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香蕉枯萎镰刀菌4号生理小种FoSP1基因功能的初步分析OA北大核心CSTPCD

Preliminary Analysis of FoSP1 Gene Function of Fusarium oxysporum f.sp.Cubense Race4

中文摘要英文摘要

香蕉枯萎病是我国香蕉生产中危害最大,且难以防治的一种真菌病害.由于香蕉枯萎病的影响,我国近年来香蕉种植面积逐年减少,给我国亚热带、热带香蕉产业带来了巨大的经济损失.该病害是由于香蕉的维管束受到病原菌尖孢镰刀菌古巴专化型(Fusarium oxysporum f.sp.cubense,Foc)入侵而发生,其繁殖速度快,引起香蕉叶片黄化和植株萎蔫.通过比较Foc1号生理小种(Foc1)N2菌株和4号生理小种(Foc4)B2菌株的基因组及转录组序列,筛选出位于基因组特异区间并且表达量较高的分泌蛋白FoSP1.结果表明:FoSP1基因开放阅读框为387 bp,编码129个氨基酸,其信号肽切割位点位于第20~21位氨基酸残基之间,且该蛋白无任何已知的结构域和功能位点,因此推断出FoSP1是一个新的分泌蛋白.为了研究该蛋白在Foc4菌株中的生物学作用,本研究利用split-marker的方法敲除了 B2菌株的FoSP1基因,并对获得的正确敲除突变体进行表型分析和致病性测定.结果表明:相比野生型B2菌株,FoSP1基因敲除突变体菌丝的营养生长无显著差异,对NaCl、山梨醇和H2O2等外源胁迫均表现不敏感,但敲除突变体的产孢量和分生孢子萌发率显著降低,且对巴西蕉苗的致病力明显减弱.由此推测分泌蛋白 FoSP1不参与Foc4菌株B2的营养生长,但在其产孢及致病的过程中发挥着重要作用.此结果为进一步研究Foc4基因组特异区间分泌蛋白的致病机制奠定基础.

Banana Fusarium wilt is a fungal disease which is the most harmful and difficult to control in banana produc-tion in China.Due to the influence of banana Fusarium wilt,the banana planting area in China has been decreasing year by year in recent years,which has brought huge economic losses to the subtropical and tropical banana industries in China.The disease occurred because the vascular bundle of banana was invaded by Fusarium oxysporum f.sp.cubense(Foc),and its propagation speed was fast,which caused the yellowing of banana leaves and plant wilting.By comparing the genome and transcriptome sequences of Foc race 1(Foc1,N2)and race 4(Foc4,B2),a secreted protein FoSP1 lo-cated in the lineage-specific region with high expression was screened out.The results showed that the open reading frame of FoSP1 gene was 387 bp,encoding 129 amino acids,and its signal peptide cleavage site was located between the 20th and 21st amino acid residues,and the protein had no known domain and functional site,so it was inferred that FoSP1 was a new secreted protein.In order to study the biological function of the protein in Foc4 strain,the split-marker method was used to knock out the FoSP1 gene of B2 strain,and the phenotype analysis and pathogenicity determination of the correctly knocked-out mutant were carried out.The results showed that compared with wild-type B2 strain,there was no significant difference in the vegetative growth of FoSP1 gene knockout mutants,and they were insensitive to exogenous stresses such as NaCl,D-sorbitol and H2O2,but the conidial yield and germination rate of knockout mutants were significantly reduced,and their pathogenicity to Brazilian banana was significantly weakened.It is inferred that the secreted protein FoSP1 does not participate in the vegetative growth of B2 strain,but plays an im-portant role in the process of sporulation and pathogenicity.This result lays a foundation for further study on the patho-genic mechanism of secreted proteins in the genome lineage-specific region of Foc4.

赵久娟;李敏;胡美姣;杨锦玉;何颖;柳志强;孙进华

中国热带农业科学院环境与植物保护研究所,海南海口 571101||海南大学生命健康学院,海南海口 570228中国热带农业科学院环境与植物保护研究所,海南海口 571101中国热带农业科学院环境与植物保护研究所,海南海口 571101||海南大学热带农林学院,海南海口 570228中国热带农业科学院环境与植物保护研究所,海南海口 571101||贵州大学精细化工研究开发中心,贵州贵阳 550025海南大学生命健康学院,海南海口 570228

生物学

尖孢镰刀菌古巴专化型FoSP1分泌蛋白基因敲除致病性

Fusarium oxysporum f.sp.cubenseFoSP1secreted proteingene knockoutpathogenicity

《热带作物学报》 2024 (007)

1332-1339 / 8

海南省自然科学基金项目(No.320RC696);国家自然科学基金项目(No.31872088);农业农村部财政专项(No.NFZX-2021).

10.3969/j.issn.1000-2561.2024.07.004

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