Genomic differences between the new Fusarium oxysporum f. sp. apii (Foa) race 4 on celery, the less virulent Foa races 2 and 3, and the avirulent on celery f. sp. coriandrii.

BACKGROUND: Members of the F. oxysporium species complex (FOSC) in the f. sp. apii (Foa) are pathogenic on celery and those in f. sp. coriandrii (Foci) are pathogenic on coriander (=cilantro). Foci was first reported in California in 2005; a new and highly aggressive race 4 of Foa was observed in 2013 in California. Preliminary evidence indicated that Foa can also cause disease on coriander, albeit are less virulent than Foci. Comparative genomics was used to investigate the evolutionary relationships between Foa race 4, Foa race 3, and the Foci, which are all in FOSC Clade 2, and Foa race 2, which is in FOSC Clade 3. RESULTS: A phylogenetic analysis of 2718 single-copy conserved genes and mitochondrial DNA sequence indicated that Foa races 3 and 4 and the Foci are monophyletic within FOSC Clade 2; these strains also are in a single somatic compatibility group. However, in the accessory genomes, the Foci versus Foa races 3 and 4 differ in multiple contigs. Based on significantly increased expression of Foa race 4 genes in planta vs. in vitro, we identified 23 putative effectors and 13 possible pathogenicity factors. PCR primers for diagnosis of either Foa race 2 or 4 and the Foci were identified. Finally, mixtures of conidia that were pre-stained with different fluorochromes indicated that Foa race 4 formed conidial anastomosis tubes (CATs) with Foci. Foa race 4 and Foa race 2, which are in different somatic compatibility groups, did not form CATs with each other. CONCLUSIONS: There was no evidence that Foa race 2 was involved in the recent evolution of Foa race 4; Foa race 2 and 4 are CAT-incompatible. Although Foa races 3 and 4 and the Foci are closely related, there is no evidence that either Foci contributed to the evolution of Foa race 4, or that Foa race 4 was the recent recipient of a multi-gene chromosomal segment from another strain. However, horizontal chromosome transfer could account for the major difference in the accessory genomes of Foa race 4 and the Foci and for their differences in host range.

Stemphylium Leaf Spot of Parsley in California Caused by Stemphylium vesicarium.

From 2009 through 2011, a previously undescribed disease occurred on commercial parsley in coastal (Ventura County) California. Symptoms of the disease consisted of circular to oval, tan to brown leaf spots and resulted in loss of crop quality and, hence, reduced yields. A fungus was consistently isolated from symptomatic parsley. Morphological and molecular data identified the fungus as Stemphylium vesicarium. When inoculated onto parsley leaves, the isolates caused symptoms that were identical to those seen in the field; the same fungus was recovered from test plants, thus completing Koch's postulates. Additional inoculation experiments demonstrated that 10 of 11 tested flat leaf and curly parsley cultivars were susceptible. The parsley isolates also caused small leaf spots on other Apiaceae family plants (carrot and celery) but not on leek, onion, spinach, and tomato. Isolates caused brown lesions to form when inoculated onto pear fruit but only when the fruit tissue was wounded. Using a freeze-blotter seedborne pathogen assay, parsley seed was found to have a low incidence (0.25%) of S. vesicarium. When inoculated onto parsley leaves, three of four isolates from seed caused the same leaf spot disease. This is the first documentation of a foliar parsley disease caused by S. vesicarium. The occurrence of S. vesicarium on parsley seed indicates that infested seed may be one source of initial inoculum. Based on the negative results in the host range experiments, it appears that this parsley pathogen differs from the S. vesicarium that causes disease on leek, garlic, onion, and pear fruit.

Biological control of Eotetranychus lewisi and Tetranychus urticae (Acari: Tetranychidae) on strawberry by four phytoseiids (Acari: Phytoseiidae).

