结球甘蓝自交系YL-1的高效遗传转化体系的建立及应用
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摘要
为了筛选结球甘蓝高效遗传转化受体材料,以高代自交系YL-1、21-3、12J35、650为试材,比较不同材料具柄子叶和下胚轴的再生频率差异。在4份供试材料中,YL-1为最优转化受体,再生频率显著高于其他3份材料;YL-1具柄子叶和下胚轴的再生频率无显著差异,均可用于遗传转化。进一步对YL-1遗传转化过程中光照强度、Basta除草剂浓度、农杆菌侵染浓度等关键影响因素进行优化,发现光照强度为4 000 lx时更有利于不定芽诱导;除草剂最适筛选浓度为8 mg·L~(-1);农杆菌侵染浓度OD600=0.3侵染8 min,YL-1抗性芽再生频率最高。基于建立的高效遗传转化体系,共获得47株抗除草剂植株,PCR检测显示其中12株为阳性,初步表明bar基因已转化到甘蓝中,转化率达到2.18%。利用转录组测序技术对2株PCR阳性材料进行基因表达分析,均检测到bar基因高效表达。甘蓝高代自交系YL-1高效遗传转化体系的建立可为今后结球甘蓝基因功能鉴定及重要农艺性状的改良提供基础。
In order to select the cabbage genotypes with high-efficiency transformation,cotyledons with petiole and hypocotyl of four inbred lines were used to compare the regeneration frequency. The results showed that YL-1 was the optimal transformation donor and its regeneration frequency was significantly higher than the other three materials. The regeneration frequency of YL-1 cotyledons with petiole and hypocotyl had no significant difference and both of them could be used for genetic transformation. The transformation factors including light intensity,concentration of herbicide Basta and Agrobacterium concentration of infection were further optimized. The results showed that the optimum culture condition for YL-1 adventitious bud regeneration was light intensity 4 000 lx + Basta 8 mg · L~(-1). When the explants were inoculated by Agrobacterium with OD600 = 0.3 and 8 min,the regeneration frequency of herbicide-resistant buds was the highest. Eventually,a total of 47 herbicide-resistant transformants were obtained and 12 of them were PCR-positive,with the transformation frequency up to 2.18%,which indicated that bar gene had been transformed into YL-1. Gene expression analysis of two PCR-positive transformants were performed by transcriptome sequencing,and bar gene was transcribed in both of them. The establishment of high-efficiency genetic transformation system for YL-1 mediated by Agrobacterium tumefaciens would provide a basis for gene function identification,the editing and improvement of important agronomic traits in the future.
In order to select the cabbage genotypes with high-efficiency transformation,cotyledons with petiole and hypocotyl of four inbred lines were used to compare the regeneration frequency. The results showed that YL-1 was the optimal transformation donor and its regeneration frequency was significantly higher than the other three materials. The regeneration frequency of YL-1 cotyledons with petiole and hypocotyl had no significant difference and both of them could be used for genetic transformation. The transformation factors including light intensity,concentration of herbicide Basta and Agrobacterium concentration of infection were further optimized. The results showed that the optimum culture condition for YL-1 adventitious bud regeneration was light intensity 4 000 lx + Basta 8 mg · L~(-1). When the explants were inoculated by Agrobacterium with OD600 = 0.3 and 8 min,the regeneration frequency of herbicide-resistant buds was the highest. Eventually,a total of 47 herbicide-resistant transformants were obtained and 12 of them were PCR-positive,with the transformation frequency up to 2.18%,which indicated that bar gene had been transformed into YL-1. Gene expression analysis of two PCR-positive transformants were performed by transcriptome sequencing,and bar gene was transcribed in both of them. The establishment of high-efficiency genetic transformation system for YL-1 mediated by Agrobacterium tumefaciens would provide a basis for gene function identification,the editing and improvement of important agronomic traits in the future.
引文
Ana L R,Miguel G L,Francisco F,Achim M L.2012.Physical methods for genetic plant transformation.Physics of Life Reviews,9:308-345.
Alicja Z.2014.Agrobacterium-mediated plant transformation:factors,applications and recent advances.Biocatalysis and Agricultural Biotechnology,3:95-102.
