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Watermelon is an important cucurbit vegetable crop grown in most of the United States. Phytophthora fruit rot of watermelon caused by Phytophthora capsici has been a major factor, limiting production for the past 15 years in the southeastern United States. The U.S. Department of Agriculture, Agricultural Research Service released five Phytophthora fruit rot-resistant germplasm lines for use in breeding programs. These lines were developed by phenotyping using a local isolate of P. capsici from South Carolina. The present study was undertaken to determine if these resistant lines had broad resistance to diverse P. capsici isolates collected from different states and crops. Five resistant germplasm lines (USVL020-PFR, USVL203-PFR, USVL782-PFR, USVL489-PFR, and USVL531-MDR) and two susceptible cultivars, Sugar Baby and Mickey Lee, used as checks were grown in a field in 2014 and 2015 to produce fruit for evaluation. Mature fruit were harvested and placed in a walk-in growth chamber and inoculated with 20 different P. capsici isolates. The chamber was maintained at 26 ± 2°C and high relative humidity (>95%) using a humidifier. All five resistant germplasm lines were significantly more resistant than the two susceptible checks to all 20 P. capsici isolates. Among the five resistant germplasm lines, USVL020-PFR, USVL782-PFR, and USVL531-MDR had broad resistance. Some P. capsici isolates induced minor lesions and rot on USVL489-PFR compared with the other resistant lines. Variation in virulence and genetic diversity among the 20 P. capsici isolates was also observed. The five watermelon germplasm lines will be useful for developing commercial watermelon cultivars with broad resistance to P. capsici.

Phytophthora capsici is an oomycete pathogen causing economically important diseases in a wide range of hosts worldwide including cucurbitaceous, solanaceous, and fabaceous crops. All plant parts, crown and roots, or only the fruit may be affected depending on the host, and symptoms can range from wilting to rot and plant death. Considered a hemibiotroph, P. capsici can be cultured in artificial media and maintained in long-term storage. In this diagnostic guide, we describe methods to identify P. capsici infection based on disease symptoms and pathogen signs. We also outline methods for molecular identification, pathogen isolation, storage of single-sporangium cultures, and pathogenicity testing.

Powdery mildew, caused by the fungus Podosphaera xanthii, is one of the most important diseases of melon. Although there are several pathogenic races of P. xanthii, race 1 is the predominant race in South Carolina and in other parts of the United States. We used a densely genotyped recombinant inbred line melon population for traditional quantitative trait loci (QTL) mapping, to identify two major (qPx1-5 and qPx1-12) and two minor (qPx1-4 and qPx1-10) QTLs (named according to race – chromosome number) associated with resistance to P. xanthii race 1. QTL mapping of disease severity in multiple tissues (hypocotyl, cotyledons, true leaves, and stems) identified the same genetic basis of resistance in all tissue types. Whole-genome resequencing of the parents was used for marker development across the major QTLs and functional annotation of single nucleotide polymorphisms (SNPs) for candidate gene analysis. Kompetitive allele-specific PCR (KASP) markers were tightly linked to the QTL peaks of qPx1-5 (pm1-5_25329892, pm1-5_25461503 and pm1-5_25625375) and qPx1-12 (pm1-12_22848920 and pm1-12_22904659) in the population and will enable efficient marker-assisted introgression of powdery mildew resistance into improved germplasm. Candidate genes were identified in both major QTL intervals that encode putative R genes with missense mutations between the parents. The candidate genes provide targets for future breeding efforts and a fundamental examination of resistance to powdery mildew in melon.

Zinc (Zn) accumulation and deficiency affect plant response to pests and diseases differently in varying pathosystems. The concentrations of Zn in plants aid in priming defense signaling pathways and help in enhanced structural defenses against plant pathogens. Studies are lacking on how concentrations of Zn in watermelon plants influence defense against two important soil-borne pathogens: Fusarium oxysporum f. sp. niveum (FON) and southern root-knot nematode (RKN, Meloidogyne incognita). In this study a comparative transcriptomics evaluation of watermelon plants in response to high (1.2 ppm) and low (0.2 ppm) levels of Zn were determined. Differential transcript-level responses differed in watermelon plants when infected with FON or RKN or both under high- and low-Zn treatment regimes in a controlled hydroponics system. Higher numbers of differentially expressed genes (DEGs) were observed in high-Zn-treated than in low-Zn-treated non-inoculated plants, in plants inoculated with FON alone and in plants inoculated with RKN alone. However, in the co-inoculated system, low-Zn treatment had higher DEGs as compared to high-Zn treatment. In addition, most DEGs were significantly enriched in hormone signal transduction and MAPK signaling pathway, suggesting an induction of systemic resistance with high-Zn concentrations. Taken together, this study substantially expands transcriptome data resources and suggests a molecular potential framework for watermelon-Zn interaction in FON and RKN.

