Leaf-associated bacteria from transgenic white poplar producing resveratrol-like compounds: isolation, molecular characterization, and evaluation of oxidative stress tolerance
Leaf-associated bacteria from transgenic white poplar producing resveratrol-like compounds: isolation, molecular characterization, and evaluation of oxidative stress tolerance
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DOI：
10.1139/w09-038
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发表时间：
2009-07-01
影响因子：
2.8
通讯作者：
Carbonera, Daniela
中科院分区：
文献类型：
--
作者：
Balestrazzi, Alma;Bonadei, Martina;Carbonera, Daniela
本研究的目的是分离和鉴定转基因白杨（Populus alba L.）叶片中的可培养细菌。‘Villafranca’)用StSy基因进行工程改造，生产白藜芦醇样化合物。少量的白藜芦醇糖苷可以从天然来源或通过昂贵的化学合成程序获得。大规模生产这些相关药物的另一种方法是使用转基因植物作为生物反应器，尽管在田间条件下生长的植物中出现的新分子可能在一定程度上干扰相关的微生物种群。从2个产生白藜芦醇糖苷的StSy转基因株系和一个在温室中生长的未转化株系的叶片中分离出附生植物和内生植物。11株分离菌株经16S rdna鉴定为芽孢杆菌属。另外，2株分离物分别属于Curtobacterium属和Kocuria属。对过氧化氢、UV-C和百草枯的耐受性进行了评估，对叶子相关细菌的游动和成群运动进行了评估。有趣的是，与从未转化的杨树系中恢复的分离株相比，从转基因组织中恢复的分离株显示出抵抗氧化应激的能力。体外生物实验表明，反式白藜芦醇抑制了所有被试细菌的群集和游泳运动。反式白藜芦醇对运动所需的鞭毛蛋白产生的影响也通过免疫印迹分析进行了研究。
The aim of this study was the isolation and characterization of the culturable bacteria inhabiting the leaves of transgenic white poplars (Populus alba L. 'Villafranca') engineered with the StSy gene for the production of resveratrol-like compounds. Resveratrol glucosides are available in small amounts from natural sources or by expensive chemical synthesis procedures. An alternative approach for the large-scale production of these relevant pharmaceuticals is the use of transgenic plants as bioreactors, although the occurrence of novel molecules in plants growing under field conditions might interfere, to some extent, with the associated microbial population. Both epiphytes and endophytes were isolated from the leaves of 2 StSy transgenic lines producing resveratrol glucosides and from an untransformed plant line grown in a greenhouse. Eleven isolates were recovered and classified as members of the genus Bacillus by 16S rDNA-based analysis. In addition, 2 isolates were classified as members of the Curtobacterium and Kocuria genera, respectively. Tolerance to hydrogen peroxide, UV-C, and paraquat was evaluated, as were the swimming and swarming motility of the leaf-associated bacteria. Interestingly, the isolates recovered from transgenic tissues showed the ability to withstand oxidative stress compared with isolates recovered from the untransformed poplar line. In vitro bioassays showed that trans-resveratrol inhibited both the swarming and swimming motilities in all the tested bacteria. The effects of trans-resveratrol on flagellin production, required for motility, were also investigated by immunoblot analysis.