The spider mite, Eotetranychus lewisi (McGregor) (Acari: Tetranychidae), is a new emerging pest in California commercial strawberries. The predatory mite Phytoseiulus persimilis (Athias-Henriot) (Acari: Phytoseiidae), typically used for biocontrol of Tetranychus urticae (Koch) (Acari: Tetranychidae), provided growers little to no control of E. lewisi. Four commonly used phytoseiid predatory mites: P. persimilis, Neoseiulus californicus (McGregor), N. fallacis (Garman), and Amblyseius andersoni (Chant), were used in lab studies to investigate which is best at managing E. lewisi populations. We als o investigated t he interactions between T. urticae and E. lewisi and in relation to phytoseiid efficiency given the potential for indirect effects of biocontrol. When E. lewisi and T. urticae are present on the same leaf, T. urticae populations increase and begin displacing E. lewisi. P. persimilis did not feed on E. lewisi, but the other three predatory mites consumed the spider mites and lowered their populations. When both E. lewisi and T. urticae are present on the same leaf, N. fallacis and A. andersoni fed on both types of mites equally and were capable of decreasing both populations. N. californicus fed on E. lewisi first and decreased its population, but allowed T. urticae populations to increase. P. persimilis may be insufficient at controlling E. lewisi and its use may instead enhance E. lewisi populations.

Distribution, Cultivar Susceptibility, and Epidemiology of Apium virus Y on Celery in Coastal California.

Apium virus Y (ApVY) is a potyvirus that was recently found to cause crop loss to celery (Apium graveolens) in California. Symptoms on leaves exhibit varying forms of chlorosis and necrosis. Depending on the cultivar, celery petioles could also exhibit extensive necrotic, sunken, elongated lesions. Severely affected plants were unmarketable. Disease incidence surveys found that a susceptible celery (cv. 414) showed 55% (2007) and 71% (2008) disease. Because it was noted that the Apiaceae weed poison hemlock (Conium maculatum) was present in almost all areas where ApVY affected celery, a 4-year survey collected overwintered hemlock from six coastal county regions and tested composite samples for ApVY using reverse transcription-polymerase chain reaction (RT-PCR) and ApVY-specific primers. These plants were consistently positive for ApVY. Seeds collected from these plants were also positive when tested with the same RTPCR method. However, when ApVY-positive hemlock seeds were germinated and the resulting seedlings tested, all results were negative. The failure of ApVY to be transmitted from hemlock seeds to seedlings was further documented by collecting newly germinated hemlock seedlings from the field and testing them with RT-PCR. All such seedlings were negative for ApVY even though large, adjacent, overwintered hemlock plants tested positive. Two crops of celery seed were produced from ApVY-positive mother plants; celery seed from these infected plants likewise tested positive for ApVY, but seedlings grown from the seed lots were negative for ApVY. Twenty-one celery and celeriac cultivars were inoculated with ApVY using viruliferous aphids, planted in a replicated field trial, and then grown to maturity. Seven cultivars remained symptomless, tested negative for ApVY, and showed signs of possible resistance. The epidemiology of disease caused by ApVY in California evidently involves poison hemlock as a common overwintering host with subsequent vectoring of the virus from hemlock to celery via aphids. ApVY was not seedborne in this weed host or in celery in our experiments. Our data suggest that growers can manage this disease by controlling poison hemlock weed populations and by planting celery cultivars that are not susceptible to ApVY.

Probabilities for survival of glassy-winged sharpshooter and olive fruit fly pests in urban yard waste piles.

Glassy-winged sharpshooter (Homolodisca coagulate) and olive fruit fly (Bactrocera oleae) were introduced into unturned, chipped yard waste piles to evaluate their survival with time and depth within the piles. In all three trials, no pests lasted more than 14 d, and in no trial did pests survive more than 4d at the 30 and 100 cm depths. No survivors were found after 14 d in any of the treatments at any depth. Neither of the pests survived 100 cm after 2d. A mathematical model for describing pest survival probabilities is described. The model modifies time according to the Arrhenius equation in order to include heat effects on pest survival and can be used to determine exposure times necessary to eliminate these pests with a determined statistical probability. Model projections suggest that for conditions similar to this study, there is 99% confidence that all glassy-winged sharpshooter eggs would be eliminated from 1000 infected leaves in 6.1d at 15 cm depth and in 4.8d at 30 cm or below. Olive fruit fly larvae at these depths would require 4.8 and 4.1d, respectively, for 1000 infected olive fruits. Projected elimination times at the surface were longer, 6.5d for sharpshooter eggs and 14.3d for fruit fly larvae.