Squash vein yellowing virus (SqVYV) causes viral watermelon vine decline. To facilitate detection of SqVYV, enzyme linked-immunosorbent assay (ELISA) and quantitative reverse-transcription polymerase chain reaction (qRT-PCR) diagnostic methods were developed. Both methods were capable of detecting SqVYV in a wide range of cucurbit hosts. ELISA was able to detect virus in infected host tissue diluted to at least 1:2,560, which was sufficient for detection in symptomatic squash and watermelon plants. The qRT-PCR method was capable of reliably detecting as few as 3.4 copies of a cloned fragment of SqVYV genomic RNA with an average cycle threshold (Ct) value of 36.4. The sensitivities and specificities for each detection method were estimated by latent class analysis for a set of inoculated squash and watermelon plants at two sampling scales. The scales were hierarchical, with individual plants representing the upper scale and samples from the plant representing the lower scale. The number of samples per plant varied from 1 to 8, and a plant was diagnosed positive if any of its samples tested positive. For all analyses, a cutoff Ct of 35 was chosen for qRT-PCR, which is approximately 2.5 cycles lower than the lowest Ct value achieved for mock-inoculated plants (presumed to be a false positive). qRT-PCR showed high sensitivities (≥0.99) at both sampling scales for squash and watermelon, whereas the sensitivities for ELISA ranged from 0.58 to 0.76. The specificities for both tests were very similar (≥0.94), with ELISA sometimes outperforming qRT-PCR. These diagnostic methods provide additional tools for the identification of SqVYV and management of SqVYV-induced watermelon vine decline.

Watermelon is an important cucurbit crop grown in 44 states in the United States of America (USA). South Carolina (SC) had the seventh largest watermelon production area in USA with 4,500 acres in 2018 valued at approximately $17 million. In June 2019, four watermelon plants displaying symptoms of virus infection including stunting, leaf crumpling, interveinal chlorosis, and necrosis of leaf margins were observed in a research trial in Charleston, SC. Whiteflies were observed on the abaxial surface of the leaves. One plant was tested for the presence of whitefly-transmitted cucurbit leaf crumple virus (CuLCrV), cucurbit yellow stunting disorder virus (CYSDV), and squash vein yellowing virus and aphid-transmitted papaya ringspot virus by RT-PCR. Amplicons of the expected size were only observed for CuLCrV (∼1 kb) and CYSDV (∼0.7 kb). Because CYSDV was not previously reported from SC, we continued to monitor the field for virus symptoms during the summer season. By mid-August 2019, 15 additional plants (1.9% incidence) were observed with similar symptoms of virus infection. Two virus identification methods (CP and Hsp70h) confirmed that CYSDV was indeed present in watermelon samples in SC. The geographic range of CYSDV and other whitefly-transmitted viruses continues to expand into and within major cucurbit-producing regions with each production season, especially in the southeastern USA. This is the first report of CYSDV infecting watermelon or any other plant in SC. These three whitefly-transmitted viruses now need to be considered in management plans by SC growers. Knowledge of the presence of CYSDV, an additional virus, infecting cucurbits, in SC is also important from a regulatory perspective.

The effect of major resistance genes (Bs1, Bs2, and Bs3) or gene combinations for resistance to bacterial spot of bell peppers (Xanthomonas campestris pv. vesicatoria) in 15 commercial cultivars on disease reduction and yield were studied during 1995 and 1996. Reaction of cultivars to specific races (races 1, 2, or 3) of the pathogen corresponded with seed company claims for resistance against these races. Races 1 to 4 were used as initial inoculum in 1995, and races 1 to 6 in 1996 field experiments. Cultivars with no known resistance genes to bacterial spot (e.g., Camelot, Jupiter, and Valiant), a single resistance gene (X3R Camelot, King Arthur), or a combination of Bs1 and Bs3 genes (Guardian, Sentinel, and Admiral) were severely diseased. Yields were reduced in all inoculated cultivars compared to non-inoculated cultivars used as controls. Although races 4 and 6 caused significant disease in cultivars with only Bs1 (King Arthur) or Bs2 (X3R Camelot) genes, cultivars with a combination of Bs1 and Bs2 (Boynton Bell, PR9300-8) had much lower levels of bacterial spot. Roger 4178, a hybrid with a combination of Bs1, Bs2, and Bs3 genes, had the lowest disease ratings. Overall, race 3 was predominant during 1995, while races 3 and 6 were recovered most frequently in 1996.

