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哈茨木霉T2-16突变体库的构建及突变株表型分析
何英,张屹,肖姬玲,朱菲莹,魏林,梁志怀
0
(湖南大学研究生院隆平分院,长沙 410125;湖南省农业生物技术研究所,长沙 410125;湖南省植物保护研究所,长沙 410125)
摘要:
为了优化高效拮抗生防菌哈茨木霉T2-16的EMS诱变条件,构建哈茨木霉T2-16突变体库,为哈茨木霉功能基因组研究奠定材料基础,以哈茨木霉T2-16为试验材料,采用单因素试验以0~0.6 mol·L-1 EMS溶液分别处理不同浓度孢子0~6 h,明确适于哈茨木霉诱变的EMS最佳浓度、处理时间和孢子浓度。以最适参数诱变处理野生型菌株,进行生长势、拮抗能力和生防效果比较。结果显示,0.4 mol·L-1 EMS溶液处理浓度为106 ·mL-1的孢子4 h,其致死率接近70%,诱变效果最佳。经生长势、拮抗能力和生防效果比较发现,生长势呈正向突变占有率达46.20%,其中,16株突变菌株拮抗作用明显增强,T42和T62的生防效果分别高出野生型菌株31.64%和27.91%,为研究哈茨木霉与生防相关的功能基因打下基础。
关键词:  哈茨木霉T2-16  EMS诱变  突变体库  表型分析
DOI:10.13610/j.cnki.1672-352x.20200113.001
基金项目:湖南农业科技创新项目(2017JC77), 国家重点研发计划( 2017YFD0200606)和国家公益性行业(农业)科研专项(201503110-03)共同资助。
Construction of Trichoderma harzianum T2-16 mutant library and phenotypic analysis of mutant
HE Ying,ZHANG Yi,XIAO Jiling,ZHU Feiying,WEI Lin,LIANG Zhihuai
(Longping Branch, Graduate School of Hunan University, Changsha 410125;Hunan Agricultural Biotechnology Research Institute, Changsha 410125;Institute of Plant Protection of Hunan Province, Changsha 410125)
Abstract:
To optimize the EMS mutagenesis conditions of Trichoderma harzianum T2-16, we constructed a mutant library of T. harzianum T2-16 to lay a material foundation for the study of the functional genome of T. harzianum. Using T. harzianum T2-16 as the experimental material, the spores were treated with 0-0.6 mol·L-1 EMS solution for 0-6 h, respectively. The optimum concentration, treatment time and spore concentration of EMS mutagenesis were determined. The wild-type strains were mutagenized with the optimum parameters, and the growth potential, antagonistic ability and biocontrol effect were compared. The results showed that spores treated with 0.4 mol·L-1 EMS solution at 106 ·mL-1 spore concentration for 4 h had the best mutagenic effect, with the lethal rate of about 70%. By comparing growth potential, antagonistic ability and biocontrol effect, it was found that the positive mutation rate in growth potential was 46.20%. Among them, the antagonistic effects of 16 mutant strains were significantly enhanced. The biocontrol effect of T42 and T62 was 31.64% and 27.91% higher than that of wild type strains, respectively.
Key words:  Trichoderma harzianum T2-16  EMS mutagenesis  mutant library  phenotypic analysis

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