白叶枯病菌与非亲和水稻IRBB4悬浮细胞互作中蛋白类激发子的纯化和鉴定

 本研究对水稻白叶枯病菌与水稻悬浮细胞非亲和互作中蛋白类激发子进行了分离纯化和鉴定.白叶枯病菌JXOV与水稻IRBB4和IR24悬浮细胞互作36 h后的上清液,经Q-Sepharose阴离子交换层析柱分离,对分离的各组分进行抗病性诱导测定,结果表明JXOV与IRBB4非亲和互作的上清液中存在蛋白类激发子.有活性的蛋白组分经阴离子交换层析柱Mono-Q进一步纯化后,SDS-PAGE分析鉴定出2个具激发活性的蛋白,其分子量分别为17.2 kD和49.2 kD,等电点分别为5.8和6.2.利用上述激发子处理水稻能减少病斑长度并诱导水稻防卫酶活性的增加.

Purification and identification of proteinous elicitor in interaction between Xanthomonas oryzae pv. oryzae and suspension-cultured incompatible rice IRBB4 cells

In this study two proteinous elicitors in extracellular supernatant during interaction between Xanthomonas oryzae pv. oryzae JXOV and suspension-cultured incompatible rice IRBB4 cells were isolated and identified. Proteins in extracellular supernatant collected 36 h after the interaction between JXOV and incompatible rice IRBB4 suspension cells or compatible rice IR24 suspension cells were fractioned by HPLC using an anion exchange column Q-Sepharose. Two fractions obtained in incompatible interaction showed elicitor effect for that length of leaf lesion on rice IR24 inoculated JXOV observably decreased after treatment with substance in that fractions. Further purification by HPLC using anion exchange column Mono-Q and preparative PAGE revealed 2 elicitors with molecular mass 17.2 kD and 49.2 kD,and pI 5.8 and 6.2. Treatment with these eli-citors at the concentration of 100 μg/mL before inoculation of JXOV not only decreased leaf lesion,but also increased the activity of defence enzymes PAL and POD in leaves of compatible rice IR24.