The new effector AbSCP1 of foliar nematode (Aphelenchoides besseyi) is required for parasitism rice

doi:10.1016/S2095-3119(21)63706-1

摘要/Abstract

摘要：

水稻干尖线虫可侵染水稻、大豆、棉花等多种作物，给农业生产造成严重损失。丝氨酸羧肽酶（SerineCarboxypeptidases，SCP）是植物寄生线虫致病的一个关键因子，但丝氨酸羧肽酶在水稻干尖线虫中的致病机制并不清楚。本研究以水稻干尖线虫为对象，利用原位杂交、qRT-PCR、瞬时表达、真核表达以及基因沉默等方法对水稻干尖线虫丝氨酸羧肽酶（AbSCP1）的功能进行研究。研究得出，AbSCP1基因全长1425 bp，编码氨基酸长度为474aa。AbSCP1编码蛋白具有信号肽、无跨膜结构域，与香蕉穿孔线虫SCP蛋白的序列相似性为67%。不同龄期水稻干尖线虫AbSCP1的qPCR分析得出，该基因在幼虫中的表达量最高，其次是雌虫、雄虫和卵。通过原位杂交证实，AbSCP1在水稻干尖线虫的食道腺中表达。使用昆虫细胞表达系统获得了AbSCP1蛋白，通过与特异性底物反应证实了该蛋白的羧肽酶活性，并得出了酶促反应最适的pH为4.5。使用烟草瞬时表达系统表达AbSCP1，在烟草细胞核中出现强烈的特异荧光信号，说明AbSCP1被定位在植物细胞核中。使用RNAi研究AbSCP1对水稻干尖线虫致病力、繁殖力的影响。结果得出，水稻干尖线虫取食AbSCP1特异的dsRNA 24 h后，AbSCP1的表达量显著下降。使用基因沉默后的线虫分别接种水稻和灰葡萄孢，分别统计水稻的发病等级和线虫数。结果表明，AbSCP1被沉默后，水稻干尖线虫的致病力、繁殖率均显著下降。本研究首次在水稻干尖线虫中明确了SCP是一类可被分泌到寄主细胞核中发挥作用的蛋白酶类效应子，在线虫寄生寄主过程中起到重要作用。本研究的成果将为以AbSCP1为靶标开发高效、安全的水稻干尖线虫防治措施奠定基础。

Abstract: Plant parasitic nematodes secrete effector proteins to parasitize hosts successfully. Of these proteins, serine carboxypeptidases have critical roles in pathogenicity. This study investigated the role of new effector AbSCP1 in Aphelenchoides besseyi pathogenicity. In situ hybridization and qRT-PCR analyses indicated that AbSCP1 was exclusively expressed in the esophageal glands and upregulated in juveniles. Subcellular localization assays indicated that the protein was expressed in the nucleus. The ability to hydrolyze C-terminal amino acid residues was proven for AbSCP1. Moreover, RNAi significantly reduced the expression of AbSCP1 and RNAi-treated nematodes’ reproductive potential. Pathogenicity assays on rice showed that RNAi-treated nematodes were less pathogenic than the untreated control groups. These results suggest the important role of AbSCP1 in the A. besseyi infection process.

Key words: Aphelenchoides besseyi , serine carboxypeptidases , effector , nematode , pathogenicity

. [J]. Journal of Integrative Agriculture, 2022, 21(4): 1084-1093.

HUANG Xin, CHI Yuan-kai, Addisie Abate BIRHAN, ZHAO Wei, QI Ren-de, PENG De-liang. The new effector AbSCP1 of foliar nematode (Aphelenchoides besseyi) is required for parasitism rice[J]. Journal of Integrative Agriculture, 2022, 21(4): 1084-1093.

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