Bi Ling,Liu Fengluan,Dong Aixiang,Zhou Xiaoyang,Ma Nan,Zhao Liangjun.2012.Establishment of genetic transformation system in protcorm like-body(plb)of Rosa canina.Journal of Nuclear Agricultural Sciences,26(2):270-274.(in Chinese)毕玲,刘凤栾,董爱香,周晓阳,马男,赵梁军.2012.狗蔷薇类原球茎遗传转化体系的建立.核农学报,26(2):270-274
Chen Yunxi,He Dandan,Liao Haoru,Ye Yanlan,Wen Zhihua,Zhang Zhisheng.2010.Factors influencing on shoot differentiation of rhizome of Cymbidium sinense×Cymbidium lancifolium.Chinese Agricultural Science Bulletin,26(9):65-69.(in Chinese)陈云喜,何丹丹,廖浩如,叶燕兰,文志华,张志胜.2010.影响墨兰×兔耳兰根状茎芽分化的因素.中国农学通报,26(9):65-69.
Cheng Z X,Hu H T,Yang L,Wang C C,Guo W D,Yang L.2017.Overexpression of EutPDS gene from Elaeagnus umbellata increases lycopene content in tomato fruit.Scientia Silvae Sinicae,53(1):62-69.
Cho M J,Banh J,Yu M,Kwan J,Jones T J.2015.Improvement of Agrobacterium-mediated transformation frequency in multiple modern elite commercial maize(Zea mays L.)inbreds by media modifications.Plant Cell,Tissue and Organ Culture,121(3):519-529.
Cui Lei,Yang Limei,Liu Nan,Lang Zhihong.Liu Yumei,Zhuang Mu,Zhang Yangyong,Zhang Youjun,Huang Dafang,Fang Zhiyuan.2009.Transformation and expressing of Bt gene cry1Ia8 in cabbage.Acta Horticulturae Sinica,36(8):1161-1168.(in Chinese)崔磊,杨丽梅,刘楠,郎志宏,刘玉梅,庄木,张扬勇,张友军,黄大昉,方智远.2009.Bt Cry1Ia8抗虫基因对结球甘蓝的转化及其表达.园艺学报,36(8):1161-1168.
Dong Zhigang,Ma Xiaohe,Wang Guoping,Zhao Qifeng,Li Xiaomei,Tan Wei,Wang Min,Tang Xiaoping.2014.Optimization of Agrobacterium-mediated transformation system of grape.Journal of Nuclear Agricultural Sciences,28(7):1169-1174.(in Chinese)董志刚,马小河,王国平,赵旗峰,李晓梅,谭伟,王敏,唐晓萍.2014.根癌农杆菌介导的葡萄遗传转化体系的优化.核农学报,28(7):1169-1174.
Han F Q,Chong Y,Fang Z Y,Yang L M,Zhuang M,Lv H H,Liu Y M,Li Z S,Liu B,Yu H L,Liu X P,Zhang Y Y.2015.Inheritance and InDel markers closely linked to petal color gene(cpc-1)in Brassica oleracea.Molecular Breeding,35:160.
Hess D,Dressler K,Nimmrichter R.1990.Transformation experiments by pipetting Agrobacterium into the spikelets of whea(tTriticum aestivum L.).Plant Science,72(2):233-244.
Horsch R B,Fry J E,Hoffmann N L,Eichholtz D,Rogers S G,Fraley R T.1985.A simple and general method for transferring genes into plants.Science,27(4691):1229-1231.
Jia Li’e.2012.Establishment of effective Agrobacterium-mediated mature embryo transformation system for Indica rice[M.D.Dessertation].Beijing:Chinese Academy of Agricultural Sciences.(in Chinese)贾丽娥.2012.农杆菌介导籼稻成熟胚高效转化体系的建立[硕士论文].北京:中国农业科学院.
Liu Jie,Ma Jian,Lei Lei,Sun Chao,Kang Jungen,Xie Jianming.2017.Construction of plant expression vector and transformation of fusarium wilt resistance gene FOC1 to Brassica oleracea.Acta Botanica Boreali-Occidentalia Sinica,26(3):429-436.(in Chinese)刘洁,马建,雷蕾,孙超,康俊根,颉建明.2017.甘蓝抗枯萎病基因FOC1植物表达载体构建及遗传转化.西北农业学报,26(3):429-436.
Liu Shuang,Guan Shuyan,Sun Su,Wei Xiaoyu.2017.Optimization of maize callus culture system and genetic transformation of ICE1.Journal of Jilin Agricultural University,39(5):518-523.(in Chinese)刘双,关淑艳,孙苏,魏晓禹.2017.玉米愈伤组织培养体系优化及ICE1基因的遗传转化.吉林农业大学学报,39(5):518-523.
