Rasche Frank, Hödl Verania, Poll Christian, Kandeler Ellen, Gerzabek Martin H, van Elsas Jan D, Sessitsch Angela
Department of Bioresources, ARC Seibersdorf Research GmbH, Seibersdorf, Austria.
FEMS Microbiol Ecol. 2006 May;56(2):219-35. doi: 10.1111/j.1574-6941.2005.00027.x.
A greenhouse experiment was performed to analyze a potential effect of genetically modified potatoes expressing antibacterial compounds (attacin/cecropin, T4 lysozyme) and their nearly isogenic, nontransformed parental wild types on rhizosphere bacterial communities. To compare plant transformation-related variations with commonly accepted impacts caused by altered environmental conditions, potatoes were cultivated under different environmental conditions, for example using contrasting soil types. Further, plants were challenged with the blackleg pathogen Erwinia carotovora ssp. atroseptica. Rhizosphere soil samples were obtained at the stem elongation and early flowering stages. The activities of various extracellular rhizosphere enzymes involved in the C-, P- and N-nutrient cycles were determined as the rates of fluorescence of enzymatically hydrolyzed substrates containing the highly fluorescent compounds 4-methylumbelliferone or 7-amino-4-methyl coumarin. The structural diversity of the bacterial communities was assessed by 16S rRNA-based terminal restriction fragment length polymorphism analysis, and 16S rRNA gene clone libraries were established for the flowering conventional and T4 lysozyme-expressing Desirée lines grown on the chernozem soil, each line treated with and without E. carotovora ssp. atroseptica. Both genetic transformation events induced a differentiation in the activity rates and structures of associated bacterial communities. In general, T4 lysozyme had a stronger effect than attacin/cecropin. In comparison with the other factors, the impact of the genetic modification was only transient and minor, or comparable to the dominant variations caused by soil type, plant genotype, vegetation stage and pathogen exposure.
进行了一项温室试验,以分析表达抗菌化合物(attacin/cecropin、T4溶菌酶)的转基因马铃薯及其近等基因、未转化的亲本野生型对根际细菌群落的潜在影响。为了将与植物转化相关的变异与环境条件改变所造成的普遍认可的影响进行比较,马铃薯在不同环境条件下种植,例如使用不同类型的土壤。此外,用黑胫病病原菌胡萝卜软腐欧文氏菌马铃薯亚种对植株进行接种。在茎伸长和初花期采集根际土壤样本。测定参与碳、磷和氮养分循环的各种细胞外根际酶的活性,以含有高荧光化合物4-甲基伞形酮或7-氨基-4-甲基香豆素的酶解底物的荧光速率表示。通过基于16S rRNA的末端限制性片段长度多态性分析评估细菌群落的结构多样性,并为在黑钙土上生长的传统Desirée品系和表达T4溶菌酶的Desirée品系建立16S rRNA基因克隆文库,每个品系分别接种和不接种胡萝卜软腐欧文氏菌马铃薯亚种。两种基因转化事件均导致相关细菌群落的活性速率和结构发生分化。总体而言,T4溶菌酶的作用比attacin/cecropin更强。与其他因素相比,基因改造的影响只是短暂和轻微的,或与土壤类型、植物基因型、植被阶段和病原菌接种所引起的主要变异相当。