Differential Life History Trait Associations of Aphids with Nonpersistent Viruses in Cucurbits.

The diversity of vectors and fleeting nature of virus acquisition and transmission renders nonpersistent viruses a challenge to manage. We assessed the importance of noncolonizing versus colonizing vectors with a 2-yr survey of aphids and nonpersistent viruses on commercial pumpkin farms. We quantified aphid alightment using pan traps, while testing leaf samples with multiplex RT-PCR targeting cucumber mosaic virus (CMV), zucchini yellow mosaic virus (ZYMV), watermelon mosaic virus (WMV), and papaya ringspot virus (PRSV). Overall, we identified 53 aphid species (3,899 individuals), from which the melon aphid, Aphis gossypii Glover, a pumpkin-colonizing species, predominated (76 and 37% of samples in 2010 and 2011, respectively). CMV and ZYMV were not detected, but WMV and PRSV were prevalent, both regionally (WMV: 28/29 fields, PRSV: 21/29 fields) and within fields (infection rates = 69 and 55% for WMV in 2010 and 2011; 28 and 25% for PRSV in 2010 and 2011). However, early-season samples showed extremely low infection levels, suggesting cucurbit viruses are not seed-transmitted and implicating aphid activity as a causal factor driving virus spread. Interestingly, neither noncolonizer and colonizer alightment nor total aphid alightment were good predictors of virus presence, but community analyses revealed species-specific relationships. For example, cowpea aphid (Aphis craccivora Koch) and spotted alfalfa aphid (Therioaphis trifolii Monell f. maculata) were associated with PRSV infection, whereas the oleander aphid (Aphis nerii Bover de Fonscolombe) was associated with WMV spread within fields. These outcomes highlight the need for tailored management plans targeting key vectors of nonpersistent viruses in agricultural systems.

Squash vein yellowing virus Identified in Watermelon (Citrullus lanatus) in Indiana.

During September 2006, moderate vine decline symptoms including vine collapse and wilt and root rot were observed on numerous watermelon plants growing in a commercial field in Sullivan County, Indiana. No symptoms were observed on the fruit. Six plants displaying typical vine decline symptoms were collected and assayed for potyvirus infection and subsequently for Squash vein yellowing virus (SqVYV) and Papaya ringspot virus type W (PRSV-W). SqVYV is a whitefly-transmitted member of the Potyviridae, recently shown to cause watermelon vine decline in Florida (1,4). Plants infected with SqVYV in Florida are also frequently infected with PRSV-W, although SqVYV is sufficient for watermelon vine decline. The six field samples harbored one or more potyviruses as determined by ELISA (Agdia, Elkhart, IN). Mechanical inoculation of squash (Cucurbita pepo) and watermelon with sap from three of the field samples induced mosaic symptoms in both that are typical of potyviruses. Vein yellowing in squash and plant death in watermelon typical of SqVYV (1) later developed in plants inoculated with one field sample. A coat protein gene fragment was amplified by reverse transcription (RT)-PCR with SqVYV primers (1) from total RNA of five of the six field samples and also from the symptomatic, inoculated plants. Nucleotide and deduced amino acid sequences of a 957-bp region of the RT-PCR product (primer sequences deleted prior to analysis) were 100% identical to SqVYV (GenBank accession No. DQ812125). PRSV-W also was identified in two of the five SqVYV-infected field samples by ELISA (Agdia) and by sequence analysis of a 3' genome fragment amplified by RT-PCR with previously described degenerate potyvirus primers (3). No evidence for infection by other potyviruses was obtained. To our knowledge, this is the first report of SqVYV in Indiana and the first report of the virus anywhere outside of Florida. The whitefly (Bemisia tabaci, B strain) vector of SqVYV is relatively uncommon in Indiana and the cold winter temperatures make it unlikely that any SqVYV-infected watermelon vines or whiteflies will overseason, necessitating reintroductions of virus and vector each season. We feel that the moderate and restricted occurrence of SqVYV in Indiana observed in September 2006 should pose little or no threat to commercial watermelon production in Indiana and should not cause growers to alter their growing practices. The occurrence of SqVYV in Indiana does not appear to explain the similar symptoms of mature watermelon vine decline (MWVD) that has been observed in Indiana since the 1980s. In contrast with the insect vectored SqVYV, MWVD seems to be caused by a soilborne biological agent (2). References: (1) S. Adkins et al. Phytopathology 97:145, 2007. (2) D. S. Egel et al. Online publication. doi:10.1094/PHP-2000-1227-01-HN. Plant Health Progress, 2000. (3) A. Gibbs and A. Mackenzie. J. Virol. Methods 63:9, 1997. (4) P. Roberts et al. Citrus Veg. Mag. December 12, 2004.

First Report of Fusarium oxysporum f. sp. niveum Race 2 as Causal Agent of Fusarium Wilt of Watermelon in Indiana.

