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| 甘蔗梢腐病病原菌Fusarium sacchari Nep1-like蛋白的筛选鉴定及功能分析 | |
| 引用本文： | 黄振,李慧雪,周宇明,暴怡雪,张木清,姚伟.甘蔗梢腐病病原菌Fusarium sacchari Nep1-like蛋白的筛选鉴定及功能分析[J].植物病理学报,2022,52(2):156-164. |
| 作者姓名： | 黄振 李慧雪 周宇明 暴怡雪 张木清 姚伟 |
| 作者单位： | 亚热带农业生物资源保护与利用国家重点实验室,南宁 530004; 广西甘蔗生物学重点实验室,南宁 530004; 广西大学农学院,南宁 530004 |
| 基金项目： | 国家自然科学基金(31760413,32001603)；;广西自然科学基金(2018JJA130113)； |
| 摘 要： | 甘蔗镰孢菌Fusarium sacchari(F. sacchari)引起的甘蔗梢腐病严重影响了甘蔗的产量和质量。解析病原菌的致病机理,对指导甘蔗抗病育种和绿色防控具有重要意义。研究发现NLPs (Nep1-like proteins)类基因在真菌侵染及定殖过程中发挥重要作用。本实验在对梢腐病病原菌基因组测序(尚未公布)基础上,通过BLASTP分析得到甘蔗梢腐病病原菌F. sacchari中4个NLP家族基因Fs_00548、Fs_03159、Fs_06646、Fs_11062。将克隆到的3个基因(Fs_00548、Fs_03159、Fs_11062;Fs_06646未克隆到)构建至PVX载体,利用农杆菌介导的烟草叶片瞬时表达系统,发现只有Fs_00548可以诱导细胞产生与Bax相似的坏死,Fs_03159和Fs_11062则不能。利用酵母信号肽分泌功能验证方法,发现Fs_00548和Fs_03159的信号肽具有分泌活性。并且Fs_00548的信号肽区域对其发挥功能具有重要作用。qRT-PCR结果显示该基因在侵染过程中均有表达,其中72 h表达量最高,是菌丝中该基因表达量的48倍。以上研究结果表明,Fs_00548在病原菌侵染过程中发挥重要作用,研究结果为进一步探究病原菌与甘蔗互作提供参考。 |
| 关 键 词： | 甘蔗 梢腐病 NLPs(Nep1-like proteins) PTI |
| 收稿时间： | 2021-02-18 |
Identification and functional analysis of Nep1-like proteins of Fusarium sacchari, the pathogen of sugarcane pokkah boeng disease |
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| HUANG Zhen,LI Huixue,ZHOU Yuming,BAO Yixue,ZHANG Muqing,YAO Wei.Identification and functional analysis of Nep1-like proteins of Fusarium sacchari, the pathogen of sugarcane pokkah boeng disease[J].Acta Phytopathologica Sinica,2022,52(2):156-164. | |
| Authors: | HUANG Zhen LI Huixue ZHOU Yuming BAO Yixue ZHANG Muqing YAO Wei |
| Institution: | State Key Lab of Conservation and Utilization of Agric-Biological Resources, Guangxi University, Nanning, Guangxi, 530004, China; Guangxi Key Lab of Sugarcane Biology, Nanning, Guangxi, 530004, China; Department of Agriculture, Guangxi University, Nanning, Guangxi, 530004, China |
| Abstract: | Pokkah boeng disease, causing agent Fusarium sacchari, is a serious disease on sugarcane and causes substantial reduction in the yield and the quality. It is of great significance to elucidate the pathogenic mechanism, which can provide insights for sugarcane disease resistance breeding. Our studies have found that NLP (Nep1-like protein) genes play important roles in fungal infection and colonization. Based on genome sequence of the pathogen, four NLP genes (Fs_00548, Fs_03159, Fs_06646, and Fs_11062) were selected out through sequence similarity using BLASTP. Among these genes, only Fs_00548 induce cell death in Nicotiana tabacum leaves by transient expression, as consistent with the Bax-induced apoptotic cell death. Moreover, yeast invertase secretion assay showed that Fs_00548 was a secreted protein with a signal peptide that played a key role in the pathogenesis. Infectivity rate of Fs_00548 was further confirmed through qRT-PCR analyses, which reported the maximum expression of Fs_00548 at 72 hours post infection. Altogether, these results suggest that Fs_00548 plays an important role in the infection process of F. sacchari and provide understanding for further research on the interaction between sugarcane and F. sacchari. |
| Keywords: | Suagrcane pokkah boeng disease NLP (Nep1-like protein) PTI |
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