瓦氏秋海棠(Begonia wallichiana Lehm.)的遗传转化
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摘要
瓦氏秋海棠(Begonia wallichiana Lehm.)由于植株个体小、生长周期短、四季均可开花结实、不易滋生病虫害等优点,是一种潜在的优良模式植物候选材料。以瓦氏秋海棠种子获得无菌苗,取其叶片为外植体,利用叶盘转化法对外植体进行不定芽诱导和植株再生。利用该再生体系将潮霉素HYG(hygromycin)基因和荷花查耳酮合成酶CHS(chalcone synthase)基因成功导入到瓦氏秋海棠基因组中。本研究通过叶盘转化法来建立瓦氏秋海棠的离体培养和再生体系,该遗传转化体系的确立对将来开展秋海棠属植物的分子生物研究及育种等提供了重要的科学依据。
Begonia wallichiana Lehm.is expected to be a potential candidate of model plants due to its advantages in small plant size,short growth cycle,flowering and fruiting throughout the year,and little vulnerability of disease and pest damage.With the leaf of aseptic seedling of B.wallichiana as explant,leaf disc transformation was used to induce the adventitious shoots from young leaf blades.Using this regeneration system,hygromycin(HYG)gene and chalcone synthase(CHS)gene from lotus(Nelumbo)were successfully integrated into genome of B.wallichiana,respectively.The in vitro culture and regeneration system of B.wallichiana was established by leaf disc transformation method,which could provide important scientific basis for the molecular research and breeding of Begonia in the future.
Begonia wallichiana Lehm.is expected to be a potential candidate of model plants due to its advantages in small plant size,short growth cycle,flowering and fruiting throughout the year,and little vulnerability of disease and pest damage.With the leaf of aseptic seedling of B.wallichiana as explant,leaf disc transformation was used to induce the adventitious shoots from young leaf blades.Using this regeneration system,hygromycin(HYG)gene and chalcone synthase(CHS)gene from lotus(Nelumbo)were successfully integrated into genome of B.wallichiana,respectively.The in vitro culture and regeneration system of B.wallichiana was established by leaf disc transformation method,which could provide important scientific basis for the molecular research and breeding of Begonia in the future.
引文
Chen G.,Liang J.L.,Chen X.W.,and Jin H.,2009,Study on tissue culture and plantlet regeneration of Begonia palmata D.Don,Beifang Yuanyi(Northern Horticulture),(6):92-94(陈刚,梁晶龙,陈雄伟,金红,2009,裂叶秋海棠组织培养及植株再生研究,北方园艺,(6):92-94)
Chen Q.,Zhang D.S.,Tian D.K.,Liu Q.Q.,Fu N.F.,Zhao X.S.,Wu X.,and Wu S.H.,2016,Tissue culture and rapid propagation of Begonia wallichiana Lehm.,Yuanyi Xuebao(Acta Horticulturae Sinica),43(S):2676(陈青,张大生,田代科,刘青青,付乃峰,赵喜双,吴璇,吴少华,2016,瓦氏秋海棠组培再生体系的建立,园艺学报,43(S):2676)
Du Q.L.,2006,Research on seedlings vitrification of Rieger begonia,Anhui Nongye Kexue(Journal of Anhui Agricultural Sciences),34(7):1337-1339(杜启兰,2006,丽佳秋海棠试管苗玻璃化现象的控制研究,安徽农业科学,34(7):1337,1339)