Bell pepper plants sprayed with the chemical acibenzolar-S-methyl (ABM, Actigard 50 WG) showed resistance to subsequent infections with the bacterial spot agent Xanthomonas axonopodis pv. vesicatoria. Induction of resistance was independent of the cultivar used, and was expressed as early as 3 days after treatment and continued for at least 2 weeks. In the field, applications of ABM every 2 weeks, alone or in combination with copper, resulted in disease control similar to the standard treatment of copper plus maneb. Yield response was variable, with certain combinations of chemical treatments and cultivars producing yields as large as the copper plus maneb treatment. In contrast, weekly applications during the entire crop season had a negative impact on yield. In plots maintained free of bacterial spot, applications of ABM every 2 weeks caused a reduction in yield for one cultivar of six tested. The use of chemical inducers for the control of bacterial spot on bell pepper, while generally promising, may result in an unpredictable loss in fruit yield.

Summary Years of selection for desirable fruit quality traits in dessert watermelon (Citrullus lanatus) has resulted in a narrow genetic base in modern cultivars. Development of novel genomic and genetic resources offers great potential to expand genetic diversity and improve important traits in watermelon. Here, we report a high‐quality genome sequence of watermelon cultivar ‘Charleston Gray’, a principal American dessert watermelon, to complement the existing reference genome from ‘97103’, an East Asian cultivar. Comparative analyses between genomes of ‘Charleston Gray’ and ‘97103’ revealed genomic variants that may underlie phenotypic differences between the two cultivars. We then genotyped 1365 watermelon plant introduction (PI) lines maintained at the U.S. National Plant Germplasm System using genotyping‐by‐sequencing (GBS). These PI lines were collected throughout the world and belong to three Citrullus species, C. lanatus, C. mucosospermus and C. amarus. Approximately 25 000 high‐quality single nucleotide polymorphisms (SNPs) were derived from the GBS data using the ‘Charleston Gray’ genome as the reference. Population genomic analyses using these SNPs discovered a close relationship between C. lanatus and C. mucosospermus and identified four major groups in these two species correlated to their geographic locations. Citrullus amarus was found to have a distinct genetic makeup compared to C. lanatus and C. mucosospermus. The SNPs also enabled identification of genomic regions associated with important fruit quality and disease resistance traits through genome‐wide association studies. The high‐quality ‘Charleston Gray’ genome and the genotyping data of this large collection of watermelon accessions provide valuable resources for facilitating watermelon research, breeding and improvement.

Bottle gourd (Lagenaria siceraria (Mol.) Standl.) is an important rootstock in watermelon production in several countries such as Japan, China, and Israel where 60 to 70% of watermelons are grafted (2). We are evaluating bottle gourds for their ability to improve disease resistance when used as rootstock for watermelon (3). In the summer of 2007, symptoms of wilting and crown necrosis appeared on bottle gourd seedlings 1 month after transplanting in a field in Charleston, SC. Infection was observed on commercial cv. Emphasis and four advanced breeding lines. In October of 2007, 35 of 85 plants examined (41%) had stem rot at the crown area just above the soil line where coarse, white mycelia and abundant sclerotia were observed. The fungus tentatively identified as Sclerotium rolfsii produced sclerotia that were white or light to dark brown and measured 0.6 to 2.5 mm in diameter (mean = 1.1 mm). Diseased tissues with sclerotia from four plants were disinfested for 1 min in 0.5% sodium hypochlorite and plated on acidified potato dextrose agar (APDA). Fungal colonies that produced white mycelia and tan-to-brown sclerotia were isolated from four wilted plants. A single PCR product of approximately 680 bp was amplified from DNA extracted from two isolates using the primers ITS1 and ITS4 (4). One PCR product was cloned into the TOPO TA cloning vector (Invitrogen, Carlsbad, CA) and sequenced (GenBank Accession No. EU338381). BLASTN analysis of the sequence in the NCBI databases revealed 99% similarity to the internal transcribed spacer (ITS) sequences of S. rolfsii and Athelia rolfsii (perfect stage of S. rolfsii), confirming that the pathogen was indeed S. rolfsii. Two S. rolfsii isolates were used to test pathogenicity. Each isolate was used to inoculate five young seedlings and five adult (10-week-old) bottle gourd plants. For inoculation, 10 sclerotia obtained from the APDA plates were placed on the surface of the potting soil 0.5 to 1 cm from the collar region of each bottle gourd plant growing in 10-cm pots. Inoculations were done carefully to ensure that the plants were not injured. After inoculation, the plants were maintained at high humidity and 25°C for 3 days and then transferred to laboratory benches. Four young seedlings and three adult noninoculated plants kept under the same conditions served as controls. The pathogenicity test was repeated once with similar results. All inoculated plants developed symptoms of southern blight. The inoculated plants developed symptoms of wilting 4 to 5 days after inoculation and completely wilted within 7 to 10 days. Symptoms of wilting were soon followed by the appearance of white-to-light brown sclerotia on the collar region. No symptoms were observed on the noninoculated plants. S. rolfsii was reisolated from the inoculated plants on APDA. Although southern blight caused by S. rolfsii has been reported on many crop plants in the southern United States, to our knowledge, this disease has not been reported previously on bottle gourd in North America. However, the disease has been reported on bottle gourd in India (1). Identifying sources of resistance to southern blight in bottle gourds may be necessary to make them suitable as rootstocks in areas where S. rolfsii is present. References: (1) K. S. Amin. Indian Phytopathol. 34:253, 1981. (2) R. Cohen et al. Plant Dis. 91:916, 2007. (3) K. S. Ling and A. Levi. HortScience 42:1124, 2007. (4) T. J. White et al. PCR Protocols: A Guide to Methods and Amplifications. Academic Press, San Diego, 1990.