Liu X P,Yang C,Han F Q,Fang Z Y,Yang L M,Zhuang M,Lv H H,Liu Y M,Li Z S,Zhang Y Y.2016.Genetics and fine mapping of a yellow-green leaf gene in cabbage(Brassica oleracea var.capitata L.).Molecular Breeding,36:82.
Liu Yanjun,Yang Jinghui,Yang Enqin,Liu Yudong,Liu Shuwu.2003.An investigation of the approaches to enhance gene transformation rate with Agrobacterium.Journal of Southwest Agricultural University,25(2):144-146.(in Chinese)刘艳军,杨静慧,杨恩芹,刘玉东,刘树梧.2003.提高农杆菌基因转化率方法的研究.西南大学学报(自然科学版),25(2):144-146.
Lv Q R,Chen C S,Xu Y J,Hu S K,Wang L,Sun K,Chen X,Li X H.2017.Optimization of Agrobacterium tumefaciens-mediated transformation systems in tea plant(Camellia sinensis).Horticultural Plant Journal,3(3):105-109.
Moloney M M,Walker J M,Sharma K K.1989.High efficiency transformation of Brassica napus using Agrobacterium vectors.Plant Cell Reports,8(4):238-242.
Murray H,Thompson W.1980.Rapid isolation of high molecular weight DNA.Nucleic Acids Research,8:4321-4325.
Ni Dexiang.1986.Regulation of light in plant tissue culture.Chinese Journal of Nature,9(3):193-198.(in Chinese)倪德祥.1986.光在植物组织培养中的调控作用.自然杂志,9(3):193-198.
Pavel K,Olga S,Ivan L,Anna D,George K,Jozef S.2015.Transient plant transformation mediated by Agrobacterium tumefaciens:principles,methods and applications.Biotechnology Advances,33:1024-1042.
Peng C J,Xu W G,Hu L,Li Y,Qi X L,Wang H W,Hua X,Zhao M Z.2018.Effects of the maize C4 phosphoenolpyruvate carboxylase(ZmPEPC)gene on nitrogen assimilation in transgenic wheat.Plant Growth Regulation,84(1):191-205.
Qiao Yahong,Zheng Yinying,Li ShiLin,Tian Guiying,Zhang Lili,Lin Tao,Xiang Benchun.2013.A regeneration system for genetic transformation of processing tomato Hongfan 3.Journal of Nuclear Agricultural Sciences,27(11):1636-1643.(in Chinese)乔亚红,郑银英,李诗林,田桂英,张丽丽,林涛,向本春.2013.红番3号加工番茄遗传转化再生体系的建立.核农学报,27(11):1636-1643.
Shen Hailong.2009.Microculture root-raising seedlings of trees in tissue culture.Beijing:China Forestry Publishing House.(in Chinese)沈海龙.2009.树木组织培养微体试管外生根育苗技术.北京:中国林业出版社.
Shi Wenhui.2017.Establishment of efficient regeneration system and genetic transformation of glyphosate resistant genes EPSPs and BnGRF2 in rape[M.D.Dissertation].Lanzhou:Gansu Agricultural University.(in Chinese)石文慧.2017.油菜高效再生体系的建立及Basta抗性基因EPSPs和BnGRF2的遗传转化[硕士论文].兰州:甘肃农业大学.
Swart V,Crampton B G,Ridenour J B,Bluhm B H,Olivier N A,Meyer J J M,Berger D K.2017.Complementation of CTB7 in the maize pathogen cercospora zeina overcomes the lack of in vitro cercosporin production.Molecular Plant-microbe Interactions,30(9):710-724.
Tang Jun,Zhang Shuning,Hou Xilin,Wang Shumin,Sun Chengzhen.2011.Establishment of high-frequency in vitro regeneration system of Chinese kale(Brassica alboglabra Bailey).Journal of Nanjing Agricultural University,34(2):29-32.(in Chinese)唐君,张蜀宁,侯喜林,王淑敏,孙成振.2011.芥蓝高频离体再生体系的建立.南京农业大学学报,34(2):29-32.
Toriyama K,Arimoto Y,Uchimiya H,Hinata K.1988.Transgenic rice plants after direct gene transfer into protoplasts.Nature Biotechnology,6(9):1072-1074.
Wang Guoliang.2004.Optimization of high-efficiency Agrobacterium-mediated BADH gene transformation system of alfalfa and detection of transgenic plants[M.D.Dissertation].Lanzhou:Gansu Agricultural University.(in Chinese)王国良.2004.农杆菌介导的紫花苜蓿BADH基因高效转化体系的优化和转基因植株的检测[硕士论文].兰州:甘肃农业大学.