Espino F.J.,Linacero R.,Rueda J.,and Vazquez A.M.,2004,Shoot regeneration in four Begonia genotypes,Biol.Plant.,48(1):101-104
Guan K.Y.,Yamaguchi H.,Li J.X.,Li H.Z.,and Ma H.,2007,Traditional uses of begonias(Begoniaceae)in China,Yunan Zhiwu Yanjiu(Acta Botanica Yunnanica),29(1):58-66(管开云,山口裕文,李景秀,李宏哲,马宏,2007,中国秋海棠属植物的传统利用,云南植物研究,29(1):58-66)
Heide O.M.,1968,Auxin level and regeneration of Begonia leaves,Planta,81(2):153-159
Hou Z.M.,Wang Z.X.,Mandula,and Enhebayaer,2001,Tissue culture and rapid propagation of Begonia masoniana and Begonia rex,Neimenggu Shida Xuebao(Journal of Inner Mongolia Normal University),30(3):260-262(侯占铭,王振兴,满都拉,恩和巴雅尔,2001,铁十字秋海棠和蟆叶秋海棠的组织培养与快速繁殖,内蒙古师大学报,30(3):260-262)
Ji C.Y.,Cui D.L.,and Ma Y.X.,2007,Tissue culture and plantlet regeneration of Begonia heracleifolia Cham.et Schlechtend.,Zhiwu Shenglixue Tongxun(Plant Physiology Communications),43(3):504(纪春艳,崔大练,马玉心,2007,枫叶秋海棠的组织培养与植株再生,植物生理学通讯,43(3):504)
Kiyokawa S.,Kikuchi Y.,Kamada H.,and Harada H.,1996,Genetic transformation of Begonia tuberhybrida by Ri rol genes,Plant Cell Rep.,15(8):606-609
Kong X.S.,and Zhang M.X.,1998,The petiole culture and plantlet regeneration of Begonia rex,Beifang Yuanyi(Northern Horticulture),3(4):102-103(孔样生,张妙霞,1998,蟆叶秋海棠的叶柄培养和植株再生,北方园艺,3(4):102-103)
Li J.X.,Guan K.Y.,and Kong F.C.,2000,Leaf culture and rapid propagation of Begonia longialata,Zhiwu Shenglixue Tongxun(Plant Physiology Communications),36(5):439-440(李景秀,管开云,孔繁才,2000,长翅秋海棠的叶片培养和快速繁殖,植物生理学通讯,36(5):439-440)
Li Y.X.,Zhang C.F.,Zong X.C.,and Liu H.L.,2002,Tissue culture and plantlet regeneration of Begonia argnia argenteo-guttata,Mudanjiang Shifan Xueyuan Xuebao(Journal of Mudanjiang Normal University),(2):24(李亚贤,张春凤,宗宪春,刘慧丽,2002,银星秋海棠组织培养及植株再生,牡丹江师范学院学报,(2):24)
Liang H.Y.,2013,The adventitious buds induction and seedlings proliferation of Begonia semperflorens,Linye Shiyong Jishu(Practical Forestry Technology),(5):59-60(梁红艳,2013,四季秋海棠不定芽诱导及试管苗的增殖,林业实用技术,(5):59-60)
Lin F.,Li J.Y.,Huang D.M.,Xu Y.,Li Y.J.,Li Y.X.,Ding Z.L.,and Tang F.L.,2015,Study of tissue culture and plant regeneration of A quilaria.sinensis,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),34(6):1296-1299(林妃,李敬阳,黄东梅,许奕,李羽佳,李艳霞,丁哲利,唐粉玲,2015,白木香组织培养技术及植株再生的研究,基因组学与应用生物学,34(6):1296-1299)
Lu Y.X.,Toshinari G.O.D.O.,Guan K.Y.,and Li H.Z.,2007,Tissue culture and plantlet regeneration of Begonia rubropunctata S.H.Huang et Shui,Zhiwu Shenglixue Tongxun(Plant Physiology Communications),43(6):1131-1132(鲁元学,神户敏成,管开云,李宏哲,2007,红斑秋海棠的组织培养和植株再生,植物生理学通讯,43(6):1131-1132)
