香蕉种苗中枯萎病菌和细菌性软腐病菌实时荧光PCR检测OA
Detection of Fusarium Wilt and Bacterial Soft Rot of Banana Seedlings by Real-Time Fluorescence Quantitative PCR
香蕉枯萎病菌和细菌性软腐病菌复合侵染且潜伏期较长,建立香蕉种苗病害早期监测的分子检测方法非常重要.本文基于尖孢镰刀菌古巴专化型1号生理小种(Fusarium oxy-sporum f.sp.cubense race 1,FOC1)基因组Contig 438区间(35631～37693 bp)(GenBank:AMGP01000438.1)设计特异扩增引物qFOC1-F/-R、4号生理小种(Fusarium oxysporum f.sp.cubense race 4,FOC4)基 因 组 Contig 195 区 间(4028～6126 bp)(GenBank:AMGQ01000195.1)设计特异扩增引物qFOC4-F/-R,同时以香蕉细菌性软腐病菌Dickeya zeae的促旋酶B 亚单位基因(Subunit B of gyrase gene)(GenBank:JQ284039)序列设计特异扩增引物qgyrB-F/-R.qPCR检测结果显示:实时荧光检测引物扩增特异性强.通过拟合曲线方程分析得到:检测香蕉枯萎病菌的DNA浓度最低限为3 pg·μL-1,细菌性软腐病菌的菌液浓度最低限灵敏度为70 cfu·mL-1,检测灵敏度较高,结果稳定可靠.因此,本研究建立的实时荧光PCR检测方法可有效应用于检测香蕉种苗中的香蕉枯萎病菌和细菌性软腐病菌,确保种苗的安全生产.
s:Establishing a molecular detection method for early monitoring of banana seedling diseases is important because of mixed infection and long incubation of banana Fusarium wilt and bacterial soft rot pathogens.The specific amplification primers of qFOC1-F/-R were designed based on the sequences of the contig 438(35631-37693 bp)(AMGP01000438.1)in the DNA of Fusarium oxysporum f.sp.cubense race 1(FOC1)and qFOC4-F/-R based on the contig 195(4028-6126 bp)(AMGQ01000195.1)in the DNA of F.oxysporum f.sp.cubense race 4(FOC4).Based on sequence of subunit B of gyrase gene of banana Fusarium wilt pathogen(GenBank:JQ284039),Dickeya zeae,the primers of qgyrB-F/-R were designed.The results of qPCR showed that the specificity of primers amplification was very strong.Curve-fitting equation analysis showed that:the minimum limit of DNA concentration for detection of F.oxysporum f.sp.cubense was 3 pg·μL-1,and the sensitivity de-tection of D.zeae was 70 cfu·mL-1;with high sensitivity if the detection,the results were stable and reliable.Therefore,the qPCR detection method developed in this study is effective to detect banana Fusarium wilt and bacterial soft rot pathogens of banana seedlings to ensure the safe production of seedlings.
梁基校;陈东;利齐欣;林鸿生;张景欣;杨祁云;王飞钊;蒲小明
阳江市阳东区农业科学研究所,广东阳江 529900博罗县供销合作联社,广东博罗 516100阳江市阳东区农业科学研究所,广东阳江 529900汕尾供销中禾农业科技服务有限公司,广东海丰 516400广东省农业科学院植物保护研究所/农业农村部华南果蔬绿色防控重点实验室/广东省植物保护新技术重点实验室,广东广州 510640广东省农业科学院植物保护研究所/农业农村部华南果蔬绿色防控重点实验室/广东省植物保护新技术重点实验室,广东广州 510640广东天禾农资股份有限公司,广东广州 510080广东省农业科学院植物保护研究所/农业农村部华南果蔬绿色防控重点实验室/广东省植物保护新技术重点实验室,广东广州 510640
植物保护学
尖孢镰刀菌古巴专化型1号菌尖孢镰刀菌古巴专化型4号菌玉米迪基氏菌实时荧光PCR检测
Fusarium.oxysporum f.sp.cubense race 1Fusarium.oxysporum f.sp.cubense race 4Dickeya zeaereal-time fluorescence quantitative PCRdetection
《农业研究与应用》 2023 (5)
8-14,7
广东省现代农业产业绿色发展共性关键技术研发创新团队建设项目(2022KJ112)广东省烟草科技项目(201802,201908,202201)广东丝苗米跨县集群产业园(汕尾市)项目(2022-2023)博罗蔬菜产业科技支撑项目(2022-2023).
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