Host-Induced Gene Silencing of an Adenylate Kinase Gene Involved in Fungal Energy Metabolism Improves Plant Resistance to Verticillium dahliae
Host-Induced Gene Silencing of an Adenylate Kinase Gene Involved in Fungal Energy Metabolism Improves Plant Resistance to Verticillium dahliae
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宿主诱导的参与真菌能量代谢的腺苷酸激酶基因沉默可提高植物对大丽黄萎病的抗性
DOI：
10.3390/biom10010127
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发表时间：
2020-01-01
期刊：
影响因子：
5.5
通讯作者：
Cheng, Hongmei
中科院分区：
文献类型：
--
作者：
Su, Xiaofeng;Lu, Guoqing;Cheng, Hongmei
由子囊菌大丽轮枝菌（Verticilliumdahliae，Vd）引起的黄萎病是许多植物物种的毁灭性疾病。然而，该真菌中的致病性/毒力相关基因，这可能是提高植物抗性的潜在目标，仍然很难阐明。对于Vd中这些基因的研究，我们使用了成熟的宿主诱导基因沉默（HIGS）方法，并确定了16个候选基因，包括一个假定的腺苷酸激酶基因（VdAK）。瞬时VdAK沉默的植物比对照植物发生了较轻的枯萎症状。VdAK基因敲除突变体对非生物胁迫更敏感，并降低了发芽率和对宿主植物的毒性。过表达VdAK dsRNA的转基因本氏烟草和拟南芥植物比野生型具有改善的Vd抗性。RT-qPCR结果表明VdAK对能量代谢也是至关重要的。重要的是，在对从转基因植物中分离的Vd菌株的总小RNA的分析中，在转基因N.本萨米亚纳我们的研究结果表明，HIGS是一个有前途的策略，有效地筛选致病性/毒力相关基因的Vd和VdAK是一个潜在的目标，以控制这种真菌。
Verticillium wilt, caused by the ascomycete fungus Verticillium dahliae (Vd), is a devastating disease of numerous plant species. However, the pathogenicity/virulence-related genes in this fungus, which may be potential targets for improving plant resistance, remain poorly elucidated. For the study of these genes in Vd, we used a well-established host-induced gene silencing (HIGS) approach and identified 16 candidate genes, including a putative adenylate kinase gene (VdAK). Transiently VdAK-silenced plants developed milder wilt symptoms than control plants did. VdAK-knockout mutants were more sensitive to abiotic stresses and had reduced germination and virulence on host plants. Transgenic Nicotiana benthamiana and Arabidopsis thaliana plants that overexpressed VdAK dsRNAs had improved Vd resistance than the wild-type. RT-qPCR results showed that VdAK was also crucial for energy metabolism. Importantly, in an analysis of total small RNAs from Vd strains isolated from the transgenic plants, a small interfering RNA (siRNA) targeting VdAK was identified in transgenic N. benthamiana. Our results demonstrate that HIGS is a promising strategy for efficiently screening pathogenicity/virulence-related genes of Vd and that VdAK is a potential target to control this fungus.