植酸酶基因phyA转化大豆品种研究

Transgenic Soybean(Glycine max.L) Plants with phyA Gene Improved Phytase Activity

土壤磷多以有机态形式存在,而植酸磷占一半以上,分解利用植酸态磷是提高土壤磷利用效率、改善植物磷素营养的新途径。利用大豆农杆菌介导子叶节转化与花粉管通道转化技术,将含有根特异启动子pyk10、信号肽S和植酸酶phyA的嵌和基因(KSA)转入冀豆12、冀豆16、五星1号和吉林35中。经PCR检测,共获得T0阳性植株114个,T1阳性植株101个,T2阳性材料28个。通过将T4代转基因株系和野生型对照在仅含植酸磷的营养液中进行培养发现,转基因植株在植酸磷条件下生长状况优于对照,并且3个转基因株系根系分泌型植酸酶活性分别比野生型提高5%、13%和24%。

Organic phosphorus（P） is the main form of phosphorus in soil,of which the phytate accounted for more than 50%.The available way that improves plant P nutrition is to hydrolyze phytate into inorganic phosphorus（Pi） to increase the soil P utilization efficiency.In this study,a phytase gene（phyA）,driven by the root-specific pyk10 promoter and directed by a carrot extracellular targeting peptide,was introduced into different soybean varieties（Jidou 12,Jidou 16,Wuxing 1 and Jilin 35） by Agrobacterium-mediated and pollen tube pathway transformation method.PCR analysis results showed that the phyA was successfully integrated into the soybean genome.Total 114,101 and 28 PCR positive plants were obtained from T0,T1 and T2 generation,respectively.The results also showed that the T4 generation plants could grow better than wild-type when cultured under the condition in which phytate was the sole source of phosphorus.And the secreted phytase activities of roots also indicated that three transgenic lines were increased by 5%,13% and 24% compared to the wild-type plants,respectively.