Wang K,Liu H Y,Du L P,Ye X G.2017.Generation of marker-free transgenic hexaploid wheat via an Agrobacterium-mediated co-transformation strategy in commercial Chinese wheat varieties.Plant Biotechnology Journal,15:614-623.
Wang Li,Yi Dengxia,Yang Limei,Liu Jianping,Fang Zhiyuan,Cheng Fei,Liu Yumei,Zhuang Mu,Zhang Yangyong,Sun Peitian.2014.Obtaining of insect-resistant material with Bt in early spring cabbage.China Vegetables,(10):12-17.(in Chinese)王丽,仪登霞,杨丽梅,刘建萍,方智远,程斐,刘玉梅,庄木,张扬勇,孙培田.2014.转Bt基因早熟春甘蓝抗虫材料的获得.中国蔬菜,(10):12-17.
Wang Q J.2012.Functional verification of drought tolerance gene PC2300AF1 in japanica rice in cold regions.Acta Agriculturae Universitatis Jiangxiensis,34(3):434-438.
Wang Xiao-hong,Zeng Ai-song,Gao Bing,Song Li-xiao,Yan Ji-yong.2013.Resistance to salt in cabbage(Brassica oleracea L.var.capitata L.)transformed with StP5CS gene.Acta Botanica Boreali-Occidentalia Sinica,33(5):885-891.(in Chinese)王肖红,曾爱松,高兵,宋立晓,严继勇.2013.StP5CS转化结球甘蓝的研究.西北植物学报,33(5):885-891.
Xu Lingling,Ding Duoduo,Tao Guirong,Dang Yuefang,Guo Bin.2013.Effects of different light intensity and light quality on adventitious buds induction and growth of Clematis‘Nelly Moser’.Acta Photonica Sinica,42(6):715-720.(in Chinese)徐玲玲,丁朵朵,陶贵荣,党悦方,郭斌.2013.不同光照强度和光质对铁线莲品种繁星(Clematis‘Nelly Moser’)不定芽诱导和生长的影响.光子学报,42(6):715-720.
Yang Chong,Zhang Yangyong,Fang Zhiyuan,Liu Yumei,Yang Limei,Zhuang Mu,Sun Peitian.2014.Photosynthetic physiological characteristics and chloroplast ultrastructure of yellow leaf mutant YL-1 in cabbage.Acta Horticulturae Sinica,41(6):1133-1144.(in Chinese)杨冲,张扬勇,方智远,刘玉梅,杨丽梅,庄木,孙培田.2014.甘蓝叶色黄化突变体YL-1的光合生理特性及其叶绿体的超微结构.园艺学报,41(6):1133-1144.
Yang Weijie.2016.Transformation of Bt Cry1Ia8/Cry1C genes and study of the resistance to pests in transgenic high generation of inbred lines in cabbage[M.D.Dissertation].Beijing:Chinese Academy of Agricultural Sciences.(in Chinese)杨卫杰.2016.Bt Cry1Ia8/Cry1 AC基因转化甘蓝及高代转基因甘蓝的抗虫型研究[硕士论文].北京:中国农业科学院.
Yang Yue,Zhang Zhihong,Liu Yuexue,He Ping,Zhou Yu,Dai Yanhong,Zhao Guiling.2010.Establishment of Agrobacterium tumefaciens-mediated transformation system for Hanfu apple.Journal of Fruit Science,27(2):174-178.(in Chinese)杨越,张志宏,刘月学,何平,周宇,代红艳,赵桂玲.2010.根癌农杆菌介导寒富苹果转化体系的建立.果树学报,27(2):174-178.
Ying Shan,He Xiaowei,Wang Xiurong.2008.Assessment of factors affecting the transformation efficiency of soybean cotyledonary-node Agrobacterium-mediated transformation system.Molecular Plant Breeding,6(1):32-40.(in Chinese)应珊,何晓薇,王秀荣.2008.影响农杆菌介导的大豆转化效率的因素研究.分子植物育种,6(1):32-40.
Yu Shou-jiang.2007.Cloning CryⅠAC gene and the study of transformation of cabbage[M.D.Dissertation].Harbin:Northeast Agricultural University.(in Chinese)于守江.2007.CryⅠAC基因的克隆及转化结球甘蓝的研究[硕士论文].哈尔滨:东北农业大学.
