尖孢镰刀菌ATMT体系的构建及其对甘草的促生效果评价  

Construction of ATMT system of Fusarium oxysporum and evaluation of its promoting effect on Glycyrrhiza uralensis

作　　者：陈嘉欣 朱泓宇 任广喜[1] 左鑫 姜丹[1] 刘春生[1] CHEN Jia-xin;ZHU Hong-yu;REN Guang-xi;ZUO Xin;JIANG Dan;LIU Chun-sheng(School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 102488,China)

机构地区：[1]北京中医药大学中药学院,北京102488

出　　处：《中国中药杂志》2024年第11期2882-2888,共7页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(82173921)。

摘　　要：该研究旨在通过耐盐能力、产吲哚乙酸(IAA)能力、溶磷能力、产铁载体能力等指标评价一株甘草内生尖孢镰刀菌的体内功能,通过根癌农杆菌介导的遗传转化技术建立该菌的稳定遗传转化体系,并通过标记基因绿色荧光蛋白(GFP)的克隆和β-葡萄糖苷酸酶(GUS)染色的效率检测转化子的稳定性与染色高效性,筛选出高效稳定的转化子用于回染甘草,评价其对甘草幼苗生长的影响。研究结果表明该菌耐盐性较好,在含7%氯化钠的马铃薯葡萄糖琼脂(PDA)培养基上仍能生长,但生长速度随着PDA培养基中氯化钠含量的增高而减缓;该菌具有产吲哚乙酸的功能,其发酵液中IAA的质量浓度约为3.32mg·m L^(-1)。该研究成功构建了该菌的遗传转化体系,且其根癌农杆菌介导的遗传转化(ATMT)体系高效而稳定。筛选获得1株兼具染色高效性和遗传稳定性的转化子,该转化子在甘草植株中的回染率为76.92%,能够显著提高一月龄甘草幼苗的主根长,促进甘草幼苗根部的生长发育。该研究结果可以为生物菌肥的开发及优质甘草的生长调控奠定基础。This study aims to evaluate the in vivo function of Fusarium oxysporum in Glycyrrhiza uralensis by salt tolerance,indoleacetic acid(IAA)production capacity,phosphate-dissolving capacity,and iron carrier production capacity.The stable genetic transformation system of the F.oxysporum was established by Agrobacterium tumefaciens-mediated genetic transformation(ATMT)technology,and the stability and staining efficiency of transformants were detected by the cloning of the marker gene green fluorescent protein(GFP)and the efficiency ofβ-glucuronidase staining(GUS).Efficient and stable transformants were selected for restaining G.uralensis and evaluating its influence on the growth of the G.uralensis seedlings.The results show that F.oxysporum has good salt tolerance and could still grow on potato glucose agar(PDA)medium containing 7%sodium chloride,but the growth rate slows down with the increase in sodium chloride content in PDA medium.F.oxysporum has the function of producing indoleacetic acid,and the concentration of IAA in its fermentation broth is about 3.32 mg·mL-1.In this study,the genetic transformation system of F.oxysporum is successfully constructed,and the ATMT system is efficient and stable.One transformant with both high staining efficiency and genetic stability is selected,and the restaining rate of the transformant in G.uralensis is 76.92%,which could significantly improve the main root length of one-month-old G.uralensis seedlings and promote the growth and development of G.uralensis seedlings.The results of this study can lay the foundation for the development of biological bacterial fertilizer and the growth regulation of high-quality G.uralensis.

关 键 词：尖孢镰刀菌 根癌农杆菌介导的遗传转化(ATMT) 体内功能 转化子 甘草 促生 

分 类 号：S567.71[农业科学—中草药栽培]