南方农业学报2023，Vol.54Issue(8)：2352-2359,8.DOI:10.3969/j.issn.2095-1191.2023.08.016
大蕉枯萎病菌二重PCR分子快速检测技术的建立
Development of duplex PCR molecular rapid detection for fusarium wilt of Dajiao
摘要
Abstract
[Objective]The objective of this study was to establish duplex PCR molecular detection for fusarium wilt strain of Dajiao based on gene sequences specific to 2 different differentiation lineages of Dajiao fusarium wilt strain,which could provide theoretical basis for prevention and control of banana fusarium wilt.[Method]Multiple pairs of PCR primers were designed based on rDNA intergene region(IGS)unique to pathogen of fusariun wilt pathogen of Dajiao,DNAs of 8 fusariun wilt strains of Dajiao,3 fusarium wilt strains of Fenjiao from Guangdong,2 fusarium wilt strains of Fenjiao from Guangxi and Hainan respectively,5F.oxysporumstrains and 5 outgrouped strains were amplified by PCR as templates to screen the specific primers for the fusarium wilt strain of Dajiao.The specificity and sensitivity of the primers were tested,and the lower limit of strain content detection of duplex PCR system was determined.[Result]Several pairs of PCR primers were designed based on the specific gene sequence of fusarium wilt strain of Dajiao.After screening and verification,2 pairs of specific primers were obtained that could be used to detect 2 different lineages of fusarium wilt strains(Lineage Ⅰ and Lineage Ⅱ),which could be directly applied to the detection of Dajiao fusarium wilt strains in Dajiao planting soil and Dajiao plants.The amplified fragments were 755 and 590 bp,respectively.When the amplified band was 755 bp,it indicated that Dajiao fusarium wilt strain was contained in the sample.When the amplified band was 590 bp,it indicated that the sample contained the strains of Fenjiao or Dajiao fusarium wilt.The sensitivity of the duplex PCR detection system for Dajiao fusarium wilt strain hypha was up to 0.4 μg,and the lower limit of soil strain content was 40 spores per gram of soil.[Conclusion]The established duplex PCR detection technique can simultaneously detect and distinguish two lineages of fusariun wilt strains of Dajiao and the dominant group of fusarium wilt of Fenjiao in Guangdong area in one PCR reaction,which can be used for the detection and screening of disease-free Fenjiao and Dajiao seedlings and planting soil.关键词
大蕉/枯萎病菌/尖孢镰孢菌古巴专化型/二重PCR检测Key words
Dajiao/fusarium wilt strain/Fusarium oxysporum f.sp.cubense/duplex PCR detection分类
农业科技引用本文复制引用
曾莉莎,王芳,周海琪,伍泽星,赖永超,傅长根,吕顺,梁少丽,刘丽琴..大蕉枯萎病菌二重PCR分子快速检测技术的建立[J].南方农业学报,2023,54(8):2352-2359,8.基金项目
广东省基础与应用基础研究基金项目(2022A1515140114) （2022A1515140114）
广东省现代农业产业技术体系创新团队项目(2022KJ109) （2022KJ109）
东莞市2021年度省乡村振兴战略专项(20211800400052) （20211800400052）
东莞市科技特派员项目(20221800500062) Guangdong Basic and Applied Basic Research Fund Project(2022A1515140114) （20221800500062）
Guangdong Modern Agricultural Industrial Technology System Innovation Team Project(2022KJ109) （2022KJ109）
Dongguan 2021 Provincial Ru-ral Revitalization Strategy Special Project(20211800400052) （20211800400052）
Dongguan Science and Technology Special Commissioner Project(20221800500062) （20221800500062）
