Abbreviations: Cb, carbenicillin; CMV, cucumber mosaic virus; GUS, (β -glucuronidase; Km, kanamycin; MS, Murashige and Skoog NPT II, neomycin phosphotransferase II; NOS, nopaline synthase. We thank Krystal A. Fober for plant care in the greenhouse
Paula P. Chee and Jerry L. Slightom
Axel O. Ramírez-Madera and Michael J. Havey
The potyviruses (Potyviridae) are especially destructive for cucumber ( Cucumis sativus L.) and production is often negatively affected by Watermelon mosaic virus (WMV), the watermelon strain of the Papaya ringspot virus (PRSV-W), and ZYMV
S. Alan Walters
The author wishes to thank Dena Fiacchino, Harry Riddle and Joyce Swenson for their technical assistance in this study, and Houston Hobbs of the University of Illinois-Dept. of Crop Sciences for virus determination. The use of trade names in this
Cucumber mosaic virus (CMV) is an aphid-transmitted virus that infects snap bean growing regions in New York State and Wisconsin. The core collection of common bean accessions (Phaseolus vulgaris), the complete collection of scarlet runner bean accessions (Phaseolus coccineus) and snap/dry bean cultivars were screened for resistance to CMV. Although variation in foliar symptom expression was observed, no resistance was observed in 93 snap bean and16 dry bean cultivars tested, and only one of the 406 accessions from the core collection (PI 309881) was symptomless. PI 309881 did not have common bean characteristics, and was later identified as a tepary bean (Phaseolus acutifolius) accession based on morphology and PCR-RFLP of chloroplast DNA. Screening of 260 P. coccineus accessions was inaccurate when a visual rating of foliar symptoms was used. It was necessary to determine infection using ELISA and test plant screening with grey zuccini. Using this approach it was determined that 80 P. coccineus accessions were susceptible to CMV; however, the remaining accessions provided possible sources for transfer of CMV resistance to snap bean. Crosses of P. coccineus accessions were made to breeding line 5-593 and backcrossed to 5-593 and snap bean cultivar `Hystyle'. PI 309881 was crossed with ICA Pijao in order to develop interspecific hybrids. Populations were developed from the interspecific crosses/backcrosses and evaluated for CMV resistance using ELISA and visual ratings of foliar symptoms.
P.B. McGarvey, M.S. Montasser and J.M. Kaper
Transgenic tomato plants (Lycopersicon esculentum Mill.) expressing cucumber mosaic virus (CMV) satellite RNA fused to a gene for β-glucuronidase were produced using Agrobacterium-mediated transformation. The R1 progeny of self-crossed R0 plants were challenge-inoculated with virion or RNA preparations of CMV or tomato aspermy virus (TAV). The transgenic plants challenged with CMV-1 showed mild disease symptoms in the first 2 weeks postchallenge followed by a decrease in symptoms, resulting in little difference between the transgenic and uninfected control group by the fourth week. Enzyme-linked immunosorbent assay results showed about a 10-fold decrease in virus accumulation in the transgenic plants compared to controls. Tolerance was evident only in plants that contained the recombinant insert and produced mature unit-length satellite RNA after CMV infection. Plants challenged with TAV showed no significant tolerance to virus-induced symptoms.
Eileen Kabelka and Rebecca Grumet
Potyviruses cause severe loss in cucurbit crops. Inbred lines derived from the Chinese cucumber cultivar, Taichung Mau Gua (TMG), have been identified to be resistant to several potyviruses including zucchini yellow mosaic virus (ZYMV), zucchini yellow fleck virus, watermelon mosaic virus, and the watermelon strain of papaya ringspot virus. Recently, an additional virus that infects cucurbits, the Moroccan watermelon mosaic virus (MWMV), has been identified to be a distinct member of the potyvirus group. In this study, we sought to determine if TMG-1 is resistant to MWMV and, if so, examine whether a relationship exists between resistance to MWMV and resistance to ZYMV. Progeny analyses show that TMG-1 is resistant to MWMV and, like resistance to ZYMV, MWMV resistance is conferred by a single recessive gene. Sequential inoculation of progeny possessing resistance to ZYMV followed by MWMV (or MWMV followed by ZYMV) suggests that both resistances are conferred by the same gene, or two tightly linked genes. Additionally, all F3 families derived from F2 individuals selected for resistance to ZYMV, were resistant to MWMV. A second source of resistance to ZYMV, allelic to the TMG-1 source, has been incorporated into the Dutch hybrid Dina. Progeny analyses show Dina to posses a single recessive gene for MWMV resistance. As with TMG-1, no segregation of resistances was observed when ZYMV resistant progeny were inoculated with MWMV (or MWMV followed by ZYMV). Collectively, these results suggest that a single gene, or two tightly linked genes, control resistance to the potyviruses ZYMV and MWMV.
