抗除草剂转基因普通白菜的获得和检测
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摘要
为了更加便捷快速地选育抗除草剂的普通白菜,试验对影响普通白菜蘸花法转化效率的因素进行探究。结果表明,在花蕾期进行蘸花处理,较短的蘸花时间、较长的蘸花间隔时长以及蘸花后喷水的组合能获得数量与质量更优的种子,但只有采用蘸花2.5 min,间隔7 d后再次蘸花,且蘸花后不喷水的方法能够获得转基因植株。研究结果将对普通白菜蘸花法遗传转化提供一定指导。
To select herbicide-resistant Pak choi(Brassica campestris L. ssp. chinensis Makino)more quickly, this experiment investigated the factors that affected the efficiency of the floral-dip method. The result showed that the floral-dip treatment was carried out in the bud stage of Pak choi, the combination of shorter dip times with longer interval and spray water after dipping could obtain better seeds in both quantity and quality. But the transgenic plants could be obtained only by dipping 2.5 min, dipping 7 d after the interval, and not spraying water after the dipping. This study results will provide some guidance for the genetic transformation of Pak choi by floral-dip method.
To select herbicide-resistant Pak choi(Brassica campestris L. ssp. chinensis Makino)more quickly, this experiment investigated the factors that affected the efficiency of the floral-dip method. The result showed that the floral-dip treatment was carried out in the bud stage of Pak choi, the combination of shorter dip times with longer interval and spray water after dipping could obtain better seeds in both quantity and quality. But the transgenic plants could be obtained only by dipping 2.5 min, dipping 7 d after the interval, and not spraying water after the dipping. This study results will provide some guidance for the genetic transformation of Pak choi by floral-dip method.
引文
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[3]屈聪玲,贺榆婷,王瑞良,等.植物转基因技术的过去、现在和未来[J].山西农业科学,2017,45(8):1376-1380,1383.
[4] TRIEU A T,BURLEIGH S H,KARDAILSKY I V,et al. Transformation of Medicago truncatula via infiltration of seedlings or flowering plants with Agrobacterium[J]. Plant Journal,2000,22(6):531-541.
[5]茹京娜,于太飞,陈隽,等.小麦锌指转录因子Ta Di19A对低温的响应及其互作蛋白的筛选[J].中国农业科学,2017,50(13):2411-2422.
[6] CLOUGH S J,BENT A F. Floral dip:a simplified method for Agrobacterium mediated transformation of Arabidopsis thaliana[J].Plant Journal for Cell&Molecular Biology,1998,16(6):735-743.
[7]许红梅,张立军,刘淳.农杆菌蘸花法侵染拟南芥的研究[J].北方园艺,2010(14):143-146.
[8] CURTIS I S,NAM H G. Transgenic radish(Raphanus sativus L.longipinnatus Bailey)by floral-dip method-plant development and surfactant are important in optimizing transformation efficiency[J].Transgenic Research,2001,10(4):363-371.
[9]付绍红,牛应泽,杨洪全,等.表面活性剂SilwetL-77对floral-dip转化甘蓝型油菜效果的影响[J].分子植物育种,2004(5):661-666.
[10]王翠艳,丁东风,于晓菊,等. Floral dip法在大豆遗传转化中的应用研究[J].南开大学学报(自然科学版),2010,43(1):34-38.
[11] BASTAKI N K,CULLIS C A. Floral-dip transformation of flax(Linum usitatissimum)to generate transgenic progenies with a high transformation rate[J]. Journal of Visualized Experiments Jove,2014,94:e52189.
[12]王翠艳,丁东风,于小菊,等.用floral dip法对黄瓜遗传转化的初步研究[C]//纪念天津科润黄瓜研究所建所二十五周年论文摘要集.天津:中国园艺学会,2010.
[13]林良斌,官春云,李恂,等.子房注射法与农杆菌介导法转化甘蓝型油菜的比较研究[J].生命科学研究,2000,4(3):231-236.
[14]曹传增.抗虫基因载体的构建及白菜转基因研究[D].北京:首都师范大学,2003.
[15] YANG G,REN X,RUAN S,et al. Genetic transformation of Chinese cabbage(Brassica rapa pekinensis)with floral-dip method[J]. Agricultural Biotechnology,2012,1(4):22-24,27.
[16]陈张彬,张振乾,陈浩,等.农杆菌介导抗除草剂基因bar和抗虫基因cry1ab/ac转化甘蓝型油菜的研究[J].华北农学报,2018,33(5):76-81.
[17]任伟,王嘉莹,任华华,等. PuCAT/Bar抗逆抗除草剂基因共转化紫花苜蓿的研究[J].华北农学报,2017,32(Z):1-6.
[18]韩笑.白菜浸花法转基因技术研究[D].曲阜:曲阜师范大学,2012.
[19] YONG Z,HUANG W,LI L,et al. Identification and functional characterization of a rice NAC gene involved in the regulation of leaf senescence[J]. Bmc Plant Biology,2013,13(1):132-145.
