T. A. Turini

A new bipartite Geminivirus (Begomovirus) causing leaf curl and crumpling in cucurbits in the Imperial Valley of California.

During fall 1998, volunteer watermelons (Citrullus lunatus L. (Thunb.) Matsum. & Nakai) showing leaf curl, crumpling, and yellowing symptoms were found in a commercial honeydew melon (Cucumis melo L. subsp. melo Inodorus group) field in the Imperial Valley of California. The plants were infected with a begomovirus (family Geminiviridae, genus Begomovirus) based on (i) a positive response in squash blots probed with a general begomovirus DNA probe (1) and (ii) amplification of DNA-A (≈1.2 kb) and DNA-B (≈1.4 kb) fragments by polymerase chain reaction (PCR) with degenerate DNA-A (PAL1v1978/PAR1c496) and DNA-B (PBL1v2040/PBR1c970) primers, respectively (3). The DNA-A and -B fragments were cloned and sequenced (GenBank accession nos. AF224760 [DNA-A] and AF224761 [DNA-B]). The DNA-A and -B fragments had a nearly identical (99.5%) common region (CR) of 186 (DNA-A) and 187 (DNA-B) nucleotides, indicating they were from the same begomovirus. Database searches conducted with these sequences revealed no high degree of sequence identity (i.e., >90%) with other begomoviruses, including Squash leaf curl virus (SqLCV [2]) from southern California. The partial AC1 sequence (669 nt) was most identical to Tomato severe leaf curl virus (ToSLCV) from Guatemala (83%) and SqLCV (81%), the partial AV1 sequence (135 nt) was most identical to Tomato golden mosaic virus from Brazil (84%) and SqLCV (81%), and the CR was most identical to Squash yellow mottle virus from Costa Rica (81%), ToSLCV (81%), and SqLCV (77%). The partial BV1 sequence (465 nt) was most identical to Bean calico mosaic virus and SqLCV (72%), and the partial BC1 sequence (158 nt) was most identical to SqLCV (75%). Watermelon seedlings bombarded with a DNA extract from infected watermelon volunteers developed crumpling and distortion symptoms, whereas seedlings bombarded with gold particles alone developed no symptoms. Geminivirus infection in symptomatic seedlings was confirmed by PCR. These results suggest a new begomovirus caused the disease symptoms in the watermelon volunteers. Leaf crumpling and curling symptoms were not observed in spring melons in the Imperial Valley in 1999, but on 2 July and 17 August 1999, cantaloupe (C. melo L. subsp. melo Cantalupensis group), muskmelon (C. melo L. subsp. melo Cantalupensis group), and watermelon plants with leaf crumpling and yellowing were found. These plants were infected with the new begomovirus based on sequence analysis of PCR-amplified DNA-A fragments (97 to 98% identity for CR and partial AC1 sequence). A survey of fall melons, conducted 23 to 24 September 1999, revealed widespread symptoms of leaf curl and crumpling on new growth of muskmelon plants in all seven commercial fields examined (estimated incidence 25 to 50%) and on watermelon volunteers. No such symptoms were observed on leaves of honeydew melons. Symptomatic muskmelon and watermelon leaves, collected from eight locations throughout the Imperial Valley, were infected with the new begomovirus based on sequence analysis of PCR-amplified DNA-A fragments. Thus, a new begomovirus has emerged in the Imperial Valley; the name Cucurbit leaf crumple virus (CuLCrV) is proposed. References: (1) R. L. Gilbertson et al. Plant Dis. 75: 336, 1991. (2) S. G. Lazarowitz and I. B. Lazdins. Virology 180:58, 1991. (3) M. R. Rojas et al. Plant Dis. 77:340, 1993.

