Watrud LS, Seidler RJ, Widmer F
National Research Council Research Associate, US EPA National Health and Environmental Effects Research Laboratory-Western Ecology Division, 200 SW 35th Street, Corvallis, OR 97333, USA
Microb Ecol. 1999 Feb;37(2):129-139. doi: 10.1007/s002489900137.
Abstract Rhizosphere bacterial communities of parental and two transgenic alfalfa (Medicago sativa L.) of isogenic background were compared based on metabolic fingerprinting using Biolog GN microplates and DNA fingerprinting of bacterial communities present in Biolog GN substrate wells by enterobacterial repetitive intergenic consensus sequence-PCR (ERIC-PCR). The two transgenic alfalfa expressed either bacterial (Bacillus licheniformis) genes for alpha-amylase or fungal (Phanerochaete chrysosporium) genes for Mn-dependent lignin peroxidase (Austin S, Bingham ET, Matthews DE, Shahan MN, Will J, Burgess RR, Euphytica 85:381-393). Cluster analysis and principal components analysis (PCA) of the Biolog GN metabolic fingerprints indicated consistent differences in substrate utilization between the parental and lignin peroxidase transgenic alfalfa rhizosphere bacterial communities. Cluster analysis of ERIC-PCR fingerprints of the bacterial communities in Biolog GN substrate wells revealed consistent differences in the types of bacteria (substrate-specific populations) enriched from the rhizospheres of each alfalfa genotype. Comparison of ERIC-PCR fingerprints of bacterial strains obtained from substrate wells to substrate community ERIC-PCR fingerprints suggested that a limited number of populations were responsible for substrate oxidation in these wells. Results of this study suggest that transgenic plant genotype may affect rhizosphere microorganisms and that the methodology used in this study may prove a useful approach for the comparison of bacterial communities.
基于使用Biolog GN微孔板的代谢指纹分析以及通过肠杆菌重复基因间共有序列-PCR（ERIC-PCR）对Biolog GN底物孔中存在的细菌群落进行DNA指纹分析，比较了亲本苜蓿和两个同基因背景的转基因苜蓿（紫花苜蓿）根际细菌群落。这两种转基因苜蓿分别表达了用于α-淀粉酶的细菌（地衣芽孢杆菌）基因或用于锰依赖性木质素过氧化物酶的真菌（黄孢原毛平革菌）基因（奥斯汀S、宾厄姆ET、马修斯DE、沙汉MN、威尔J、伯吉斯RR，《植物遗传资源学报》85:381 - 393）。Biolog GN代谢指纹的聚类分析和主成分分析（PCA）表明，亲本苜蓿和木质素过氧化物酶转基因苜蓿根际细菌群落在底物利用方面存在一致差异。对Biolog GN底物孔中细菌群落的ERIC-PCR指纹进行聚类分析，揭示了从每种苜蓿基因型根际富集的细菌类型（底物特异性种群）存在一致差异。将从底物孔中获得的细菌菌株的ERIC-PCR指纹与底物群落ERIC-PCR指纹进行比较，表明在这些孔中有限数量的种群负责底物氧化。本研究结果表明，转基因植物基因型可能影响根际微生物，并且本研究中使用的方法可能被证明是比较细菌群落的一种有用方法。
