Enhanced tolerance of transgenic grapevines expressing chitinase and beta-1,3-glucanase genes to downy mildew
Title | Enhanced tolerance of transgenic grapevines expressing chitinase and beta-1,3-glucanase genes to downy mildew |
Publication Type | Journal Article |
Year of Publication | 2012 |
Authors | Nookaraju, A, Agrawal, DC |
Journal | Plant Cell Tissue and Organ Culture |
Volume | 111 |
Issue | 1 |
Pagination | 15-28 |
Date Published | OCT |
ISSN | 0167-6857 |
Keywords | Agrobacterium tumefaciens, Anti-oxidants, Downy mildew tolerance, Grapevine, Sonication |
Abstract | An Agrobacterium-mediated transformation protocol for grapevine cv. Crimson Seedless using sonication and anti-necrotic agents has been optimized, and transgenic lines carrying wheat chitinase and beta-1,3-glucanase genes have exhibited enhanced tolerance to downy mildew incited by Plasmopara viticola. cDNA clones encoding chitinase and beta-1,3-glucanase have been isolated from a cDNA library, constructed from scab-infected Sumai-3 wheat, and introduced into a plant cloning vector to generate the plasmids pCAMBAR.chi.11 and pCAMBAR.638. Embryogenic cultures, established from in vitro-derived leaves, of Crimson Seedless were used as explants for Agrobacterium tumefaciens-mediated transformation studies. Sonication of somatic embryos in a bacterial suspension of A. tumefaciens and incorporation of anti-necrotic agents in the co-cultivation medium significantly enhanced transformation efficiency. Transformation efficiency of embryos with either chitinase or beta-1,3-glucanase gene was highest when embryos were suspended in a bacterial cell suspension at 0.5 OD600 and sonicated for 2 or 3 s at 60 kHz. Transformation efficiency with chitinase was highest on incorporation of 2 or 3 mg l(-1) phenylalanine, 1 or 2 mg l(-1) silver nitrate or 400 mg l(-1) l-cysteine in co-cultivation medium while incorporation of 20 mg l(-1) sodium thiosulphate produced highest transformation efficiency with beta-1,3-glucanase. Confirmed transgenic grapevine lines harboring anti-fungal genes exhibited higher levels of chitinase and beta-1,3-glucanase transcripts as well as enzymatic activities. Moreover, transgenic lines showed enhanced tolerance to P. viticola infection following detached leaf assays. |
DOI | 10.1007/s11240-012-0166-1 |
Type of Journal (Indian or Foreign) | Foreign |
Impact Factor (IF) | 3.633 |