Yu Fengli.2005.Study on transferring of coat pritein gene potato virus a(PVA CP)into potato[M.D.Dissertation].Harbin:Northeast Agricultural University.(in Chinese)于凤丽.2005.马铃薯A病毒外壳蛋白(PVA CP)基因的遗传转化研究[硕士论文].哈尔滨:东北农业大学.
Zhao Xiushu,Li Mingyang,Zhang Wenling,Liu Fan.2009.Establishment of high adventitious shoot regeneration system of ornamental kale.Genomics&Applied Biology,28(1):141-148.(in Chinese)赵秀枢,李名扬,张文玲,刘凡.2009.观赏羽衣甘蓝高频再生体系的建立.基因组学与应用生物学,28(1):141-148.
Zhu W,Zuo Y,Zhou R F,Huang J Y,Tang M Q,Cheng X H,Liu Y Y,Tong C B,Xiang Y,Dong C H,Liu S Y.2016.Vacuolar iron transporter BnMEB2 is involved in enhancing iron tolerance of Brassica napus.Frontiers in Plant Science,7(9):1353.
Zimmermann U,Vienken J.1982.Stable transformation of maize after gene transfer by electroporation.The Journal of Membrane Biology,67:165-182.
Zou Zhi,Lu Changming.2008.An overview of plant factors influencing Agrobacterium-mediated transformation.Biotechnology Bulletin,18(1):1-9.(in Chinese)邹智,卢长明.2008.影响农杆菌介导遗传转化的植物因子研究进展.生物技术通报,18(1):1-9.
Zhai Rui,Gao Le,Ding Xueni,Liao Wenlin,Zhen Mingjie,Lu Zhiwen,Zhi Haijian.2015.Optimization of cotyledonary-node Agrobacteriummediated soybean transformation system.Soybean Science,34(5):768-775.(in Chinese)翟锐,高乐,丁雪妮,廖文林,郑明洁,陆智文,智海剑.2015.农杆菌介导大豆子叶节遗传转化体系的优化.大豆科学,34(5):768-775.
Alicja Z.2014.Agrobacterium-mediated plant transformation:factors,applications and recent advances.Biocatalysis and Agricultural Biotechnology,3:95-102.
Bi Ling,Liu Fengluan,Dong Aixiang,Zhou Xiaoyang,Ma Nan,Zhao Liangjun.2012.Establishment of genetic transformation system in protcorm like-body(plb)of Rosa canina.Journal of Nuclear Agricultural Sciences,26(2):270-274.(in Chinese)毕玲,刘凤栾,董爱香,周晓阳,马男,赵梁军.2012.狗蔷薇类原球茎遗传转化体系的建立.核农学报,26(2):270-274
Chen Yunxi,He Dandan,Liao Haoru,Ye Yanlan,Wen Zhihua,Zhang Zhisheng.2010.Factors influencing on shoot differentiation of rhizome of Cymbidium sinense×Cymbidium lancifolium.Chinese Agricultural Science Bulletin,26(9):65-69.(in Chinese)陈云喜,何丹丹,廖浩如,叶燕兰,文志华,张志胜.2010.影响墨兰×兔耳兰根状茎芽分化的因素.中国农学通报,26(9):65-69.
Cheng Z X,Hu H T,Yang L,Wang C C,Guo W D,Yang L.2017.Overexpression of EutPDS gene from Elaeagnus umbellata increases lycopene content in tomato fruit.Scientia Silvae Sinicae,53(1):62-69.
Cho M J,Banh J,Yu M,Kwan J,Jones T J.2015.Improvement of Agrobacterium-mediated transformation frequency in multiple modern elite commercial maize(Zea mays L.)inbreds by media modifications.Plant Cell,Tissue and Organ Culture,121(3):519-529.
Cui Lei,Yang Limei,Liu Nan,Lang Zhihong.Liu Yumei,Zhuang Mu,Zhang Yangyong,Zhang Youjun,Huang Dafang,Fang Zhiyuan.2009.Transformation and expressing of Bt gene cry1Ia8 in cabbage.Acta Horticulturae Sinica,36(8):1161-1168.(in Chinese)崔磊,杨丽梅,刘楠,郎志宏,刘玉梅,庄木,张扬勇,张友军,黄大昉,方智远.2009.Bt Cry1Ia8抗虫基因对结球甘蓝的转化及其表达.园艺学报,36(8):1161-1168.