Squash vein yellowing virus (SqVYV) is the cause of viral watermelon vine decline. The virus is whitefly-transmitted, induces a systemic wilt of watermelon plants, and causes necrosis and discoloration of the fruit rind. In the field, SqVYV is often detected in watermelon in mixed infections with other viruses including the aphid-transmitted Papaya ringspot virus type W (PRSV-W). In this study, watermelon plants of different ages were inoculated with SqVYV or SqVYV+PRSV-W in the greenhouse or SqVYV in the field to characterize the physiological response to infection. Symptoms of vine decline appeared about 12 to 16 days after inoculation with SqVYV regardless of plant age at time of inoculation, plant growth habit (trellised or nontrellised), and location (greenhouse or field). However, the presence of PRSV-W delayed the appearance of vine decline symptoms by 2 to 4 days, and vine decline did not develop on plants with no fruit. For all inoculation treatments, more severe symptoms were observed in younger watermelon plants. Physiological responses to SqVYV infection included reduction in plant and fruit weights, alterations in fruit rind and flesh color, reduction in fruit sucrose content, increase in fruit acid content, and changes in plant nutrient composition, particularly increases in Ca, Mg, B, Mn, and Zn and decreases in K and N. These results demonstrate wide-ranging physiological effects of SqVYV infection and provide new insights into watermelon vine decline.

The responses of a diverse group of vining cucurbits to inoculation with Squash vein yellowing virus (SqVYV) were determined. For the first time, Cucurbita maxima, Cucumis dipsaceus, and Cucumis metuliferus were observed to develop necrosis and plant death similar to the SqVYV-induced vine decline in watermelon (Citrullus lanatus var. lanatus). The majority of cucurbits inoculated, however, either exhibited no symptoms of infection, or developed relatively mild symptoms such as vein yellowing of upper, noninoculated leaves. All inoculated plants were sectioned and tested for the presence of SqVYV. The virus was widely distributed in mature, fruit-bearing cucurbits with over 72% of plant sections testing positive for SqVYV by tissue-blot and/or reverse transcription-polymerase chain reaction. Plants of several cucurbits, including a wild citron (Citrullus lanatus var. citroides), were symptomless and had a decreased frequency of virus infection of vine segments compared to susceptible vining cucurbits, indicating a higher level of resistance. However, no significant relationship between the frequency of infection or virus distribution within plants and the symptom response was observed. These results demonstrate that a diverse group of cucurbits may decline when infected with SqVYV, and suggest that widespread distribution of virus within the plant is not the sole cause of decline.

Watermelon is an important crop grown in 44 states in the United States. Phytophthora fruit rot caused by Phytophthora capsici is a serious disease in the southeastern U.S.A., where over 50% of the watermelons are produced. The disease has resulted in severe losses to watermelon growers, especially in Georgia, South Carolina, and North Carolina during the past few years. Several fruit rot-resistant watermelon germplasm lines have been developed for use in breeding programs. To evaluate the development of Phytophthora fruit rot on fruit of different ages, plants of fruit rot-resistant and susceptible lines were planted at weekly intervals for five consecutive weeks in experiments conducted over three years (2011 to 2013). Flowers were routinely inspected and hand pollinated to ensure having fruit of different ages. In each year, different aged fruit were harvested on the same day and inoculated with a 5-mm agar plug from an actively growing colony of P. capsici. Inoculated fruit were maintained in a room set to conditions conducive for disease development (>95% relative humidity, 26 ± 2°C). After 5 days, lesion diameter and intensity of sporulation was recorded for each fruit. Lesion diameter and sporulation intensity were significantly greater on fruit of susceptible lines compared with resistant lines. Fruit age did not have an effect on either measurement on susceptible (Sugar Baby) or resistant lines (PI 560020 and PI 595203). Our results showed that resistance to Phytophthora fruit rot in watermelon was not correlated with fruit age.

