|Satyavathi, Venkata - PLNT SCI, NDSU, FARGO ND|
Submitted to: Plant and Animal Genome VX Conference Abstracts
Publication Type: Abstract Only
Publication Acceptance Date: October 10, 2003
Publication Date: January 10, 2004
Citation: Satyavathi, V.V., Jauhar, P.P. 2004. Transformation of a commercial durum cultivar Maier with antifungal genes. 12th Plant and Animal Genome Abstracts. p. 96. Technical Abstract: Scab or Fusarium head blight (FHB), caused by the fungus Fusarium graminearum Schwabe, is a serious disease of durum wheat (Triticum turgidum L.), an important cereal used for pasta and semolina for human consumption worldwide. There is no reliable source of scab resistance in durum cultivars. It would be advisable to apply transgenic approach to control this ravaging disease. An important limitation to genetic transformation of durum wheat has been the availability of an efficient in vitro regeneration protocol applicable to current commercial cultivars. Therefore, we standardized the regeneration protocol for four commercial durum cultivars, Ben, Maier, Munich, and Lebsock, grown in North Dakota. Earlier, we standardized the transgenic technology for durum wheat. In an attempt to enhance resistance to FHB, we engineered Maier with pathogenesis-related gene (tlp) in combination with antitoxin gene like Tri101 (with bar gene as a selectable marker). Two week-old calli from isolated scutella of the cultivar Maier were co-bombarded with plasmid (pAHRC-TLP) containing bar and rice tlp genes along with plasmid pUBK containing modified Tri101 (mTri101). The regenerants were selected on medium containing 5.0 mg L-1 bialaphos. Putative transformants were hardened in a growth chamber and established in the greenhouse. The presence of the transgenes was tested initially by PCR analysis using primers specific to bar / Tri101 genes. PCR positive plants were selected for further molecular analyses. Western blot analysis using specific anti-Tlp-antibody confirmed the expression of the tlp gene. The results of Southern and Northern blot analyses will be discussed.