Situ L.L.,Zhang X.J.,Zong X.C.,and Ren R.Y.,2007,Callus induction and plantlet regeneration of Begonia imperialis Lem.,Zhiwu Shenglixue Tongxun(Plant Physiology Communications),43(1):117(司徒琳莉,张小军,宗宪春,任如意,2007,帝王秋海棠愈伤组织的诱导和植株再生,植物生理学通讯,43(1):117)
Tang F.L.,Huang N.Z.,Jiang N.,Xian K.H.,and Huang X.M.,2013,Tissue culture and rapid propagation of Begonia retinervia,Xinan Nongye Xuebao(Southwest China Journal of Agricultural Sciences),26(2):762-765(唐凤鸾,黄宁珍,蒋能,冼康华,黄雪梅,2013,突脉秋海棠组织培养及快速繁殖研究,西南农业学报,26(2):762-765)
Tian D.K.,Guan K.Y.,Guo R.X.,and Li J.X.,2001,Propagation and cultivation of Begonia versicolor,Guangxi Zhiwu(Guihaia),21(4):375-380(田代科,管开云,郭瑞贤,李景秀,2001,变色秋海棠的繁殖栽培,广西植物,21(4):375-380)
Tian D.K.,Li C.,Xiao Y.,Fu N.F.,Tong Y.,and Wu R.J.,2017,Occurrence and characteristics of natural hybridization in Begonia in China,Shengwu Duoyangxing(Biodiversity Science),25(6):654-674(田代科,李春,肖艳,付乃峰,童毅,吴瑞娟,2017,中国秋海棠属植物的自然杂交发生及其特点,生物多样性,25(6):654-674)
Xing G.M.,Wu H.H.,Li D.,and Xu X.W.,2013,Influencing factors of regeneration in vitro on Begonia tuberhybrida Voss cv Jinzhengri,Heilongjiang Nongye Kexue(Heilongjiang Agricultural Sciences),12(3):9-12(邢桂梅,吴海红,李丹,徐兴伟,2013,球根秋海棠金正日离体再生影响因素研究,黑龙江农业科学,12(3):9-12)
Xu Q.L.,Dong J.L.,Gao N.,Ruan M.Y.,Jia H.Y.,Zhang L.,and Wang C.Y.,2011,Transgenic lines of Begonia maculata generated by ectopic expression of PttKN1,Biologia,66(2):251-257
Xu Q.L.,Xie Y.H.,Dong J.L.,Ruan M.Y..,Hu X.,and Wang C.Y.,2011,Altered leaf morphology generated by overexpression of PttKN1 gene in Begonia maculata,Beifang Yuanyi(Northern Horticulture),5(21):98-102(徐全乐,谢亚红,董江陵,阮美煜,胡鑫,王崇英,2011,PttKN1基因过表达对斑叶竹节秋海棠叶型的影响,北方园艺,5(21):98-102)
Zheng R.X.,1985,Rapid propagation of Begonia rex by using the tissue culture of leaf blade,Yunan Zhiwu Yanjiu(Acta Botanica Yunnanica),7(1):125-127(郑若仙,1985,毛叶秋海棠叶片组织培养快速繁殖,云南植物研究,7(1):125-127)
Zong X.C.,and Zong C.H.,2010,Study on the rapid propagation with tissue culture of Begonia heracleifolia,Anhui Nongye Kexue(Journal of Anhui Agricultural Sciences),38(24):13561-13562(宗宪春,宗灿华,2010,枫叶秋海棠组织培养与快繁技术的研究,安徽农业科学,38(24):13561-13562)
Chen Q.,Zhang D.S.,Tian D.K.,Liu Q.Q.,Fu N.F.,Zhao X.S.,Wu X.,and Wu S.H.,2016,Tissue culture and rapid propagation of Begonia wallichiana Lehm.,Yuanyi Xuebao(Acta Horticulturae Sinica),43(S):2676(陈青,张大生,田代科,刘青青,付乃峰,赵喜双,吴璇,吴少华,2016,瓦氏秋海棠组培再生体系的建立,园艺学报,43(S):2676)
Du Q.L.,2006,Research on seedlings vitrification of Rieger begonia,Anhui Nongye Kexue(Journal of Anhui Agricultural Sciences),34(7):1337-1339(杜启兰,2006,丽佳秋海棠试管苗玻璃化现象的控制研究,安徽农业科学,34(7):1337,1339)