Since the 1950s, research on the animal neurohormone, melatonin, has focused on its multiregulatory effect on patients suffering from insomnia, cancer, and Alzheimer's disease. In plants, melatonin plays major role in plant growth and development, and is inducible in response to diverse biotic and abiotic stresses. However, studies on the direct role of melatonin in disease suppression and as a signaling molecule in host-pathogen defense mechanism are lacking. This study provides insight on the predicted biosynthetic pathway of melatonin in watermelon (Citrullus lanatus), and how application of melatonin, an environmental-friendly immune inducer, can boost plant immunity and suppress pathogen growth where fungicide resistance and lack of genetic resistance are major problems. We evaluated the effect of spray-applied melatonin and also transformed watermelon plants with the melatonin biosynthetic gene SNAT (serotonin N-acetyltransferase) to determine the role of melatonin in plant defense. Increased melatonin levels in plants were found to boost resistance against the foliar pathogen Podosphaera xanthii (powdery mildew), and the soil-borne oomycete Phytophthora capsici in watermelon and other cucurbits. Further, transcriptomic data on melatonin-sprayed (1 mmol/L) watermelon leaves suggest that melatonin alters the expression of genes involved in both PAMP-mediated (pathogen-associated molecular pattern) and ETI-mediated (effector-triggered immunity) defenses. Twenty-seven upregulated genes were associated with constitutive defense as well as initial priming of the melatonin-induced plant resistance response. Our results indicate that developing strategies to increase melatonin levels in specialty crops such as watermelon can lead to resistance against diverse filamentous pathogens.

Watermelon vine decline (WVD) caused by the whitefly-transmitted Squash vein yellowing virus (SqVYV) has been a serious limiting factor in watermelon production in southwest and west-central Florida over the past few years. Symptoms of WVD typically appear as sudden decline of vines a few weeks before harvest or just after the first harvest. Fruit symptoms include rind necrosis and flesh discoloration that affects fruit quality and marketability. The combination of insecticide treatments consisting of an imidacloprid drench (Admire Pro, 560 ml/ha) at transplanting followed by two foliar applications of spiromesifen (Oberon, 2SC, 490 ml/ha) and reflective plastic mulch was evaluated for management of WVD during fall growing seasons of 2006, 2007, and 2009. Virus inoculum source was introduced by planting SqVYV-infected squash plants at the ends of each plot. In all three experiments, the insecticide-treated plots had significantly lower levels of WVD on foliage and fruit compared to non-treated plots. In 2007, the reflective plastic mulch was effective in reducing foliar WVD compared to non-reflective mulch, but not in 2006 and 2009. No significant interaction between plastic mulch and chemical treatments was observed on WVD development on foliage or fruit. Our results suggest that application of insecticides for managing whiteflies can help manage SqVYV-caused WVD. Accepted for publication 13 January 2015. Published 25 March 2015.

Virus-like symptoms of curled and crumpled leaves, bright yellow chlorosis, and marginal leaf necrosis typical of begomovirus infection were observed on watermelon, summer squash, and muskmelon in three counties in South Carolina in August 2017. Genus-specific primers for begomovirus-A and begomovirus-B components produced amplicons of the expected sizes. Sanger sequencing of amplicons from both A and B components and BLASTn analysis revealed a high nucleotide sequence identity in seven watermelon samples from Beaufort and Charleston counties and five muskmelon samples from Charleston County with cucurbit leaf crumple virus.

Resistance to boscalid, one of the older succinate-dehydrogenase inhibitors (SHDI) in Fungicide Resistance Action Committee (FRAC) code 7, was detected in Podosphaera xanthii, the cucurbit powdery mildew fungus, in South Carolina in July 2017. Resistance to the field rate (682 ppm) of boscalid was confirmed in greenhouse experiments and laboratory bioassays conducted on summer squash plants and cotyledons, respectively, that had been treated with a range of boscalid concentrations. This report is the first documentation of resistance to boscalid in P. xanthii in the southern United States.

Crown rot caused by Phytophthora capsici is becoming an increasingly important disease of vegetable crops in southeastern U.S. In recent years, the practice of grafting watermelon onto rootstocks belonging to other Cucurbitaceae genera has also been slowly gaining adoption in the U.S. However, it is not known how these rootstocks, developed mainly in Asia, will respond to diseases prevalent in local production areas. We evaluated the available commercial watermelon rootstocks for resistance to Phytophthora crown rot by inoculating them with a zoospore suspension of P. capsici in four different trials. Disease development on rootstocks was rated on a 1–9 scale (1 = no symptoms, 9 = plant dead). Based on all the four trials, the commercial Lagenaria siceraria (bottle gourd) rootstocks, FR-Strong, Emphasis, Macis and WMXP-3938 were resistant (Mean rating ≤3) to crown rot when compared to susceptible watermelon checks (mean rating >8). All Cucurbita inter-specific hybrid rootstocks and a watermelon rootstock Ojakkyo, were highly susceptible to crown rot (mean rating >8). Real-time quantitative polymerase chain reaction (qPCR) using two different P. capsici specific primers (ITS and β-tubulin) indicated the presence of significantly ( P P. capsici DNA g −1 plant tissue in susceptible Cucurbita inter-specific hybrid rootstocks (β-tubulin, mean = 2895 ng) and watermelon (2665 ng) compared to the L. siceraria rootstocks (357 ng). Crown rot resistant bottle gourd rootstocks may be useful in areas where P. capsici is a recurring problem. The present study identified several commercial bottle gourd rootstocks with resistance to Phytophthora crown rot, and confirmed their levels of resistance using qPCR.

