| Sclerotinia sclerotiorum is a ubiquitous phytopathogenic Ascomycete fungus capable of infecting a wide range of plants. It caused some economic plants stem rot at mature stage and has become a limiting factor for production. Arabidopsis has good characteristics of small genome, simple structure, small plant, short life period, high propagating rate, self-pollinating etc. And the whole genome sequencing has been completed in December 2000. So it is the model plant for studying plant and pathogen interaction. We have established methods for screening for mutants of Arabidopsis with enhanced resisatnce to S.sclerotiorum and screened more than 50,000 activated tagging mutants and achieved some resistant mutant lines. A muatant of loss-of-function in CJPK23 with enhanced resistance to S.sclerotiorum, but more sensitive to the salt stress. Then we cloned the gene and tested its function. The major achievements are summarized as in the following.1. We tried different screening methods, such as spraying oxalic acid or S. sclerotiorum mycelium suspensions or sticking the mycelium agar peg of S. sclerotiorum on leaves, or culturing seeds with oxalic acid. The effect of sticking the mycelium agar pegs of S. sclerotiorum on leaves is the best, so we used the method to screen resistant mutants.2. We screened 50,000 activation-tagged mutants and obtained 50 A. thaliana mutants with enhanced resistance to S .sclerotiorum. We further amplified T-DNA flanking sequences by TAIL-PCR in 44 mutants, then blasted the sequences at the TAIR website or the MIPS website. We got 34 effcient sequences and located the insertion site. Among them, five mutants(M150, M138, M12, M833, Mc140) inserted in the same locus At1g30270(CBL-interacting protein kinase 23 (CIPK23)) intron regions, then named as CIPK23 mutant.3. We then studied the regulation effects of enhancer on its adjacent genes located 12 kb upstream and downstream of enhancers in mutant line M150 by RT-PCR. The enhancer...
|
PDF Full Text Request