PCR快速检测种子携带绿豆晕疫病菌方法的应用

doi:10.12190/j.issn.2096-1197.2021.05.15

北方农业学报 ›› 2021, Vol. 49 ›› Issue (5): 106-111.doi: 10.12190/j.issn.2096-1197.2021.05.15

PCR快速检测种子携带绿豆晕疫病菌方法的应用

田晓燕¹, 贺小勇¹, 孔庆全¹, 赵存虎¹, 陈文晋¹, 李志栋², 贾转青³

1.内蒙古自治区农牧业科学院植物保护研究所,内蒙古呼和浩特 010031;
2.呼和浩特市农业技术推广中心,内蒙古呼和浩特 010030;
3.呼和浩特市玉泉区教育教学研究中心,内蒙古呼和浩特 010031

收稿日期:2021-08-23 发布日期:2022-01-06
通讯作者: 贺小勇（1969—）,男,研究员,硕士,主要从事作物病虫害防治技术的研究工作。
作者简介:田晓燕（1984—）,女,助理研究员,硕士,主要从事农作物病虫害防控技术的研究工作。
基金资助:
财政部和农业农村部：国家现代农业产业技术体系-食用豆项目（CARS-08-Z06）; 内蒙古农牧业科技创新基金项目（2016CXJJN06）

Rapid detection of seed carrying Pseudomonas savastanoi pv. phaseolicola in mung beans using PCR

TIAN Xiaoyan¹, HE Xiaoyong¹, KONG Qingquan¹, ZHAO Cunhu¹, CHEN Wenjin¹, LI Zhidong², JIA Zhuanqing³

1. Institute of Plant Protection,Inner Mongolia Academy of Agricultural and Animal Husbandry Sciences,Hohhot 010031,China;
2. Hohhot Agricultural Technology Extension Center,Hohhot 010030,China;
3. Education and Teaching Research Center of Yuquan District of Hohhot,Hohhot 010031,China

Received:2021-08-23 Published:2022-01-06

摘要/Abstract

摘要： 【目的】通过PCR快速检测方法筛选对种子表面携带绿豆晕疫病菌灵敏度较高的引物,为绿豆晕疫病菌的检测工作提供技术支持。【方法】选择7对特异性引物,对绿豆晕疫病菌不同浓度菌悬液进行一系列的PCR反应,并验证其灵敏度。【结果】最终筛选出灵敏度较高的3对引物;模拟种子表面带菌检测结果表明,引物B4-1-F/B4-1-R和B4-2-F/B4-2-R检测的最小菌悬液浓度均为1×10⁴ CFU/mL,引物HB14F/HB14R检测的最小菌悬液浓度为1×10⁶ CFU/mL;种子带菌率检测结果表明,引物B4-1-F/B4-1-R和B4-2-F/B4-2-R振荡洗涤1 h即可检出带菌率为0.062 5%的种子样品,引物HB14F/HB14R振荡洗涤4 h后可检出带菌率为0.125%的种子样品。【结论】 PCR快速检测方法可直接对种子表面携带绿豆晕疫病菌进行检测,方便快捷。

关键词: 绿豆, 晕疫病, PCR, 种子带菌, 引物

Abstract: 【Objective】 Using PCR to rapidly screen primers with high sensitivity to the Pseudomonas savastanoi pv. phaseolicola on mung bean seed surface. Provide technical support for the detection of seed-carrying Pseudomonas savastanoi pv. phaseolicola in mung bean. 【Methods】 Seven pairs of specific primers were selected to perform a series of PCR reactions on Pseudomonas savastanoi pv. phaseolicola suspensions at different concentrations,and their sensitivity was verified. 【Results】 Three pairs of primers with high sensitivity were selected. The simulated seed surface bacteria detection results showed that the minimum concentration of bacterial suspension detected by primer B4-1-F/B4-1-R and B4-2-F/B4-2-R was 1×10⁴ CFU/mL,and the minimum concentration of bacterial suspension detected by primer HB14F/HB14R was 1×10⁶ CFU/mL. The results showed that primer B4-1-F/B4-1-R and B4-2-F/B4-2-R could detect the seed sample with seed-carrying rate of 0.062 5% after shaking and washing for 1 h,and primer HB14F/HB14R could detect the seed sample with seed-carrying rate of 0.125% after shaking and washing for 4 h. 【Conclusion】 PCR could be used to rapidly detect seed carrying the Pseudomonas savastanoi pv. phaseolicola on mung bean seed surface,which was convenient and quick.

Key words: Mung bean, Halo blight, PCR, Seed-carrying, Primer

中图分类号:

S432.4

田晓燕, 贺小勇, 孔庆全, 赵存虎, 陈文晋, 李志栋, 贾转青. PCR快速检测种子携带绿豆晕疫病菌方法的应用[J]. 北方农业学报, 2021, 49(5): 106-111.

TIAN Xiaoyan, HE Xiaoyong, KONG Qingquan, ZHAO Cunhu, CHEN Wenjin, LI Zhidong, JIA Zhuanqing. Rapid detection of seed carrying Pseudomonas savastanoi pv. phaseolicola in mung beans using PCR[J]. Journal of Northern Agriculture, 2021, 49(5): 106-111.

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PCR快速检测种子携带绿豆晕疫病菌方法的应用

Rapid detection of seed carrying Pseudomonas savastanoi pv. phaseolicola in mung beans using PCR

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