小麦内生固氮菌分离及其ACC脱氨酶测定-中国农业科学院农业资源.PDF

2012,45(6):1066-1073 Scientia Agricultura Sinica doi: 10.3864/j.issn.0578-1752.2012.06.004 / 100081 · · µα· ·µα· · Isolation of Wheat Endophytic Diazotrophs and Determination of 1-Aminocyclopropane-1-Carboxylate Deaminase QIN Bao-jun, LUO Qiong, GAO Miao, HU Hai-yan, XU Jing, ZHOU Yi-qing, SUN Jian-guang (Institute of Agricultural Resources and Regional Planning, Chinese Academy of Agricultural Sciences/Key Laboratory of Crop Nutrition and Fertilization of Ministry of Agriculture, Beijing 100081) Abstract: Objective The objective of this study is to determine the ammount of wheat endophytic diazotrophs and screen for ACC (1-aminocyclopropane-1-carboxylate) deaminase activity from the diazotrophs, determine the phylogenetic and classific position of selected strains and prepare strains for microbial fertilizer production. Method Surface sterilization and nitrogen-free medium were used to isolate diazotroph and ACC was used as sole nitrogen source to screen strains with ACC deaminase activity. Nitrogenase activity was determined with acetylene reduction assay. 16S rDNA was amplified with PCR and analysed with MEGA software. Strain identification was carried out based on the morphology, physiology, biochemical test results and 16S rDNA analysis. Result The ammount of endophytic diazotrophs at jointing stage of wheat was (0.2-17.8)×105 cfu·g-1 fresh weight. Sixty endophytic diazotrophs with nitrogenase activity ranging 1-36 nmol C H /h·mg protein