农杆菌介导大豆遗传转化优化的初步研究
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摘要
利用农杆菌对大豆成熟种子子叶节进行转化,探究了不同种植物提取物、植物提取物浓度、大豆基因型、萌发天数以及共培养天数对提高农杆菌遗传转化效率的影响。结果表明:含有植物提取物的共培养菌液侵染子叶节均能够明显提高抗性芽的再生率,但不同基因型的大豆所需的最适提取物浓度不同,‘豫豆22’最适浓度为7mL/L,‘中黄42’最适浓度为14mL/L;‘豫豆22’是本研究中最适宜转化的大豆基因型;萌发5d的大豆子叶节转化率最高;共培养10d获得阳性转化植株率最高。
In this study,the soybean cotyledon nodes were used for genetic transformation by Agrobacterium tumefaciens to study the transformation efficiency affected by different kinds of plant extractions,the concentration of plant extractions,the soybean genotypes,and the days of germination and co-cultivation.The results showed that the co-cultured bacteria liquid that contained different concentrations of plant extractions significantly improved the frequency of regenerated shoots,but the optimal extraction concentrations for different genotypes of soybeans were different.The best-fit concentrations for ‘Yudou22'and ‘Zhonghuang42'were 7.14 and 14.3mL/L,respectively.The soybean ‘Yudou22'was the first-rate genotype for transformation in this study.The highest transformation efficiency was found when soybean cotyledon nodes germinated for 5days,and the highest efficiency of positive transformation appeared at the 10 th day of co-cultivation.
In this study,the soybean cotyledon nodes were used for genetic transformation by Agrobacterium tumefaciens to study the transformation efficiency affected by different kinds of plant extractions,the concentration of plant extractions,the soybean genotypes,and the days of germination and co-cultivation.The results showed that the co-cultured bacteria liquid that contained different concentrations of plant extractions significantly improved the frequency of regenerated shoots,but the optimal extraction concentrations for different genotypes of soybeans were different.The best-fit concentrations for ‘Yudou22'and ‘Zhonghuang42'were 7.14 and 14.3mL/L,respectively.The soybean ‘Yudou22'was the first-rate genotype for transformation in this study.The highest transformation efficiency was found when soybean cotyledon nodes germinated for 5days,and the highest efficiency of positive transformation appeared at the 10 th day of co-cultivation.
引文
[1]邓小莉,黄建新.转基因植物的研究概况及应用领域[J],生物学杂志,2003,20(6):14-16.
[2]Hinehee MAW,Connor Ward D V,Newell C A,et a1.Production of transgenic soybean plant using Agraobacteriunm-mediated DNA transfer[J].Bio Technolog,1988,6:915-922.
[3]崔姗姗.农杆菌介导大豆子叶节转化体系优化及转植酸酶基因大豆功能鉴定[D].秦皇岛:河北科技师范学院,2012.
[4]王振东,孙仓,王惠,等.不同方法从大豆不同组织中提取基因组DNA效果的比较[J].大豆科学,2008,27(1):42-46.
[5]刘艳军,杨静慧,杨恩芹,等.提高农杆菌基因转化率方法的研究[J].西南农业大学学报,2003,25(2):144-146,171.
[6]李茂福,李睿,傅永福,等.农杆菌介导大豆遗传转化的影响因素[J].山地农业生物学报,2006,25(4):283-286,292.
[7]乔利仙,于新玲,隋炯明,等.农杆菌介导的花生遗传转化体系的优化[J].核农学报,2012,26(9):1244-1248.
[8]刘峰,单世华,闫彩霞,等.花生子叶遗传转化再生体系影响因素的研究[J].花生学报,2007(01):13-19.
[9]王静,毛自文,范黎明,等.30种植物提取物抑菌活性研究[J].安徽农业科学,2012(10):5918-5919,6185.
[10]Trick HN,Dinkins RD,Santarem ER,et al.Recentadvances in soybean transformation[J].Plant Tissue Culture and Biotechnology,1997,3:9-26.
[11]刘旭娇.农杆菌介导大豆转化体系优化及报告基因DsRed2表达效率评估[D].哈尔滨:哈尔滨师范大学,2014.
[12]贾钰莹,蒋滢,赵强,等.农杆菌介导超高产大豆子叶节遗传转化研究[J].大豆科学,2014(5):634-637.
[13]应珊,何晓薇,王秀荣,等.影响农杆菌介导的大豆转化效率的因素研究[J].分子植物育种,2008(1):32-40.
[2]Hinehee MAW,Connor Ward D V,Newell C A,et a1.Production of transgenic soybean plant using Agraobacteriunm-mediated DNA transfer[J].Bio Technolog,1988,6:915-922.
[3]崔姗姗.农杆菌介导大豆子叶节转化体系优化及转植酸酶基因大豆功能鉴定[D].秦皇岛:河北科技师范学院,2012.
[4]王振东,孙仓,王惠,等.不同方法从大豆不同组织中提取基因组DNA效果的比较[J].大豆科学,2008,27(1):42-46.
[5]刘艳军,杨静慧,杨恩芹,等.提高农杆菌基因转化率方法的研究[J].西南农业大学学报,2003,25(2):144-146,171.
[6]李茂福,李睿,傅永福,等.农杆菌介导大豆遗传转化的影响因素[J].山地农业生物学报,2006,25(4):283-286,292.
[7]乔利仙,于新玲,隋炯明,等.农杆菌介导的花生遗传转化体系的优化[J].核农学报,2012,26(9):1244-1248.
[8]刘峰,单世华,闫彩霞,等.花生子叶遗传转化再生体系影响因素的研究[J].花生学报,2007(01):13-19.
[9]王静,毛自文,范黎明,等.30种植物提取物抑菌活性研究[J].安徽农业科学,2012(10):5918-5919,6185.
[10]Trick HN,Dinkins RD,Santarem ER,et al.Recentadvances in soybean transformation[J].Plant Tissue Culture and Biotechnology,1997,3:9-26.
[11]刘旭娇.农杆菌介导大豆转化体系优化及报告基因DsRed2表达效率评估[D].哈尔滨:哈尔滨师范大学,2014.
[12]贾钰莹,蒋滢,赵强,等.农杆菌介导超高产大豆子叶节遗传转化研究[J].大豆科学,2014(5):634-637.
[13]应珊,何晓薇,王秀荣,等.影响农杆菌介导的大豆转化效率的因素研究[J].分子植物育种,2008(1):32-40.