[20]李韩帅,周秒依,杨凤玲,等.农杆菌介导的玉米自交系Z21遗传转化体系的研究[J].山西农业科学,2018,46(4):517-521,547.
[21]齐雯雯,宫晓琳,王洋,等.蘸花法在植物遗传转化上的应用研究进展[J].现代农业科技,2014(24):9-10.
[22] CHUNG M H,CHEN M K,PAN S M. Floral spray transformation can efficiently generate Arabidopsis transgenic plants[J]. Transgenic Research,2000,9(6):471-486.
[23]党智笙,肖成斌,马燕林,等.农杆菌介导的浸花法(Floral-dip)存在的问题及对策[J].中国食品工业,2014(6):58.
[24]韩笑.浸花转化技术中不结球白菜与拟南芥的比较研究[J].分子植物育种,2012,10(1):1133-1137.
[2]高丽,崔崇士,屈淑平.白菜类蔬菜转基因研究进展[J].中国蔬菜,2010(12):7-13.
[3]屈聪玲,贺榆婷,王瑞良,等.植物转基因技术的过去、现在和未来[J].山西农业科学,2017,45(8):1376-1380,1383.
[4] TRIEU A T,BURLEIGH S H,KARDAILSKY I V,et al. Transformation of Medicago truncatula via infiltration of seedlings or flowering plants with Agrobacterium[J]. Plant Journal,2000,22(6):531-541.
[5]茹京娜,于太飞,陈隽,等.小麦锌指转录因子Ta Di19A对低温的响应及其互作蛋白的筛选[J].中国农业科学,2017,50(13):2411-2422.
[6] CLOUGH S J,BENT A F. Floral dip:a simplified method for Agrobacterium mediated transformation of Arabidopsis thaliana[J].Plant Journal for Cell&Molecular Biology,1998,16(6):735-743.
[7]许红梅,张立军,刘淳.农杆菌蘸花法侵染拟南芥的研究[J].北方园艺,2010(14):143-146.
[8] CURTIS I S,NAM H G. Transgenic radish(Raphanus sativus L.longipinnatus Bailey)by floral-dip method-plant development and surfactant are important in optimizing transformation efficiency[J].Transgenic Research,2001,10(4):363-371.
[9]付绍红,牛应泽,杨洪全,等.表面活性剂SilwetL-77对floral-dip转化甘蓝型油菜效果的影响[J].分子植物育种,2004(5):661-666.
[10]王翠艳,丁东风,于晓菊,等. Floral dip法在大豆遗传转化中的应用研究[J].南开大学学报(自然科学版),2010,43(1):34-38.
[11] BASTAKI N K,CULLIS C A. Floral-dip transformation of flax(Linum usitatissimum)to generate transgenic progenies with a high transformation rate[J]. Journal of Visualized Experiments Jove,2014,94:e52189.
[12]王翠艳,丁东风,于小菊,等.用floral dip法对黄瓜遗传转化的初步研究[C]//纪念天津科润黄瓜研究所建所二十五周年论文摘要集.天津:中国园艺学会,2010.
[13]林良斌,官春云,李恂,等.子房注射法与农杆菌介导法转化甘蓝型油菜的比较研究[J].生命科学研究,2000,4(3):231-236.
[14]曹传增.抗虫基因载体的构建及白菜转基因研究[D].北京:首都师范大学,2003.
[15] YANG G,REN X,RUAN S,et al. Genetic transformation of Chinese cabbage(Brassica rapa pekinensis)with floral-dip method[J]. Agricultural Biotechnology,2012,1(4):22-24,27.
[16]陈张彬,张振乾,陈浩,等.农杆菌介导抗除草剂基因bar和抗虫基因cry1ab/ac转化甘蓝型油菜的研究[J].华北农学报,2018,33(5):76-81.
[17]任伟,王嘉莹,任华华,等. PuCAT/Bar抗逆抗除草剂基因共转化紫花苜蓿的研究[J].华北农学报,2017,32(Z):1-6.
[18]韩笑.白菜浸花法转基因技术研究[D].曲阜:曲阜师范大学,2012.
[19] YONG Z,HUANG W,LI L,et al. Identification and functional characterization of a rice NAC gene involved in the regulation of leaf senescence[J]. Bmc Plant Biology,2013,13(1):132-145.
[20]李韩帅,周秒依,杨凤玲,等.农杆菌介导的玉米自交系Z21遗传转化体系的研究[J].山西农业科学,2018,46(4):517-521,547.
[21]齐雯雯,宫晓琳,王洋,等.蘸花法在植物遗传转化上的应用研究进展[J].现代农业科技,2014(24):9-10.
[22] CHUNG M H,CHEN M K,PAN S M. Floral spray transformation can efficiently generate Arabidopsis transgenic plants[J]. Transgenic Research,2000,9(6):471-486.
[23]党智笙,肖成斌,马燕林,等.农杆菌介导的浸花法(Floral-dip)存在的问题及对策[J].中国食品工业,2014(6):58.
[24]韩笑.浸花转化技术中不结球白菜与拟南芥的比较研究[J].分子植物育种,2012,10(1):1133-1137.