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一种新型效应蛋白 MJ-NULG1a,靶向大细胞核,在根结线虫寄生中发挥作用。

A novel effector protein, MJ-NULG1a, targeted to giant cell nuclei plays a role in Meloidogyne javanica parasitism.

机构信息

South China Agricultural University, Guangzhou, China.

出版信息

Mol Plant Microbe Interact. 2013 Jan;26(1):55-66. doi: 10.1094/MPMI-05-12-0114-FI.

Abstract

Secretory effector proteins expressed within the esophageal glands of root-knot nematodes (Meloidogyne spp.) are thought to play key roles in nematode invasion of host roots and in formation of feeding sites necessary for nematodes to complete their life cycle. In this study, a novel effector protein gene designated as Mj-nulg1a, which is expressed specifically within the dorsal gland of Meloidogyne javanica, was isolated through suppression subtractive hybridization. Southern blotting and BLAST search analyses showed that Mj-nulg1a is unique for Meloidogyne spp. A real-time reverse-transcriptase polymerase chain reaction assay showed that expression of Mj-nulg1a was upregulated in parasitic second-stage juveniles and declined in later parasitic stages. MJ-NULG1a contains two putative nuclear localization signals and, consistently, in planta immunolocalization analysis showed that MJ-NULG1a was localized in the nuclei of giant cells during nematode parasitism. In planta RNA interference targeting Mj-nulg1a suppressed the expression of Mj-nulg1a in nematodes and attenuated parasitism ability of M. javanica. In contrast, transgenic Arabidopsis expressing Mj-nulg1a became more susceptible to M. javanica infection than wild-type control plants. These results depict a novel nematode effector that is targeted to giant cell nuclei and plays a critical role in M. javanica parasitism.

摘要

根结线虫(Meloidogyne spp.)食管腺中表达的分泌效应蛋白被认为在根结线虫入侵宿主根系和形成必需的取食部位方面发挥着关键作用,这些部位对于线虫完成其生命周期是必要的。在这项研究中,通过抑制性消减杂交分离到一个新的效应蛋白基因,命名为 Mj-nulg1a,该基因在爪哇根结线虫的背腺中特异性表达。Southern 印迹和 BLAST 搜索分析表明,Mj-nulg1a 是 Meloidogyne spp. 的特有基因。实时逆转录聚合酶链反应分析显示,Mj-nulg1a 的表达在寄生的第二阶段幼虫中上调,在后期寄生阶段下降。MJ-NULG1a 包含两个假定的核定位信号,因此,植物体内免疫定位分析表明,在根结线虫寄生过程中,MJ-NULG1a 定位于巨型细胞的核内。针对 Mj-nulg1a 的植物体内 RNA 干扰靶向抑制了线虫中 Mj-nulg1a 的表达,并减弱了爪哇根结线虫的寄生能力。相比之下,表达 Mj-nulg1a 的转基因拟南芥比野生型对照植物更容易感染爪哇根结线虫。这些结果描绘了一种新型的线虫效应蛋白,该蛋白靶向巨型细胞核,在爪哇根结线虫寄生中发挥关键作用。

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