植物营养与肥料学报2017,23(5):1276-1285 Journal of Plant Nutrition and Fertilizer
doi: 10.11674/zwyf.17078 http:/www.plantnutrifert.org
钩状木霉ACCC31649的GFP标记及其对辣椒
定殖和促生作用
赵兴丽，陶刚*，赵琳，卯婷婷，王廿，顾金刚3（1贵州师范大学生命科学学院，贵阳550000；2贵州省植物保护研究所，贵阳550006
3中国农业科学院农业资源与农业区划研究所，北京100081）
摘要：【目的】旨在建立农杆菌介导的绿色荧光蛋白（GFP）基因转化钩状木霉（Trichodermahamatum）ACCC 31649的技术方法，筛选遗传稳定的GFP标记转化子，并研究该菌株在辣椒植株中定殖和对辣椒的促生作用，为进一步阐明霉在辣椒根际定殖及其与辣概病原菌在辣椒根际和植株中的定殖、互作和生防作用奠定基础。【方法】通过根瘤农杆菌介导的遗传转化方法筛选遗传稳定的GFP标记转化子，通过灌根接种方法和组织切片的水玻片荧光观紧研究了钩状本毒在辣椒植株中定殖过程和对辣椒的促生作用。【结果获得了遗传稳定 GFP标记的钩状本毒转化子。荧光显微观察表明，辣椒根、茎、叶组织中都检测到GFP标记菌株的定殖。标记菌株首先在根部定殖，然后通过根部维管束逐步定殖到茎和叶片组织中。野生型菌株和GFP标记菌株谨根接种4叶期辣椒幼苗，30天后，GFP标记菌株与水处理对照相比，辣的株高增长13.5%，根长增长16.2%，鲜重和干重分别增加了43.8%和45.3%，而且野生型菌株与GFP标记菌株对辣椒的促生作用没有显著差异。【结论】钩状木霉能够在辣概植株根、茎和叶组织中定殖，并且对辣椒其有显著的促生作用。同时，GFP标记
的钩状木霉将在进一步阐明该菌株在辣椒根际定殖及其对病原菌措抗和互作研究中发挥重要作用。关键词：绿色荧光蛋白；遗传转化；钩状木霉；辣椒；促生作用；定殖
GFP-labeledtransformationofTrichodermahamatumACCC31649andits
promotiononcolonizationandgrowthofpepperplants
ZHAO Xing-li', TAO Gangt, ZHAO Dai-lin', MAO Ting-ting', WANG Nian', GU Jin-gang
( I Guizhou Normal University, Guiyang 550000, China,; 2 Guizhou Institute of Plant Protection, Guiyang 550006, Chind,
3 Institute of.Agricultural Resources and Regional Planning, CAAS, Beijing J00081, China)
Abstract:[Objectives]The greenfluorescent protein (GFP)gene was transferred into Trichoderma hamatum ACCC31649 by the Agrobacterium tumefaciens-mediated transformation. The growth-promoting effect and the colonizing patterns in pepper plants of this strain were investigated in this paper, which would provide a technical basis for the research of the rhizosphere colonization of T. hamatum in pepper plants and biocontrol mechanism of antagonistic fungus against pathogen. [ Methods J The Agrobacterium tumefaciens-mediated transformation was used to select the GFP-labeled strain T. hamatum-GF21 which inherited stably, and the inoculation of T. hamatum by pouring root treatment and the observation of water slides of pepper tissue sections by fluorescence microscopy were used for the growth- promoting effect of pepper plants and its colonizing patterns. [ Results ] The stable GFP-labeled strain T. hamatum-GF21 was obtained. Fluorescence microscopy observation confirmed that the GFP-labeled strain T. hamatum-GF21 colonized in the roots, stems and leaves of the pepper. It colonized on the
收稿日期：2017-03-03
接受日期：2017-0322
网路出版日期：2017-06-01
基金项目：责州省优秀青年科技人才培养对象专项资金（黔科合人字（2015】27号）；责州省科学技术联合基金（黔科合LH字【
2015）7082号）；贵州农科院专项（黔农科院院专项【2016）029号）；贵州省农科院自主创新能力项目（黔农科院自主创新科研专项字（2014）002）资助。
作者简介：赵兴丽（1989—），女，责州六盘水人，颈士研究生，主要从事植物真菌病害生物防治研究。
E-mail:18786037305@163.com。*通信作者 E-mail;ttg729@sina.com; gujingang@caas.cn
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