Isolation of Molecular Markers Linked to the <Emphasis Type="Italic">Cry</Emphasis> Locus Conferring Resistance to <Emphasis Type="Italic">Cucumber mosaic cucumovirus </Emphasis>Infection in Cowpea |
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| Authors: | Yutaka CHIDA Kazuyuki OKAZAKI Akira KARASAWA Kayoko AKASHI Yoshiko NAKAZAWA-NASU Shuu HASE Hideki TAKAHASHI Yoshio EHARA |
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| Institution: | (1) Laboratory of Plant Pathology, Department of Applied Life Science, Graduate School of Agricultural Science, Tohoku University, 1–1, Tsutsumidori-Amamiyamachi, Aoba-ku, Sendai 981–8555, Japan, JP |
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| Abstract: | We previously demonstrated that cowpea Vigna unguiculata (L.) Walp.] cultivar Kurodane-Sanjaku contains the Cry gene, which confers resistance against Cucumber mosaic cucumovirus infection. In this paper, randomly amplified polymorphic DNA (RAPD) analysis was carried out to tag the Cry locus. Bulked segregant analysis for RAPD resulted in many polymorphisms in amplified DNA patterns. Candidates were further
screened using parental and/or F2 cowpea DNAs. As a result, we obtained three RAPD markers, D13/E14-350, WA3-850 and OPE3-500, flanking the Cry locus. In addition, we amplified cowpea sequences coding for the putative nucleotide-binding site (NBS). Degenerate primers
based on NBS sequences of tobacco N and Arabidopsis RPS2 disease resistance genes were used for polymerase chain reaction, and resultant products were cloned and sequenced. Among
eight independent clones, cowpea resistance gene analog (CRGA) 5 showed a distinct polymorphism when used as a probe for restriction
fragment length polymorphism analysis against the susceptible cowpea cultivar PI 189375 and a near-isogenic line for the Cry. Linkage analyses of these molecular markers showed that genetic distances of CRGA5, D13/E14-350, WA3-850 and OPE3-500 to
the Cry locus were 0.7, 5.2, 11.5 and 24.5 cM, respectively.
Received 16 December 1999/ Accepted in revised form 22 March 2000 |
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| Keywords: | : CMV cowpea RAPD RFLP resistance gene analog linkage analysis |
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