Aphid effector Me10 interacts with tomato TFT7, a 14-3-3 isoform involved in aphid resistance
Aphid effector Me10 interacts with tomato TFT7, a 14-3-3 isoform involved in aphid resistance
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DOI：
10.1111/nph.15475
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发表时间：
2019-02-01
期刊：
影响因子：
9.4
通讯作者：
Kaloshian, Isgouhi
中科院分区：
文献类型：
--
作者：
Chaudhary, Ritu;Peng, Hsuan-Chieh;Kaloshian, Isgouhi
我们之前证明了大管蚜唾液蛋白Me 10的表达增强了蚜虫在其宿主番茄（番茄）上的繁殖。然而，Me 10的作用机制仍然是难以捉摸的。为了证实Me 10由蚜虫分泌到植物组织中，我们制备了Me 10多克隆抗体。为了鉴定Me 10的植物靶基因，构建了番茄免疫诱导互补DNA酵母双杂交文库，并以Me 10为诱饵进行筛选。通过免疫沉淀和双分子荧光互补（BiFC）实验验证了Me 10在植物中的相互作用，并利用病毒诱导的基因沉默对Me 10在番茄中的功能进行了研究，证明Me 10在酵母中分泌到植物组织中并与番茄14-3-3异构体7（TFT 7）相互作用。免疫沉淀试验证实Me 10及其在棉蚜中的同源物Ag 10 k与植物中的TFT 7相互作用。此外，BiFC揭示了Me 10与TFT 7的相互作用发生在植物细胞质中。虽然TFT 7基因在番茄叶片中的沉默并不影响番茄对棉蚜的敏感性，但它提高了非寄主棉蚜的寿命和繁殖力。我们的研究结果表明，TFT 7在番茄抗蚜中发挥作用，Me 10/Ag 10 k家族的效应子在蚜虫侵染期间干扰TFT 7的功能。
We demonstrated previously that expression of Macrosiphum euphorbiae salivary protein Me10 enhanced aphid reproduction on its host tomato (Solanum lycopersicum). However, the mechanism of action of Me10 remained elusive.To confirm the secretion of Me10 by the aphid into plant tissues, we produced Me10 polyclonal antibodies. To identify the plant targets of Me10, we developed a tomato immune induced complementary DNA yeast two-hybrid library and screened it with Me10 as bait. Immunoprecipitation and bimolecular fluorescence complementation (BiFC) assays were performed to validate one of the interactions in planta, and virus-induced gene silencing was used for functional characterization in tomato.We demonstrated that Me10 is secreted into the plant tissues and interacts with tomato 14-3-3 isoform7 (TFT7) in yeast. Immunoprecipitation assays confirmed that Me10 and its homologue in Aphis gossypii, Ag10k, interact with TFT7 in planta. Further, BiFC revealed that Me10 interaction with TFT7 occurs in the plant cell cytoplasm. While silencing of TFT7 in tomato leaves did not affect tomato susceptibility to M.euphorbiae, it enhanced longevity and fecundity of A.gossypii, the non-host aphid.Our results suggest the model whereby TFT7 plays a role in aphid resistance in tomato and effectors of the Me10/Ag10k family interfere with TFT7 function during aphid infestation.