A variety of fresh market vegetables, including watermelon and tomato are economically important crops in Florida. Whitefly-transmitted Squash vein yellowing virus (SqVYV) was first identified in squash and watermelon in Florida in 2005 and shown to cause a severe decline of watermelon vines as crops approach harvest. Florida is most economically impacted by SqVYV, although the virus has been detected more recently in Indiana and South Carolina. The origin and evolutionary history of SqVYV, one of the few members of the genus Ipomovirus within the family Potyviridae, are not known. Sequence diversity of SqVYV isolates collected at different times, from different locations and from different plant species is being analyzed for insights into the origin of the virus. More recently, Cucurbit leaf crumple virus (CuLCrV) and Cucurbit yellow stunting disorder virus (CYSDV), also whitefly-transmitted, have been detected in watermelon in Florida. Tomato yellow leaf curl virus (TYLCV) was first detected in south Florida tomato crops in 1997. Several surveys have been conducted in the region to identify alternative hosts for these four viruses. Cucurbit weeds including Balsam-apple (Momordica charantia), creeping cucumber (Melothria pendula) and smellmelon (Cucumis melo var. dudaim) provide reservoirs for SqVYV, CuLCrV and/or CYSDV. Green bean (Phaseolus vulgaris) also can be a reservoir for CuLCrV. No wild hosts of TYLCV have been reported in Florida. The effectiveness of insecticides and silver plastic mulch to manage whiteflies and mitigate TYLCV has been demonstrated and is currently being evaluated for SqVYV, CuLCrV and CYSDV. In addition, potential sources of SqVYV resistance have been identified in greenhouse and field screening of watermelon germplasm. Further studies to refine these sources of resistance are underway. Lastly, a comprehensive map of 33,560 hectares (82,928 acres) of vegetable fields in the three counties comprising the majority of the southwest Florida vegetable production area has been developed to identify 'hot spots' and reservoir crops for viruses and whiteflies, and will be useful in evaluation of management strategies to decrease virus incidence in commercial fields.

Whitefly-transmitted Squash vein yellowing virus (SqVYV) and Cucurbit leaf crumple virus (CuLCrV) and aphid-transmitted Papaya ringspot virus type W (PRSV-W) have had serious impact on watermelon production in southwest and west-central Florida in the past 5 years. Tissue-blot nucleic acid hybridization assays were developed for simple, high-throughput detection of these three viruses as well as Cucurbit yellow stunting disorder virus (CYSDV), which was first reported in Florida in 2008. To determine virus distribution within plants, we collected 80 entire plants just before or during the harvest period in a systematic sample, 20 each on 11 April, 18 April, 26 April, and 3 May 2007, from a fruiting commercial watermelon field near Immokalee, FL showing symptoms of infection by SqVYV, CuLCrV, and PRSV-W and, possibly, CYSDV. This was followed by a sampling of five plants collected at harvest showing symptoms of virus infection on 11 October 2007 in a different commercial planting located in Duette, FL. Tissue prints were made from cross sections of watermelon plants from the crowns through the tips at 0.6-m intervals on nylon membranes and nucleic acid hybridization assays were used for virus detection. Results from testing crown tissue showed that SqVYV, CuLCrV, and PRSV-W were present in ≈37, 44, and 54%, respectively, of the 80 plants collected over the four sampling dates from the first field. For individual vines diagnosed with SqVYV, the distribution of SqVYV in vine tissue decreased proportionately with distance from the crown. The probability of detecting SqVYV was 70% at the base of the vine compared with 23% at the tip of the vine. In contrast, CuLCrV tended to be more evenly distributed throughout the plant, with ≈10% higher probability of detection at the growing tip relative to the crown of the plant. The distribution of PRSV-W resembled that of SqVYV but with ≈20% higher probability of detection at the tip of the vine. Similar trends were detected in the smaller sampling; however, CYSDV was also detected in three of the plants. Overall, the results indicated that SqVYV and PRSV-W were distributed differently than CuLCrV in watermelon plants, and this difference has implications on how samples should be collected and may affect vector acquisition and transmission of these viruses.

