Identification of Arabidopsis Mutants with Enhanced Resistance to Sclerotinia Stem Rot Disease from an Activation-tagged Library
2009
Wang, Ai-Rong | Zhang, Chun-Hua | Zhang, Li Li | Lin, Wenwei | Lin, De-Shu | Lu, Guo-Dong | Zhou, Jie | Wang, Zong-Hua
Sclerotinia sclerotiorum causes severe stem rot disease in many crops. Little is known about the genetic mechanism of pathogenesis, making it difficult to breed new, disease-resistant crop varieties. Here, we report a screen for Arabidopsis mutants with enhanced resistance to S. sclerotiorum through a functional genomics approach. First, an activation-tagged library of Arabidopsis thaliana Col-4 was generated, followed by screening of more than 50 000 T1 lines for enhanced resistance by inoculating detached leaves. Second, 50 lines with enhanced resistance were further characterized to identify the responsible genes by analyzing T-DNA flanking regions via a sequence BLAST analysis against the Arabidopsis genomic database. Third, the expression of candidate genes expression was confirmed by reverse transcription-polymerase chain reaction. Finally, SALK T-DNA insertion mutants were generated from Arabidopsis Biological Resource Center, which contain loss-of-function mutations for these candidate genes, and inoculated with S. sclerotiorum to determine the effect on infection. Our results suggest that a functional genomics approach is effective in dissecting the molecular mechanism of plant pathosystems that do not show a gene-for-gene relationship.
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