Fusarium oxysporum f. sp. niveum race 1 is uniformly distributed throughout watermelon (Citrullus lanatus (Thunb.) Matsum. & Nakai) growing regions, but F. oxysporum f. sp. niveum race 2 has a limited known distribution in the United States (Texas, Florida, Oklahoma, Maryland, and Delaware) (3,4). Since the spring of 2001, commercial watermelon fields in Knox and Gibson counties in southwestern Indiana have been observed with symptoms of one-sided wilt and vascular discoloration typical of Fusarium wilt. Race 2 of F. oxysporum f. sp. niveum was suspected as the casual agent since the diseased watermelon cultivars are considered resistant to races 0 and 1. Two isolates of F. oxysporum obtained from wilted watermelon plants in two different commercial fields and one isolate obtained from a wilted seedling in a transplant house were compared for pathogenicity in a greenhouse assay. Known isolates of F. oxysporum f. sp. niveum races 0, 1, and 2 were obtained from Don Hopkins (University of Florida, Apopka), Kate Everts (University of Maryland/University of Delaware, Salisbury, MD), and Ray Martyn (Purdue University, West Lafayette, IN), respectively, and were used for comparison. All isolates were grown in shake cultures in a mineral salts liquid medium. (1). After 72 hr, the predominately microconidal suspensions were filtered through cheesecloth and adjusted to 1 × 105 conidia/ml with the aid of a hemacytometer. A concentration of 1 × 105 condia/ml was shown previously to cause the desired disease reaction in the standard cultivars. Seedlings of the differential cvs, Black Diamond (universal susceptible), Charleston Gray (race 0 resistant), and Calhoun Gray (race 0 and 1 resistant) were grown in a 1:1, (v:v) sand/ vermiculite mixture to the first true-leaf stage after which the plants were uprooted and the roots carefully washed prior to root dip inoculation. Subsequent to inoculation, seedlings were planted in a sand/vermiculite/ peat mixture (4:1:1, [v:v:v]) with four seedlings to a 15-cm-diameter pot. The experimental design was a randomized complete block with five replications. Two isolates from the commercial field plants caused an average of 100% wilt on cv. Black Diamond, 95% wilt on cv. Charleston Gray, and 80% wilt on cv. Calhoun Gray, resulting in a designation of race 2. The isolate from a commercial transplant house resulted in 100, 60, and 15% wilt, respectively, on the three standard cultivars resulting in a race 1 designation. The presence of F. oxysporum f. sp. niveum race 2 in Indiana is significant because Indiana currently ranks fifth in the United States in watermelon production and there are no commercially available cultivars that possess resistance to race 2. To our knowledge, this is the first report of F. oxysporum f. sp. niveum race 2 in Indiana and the first report of race 2 from the Midwest region of the United States. Race 2, first described from the United States in 1985 (2), has now been confirmed in six states. References: (1) R. Esposito and A. Fletcher. Arch. Biochem. Biophys. 93:369, 1961. (2) R. Martyn, Plant Dis. 69:1007, 1985. (3) R. Martyn, Plant Dis. 71:233, 1987. (4) X. Zhou and K. Everts. Plant Dis. 87:692, 2003.

Genetic analysis of hrp-related DNA sequences of Xanthomonas campestris strains causing diseases of citrus.

The hrp gene cluster of strains of Xanthomonas campestris that cause diseases of citrus was examined by Southern hybridization of genomic DNA and by restriction endonuclease analysis of enzymatically amplified DNA fragments of the hrp gene cluster. The hrp genes were present in all strains of the pathovars of X. campestris tested in this study, including strains of the three aggressiveness groups of the citrus bacterial spot pathogen, X. campestris pv. citrumelo. X. campestris pv. citri strains in groups A, B, and C, which cause citrus canker A, B, and C, respectively, each produced characteristic restriction banding patterns of amplified hrp fragments. The restriction banding patterns of all strains within each group were identical. In contrast, restriction fragment length polymorphism was evident among strains of the moderately and weakly aggressive groups of X. campestris pv. citrumelo. X. campestris pv. citrumelo strains in the highly aggressive group had a homogeneous restriction banding pattern. The characteristic banding patterns obtained for each bacterial group indicate that X. campestris strains causing disease in citrus can be reliably differentiated and identified by restriction analysis of amplified DNA fragments of the hrp gene cluster. In addition, the phylogenetic analysis based on the restriction banding patterns of amplified fragments suggests a polyphyletic relationship of the hrp genes among the strains of X. campestris that cause disease in citrus.

Genomic Relatedness of Xanthomonas campestris Strains Causing Diseases of Citrus.

Xanthomonas campestris strains that cause disease in citrus were compared by restriction endonuclease analysis of DNA fragments separated by pulsed-field gel electrophoresis and by DNA reassociation. Strains of X. campestris pv. citrumelo, which cause citrus bacterial spot, were, on average, 88% related to each other by DNA reassociation, although these strains exhibited diverse restriction digest patterns. In contrast, strains of X. campestris pv. citri groups A and B, which cause canker A and canker B, respectively, had relatively homogeneous restriction digest patterns. The groups of strains causing these three different citrus diseases were examined by DNA reassociation and were found to be from 55 to 63% related to one another. Several pathovars of X. campestris, previously shown to cause weakly aggressive symptoms on citrus, ranged from 83 to 90% similar to X. campestris pv. citrumelo by DNA reassociation. The type strain of X. campestris pv. campestris ranged from 30 to 40% similar in DNA reassociation experiments to strains of X. campestris pv. citrumelo and X. campestris pv. citri groups A and B. Whereas DNA reassociation quantified the difference between relatively unrelated groups of bacterial strains, restriction endonuclease analysis distinguished between closely related strains.