Yiping Zhang, Vince Lackney and Molly Kyle
We report the detection of molecular markers linked to polygenic recessive resistance to cucumber mosaic virus (CMV) in pepper using distributional extreme analysis. A total of 132 mapped tomato genomic, cDNA, and pepper genomic clones from different linkage groups in two existing pepper maps, especially from the ones showing tentative CMVR QTLs, were selected as hybridization probes for Southern blots, in which DNA from the 12 most resistant and 15 most susceptible individuals from a large segregating F2 were probed. Several clones appeared to cosegregate with CMV resistance phenotype. Further analysis is being done to place these markers on existing molecular linkage maps. The precise locations of resistance genes can be defined by examining additional markers within the region. The availability of closely linked DNA markers may facilitate marker-aided selection in pepper CMV resistance breeding programs.
J.R. Fisher and S.T. Nameth
Ajuga reptans L. is an herbaceous ornamental mint grown in borders or as a groundcover, and is commonly propagated vegetatively and by seed. Three hundred and fifty-six A. reptans samples were obtained from growers in Washington, Michigan, Iowa, and Ohio, and screened for alfalfa mosaic virus (AMV), tobacco streak ilarvirus (TSV), cucumber mosaic cucumovirus (CMV), tomato aspermy cucumovirus (TAV), tomato spotted wilt tospovirus (TSWV), impatiens necrotic spot tospovirus (INSV), tobacco mosaic tobamovirus (TMV), potato virus × potexvirus (PVX), and 80 potyviruses, using direct antibody sandwich (DAS) and indirect enzyme-linked immunosorbent assay (ELISA). Viral-associated double-stranded ribonucleic acid (dsRNA) analysis was used to detect an apparent satellite (sat) RNA, and northern hybridization using a digoxigenin (DIG) labeled (S) CARNA-5 cDNA probe was used to confirm the identity of the apparent satRNA. No incidences of TAV, TMV, TSWV, INSV, PVX, or potyviruses were detected. CMV was detected in 11%, AMV in 22.2%, TSV in 3.7%, and mixed infections of CMV and AMV in 1.1% of the samples. SatRNA was detected in 36 A. reptans `Royalty', two `Rainbow', and two `Burgundy Glow' samples by dsRNA analysis, and confirmed by hybridization in 29 `Royalty' and one `Burgundy Glow' samples. Sixteen A. reptans `Royalty' seedlings grown from seed harvested from CMV-infected plants were tested by ELISA for CMV, AMV, and TSV. All were positive for CMV, and two were positive for a mixed infection of CMV and AMV. SatRNA was detected in all 16 seedlings by RT-PCR.
Cowpea cultivars Early Ramshorn and Dixie Queen, reported to be resistant to cucumber mosaic virus (CMV) by Sinclair and Walker in 1955, were reexamined for the existence of individual CMV-resistant genotypes within seedling populations. CMV-inoculated populations of these two cultivars became CMV-infected at rates of 60% and 80%, respectively, as determined by DAS-ELISA. CMV-free plants were grown to maturity, seeds from these sources were planted, and second-generation seedlings were inoculated again with CMV. Rates at which these seedlings became CMV-infected were 10% and 24%, respectively. The complementary 90% and 76% of these populations remained completely free of ELISA-detectable CMV and were saved to maturity. Evaluations of third-generation populations are being performed. Twenty additional cultivars and breeding lines also are being evaluated. Seedlings of 14 of the 22 cultivars/lines became 95% to 100% infected when inoculated with CMV. Results from eight of the 22 suggested that seedling populations contain CMV-susceptible and -resistant plants. Near isogeneic sublines of susceptible and resistant plants per cultivar/line are being conveyed to cowpea breeders for genetic analyses and breeding purposes.
R.C. Grube, Y. Zhang, B. Huang and M.M. Kyle
Resistance to cucumber mosaic virus (CMV) in Capsicum from two sources is being transferred into three commercial types (bell, jalapeno, and Anaheim) using a backcross breeding scheme. We have optimized our CMV seedling screening protocol, which involves multiple inoculations beginning at the cotyledon stage with a severe CMV serogroup I isolate. Both sources of resistance, C. annuum `French Perennial' and a C. frutescens accession (BG2814-6), exhibit oligogenic recessive inheritance and share some but not all resistance alleles. Selection for type in the BCF1 generation had no effect on the frequency of resistant individuals in the BCF2 generation. We have determined that it is necessary to self-pollinate every other backcross generation to screen for resistance. Occasionally disease symptoms appear in adult plants that were initially resistant to multiple inoculations at the seedling stage, and we are investigating the correlation between seedling resistance and adult plant resistance. We are also exploring the extent to which the different sources of resistance behave differently as a function of genetic background. Additionally, we are mapping quantitative trait loci (QTLs) for CMV resistance in pepper with the goal of converting RFLP and/or RAPD markers into PCR-based markers to facilitate molecular marker-assisted selection for CMV resistance.