Abstract:A Receptor-like kinase(RLK)gene, OsSIK1,plays important roles in drought stress-tolerance in rice,through the activation of the antioxidative system.To make maize plants have much more drought resistance genes and further to obtain drought tolerance maize germplasm.In this study, OsSIK1 gene of rice was transformated into maize inbred Zheng 58 plants by pollen-mediated method.First,transgenic plants in T1,T2 and T3 were detected by Kanamycin resistance screening,PCR and Southern Blotting,transgenic plants were obtained from T1 and pure transgenic lines was obtained from T3.Next,drought resistance analysis to transgenic maize plants and non-transformation control plants were conducted under the condition of 16.1% PEG drought stress.The results showed that compared with non-transgenic plants,the seedling leaf relative water content,chlorophyll content and SOD activity of transgenic plants were increased by 7.4%-19.8%,11.3%-106.9% and 45.8%-93.4%,respectively;furthermore,the relative conductivity and MDA content were decreased by 35.4%-58.1%,and 25.7%-50.4%,respectively.All the physiological indexes under the drought stress proved that transgenic OsSIK1 gene improved the drought resistance of transgenic maize plants,further analysis found that there were significant differences on drought tolerance between 5 transformed lines and their control groups,and their performance of field were superior to that of non-transgenic maize seedlings.At last,5 transgenic maize inbred lines were obtained,which suggested that genetically modified maize has improved the drought resistance by introducing foreign OsSIK1 gene of rice.
[1] 国家统计局.国家统计局关于2015年粮食产量的公告[EB/OL].[2015-12-08].http://www.yumi.com.cn/html/2015/12/20151208165617184726.html.
[2] 韦正乙,张玉英,王云鹏,等.基因工程在玉米抗旱育种中的应用[J].玉米科学,2014,22(4):1-7.
[3] 徐蕊,王启柏,张春庆,等.玉米自交系抗旱性评价指标体系的建立[J].中国农业科学,2009,42(1):72-84.
[4] 王小丽,杜建中,郝曜山,等.转BADH 基因玉米植株的获得及其耐盐性分析[J].作物学报,2014,40(11):1973-1979.
[5] 王小丽,赵欣梅,张欢欢,等.盐胁迫对转TaNHX2 基因玉米幼苗生长的影响[J].华北农学报,2014,29(2):165-169.
[6] 程继东,安玉麟,孙瑞芬,等.抗旱、耐盐基因类型及其机理的研究进展[J].华北农学报,2006,21(专辑):116-120.
[7] Liu X X,Zhai S M,Zhao Y J,et al.Over expression of the phosphatidylinositol synthase gene(ZmPIS) conferring drought stress tolerance by altering membrane lipid composition and increasing ABA synthesis in maize[J].Plant Cell and Environment,2013,36(5):1037-1055.
[8] 石薇,黄丛林,张秀海,等.采用花粉管通道法将蛋白激酶基因导入玉米自交系的研究[J].华北农学报,2011,21(4):46-49.
[9] He C M,He Y,Liu Q,et al.Co-expression of genes ApGSMT2 and ApDMT2 for glycinebetaine synthesis in maize enhances the drought tolerance of plants[J].Mol Breeding,2013,31(3):559-573.
[10] 杨政伟,胡银岗,王琨,等.转SAMS 基因玉米自交系获得及抗旱性分析[J].西北植物学报,2011,31(6):1084-1089.
[11] Zhang S J,Li N,Gao F,et al.Over-expression of TsCBF1 gene confers improved drought tolerance in transgenic maize[J].Molecular Breeding,2010,26:455-465.
[12] Nguyen T X,Sticklen M.Barley HVA1 gene confers drought and salt tolerance in transgeic maize(Zea may L.)[J].Advances in Crop Science and Technology,2013,1(1):105.
[13] Ouyang S Q.Receptor-like kinase OsSIKl improves drought and salt stress tolerance in rice(Oryza sativa) plants[J].The Plant Journal,2010,62(2):316-329.
[14] 张婷婷,王铭,杜建中,等.花粉介导法将水稻OsSIK1 基因导入玉米自交系的研究[J].山西农业科学,2011,39(5):395-399.
[15] 杨慧珍,车丽,任志强,等.抗草丁膦转基因玉米不同DNA提取方法及PCR检测[J].山西农业科学,2011,39(9):925-927,1028.
[16] 王关林,方宏筠.植物基因工程[M].2版.北京:科学出版社,2004:742-744.
[17] Huang P T,Wang H L,Zhou X W,et al.Molecular cloning:a laboratory manual[M].3rded.Beijing:Science Press,2002:212-219.
[18] 高俊凤.植物生理学实验指导[M].西安:世界地图出版社,2000.
[19] 郝曜山,孙毅,杜建中,等.转双价抗虫基因BmkIT-Chitinase 玉米株系的获得[J].分子植物育种,2012,10(2):147-154.
[20] 姚正培,孟君,李冠.玉米自交系芽苗期耐盐性的鉴定与筛选[J].华北农学报,2007,22(5):27-30.
[21] 王彦玲,卫文星,铁双贵,等.郑58和掖478玉米自交系基因组差异性分析[J].玉米科学,2010,18(3):57-60.
[22] 任小燕,杜建中,孙毅.转AhCMO 基因玉米后代的获得及耐盐性鉴定[J].分子植物育种,2013,11(3):332-338.
[23] 柳斌辉,张文英,栗雨勤.玉米耐盐性的鉴定评价[J].华北农学报,2012,27(S1):101-105.