新疆外来入侵杂草豚草属LAMP快速检测技术的建立  

Establishment of loop-mediated isothermal amplification for rapid detection of the exotic invasive weed Ambrosia L.in Xinjiang

作　　者：于海鑫 付开赟 丁新华[2] 贾尊尊 吐尔逊·阿合买提[2] 王兰[1] 郭文超[2] YU Haixin;FU Kaiyun;DING Xinhua;JIA Zunzun;TURERXUN·Ahemaiti;WANG Lan;GUO Wenchao(College of Agronomy,Tarim University,Alar,Xinjiang 843300,China;Key Laboratory of Integrated Management of Harmful Crop Vermin of China Northwestern Oasis,Ministry of Agriculture and Rural Affairs of the P.R.C./Institute of Plant Protection,Xinjiang Academy of Agricultural Sciences,Urumqi,Xinjiang 830091,China)

机构地区：[1]塔里木大学农学院,新疆阿拉尔843300 [2]新疆农业科学院植物保护研究所/农业农村部西北荒漠绿洲作物有害生物综合治理重点实验室,新疆乌鲁木齐830091

出　　处：《生物安全学报》2023年第3期277-281,共5页Journal of biosafety

基　　金：新疆外来入侵生物监测检测数据共享平台(PT2028);新疆农业科学院科技创新重点培育专项(xjkcpy-002);新疆农业科学院科技创新重点培育专项(2022-2024)。

摘　　要：【目的】开发外来入侵生物三裂叶豚草和豚草不同生育期、不同部位的环介导等温扩增(loop-mediated isothermal amplification,LAMP)技术,以达到田间快速、准确和高效识别的目的。【方法】以SYBR Green Ⅰ为指示剂,分别针对三裂叶豚草和豚草不同发育阶段(幼苗期、生长期、种子期)开展LAMP技术开发。【结果】特异性验证结果显示,所检测杂草的LAMP产物均呈阳性(产生白色沉淀),而与其对照的其他2种杂草的LAMP产物均为阴性(无白色沉淀)。灵敏度检测结果显示,该体系的DNA最低检测限为10^(-10) ng·μL-1,比常规聚合酶链式反应灵敏度高。【结论】本研究建立的LAMP检测体系能有效应用于三裂叶豚草和豚草样本的快速检测,为其快速、高效识别提供技术支撑。【Aim】This study aimed to develop a loop-mediated isothermal amplification(LAMP)technique for rapid,accurate,and efficient identification of the invasive alien species ragweed(Ambrosia trifida L.)and giant ragweed(A.artemisiifolia L.)at different growth stages using various plant parts.【Method】SYBR Green I was used as an indicator to develop the LAMP method for identifying exotic invasive weed at difficult-to-distinguish periods,such as seeds,seedling,and growth stages.【Result】Specific validation results showed that all LAMP products were positive(white precipitation)in the ragweed and giant ragweed testing groups,whereas all LAMP products pertaining to the other tested weeds were negative(transparent).Sensitivity test results revealed that the minimum DNA detection threshold in the system was 10^(-10) ng·μL^(-1),which was more sensitive than that revealed by conventional polymerase chain reaction(PCR)techniques.【Conclusion】Thus,the LAMP detection system established in this study can provide effective technical support for the rapid detection of ragweed and giant ragweed samples.

关 键 词：三裂叶豚草 豚草 环介导等温扩增(LAMP) 快速检测技术 

分 类 号：S451[农业科学—植物保护]