Dong Zhigang,Ma Xiaohe,Wang Guoping,Zhao Qifeng,Li Xiaomei,Tan Wei,Wang Min,Tang Xiaoping.2014.Optimization of Agrobacterium-mediated transformation system of grape.Journal of Nuclear Agricultural Sciences,28(7):1169-1174.(in Chinese)董志刚,马小河,王国平,赵旗峰,李晓梅,谭伟,王敏,唐晓萍.2014.根癌农杆菌介导的葡萄遗传转化体系的优化.核农学报,28(7):1169-1174.
Han F Q,Chong Y,Fang Z Y,Yang L M,Zhuang M,Lv H H,Liu Y M,Li Z S,Liu B,Yu H L,Liu X P,Zhang Y Y.2015.Inheritance and InDel markers closely linked to petal color gene(cpc-1)in Brassica oleracea.Molecular Breeding,35:160.
Hess D,Dressler K,Nimmrichter R.1990.Transformation experiments by pipetting Agrobacterium into the spikelets of whea(tTriticum aestivum L.).Plant Science,72(2):233-244.
Horsch R B,Fry J E,Hoffmann N L,Eichholtz D,Rogers S G,Fraley R T.1985.A simple and general method for transferring genes into plants.Science,27(4691):1229-1231.
Jia Li’e.2012.Establishment of effective Agrobacterium-mediated mature embryo transformation system for Indica rice[M.D.Dessertation].Beijing:Chinese Academy of Agricultural Sciences.(in Chinese)贾丽娥.2012.农杆菌介导籼稻成熟胚高效转化体系的建立[硕士论文].北京:中国农业科学院.
Liu Jie,Ma Jian,Lei Lei,Sun Chao,Kang Jungen,Xie Jianming.2017.Construction of plant expression vector and transformation of fusarium wilt resistance gene FOC1 to Brassica oleracea.Acta Botanica Boreali-Occidentalia Sinica,26(3):429-436.(in Chinese)刘洁,马建,雷蕾,孙超,康俊根,颉建明.2017.甘蓝抗枯萎病基因FOC1植物表达载体构建及遗传转化.西北农业学报,26(3):429-436.
Liu Shuang,Guan Shuyan,Sun Su,Wei Xiaoyu.2017.Optimization of maize callus culture system and genetic transformation of ICE1.Journal of Jilin Agricultural University,39(5):518-523.(in Chinese)刘双,关淑艳,孙苏,魏晓禹.2017.玉米愈伤组织培养体系优化及ICE1基因的遗传转化.吉林农业大学学报,39(5):518-523.
Liu X P,Yang C,Han F Q,Fang Z Y,Yang L M,Zhuang M,Lv H H,Liu Y M,Li Z S,Zhang Y Y.2016.Genetics and fine mapping of a yellow-green leaf gene in cabbage(Brassica oleracea var.capitata L.).Molecular Breeding,36:82.
Liu Yanjun,Yang Jinghui,Yang Enqin,Liu Yudong,Liu Shuwu.2003.An investigation of the approaches to enhance gene transformation rate with Agrobacterium.Journal of Southwest Agricultural University,25(2):144-146.(in Chinese)刘艳军,杨静慧,杨恩芹,刘玉东,刘树梧.2003.提高农杆菌基因转化率方法的研究.西南大学学报(自然科学版),25(2):144-146.
Lv Q R,Chen C S,Xu Y J,Hu S K,Wang L,Sun K,Chen X,Li X H.2017.Optimization of Agrobacterium tumefaciens-mediated transformation systems in tea plant(Camellia sinensis).Horticultural Plant Journal,3(3):105-109.
Moloney M M,Walker J M,Sharma K K.1989.High efficiency transformation of Brassica napus using Agrobacterium vectors.Plant Cell Reports,8(4):238-242.
Murray H,Thompson W.1980.Rapid isolation of high molecular weight DNA.Nucleic Acids Research,8:4321-4325.
Ni Dexiang.1986.Regulation of light in plant tissue culture.Chinese Journal of Nature,9(3):193-198.(in Chinese)倪德祥.1986.光在植物组织培养中的调控作用.自然杂志,9(3):193-198.
Pavel K,Olga S,Ivan L,Anna D,George K,Jozef S.2015.Transient plant transformation mediated by Agrobacterium tumefaciens:principles,methods and applications.Biotechnology Advances,33:1024-1042.
Peng C J,Xu W G,Hu L,Li Y,Qi X L,Wang H W,Hua X,Zhao M Z.2018.Effects of the maize C4 phosphoenolpyruvate carboxylase(ZmPEPC)gene on nitrogen assimilation in transgenic wheat.Plant Growth Regulation,84(1):191-205.
