing hybrids were evaluated in 1996 and 1997 for reactions to Gibberella zeae following one of two methods of inoculation: inserting an infested toothpick or injecting a spore suspension in the silk channel 7 days after the mid-silk growth stage. Ratings of ear rot severity ranged from 2.4 to 8 on a scale of 1 to 9. Severity of ear rot differed among hybrids, but none of the hybrids was highly resistant, nor did the hybrids display a great degree of variation for reaction to G. zeae. Severity of ear rot was consistently lower for cvs. Sprint and HMX 5372 than for the other hybrids. Ear rot ratings also were lower for cvs. GH 1834 and GH 2690 in three of four comparisons. Severity of ear rot was consistently higher for cv. GH 2628 in all comparisons and for cvs. Empire, Excalibur, GG 42, GH 1861, Jubilee, Rival, and WSS 3680 in three of four comparisons. Ranks of some hybrids changed between inoculation methods, years, or both, but most hybrids were ranked similarly in at least three of four comparisons. Coefficients of variation were approximately 5% higher for the injection method of inoculation in 1996 than for the other trials. The toothpick method of inoculation required less labor and ranked hybrids as consistently or more consistently than the injection method.