Expression of the stilbene synthase (StSy) gene from grapevine in transgenic white poplar results in high accumulation of the antioxidant resveratrol glucosides
Expression of the stilbene synthase (StSy) gene from grapevine in transgenic white poplar results in high accumulation of the antioxidant resveratrol glucosides
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DOI：
10.1023/b:trag.0000034658.64990.7f
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发表时间：
2004-06-01
影响因子：
3
通讯作者：
Confalonieri, M
中科院分区：
文献类型：
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作者：
Giorcelli, A;Sparvoli, F;Confalonieri, M
当存在时，芪合酶导致白藜芦醇化合物的产生，白藜芦醇化合物是对抗植物的真菌病原体的植物抗毒素应答的主要组分，并且是具有药用价值的高度生物活性物质。白色白杨（Populus alba L.）用含有编码来自葡萄藤（葡萄属葡萄属L.）的芪合酶的cDNA插入片段的构建体转化，在花椰菜花叶病毒（CaMV）35 S启动子的控制下，和嵌合卡那霉素抗性基因。Southern杂交分析表明外源DNA序列已整合到白杨基因组中。通过Western blot和北方分析证实了芪苷酶编码基因在不同转基因株系中的表达。通过高效液相色谱（HPLC）、紫外分光光度法、电喷雾质谱（HPLC-ESI-MS）和酶解鉴定，发现转基因植株中有2个新化合物，分别为白藜芦醇苷（Piceid）的顺式和反式异构体。由于白杨是一个很好的生物质生产者和云杉积累了大量的（高达615.2 μ g/g叶鲜重），转基因植物代表了一个潜在的替代来源，用于生产这些化合物具有高药理价值。尽管白藜芦醇苷的存在下，在我们的实验条件下，没有增加对病原体Melampsora pulcherrima，这会导致锈病的抗性，在体外生物测定时进行了观察。
When present, stilbene synthase leads to the production of resveratrol compounds, which are major components of the phytoalexin response against fungal pathogens of the plant and are highly bioactive substances of pharmaceutical interest. White poplar ( Populus alba L.) was transformed with a construct containing a cDNA insert encoding stilbene synthase from grapevine ( Vitis vinifera L.), under the control of the cauliflower mosaic virus ( CaMV) 35S promoter, and a chimeric kanamycin resistance gene. Southern blot hybridization analysis demonstrated the presence and integration of exogenous DNA sequences in the poplar genome. Expression of the stilbene synthase-encoding gene in different transgenic lines was confirmed by Western blot and Northern analyses. Compared to the controls, in the transgenic plants two new compounds were detected and were identified as the trans- and cis-isomers of resveratrol-3-glucoside (piceid) by high-pressure liquid chromatography (HPLC), UV spectrophotometry, electrospray mass spectrometry (HPLC-ESI-MS) and enzymatic hydrolysis. Since poplar is a good biomass producer and piceids are accumulated in substantial amounts ( up to 615.2 mug/g leaf fresh weight), the transgenic plants represent a potential alternative source for the production of these compounds with high pharmacological value. Despite the presence of piceid, in our experimental conditions no increased resistance against the pathogen Melampsora pulcherrima, which causes rust disease, was observed when in vitro bioassays were performed.