TITLE	Segregation patterns for integration and expression of Coniothyrium minitans xylanase gene in Arabidopsis thaliana transformants
AUTHOR	Z.-X. Lu Agriculture and Agri-Food Canada, Lethbridge Research Centre, P.O. Box 3000, Lethbridge, Alberta T1J 4B1, Canada A. Laroche* Agriculture and Agri-Food Canada, Lethbridge Research Centre, P.O. Box 3000, Lethbridge, Alberta T1J 4B1, Canada H.C. Huang Agriculture and Agri-Food Canada, Lethbridge Research Centre, P.O. Box 3000, Lethbridge, Alberta T1J 4B1, Canada
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ABSTRACT	A xylanase gene cxy1, isolated from the mycoparasitic fungus Coniothyrium minitans, has been transferred into Arabidopsis thaliana by Agrobacterium-mediated transformation. Ten A. thaliana transformants with herbicide resistance were selected, and the transformation efficiency of the modified "Floral Dip" method was about 0.08 %. Most transformants had one or two copies of T-DNA inserts with 3R:1S or 15R:1S segregation ratios while high proportions of susceptible progenies were also observed in some transgenic plants. The pMB4-2 transformant segregated as 1R:2S ratio in its T2 and T3 generations, suggesting that only one copy of T-DNA was integrated into this line's genome and the insertion likely interrupted a gene important or essential to its gametophytic development or alternatively, affected the transmission of such gene. The genetic analysis of the pMB4/cxy1-6 transformant indicated that two copies of T-DNAs were inserted independently into this line's genome, in which the resistant homozygotes on one locus were non-viable. The RBB-Xylan assay for six pMB4/cxy1 transformants indicated that two lines (pMB4/cxy1-3 and pMB4/cxy1-6) expressed the cxy1 gene, and variations for the xylanase activity were observed among T2 progenies of pMB4/cxy1-6 transformant.
KEYWORD	Agrobacterium-mediated transformation; Arabidopsis thaliana; Coniothyrium minitans; Xylanase gene;
ARTICLE INFO	Botanical Bulletin of Academia Sinica, Volume 45 Number 1 January 2004, page 23-31, 9 pages
PUBLISHER	Institute of Plant and Microbial Biology, Academia Sinica, Taipei, Taiwan, Republic of China