Qiao Yahong,Zheng Yinying,Li ShiLin,Tian Guiying,Zhang Lili,Lin Tao,Xiang Benchun.2013.A regeneration system for genetic transformation of processing tomato Hongfan 3.Journal of Nuclear Agricultural Sciences,27(11):1636-1643.(in Chinese)乔亚红,郑银英,李诗林,田桂英,张丽丽,林涛,向本春.2013.红番3号加工番茄遗传转化再生体系的建立.核农学报,27(11):1636-1643.
Shen Hailong.2009.Microculture root-raising seedlings of trees in tissue culture.Beijing:China Forestry Publishing House.(in Chinese)沈海龙.2009.树木组织培养微体试管外生根育苗技术.北京:中国林业出版社.
Shi Wenhui.2017.Establishment of efficient regeneration system and genetic transformation of glyphosate resistant genes EPSPs and BnGRF2 in rape[M.D.Dissertation].Lanzhou:Gansu Agricultural University.(in Chinese)石文慧.2017.油菜高效再生体系的建立及Basta抗性基因EPSPs和BnGRF2的遗传转化[硕士论文].兰州:甘肃农业大学.
Swart V,Crampton B G,Ridenour J B,Bluhm B H,Olivier N A,Meyer J J M,Berger D K.2017.Complementation of CTB7 in the maize pathogen cercospora zeina overcomes the lack of in vitro cercosporin production.Molecular Plant-microbe Interactions,30(9):710-724.
Tang Jun,Zhang Shuning,Hou Xilin,Wang Shumin,Sun Chengzhen.2011.Establishment of high-frequency in vitro regeneration system of Chinese kale(Brassica alboglabra Bailey).Journal of Nanjing Agricultural University,34(2):29-32.(in Chinese)唐君,张蜀宁,侯喜林,王淑敏,孙成振.2011.芥蓝高频离体再生体系的建立.南京农业大学学报,34(2):29-32.
Toriyama K,Arimoto Y,Uchimiya H,Hinata K.1988.Transgenic rice plants after direct gene transfer into protoplasts.Nature Biotechnology,6(9):1072-1074.
Wang Guoliang.2004.Optimization of high-efficiency Agrobacterium-mediated BADH gene transformation system of alfalfa and detection of transgenic plants[M.D.Dissertation].Lanzhou:Gansu Agricultural University.(in Chinese)王国良.2004.农杆菌介导的紫花苜蓿BADH基因高效转化体系的优化和转基因植株的检测[硕士论文].兰州:甘肃农业大学.
Wang K,Liu H Y,Du L P,Ye X G.2017.Generation of marker-free transgenic hexaploid wheat via an Agrobacterium-mediated co-transformation strategy in commercial Chinese wheat varieties.Plant Biotechnology Journal,15:614-623.
Wang Li,Yi Dengxia,Yang Limei,Liu Jianping,Fang Zhiyuan,Cheng Fei,Liu Yumei,Zhuang Mu,Zhang Yangyong,Sun Peitian.2014.Obtaining of insect-resistant material with Bt in early spring cabbage.China Vegetables,(10):12-17.(in Chinese)王丽,仪登霞,杨丽梅,刘建萍,方智远,程斐,刘玉梅,庄木,张扬勇,孙培田.2014.转Bt基因早熟春甘蓝抗虫材料的获得.中国蔬菜,(10):12-17.
Wang Q J.2012.Functional verification of drought tolerance gene PC2300AF1 in japanica rice in cold regions.Acta Agriculturae Universitatis Jiangxiensis,34(3):434-438.
Wang Xiao-hong,Zeng Ai-song,Gao Bing,Song Li-xiao,Yan Ji-yong.2013.Resistance to salt in cabbage(Brassica oleracea L.var.capitata L.)transformed with StP5CS gene.Acta Botanica Boreali-Occidentalia Sinica,33(5):885-891.(in Chinese)王肖红,曾爱松,高兵,宋立晓,严继勇.2013.StP5CS转化结球甘蓝的研究.西北植物学报,33(5):885-891.
Xu Lingling,Ding Duoduo,Tao Guirong,Dang Yuefang,Guo Bin.2013.Effects of different light intensity and light quality on adventitious buds induction and growth of Clematis‘Nelly Moser’.Acta Photonica Sinica,42(6):715-720.(in Chinese)徐玲玲,丁朵朵,陶贵荣,党悦方,郭斌.2013.不同光照强度和光质对铁线莲品种繁星(Clematis‘Nelly Moser’)不定芽诱导和生长的影响.光子学报,42(6):715-720.
