转MINAC2基因芒的获得及其在NaCl胁迫下的表达

Transgenic MINAC2 Miscanthus sinensis and Its Expression under NaCl Stress

芒(Miscanthus sinensis)隶属于多年生高大禾草--芒属(Miscanthus),其生态效应和经济效益备受国内外研究者的关注,是中国广阔边际土地种植与利用的能源植物。为获得芒的转基因株系,同时对MINAC2基因在盐胁迫下的表达特征进行研究,本研究利用农杆菌介导法将MINAC2基因导入芒的基因组,并采用RT-qPCR技术测定了0.6%Na Cl溶液胁迫下转基因株系0、1 h、3 h、9 h、12 h、24 h、48 h的MINAC2相对表达量。获得的主要研究结果如下:(1)经PCR鉴定确定了转MINAC2基因阳性株系;(2)未经NaCl胁迫时转MINAC2基因株系的表达量显著低于对照组,然而在0.6%NaCl处理达到一定时长后转MINAC2基因株系的表达量是显著高于对照组(p<0.05),为对照组的5.93倍。通过本研究,建立了芒的农杆菌转化体系,并对MINAC2基因在芒的耐盐能力提高中所起的作用进行了初步探讨。

Miscanthus sinensis belongs to perennial giant grass Miscanthus as a renewable energy plant with huge biomass yield.Because of ecological effects and economic benefits,they were super candidates to be planted on the marginal lands of China.In order to obtain a new transgenic germplasm and to demonstrate the expression characteristics of MINAC2 gene under salt stress,the MINAC2 gene has been transferred into the embryogenic callus of M.sinensis mediated with agrobacterium,and the transgenic lines were treated with 0.6%NaCl solution0,1 h,3 h,9 h,12 h,24 h or 48 h,and the relative expression level of MINAC2 has been measured by RT-qPCR.The main results were as follows:(1)Transgenic lines via PCR appraisal had been obtained.(2)Expression level of MINAC2 in the MINAC2 gene line was significantly lower than that in the control group without NaCl stress.However,the expression of MINAC2 in the MINAC2 gene line was significantly higher than that in the control group after 0.6%NaCl treatment for a certain period of time(p<0.05).The results showed that the salt tolerance of awns was significantly increased after transfer into the MINAC2 gene.In this work,the transformation system of Agrobacterium tumefaciens was established,and the role of MINAC2 gene in improving salt tolerance of Miscanthus sinensis was discussed.