Secondary amplification of siRNA machinery limits the application of spray-induced gene silencing
Secondary amplification of siRNA machinery limits the application of spray-induced gene silencing
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siRNA机器的二次扩增限制了喷雾诱导的基因沉默的应用
DOI：
10.1111/mpp.12728
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发表时间：
2018-12-01
影响因子：
4.9
通讯作者：
Zhou, Ming-Guo
中科院分区：
文献类型：
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作者：
Song, Xiu-Shi;Gu, Kai-Xin;Zhou, Ming-Guo
喷雾诱导的基因沉默（SIGS）是一种创新的作物保护策略。然而，SIGS的机制尚不清楚。在这里，我们首先证明了二级小干扰RNA（siRNA）扩增限制了SIGS的应用。选择肌球蛋白5基因（Myo 5）作为农学上重要病原体-亚洲镰孢的SIGS靶点。发现对应于Myo 5基因的不同区域的五个片段有效地沉默Myo 5，导致细胞壁缺陷、生命周期破坏和毒力降低。Myo 5 -8（Myo 5片段之一）在F. asiaticum、F.禾谷镰孢F. tricinctum和F.尖孢菌，但不是在其他真菌，在体外。值得注意的是，Myo 5的沉默仅持续9小时，除非双链RNA（dsRNA）的持续供应，因为F。asiaticum不能维持siRNA扩增。在对植物喷洒后，dsRNA通过受伤表面更有效地被吸收。植物细胞吸收的双链RNA的抗真菌活性比干燥到植物表面的双链RNA更高且持续时间更长。与真菌中的dsRNA相比，植物细胞中的dsRNA可以通过二级扩增机制有效地转化为大量的siRNA。我们的研究结果提供了新的意义，发展SIGS作为一个主流的疾病控制策略，对镰刀菌和其他真菌。
Spray-induced gene silencing (SIGS) is an innovative strategy for crop protection. However, the mechanism of SIGS is not known. Here, we first demonstrate that secondary small interfering RNA (siRNA) amplification limits the application of SIGS. A myosin5 gene (Myo5) was chosen as the target of SIGS in an agronomically important pathogen-Fusarium asiaticum. Five segments corresponding to the different regions of the Myo5 gene were found to efficiently silence Myo5, resulting in cell wall defects, life cycle disruption and virulence reduction. Myo5-8 (one of the Myo5 segments) induced sequence-specific RNA interference (RNAi) activity in F. asiaticum, F. graminearum, F. tricinctum and F. oxysporum, but not in other fungi, in vitro. Remarkably, the silencing of Myo5 lasted for only 9 h unless the double-stranded RNA (dsRNA) was continuously supplied, because F. asiaticum is unable to maintain siRNA amplification. After spraying on plants, dsRNAs were more efficiently taken up via the wounded surface. The antifungal activity of dsRNAs taken up by plant cells was higher and longer lasting than that dried onto the plant surface. In contrast with dsRNAs in fungi, dsRNAs in plant cells could efficiently turn into substantial siRNAs via secondary amplification machinery. Our findings provide new implications to develop SIGS as a mainstream disease control strategy against Fusarium and other fungi.