稻瘟菌为了实现对水稻的有效侵染, 在侵染水稻时可能通过表达和转运一定数量的效应蛋白进入到水稻细胞, 抑制和干扰水稻的先天免疫机制。文章利用Solexa第二代测序技术, 通过开展水稻和稻瘟菌互作早期转录组的测定和分析, 克隆和鉴定在互作早期表达的稻瘟菌效应蛋白基因。利用序列同源比对, 我们从总计约12.5 M条序列标签中, 分离和鉴定了338 942条来源于稻瘟菌的序列, 并最终定位到779个稻瘟菌预测基因。其中108个基因很可能参与了水稻和稻瘟菌互作过程, 42个基因为预测的分泌蛋白基因。通过RT-PCR分析, 最终确认了42个预测分泌蛋白基因中有12个基因在侵染水稻早期有显著的表达, 而其中有4个基因表现为侵染早期特异表达。文章尝试利用第二代测序技术实现稻瘟菌侵染早期特异表达基因, 尤其是分泌蛋白基因的快速克隆和鉴定, 为稻瘟菌效应蛋白基因的克隆和功能鉴定提供了较为有意义的探索。
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