园艺学报 ›› 2023, Vol. 50 ›› Issue (2): 279-294.doi: 10.16420/j.issn.0513-353x.2021-1081

• 研究论文 • 上一篇    下一篇


张欣, 漆艳香, 曾凡云, 王艳玮, 谢培兰, 谢艺贤*(), 彭军*()   

  1. 中国热带农业科学院环境与植物保护研究所,农业农村部热带作物有害生物综合治理重点实验室,海南省热带农业有害生物监测与控制重点实验室,海口 571101
  • 收稿日期:2022-06-16 修回日期:2022-09-27 出版日期:2023-02-25 发布日期:2023-03-06
  • 通讯作者: *(
  • 基金资助:

Functional Analysis of Dicer-like Genes in Fusarium oxysporum f. sp. cubense Race 4

ZHANG Xin, QI Yanxiang, ZENG Fanyun, WANG Yanwei, XIE Peilan, XIE Yixian*(), PENG Jun*()   

  1. Key Laboratory of Integrated Pest Management on Tropical Crops,Ministry of Agriculture and Rural Affairs,Hainan Key Laboratory for Monitoring and Control of Tropical Agricultural Pests,Environment and Plant Protection Institute,Chinese Academy of Tropical Agricultural Sciences,Haikou 571101,China
  • Received:2022-06-16 Revised:2022-09-27 Online:2023-02-25 Published:2023-03-06
  • Contact: *(


香蕉枯萎病病原菌尖孢镰刀菌古巴专化型(Fusarium oxysporum f. sp. cubense)4号生理小种(Foc4)含有两个进化上高度保守的Dicer-like基因FocDCL1FocDCL2。为了探究该基因的功能及其在RNAi中的作用机制,利用同源重组的方法获得ΔFocDCL1ΔFocDCL2ΔFocDCL1/2基因敲除突变体。表型分析显示,与Foc4野生型相比,突变体菌丝的营养生长无显著差异,但产孢量下降;ΔFocDCL2突变体在非生物胁迫刚果红处理后菌落明显变小且气生菌丝增多;ΔFocDCL2ΔFocDCL1/2的致病力下降。miRNA深度测序数据显示,与Foc4野生型相比,ΔFocDCL1ΔFocDCL2ΔFocDCL1/2敲除突变体的miRNA长度分布和5′-端首位碱基出现频率都发生了改变,都能产生自身特异miRNA。DCL功能存在交叉和冗余,miRNA可以通过依赖独立的、交叉的或者联合的DCL发生;此外,鉴定到不依赖DCL形成的miRNA。这些结果表明FocDCL在产孢量、非生物胁迫、致病力以及小RNA发生中发挥作用。

关键词: 香蕉, 枯萎病, Fusarium oxysporum f. sp. cubense, Dicer-like基因, 致病力测试, miRNA


The fungal pathogen Fusarium oxysporum f. sp. cubense(Foc)is an important soil-borne pathogenic fungus which causes Banana Fusarium wilt. The Foc race 4(Foc 4)attacking almost all Cavendish cultivars is currently noteworthy in global banana production. In order to explore the function of DCL-like genes in Foc4,the gene-knockout mutants ΔFocDCL1ΔFocDCL2 and ΔFocDCL1/2 were obtained by homologous recombination technique. The phenotype analysis showed the mutants showed no significant differences on vegetative growth but significant reduced in conidia production compared to the Foc4. The ΔFocDCL2 mutant increased sensitive to Congo red,and the mutant significantly reduced the size of the colony and increased the number of aerial hyphae after Congo red treatment. Next,the pathogenicity test showed that virulence of ΔFocDCL2 and ΔFocDCL1/2 were significantly reduced compared with Foc4,respectively. Furthermore,miRNA deep sequencing data showed the length distribution of the total reads,and the frequency of the 5′ first nucleotide bias of the mutants were markedly changed compared with Foc4,respectively. The miRNA could be generated depending on individual,alternative or joint DCL manners,indicating the FocDCL with overlapped and redundant function in small RNA biogenesis. In addition,DCLs-independent miRNA also be identified in the sequencing data. These results indicated that FocDCL function involved in the stress response,conidia,pathogenicity and generation of miRNAs in Foc4.

Key words: banana, Fusarium wilt, Fusarium oxysporum f. sp. cubense, Dicer-like gene, pathogenicity test, miRNA