Biology and Molecular Characterization of Cucurbit leaf crumple virus, an Emergent Cucurbit-Infecting Begomovirus in the Imperial Valley of California

Cucurbit leaf crumple virus (CuLCrV) is an emergent and potentially economically important bipartite begomovirus first identified in volunteer watermelon plants in the Imperial Valley of southern California in 1998. Field surveys indicated that CuLCrV has become established in the Imperial Valley; and field plot studies revealed that CuLCrV primarily infects cucurbits, including cantaloupe, squash, and watermelon. Full-length DNA-A and DNA-B clones of an Imperial Valley isolate of CuLCrV were obtained by polymerase chain reaction (PCR) with overlapping primers. These clones were infectious in various cucurbits and common bean (cv. Topcrop); symptoms included stunted growth and leaf crumple, curl, and chlorosis. CuLCrV was not sap-transmissible, and immunolocalization and DNA in situ hybridization studies revealed that it is phloem-limited. A CuLCrV agroinoculation system was generated, and host range studies revealed differential susceptibility in cucurbits, with squash, watermelon, cantaloupe, and honeydew melon being most to least susceptible, respectively. Germplasm screening studies identified a number of resistant cantaloupe and honeydew melon cultivars. The genome organization of this CuLCrV isolate (CuLCrV-CA) is similar to other bipartite begomoviruses, and phylogenetic analysis placed CuLCrV in the Squash leaf curl virus (SLCV) cluster of New World bipartite begomoviruses. A CuLCrV-specific PCR test was developed which allows for differentiation from other begomoviruses, including SLCV.

Application of preplant fumigants via drip irrigation systems for the management of root rot of melons caused by Monosporascus cannonballus

Root rot and vine decline, caused by Monosporascus cannonballus, is a destructive disease of melons in the desert production regions of southern California. In 1998, we initiated studies on the use of preplant fumigation to reduce resident pathogen populations in soil. Preplant fumigation with methyl iodide injected as a hot gas at 448.4 kg/ha through drip irrigation tape in preformed, tarped beds consistently provided significant (P < 0.05) reductions in the percentage of roots infected compared with the nonfumigated controls; these reductions were equal to or better than those achieved with an equivalent rate (448.4 kg/ha) of methyl bromide. Chloropicrin applied in water at 249.0 kg/ha through buried drip irrigation tape to either tarped or nontarped beds significantly (P < 0.05) reduced the percentages of both roots infected and roots on which perithecia were produced compared with nonfumigated controls.

Effect of Sclerotium Density and Irrigation on Disease Incidence and on Efficacy of Coniothyrium minitans in Suppressing Lettuce Drop Caused by Sclerotinia sclerotiorum

Field experiments were conducted over 2 years in Yuma, AZ, and Holtville, CA, to establish the relationship between soil sclerotium density of Sclerotinia sclerotiorum and the incidence of lettuce drop on different lettuce (Lactuca sativa) types under different irrigation systems, and to determine the efficacy of the biocontrol agent Coniothyrium minitans (Contans) against S. sclerotiorum on crisphead lettuce at varied sclerotium densities under different irrigation systems. There was no significant interaction of irrigation (overhead sprinkler versus furrow) with either sclerotium density or with biocontrol treatment. Lettuce drop incidence was lowest in romaine lettuce compared with crisphead or leaf lettuce at all soil sclerotium densities. There was a significant positive correlation between the sclerotial density and the percent disease incidence. Disease incidence in plots infested with 2 sclerotia/m2 of bed was not significantly higher than in control plots regardless of lettuce type. However, plots infested with 40 or 100 sclerotia/m2 of bed revealed a significantly higher disease incidence over the control in all lettuce types. A single application of Contans at planting significantly reduced the incidence of lettuce drop in all lettuce types even under high disease pressure. There were no significant differences between recommended (2.2 kg/ha) and high (4.4 kg/ha) application rates of Contans or between one or two applications of the product.

Characterization and Epidemiology of Outbreaks of Impatiens necrotic spot virus on Lettuce in Coastal California

