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黄芪甲苷(Astragaloside IV,ASI)是药材黄芪中的活性成分之一。由于目前对于黄芪皂苷代谢途径知之甚少,使得通过直接转入皂苷母核合成相关基因从而提高黄芪毛状根中黄芪甲苷含量的方法不能施行。考虑到皂苷中的糖基侧链,增加活性葡萄糖代谢流入也许可以增加黄芪甲苷的最终转化效率。为了验证这个假说,该文将ADP-葡萄糖代谢相关基因颗粒结合型淀粉合成酶(Granule-bound starch synthase,GBSS)转入黄芪毛状根中,分子鉴定表明其基因已经整合进黄芪毛状根基因组,并且在mRNA水平增加了表达;GBSS酶活性分析表明,转基因株系平均酶活增加了约21.8倍;PAS染色结果表明,转基因株系中的多糖成分显著增加;与此同时,ASI含量亦提高了4倍。研究结果表明,通过转入活性糖代谢相关基因方法提高皂苷产量的方法切实可行。
Astragaloside IV (ASI) is one of the active ingredients in Radix Astragali. Due to the poor knowledge of the current astragaloside metabolic pathway, the method of increasing the content of Astragaloside in the hairy roots of Astragalus can not be implemented by directly transferring the gene into the saponin core. Given the glycosyl side chains in saponins, increasing the activity of glucose metabolism may increase the final conversion efficiency of astragaloside IV. In order to verify this hypothesis, we transferred Granule-bound starch synthase (GBSS), an ADP-glucose metabolism-related gene, into the hairy root of Astragalus. Molecular identification showed that its gene had been integrated into the hairy root genome of Astragalus , And increased at mRNA level. The analysis of GBSS activity showed that the average enzyme activity of transgenic lines increased about 21.8 times. PAS staining showed that the polysaccharide content of transgenic lines increased significantly. At the same time, ASI content also increased 4 times. The results show that it is feasible to improve the yield of saponin by transferring the gene related to active glucose metabolism.