Effects of P-loop, Bet v 1 domain deletion and mutation on ability of GmPR10 and Gly m 4l to inhibit Phytophthora sojae.
Effects of P-loop, Bet v 1 domain deletion and mutation on ability of GmPR10 and Gly m 4l to inhibit Phytophthora sojae.
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DOI：
10.19802/j.issn.1007-9084.2019235
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发表时间：
2020-01-01
期刊：
影响因子：
--
通讯作者：
Xu, P. F.
中科院分区：
文献类型：
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作者：
Fang, Xin;Fan, Su-Jie;Xu, P. F.
大豆疫霉根腐病是由卵菌类病原体大豆疫霉引起的一种严重影响大豆生长的病害。我们前期的研究表明，具有P-loop结构域的GmPR10和同时具有p-loop和Bet v1结构域的Gly m 4l可以抑制大豆疫霉的生长，并且过表达GmPR10和Gly m 4l的转基因大豆植物可以增加对大豆疫霉的抗性。为了进一步研究GmPR10和Gly m 4l的病原抑制机制，GmPR10的P环结构域突变体(Gly48/Thr48和Gly51/Arg51)、Gly m 4l的P环结构域突变体(Gly49/Ile49和Lys55/Pro55)、GmPR10和Gly m 4l的P环结构域缺失突变体，利用点突变技术获得了Gly m 4l的Bet v1结构域缺失突变体，并纯化了相应突变体的蛋白，对大豆疫霉进行了体外抑制。结果表明，具有P-loop或Bet v1结构域突变或缺失的GmPR10和Gly m 4l失去了抑制大豆疫霉（小种1）生长的能力，表明P-loop和Bet v1结构域对于GmPR10和Gly m 4l发挥抗病原体功能至关重要。
Phytophthora root rot caused by the ommycete pathogen Phytophthora sojae is a serious disease affecting soybean growth. In our previous study, it showed that GmPR10 with P-loop domain and Gly m 4l with both p-loop and Bet v1 domains could inhibit the growth of Phytophthora sojae, and transgenic soybean plants of overexpressing GmPR10 and Gly m 4l could increase resistance to Phytophthora sojae. In order to further study the pathogen inhibition mechanism of GmPR10 and Gly m 4l, the P-loop structure domain mutant of GmPR10 (Gly48/Thr48 and Gly51/Arg51), the P-loop structure domain mutant of Gly m 4l (Gly49/Ile49 and Lys55/Pro55), the P-loop structure domain deletion mutant of GmPR10 and Gly m 4l, the Bet v1 structure domain deletion mutant of Gly m 4l by using the point mutation technique were obtained and the purified proteins of corresponding mutations to inhibit Phytophthora sojae in vitro were carried out. The results demonstrated that GmPR10 and Gly m 4l with P-loop or Bet v1 domain mutation or deletion lost ability to inhibit the growth of Phytophthora sojae (Race 1), indicating that P-loop and Bet v1 domain were crucial for GmPR10 and Gly m 4l to perform anti-pathogen functions.