Watermelon [Citrullus lanatus v. lanatus (Thunb.) Matsum & Nakai] seed exudates are inhibitory to germination and seedling growth of other plant species. A miniature bioassay experiment that measured proso millet (Panicum miliaceum L.) radicle growth was used to assess the inhibition caused by seed exudates of 125 genotypes of watermelon and related Citrullus species. Exudates of most genotypes were not inhibitory; however, exudates of 53 accessions reduced radicle growth in comparison with the control. In subsequent proso millet radicle growth experiments, genotypes were found to vary in inhibitory potential, and concentration response curves generated using filtered, pasteurized exudates were different among genotypes. Filter-sterilized seed exudates of Citrullus accessions also varied in the level of inhibition in a bioassay that measured their effect on sporangia formation by the watermelon pathogen, Phytophthora capsici. These observations suggest that constituents in Citrullus seed exudates affect organisms in the spermosphere and that the inhibitory potential of seed exudates varies among genotypes. Differences in concentration response curves in the millet bioassay and differences in the relative inhibition of genotypes in the millet and fungus bioassays indicate that the inhibitory constituents in seed exudates vary among genotypes.

Powdery mildew (PM) disease causes significant loss in watermelon. Due to the unavailability of a commercial watermelon variety that is resistant to PM, grafting susceptible cultivars on wild resistant rootstocks is being explored as a short-term management strategy to combat this disease. Nuclear magnetic resonance-based metabolic profiles of susceptible and resistant rootstocks of watermelon and their corresponding susceptible scions (Mickey Lee) were compared to screen for potential metabolites related to PM resistance using multivariate principal component analysis. Significant score plot differences between the susceptible and resistant groups were revealed through Mahalanobis distance analysis. Significantly different spectral buckets and their corresponding metabolites (including choline, fumarate, 5-hydroxyindole-3-acetate, and melatonin) have been identified quantitatively using multivariate loading plots and verified by volcano plot analyses. The data suggest that these metabolites were translocated from the powdery mildew resistant rootstocks to their corresponding powdery mildew susceptible scions and can be related to PM disease resistance.

The genus Ipomovirus is a small group of whitefly-transmitted viruses within the family Potyviridae, the largest group of RNA plant viruses, which are mostly aphid transmitted (Berger et al. 2005). Under current taxonomic guidelines there are three accepted members [Cucumber vein yellowing virus (CVYV), Cassava brown streak virus (CBSV) and Sweet potato mild mottle virus (SPMMV)] and one tentative member [Sweet potato yellow dwarf virus (SPYDV)] in the genus Ipomovirus (Berger et al. 2005; Colinet et al. 1996, 1998; Janssen et al. 2005; Lecoq et al. 2000).

When bacterial spot-resistant pepper plants carrying resistance gene Bs2 and infiltrated with incompatible strains of Xanthomonas axonopodis pv. vesicatoria carrying a functional avrBs2 gene (races P1 and P3) were incubated at 32 degrees C, they exhibited an electrolyte leakage and bacterial multiplication pattern in planta similar to that obtained with a compatible strain (race P4) carrying a nonfunctional avrBs2 gene. They also developed disease-like symptoms. Pretreatment of incompatible bacteria at 32 degrees C before infiltration caused a delay in electrolyte leakage less pronounced than that caused by exposing plants to 32 degrees C. Also, plants had to be exposed to 32 degrees C for an hour prior to inoculation to increase symptom expression. These data suggest that the Bs2 gene is temperature sensitive. In other experiments, the avrBs1-Bs1 interaction appeared to be the most heat tolerant and thus the least likely to revert to compatible, whereas the avrBs3-Bs3 interaction had an intermediate sensitivity to elevated temperatures.

The bacterial plant pathogen Xanthomonas axonopodis pv. vesicatoria, also known as Xanthomonas campestris pv. vesicatoria group A, is the causal agent of bacterial spot in pepper and tomato. In order to test different models that may explain the coevolution of avrBs2 with its host plants, we sequenced avrBs2 and six chromosomal loci (total of 5.5 kb per strain) from a global sample of 55 X. axonopodis pv. vesicatoria strains collected from diseased peppers. We found an extreme lack of genetic variation among all X. axonopodis pv. vesicatoria genomic loci (average nucleotide diversity, π = 9.1 × 10 −5 ), including avrBs2 . This lack of diversity is consistent with X. axonopodis pv. vesicatoria having undergone a recent population bottleneck and/or selective sweep followed by population expansion. Coalescent analysis determined that approximately 1.4 × 10 4 to 7.16 × 10 4 bacterial generations have passed since the most recent common ancestor (MRCA) of the current X. axonopodis pv. vesicatoria population. Assuming a range of 50 to 500 bacterial generations per year, only 28 to 1,432 years have passed since the MRCA. This time frame coincides with human intervention with the pathogen's host plants, from domestication to modern agricultural practices. Examination of 19 mutated (loss-of-function) avrBs2 alleles detected nine classes of mutations. All mutations affected protein coding, while no synonymous changes were found. The nature of at least one of the avrBs2 mutations suggests that it may be possible to observe one stage of an evolutionary arms race as X. axonopodis pv. vesicatoria responds to selection pressure to alter avrBs2 to escape host plant resistance.