Yang Chong,Zhang Yangyong,Fang Zhiyuan,Liu Yumei,Yang Limei,Zhuang Mu,Sun Peitian.2014.Photosynthetic physiological characteristics and chloroplast ultrastructure of yellow leaf mutant YL-1 in cabbage.Acta Horticulturae Sinica,41(6):1133-1144.(in Chinese)杨冲,张扬勇,方智远,刘玉梅,杨丽梅,庄木,孙培田.2014.甘蓝叶色黄化突变体YL-1的光合生理特性及其叶绿体的超微结构.园艺学报,41(6):1133-1144.
Yang Weijie.2016.Transformation of Bt Cry1Ia8/Cry1C genes and study of the resistance to pests in transgenic high generation of inbred lines in cabbage[M.D.Dissertation].Beijing:Chinese Academy of Agricultural Sciences.(in Chinese)杨卫杰.2016.Bt Cry1Ia8/Cry1 AC基因转化甘蓝及高代转基因甘蓝的抗虫型研究[硕士论文].北京:中国农业科学院.
Yang Yue,Zhang Zhihong,Liu Yuexue,He Ping,Zhou Yu,Dai Yanhong,Zhao Guiling.2010.Establishment of Agrobacterium tumefaciens-mediated transformation system for Hanfu apple.Journal of Fruit Science,27(2):174-178.(in Chinese)杨越,张志宏,刘月学,何平,周宇,代红艳,赵桂玲.2010.根癌农杆菌介导寒富苹果转化体系的建立.果树学报,27(2):174-178.
Ying Shan,He Xiaowei,Wang Xiurong.2008.Assessment of factors affecting the transformation efficiency of soybean cotyledonary-node Agrobacterium-mediated transformation system.Molecular Plant Breeding,6(1):32-40.(in Chinese)应珊,何晓薇,王秀荣.2008.影响农杆菌介导的大豆转化效率的因素研究.分子植物育种,6(1):32-40.
Yu Shou-jiang.2007.Cloning CryⅠAC gene and the study of transformation of cabbage[M.D.Dissertation].Harbin:Northeast Agricultural University.(in Chinese)于守江.2007.CryⅠAC基因的克隆及转化结球甘蓝的研究[硕士论文].哈尔滨:东北农业大学.
Yu Fengli.2005.Study on transferring of coat pritein gene potato virus a(PVA CP)into potato[M.D.Dissertation].Harbin:Northeast Agricultural University.(in Chinese)于凤丽.2005.马铃薯A病毒外壳蛋白(PVA CP)基因的遗传转化研究[硕士论文].哈尔滨:东北农业大学.
Zhao Xiushu,Li Mingyang,Zhang Wenling,Liu Fan.2009.Establishment of high adventitious shoot regeneration system of ornamental kale.Genomics&Applied Biology,28(1):141-148.(in Chinese)赵秀枢,李名扬,张文玲,刘凡.2009.观赏羽衣甘蓝高频再生体系的建立.基因组学与应用生物学,28(1):141-148.
Zhu W,Zuo Y,Zhou R F,Huang J Y,Tang M Q,Cheng X H,Liu Y Y,Tong C B,Xiang Y,Dong C H,Liu S Y.2016.Vacuolar iron transporter BnMEB2 is involved in enhancing iron tolerance of Brassica napus.Frontiers in Plant Science,7(9):1353.
Zimmermann U,Vienken J.1982.Stable transformation of maize after gene transfer by electroporation.The Journal of Membrane Biology,67:165-182.
Zou Zhi,Lu Changming.2008.An overview of plant factors influencing Agrobacterium-mediated transformation.Biotechnology Bulletin,18(1):1-9.(in Chinese)邹智,卢长明.2008.影响农杆菌介导遗传转化的植物因子研究进展.生物技术通报,18(1):1-9.
Zhai Rui,Gao Le,Ding Xueni,Liao Wenlin,Zhen Mingjie,Lu Zhiwen,Zhi Haijian.2015.Optimization of cotyledonary-node Agrobacteriummediated soybean transformation system.Soybean Science,34(5):768-775.(in Chinese)翟锐,高乐,丁雪妮,廖文林,郑明洁,陆智文,智海剑.2015.农杆菌介导大豆子叶节遗传转化体系的优化.大豆科学,34(5):768-775.