Determination of key residues in tospoviral NSm required for Sw-5b recognition, their potential ability to overcome resistance, and the effective resistance provided by improved Sw-5b mutants.
Determination of key residues in tospoviral NSm required for Sw-5b recognition, their potential ability to overcome resistance, and the effective resistance provided by improved Sw-5b mutants.
复制标题
确定 Sw-5b 识别所需的拓扑病毒 NSm 中的关键残基、其克服耐药性的潜在能力以及改进的 Sw-5b 突变体提供的有效耐药性。
DOI：
10.1111/mpp.13182
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发表时间：
2022-05
影响因子：
4.9
通讯作者：
Tao X
中科院分区：
文献类型：
--
作者：
Huang H;Zuo C;Zhao Y;Huang S;Wang T;Zhu M;Li J;Tao X
Sw-5 b基因是一种有效的抗病基因，广泛应用于番茄中以控制番茄斑萎病毒（TSWV），该病毒在世界范围内造成严重的作物损失。Sw-5 b通过识别病毒运动蛋白NSm（NSm 21，氨基酸115-135）的21个氨基酸的肽区来赋予抗性。然而，NSm的该肽区域内的C118 Y或T120 N突变引起了田间耐药性破坏（RB）TSWV分离株。为了研究TSWV破坏Sw-5 b-介导的抗性的潜在能力，我们对NSm 21上的每个氨基酸进行诱变，并确定哪些氨基酸突变会逃避Sw-5 b识别。在所有丙氨酸扫描突变体中，NSmP 119 A、NSmW 121 A、NSmD 122 A、NSmR 124 A和NSmQ 126 A当与Sw-5 b在本氏烟草叶片中共表达时未能诱导过敏反应（HR）。具有NSmP 119 A、NSmW 121 A或NSmQ 126 A突变的TSWV在病毒细胞间运动和全身感染方面存在缺陷，而携带NSmD 122 A或NSmR 124 A突变的TSWV不仅能够感染野生型N. Sw-5 b-转基因烟草不仅能够系统感染本氏烟株，而且还能够打破Sw-5 b-介导的抗性，并在Sw-5 b-转基因烟草上建立系统感染。本萨米亚那植物我们最近设计的两种改良突变体Sw-5 bL 33 P/K319 E/R927 A和Sw-5 bL 33 P/K319 E/R927 Q可有效抵抗携带NSmC 118 Y或NSmT 120 N突变的田间RB分离物，识别所有NSm 21丙氨酸取代突变体，并对新的实验性RB TSWV（NSmD 122 A或NSmR 124 A突变）赋予有效抗性。总之，我们确定了NSm识别Sw-5 b的关键残基，研究了它们潜在的RB能力，并证明了改进的Sw-5 b突变体可以对田间和潜在的RB TSWV分离物提供有效的抗性。两个人工改良的Sw-5 b抗性基因突变体赋予了对番茄斑萎病毒的潜在抗性破坏分离物的有效抗性。
Sw‐5b is an effective resistance gene used widely in tomato to control tomato spotted wilt virus (TSWV), which causes severe losses in crops worldwide. Sw‐5b confers resistance by recognizing a 21‐amino‐acid peptide region of the viral movement protein NSm (NSm21, amino acids 115–135). However, C118Y or T120N mutation within this peptide region of NSm has given rise to field resistance‐breaking (RB) TSWV isolates. To investigate the potential ability of TSWV to break Sw‐5b‐mediated resistance, we mutagenized each amino acid on NSm21 and determined which amino acid mutations would evade Sw‐5b recognition. Among all alanine‐scan mutants, NSmP119A, NSmW121A, NSmD122A, NSmR124A, and NSmQ126A failed to induce a hypersensitive response (HR) when coexpressed with Sw‐5b in Nicotiana benthamiana leaves. TSWV with the NSmP119A, NSmW121A, or NSmQ126A mutation was defective in viral cell‐to‐cell movement and systemic infection, while TSWV carrying the NSmD122A or NSmR124A mutation was not only able to infect wild‐type N. benthamiana plants systemically but also able to break Sw‐5b‐mediated resistance and establish systemic infection on Sw‐5b‐transgenic N. benthamiana plants. Two improved mutants, Sw‐5bL33P/K319E/R927A and Sw‐5bL33P/K319E/R927Q , which we recently engineered and which provide effective resistance against field RB isolates carrying NSmC118Y or NSmT120N mutations, recognized all NSm21 alanine‐substitution mutants and conferred effective resistance against new experimental RB TSWV with the NSmD122A or NSmR124A mutation. Collectively, we determined the key residues of NSm for Sw‐5b recognition, investigated their potential RB ability, and demonstrated that the improved Sw‐5b mutants could provide effective resistance to both field and potential RB TSWV isolates. Two artificially improved Sw‐5b resistance gene mutants confer effective resistance to potential resistance‐breaking isolates of tomato spotted wilt orthotospovirus.
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影响因子：
6.7
作者：
Ishibashi K;Mawatari N;Miyashita S;Kishino H;Meshi T;Ishikawa M
通讯作者：
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影响因子：
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