T-DNA随机插入法获得甘薯蔓割病菌非致病生防菌株  

Obtaining Nonpathogenic Biological Control Strains against Sweetpotato Fusarium Wilt by Agrobacterium-mediated Transformation

作　　者：张鸿[1] 林志坚[1] 林赵淼[1] 李国良[1] 许泳清[1] 刘中华[1] 邱永祥[1] 李华伟[1] 纪荣昌[1] 罗文彬[1] 汤浩[1] 邱思鑫[1] ZHANG Hong LIN Zhijian LIN Zhaomiao LI Guoliang XU Yongqing LIU Zhonghua QIU Yongxiang LI Huawei JI Rongchang LUO Wenbin TANG Hao QIU Sixin(Institute of Crop Sciences, Fujian Academy of Agricultttral Sciences, Fuzhou 350013, China)

机构地区：[1]福建省农业科学院作物研究所,福州350013

出　　处：《中国生物防治学报》2016年第5期610-618,共9页Chinese Journal of Biological Control

基　　金：福建省省属公益类科研院所基本科研专项(2014R1026-10);现代农业产业技术体系(CARS-11-B-10-2016);福建省农业科学院科技创新团队(CXTD-1-1301);福建省农科院青年人才创新基金(2014CX-10);福建省农科院作物所青年开放基金(2015QN-2)

摘　　要：由尖孢镰刀菌甘薯专化型Fusarium oxysporum f.sp.batatas引起的甘薯蔓割病是甘薯生产上一种重要病害。本研究将带有GFP绿色荧光蛋白基因的T-DNA随机插入到蔓割病菌基因组中,获得711个突变体。经过致病力筛选,获得了3株非致病突变体,编号为50-5、55-7和103-4,其中50-5和103-4产孢能力显著下降。平板对峙试验结果表明这3株非致病突变体均可和蔓割病菌发生营养竞争作用。通过提前对感病品种＂新种花＂接种非致病突变体,再接种蔓割病菌,结果显示当蔓割病菌和非致病突变体孢子浓度同为5×10~5 CFU/m L时,103-4和55-7分别预处理感病品种8 h后,生防效果趋于稳定,分别为86.95%和83.80%;50-5预处理16 h后生防效果趋于稳定,为75.65%。以上结果表明,通过农杆菌转化筛选非致病突变体的方法,成功获得3株蔓割病生防菌株,为深入开展甘薯蔓割病的生物防治提供基础,也为其他尖孢镰刀菌创造非致病生防菌提供借鉴。Sweetpotato Fusarium wilt is an important fungal disease caused by Fusarium oxysporum f. sp. batatas. In this study, Agrobacterium-mediated transformation was applied to randomly insert the T-DNA containing GFP gene into the genome of F. oxysporum and 711 mutants were obtained. Nonpathogenic mutant strains 50-5, 55-7 and 103-4 were selected from the mutant library. The sporulation capacity of 50-5 and 103-4 was lower compared with the wild type strain. All the 3 nonpathogenic mutant strains showed antagonistic effect when being co-cultured with the wild-type strain. When the spore concentration of both nonpathogenic mutants and pathogenic strains was 5×105 CFU/m L, the control efficiency of 50-5, 55-7, 103-4 increased with the prolonging pretreatment time and then became stable of 75.65%, 83.80%, 86.95% after 16, 8, 8 h pretreatment, respectively. In conclusion, these three nonpathogenic mutants could effectively control sweetpotato Fusarium wilt and Agrobacterium-mediated transformation could be an effective method to create nonpathogenic biological control agents of other fungal species.

关 键 词：甘薯蔓割病 尖孢镰刀菌 生物防治 非致病菌株 农杆菌转化 

分 类 号：S476[农业科学—农业昆虫与害虫防治]