California is the leading producer of lettuce (Lactuca sativa) for the United States and grows 77% of the countrys supply. Prior to 2006, coastal California lettuce was only periodically and incidentally infected by a single tospoviruses species: Tomato spotted wilt virus (TSWV). However, beginning in 2006 and continuing through 2012, severe outbreaks of disease caused by Impatiens necrotic spot virus (INSV) have affected the coastal lettuce crop, though TSWV was also present. In contrast, TSWV was the only tospovirus associated with disease outbreaks in Central Valley lettuce during this period. Disease surveys conducted over two seasons (2008 and 2009) in 10 commercial fields (acreage of 6 to 20 ha) indicated that INSV was the only tospovirus associated with economically damaging disease outbreaks in lettuce in the coastal region, with incidences of 0.5 to 27% (mean = 5.7%). Molecular characterization of INSV isolates associated with these disease outbreaks revealed little genetic diversity and indicated that lettuce-infecting INSV isolates were nearly identical to those previously characterized from ornamental or other hosts from different locations in the United States and the world. Monitoring of thrips revealed moderate to large populations in all surveyed lettuce fields, and the majority of thrips identified from these fields were western flower thrips, Frankliniella occidentalis. There was significant positive correlation (r2 = 0.91, P = 0.003) between thrips populations and INSV incidence in the most commonly encountered type of commercial lettuce (romaine, direct seeded, conventional) included in this study. A reverse-transcription polymerase chain reaction assay developed for detection of INSV in thrips showed promise as a monitoring tool in the field. Surveys for INSV reservoir hosts in the coastal production area revealed that the weeds little mallow (Malva parvifolia) and shepherds purse (Capsella bursa-pastoris) were commonly infected. M. parvifolia plants infected in the field did not show obvious symptoms, whereas plants of this species inoculated in the laboratory with INSV by sap transmission developed necrotic spots and chlorosis. Eleven other weed species growing in the lettuce production areas were found to be hosts of INSV. Coastal crops found to be infected with INSV included basil (Ocimum basilicum), bell pepper (Capsicum annuum), calla lily (Zantedeschia aethiopica), faba bean (Vicia faba), radicchio (Cichorium intybus), and spinach (Spinacia oleracea). Thus, it is likely that INSV was introduced into coastal California lettuce fields via viruliferous thrips that initially acquired the virus from other local susceptible plant species. Results of this study provide a better understanding of INSV epidemiology in coastal California and may help growers devise appropriate disease management strategies.

Evaluation of Cotton Germ Plasm for Resistance to the Whitefly and Cotton Leaf Crumple (CLCr) Disease and Etiology of CLCr in California's Imperial Valley

Cotton (Gossypium hirsutum) entries were evaluated for resistance to the whitefly (Bemisia tabaci biotype B) and cotton leaf crumple (CLCr) disease during the 1999 to 2001 growing seasons in the Imperial Valley of California. Entries were evaluated for densities of whitefly adults and nymphs, and for CLCr, by visual rating and squash/dot blot hybridization analyses. Differences in whitefly densities were detected among entries, but none were highly resistant, nor was there any correlation with CLCr disease severity. Entries AP 4103 and AP 6101 had relatively low whitefly densities and were highly susceptible (high CLCr disease severity ratings and viral titers), whereas NK 2387C and DPX 1883 also had low whitefly densities but were highly resistant (no symptoms or detectable viral titers). Other entries showed moderate CLCr resistance, which was independent of whitefly density. Geminivirus DNA-A and DNA-B components were consistently detected in cotton leaves with CLCr symptoms by polymerase chain reaction (PCR) with degenerate begomovirus primers, and full-length DNA-A and DNA-B clones were obtained. Cotton seedlings inoculated with these cloned DNAs by particle bombardment developed CLCr symptoms, and progeny virus was whitefly-transmissible. Sequence analysis revealed that these clones comprised the genome of a California isolate of the bipartite begomovirus Cotton leaf crumple virus (CLCrV-CA). Thus, CLCr disease in the Imperial Valley is caused by CLCrV-CA, and cotton entries with high levels of resistance were identified.

Onion Growth, Yield, and Production Costs as Affected by Irrigation System

Onion (Allium cepa) production in California’s San Joaquin Valley (SJV) typically involves intensive tillage and sprinklers for crop establishment followed by drip irrigation (DR). Studies were conducted at Five Points, California, in 2011 and 2013 to compare minimum tillage (MT) practices under overhead irrigation (OH) and DR relative to crop growth, yield, and costs. The experimental design was a randomized complete block with four replications of each irrigation treatment. Crop growth in both systems was similar; however, there was a year X irrigation system interaction for crop yields. In 2011, yields were not affected by irrigation, but in 2013, irrigation type affected yield with the OH system having 15.4 t/ha higher production than the DR system (78.6 t/ha vs. 63.4 t/ha). Onion size was not affected by irrigation system. Cost estimates indicated that onion production could be

Crop residue destruction strategies that enhance rather than inhibit reproduction of Monosporascus cannonballus

564 per hectare lower with the OH system compared with the DR system, if yields were maintained. Because of cost savings from OH, in both years the OH system was more profitable than DR. The study showed that onions could be successfully grown with MT using the OH system in the SJV and similar amounts of water.