Pigweeds (genus Amaranthus) are problematic weeds in crop production throughout the world and are responsible for significant yield losses in many crops (2). Members of this genus can produce hundreds of thousands of seeds per plant and are also capable of supporting populations of important crop pathogens including viruses, nematodes, fungi, and oomycetes. Thirty-one pigweed samples (tentatively identified as Amaranthus lividus L. based on leaf notch and growth habit) were collected in November and December of 2009 from a watermelon field near Immokalee, FL, previously found to contain watermelon plants infected with three whitefly-transmitted viruses: Cucurbit yellow stunting disorder virus (CYSDV), Cucurbit leaf crumple virus (CuLCrV), and Squash vein yellowing virus (SqVYV). Although no obvious virus symptoms were observed on any of the pigweed plants, whiteflies (Bemisia tabaci), a known vector of CYSDV, CuLCrV, and SqVYV, were observed on leaves. Consequently, replica tissue blots were made from all pigweed samples and tested independently by tissue blot nucleic acid hybridization assay for CYSDV, CuLCrV, or SqVYV (3). Tissue blots indicated CYSDV infection in six pigweed samples. Neither CuLCrV nor SqVYV was detected. Three of the tissue blot-positive pigweed samples were further tested by reverse transcription (RT)-PCR amplification from total RNA (extracted from leaf tissue with TRIzol Reagent [Invitrogen, Carlsbad, CA]) with HSP70 and coat protein (CP) gene primers (1). HSP70 and CP gene RT-PCR products of the expected sizes (175 and 707 nt, respectively) were amplified, sequenced, and found to be 100% identical for all three pigweed samples. The partial HSP70 gene sequence from pigweed shared 98.3 to 100% nucleotide identity with CYSDV isolates from Arizona, California, and Spain (GenBank Accession Nos. FJ492808, EU596530, and NC_004810, respectively). The partial CP gene sequence from pigweed shared 88.8 to 100% nucleotide identity with CYSDV isolates from Arizona, Saudi Arabia, Texas, and Spain (GenBank Accession Nos. EF210558, AF312811, AF312806, and AF312808, respectively). To our knowledge, this is the first report of CYSDV infection of pigweed in Florida. Infection of redroot pigweed (A. retroflexus) was recently reported in California (4). These results collectively indicate that control of noncucurbit weeds may be important for effective management of CYSDV in cucurbit crops. References: (1) S. Adkins et al. Online publication. doi:10.1094/PHP-2009-1118-01-BR. Plant Health Progress, 2009. (2) L. Holm et al. World's Weeds: Natural Histories and Distributions. John Wiley and Sons, Inc. New York, NY, 1997. (3) W. W. Turechek et al. Phytopathology 100:1194, 2010. (4) W. M. Wintermantel et al. Plant Dis. 93:685, 2009.

Race shifts in Xanthomonas campestris pv. vesicatoria, (proposed: X. axonopodis pv. vesicatoria), the pepper bacterial spot pathogen, were studied in the field during 1993 and 1994. In 1993, Early Calwonder (ECW) pepper plants inoculated with rifampicin-resistant race 1 strain Xcv 33 rif , which carries avirulence gene avrBs3, were planted in a plot of ECW-30R plants, which carry the Bs3 gene for resistance to race 1 of the pathogen. Disease was observed on plants adjacent to the inoculum plants 2 weeks after inoculation, and within 10 weeks, all plants were diseased. Rifampicin-resistant single colonies isolated from diseased ECW-30R plants were screened for race reaction on pepper differentials, and all were race 3 because they were compatible on ECW-30R. Total DNA from 25% of the single colonies hybridized to an avrBs3 gene probe, and the plasmid carrying avrBs3 was not detected in the colonies that did not hybridize to avrBs3. Similarly, in 1994 the population derived from a strain of race 2 that carries avrBs1 rapidly shifted to race 3 when inoculated plants were placed in a plot of ECW-10R plants carrying the single resistance gene Bs1. Total DNA from all isolated single colonies with the race 3 phenotype hybridized with avrBs1 and insertion element IS476. Thus, a race 3 pathogen population of X. campestris pv. vesicatoria rapidly developed from race 1 and 2 populations within a growing season in the field when exposed to a resistant pepper cultivar carrying a single resistance gene. The race shift from race 1 to 3 was due to loss of the plasmid carrying the avirulence gene or to inactivation of avrBs3. Shift from race 2 to 3 apparently resulted from insertion element IS476 inactivating avirulence gene avrBs1.