Espino F.J.,Linacero R.,Rueda J.,and Vazquez A.M.,2004,Shoot regeneration in four Begonia genotypes,Biol.Plant.,48(1):101-104
Guan K.Y.,Yamaguchi H.,Li J.X.,Li H.Z.,and Ma H.,2007,Traditional uses of begonias(Begoniaceae)in China,Yunan Zhiwu Yanjiu(Acta Botanica Yunnanica),29(1):58-66(管开云,山口裕文,李景秀,李宏哲,马宏,2007,中国秋海棠属植物的传统利用,云南植物研究,29(1):58-66)
Heide O.M.,1968,Auxin level and regeneration of Begonia leaves,Planta,81(2):153-159
Hou Z.M.,Wang Z.X.,Mandula,and Enhebayaer,2001,Tissue culture and rapid propagation of Begonia masoniana and Begonia rex,Neimenggu Shida Xuebao(Journal of Inner Mongolia Normal University),30(3):260-262(侯占铭,王振兴,满都拉,恩和巴雅尔,2001,铁十字秋海棠和蟆叶秋海棠的组织培养与快速繁殖,内蒙古师大学报,30(3):260-262)
Ji C.Y.,Cui D.L.,and Ma Y.X.,2007,Tissue culture and plantlet regeneration of Begonia heracleifolia Cham.et Schlechtend.,Zhiwu Shenglixue Tongxun(Plant Physiology Communications),43(3):504(纪春艳,崔大练,马玉心,2007,枫叶秋海棠的组织培养与植株再生,植物生理学通讯,43(3):504)
Kiyokawa S.,Kikuchi Y.,Kamada H.,and Harada H.,1996,Genetic transformation of Begonia tuberhybrida by Ri rol genes,Plant Cell Rep.,15(8):606-609
Kong X.S.,and Zhang M.X.,1998,The petiole culture and plantlet regeneration of Begonia rex,Beifang Yuanyi(Northern Horticulture),3(4):102-103(孔样生,张妙霞,1998,蟆叶秋海棠的叶柄培养和植株再生,北方园艺,3(4):102-103)
Li J.X.,Guan K.Y.,and Kong F.C.,2000,Leaf culture and rapid propagation of Begonia longialata,Zhiwu Shenglixue Tongxun(Plant Physiology Communications),36(5):439-440(李景秀,管开云,孔繁才,2000,长翅秋海棠的叶片培养和快速繁殖,植物生理学通讯,36(5):439-440)
Li Y.X.,Zhang C.F.,Zong X.C.,and Liu H.L.,2002,Tissue culture and plantlet regeneration of Begonia argnia argenteo-guttata,Mudanjiang Shifan Xueyuan Xuebao(Journal of Mudanjiang Normal University),(2):24(李亚贤,张春凤,宗宪春,刘慧丽,2002,银星秋海棠组织培养及植株再生,牡丹江师范学院学报,(2):24)
Liang H.Y.,2013,The adventitious buds induction and seedlings proliferation of Begonia semperflorens,Linye Shiyong Jishu(Practical Forestry Technology),(5):59-60(梁红艳,2013,四季秋海棠不定芽诱导及试管苗的增殖,林业实用技术,(5):59-60)
Lin F.,Li J.Y.,Huang D.M.,Xu Y.,Li Y.J.,Li Y.X.,Ding Z.L.,and Tang F.L.,2015,Study of tissue culture and plant regeneration of A quilaria.sinensis,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),34(6):1296-1299(林妃,李敬阳,黄东梅,许奕,李羽佳,李艳霞,丁哲利,唐粉玲,2015,白木香组织培养技术及植株再生的研究,基因组学与应用生物学,34(6):1296-1299)
Lu Y.X.,Toshinari G.O.D.O.,Guan K.Y.,and Li H.Z.,2007,Tissue culture and plantlet regeneration of Begonia rubropunctata S.H.Huang et Shui,Zhiwu Shenglixue Tongxun(Plant Physiology Communications),43(6):1131-1132(鲁元学,神户敏成,管开云,李宏哲,2007,红斑秋海棠的组织培养和植株再生,植物生理学通讯,43(6):1131-1132)
Situ L.L.,Zhang X.J.,Zong X.C.,and Ren R.Y.,2007,Callus induction and plantlet regeneration of Begonia imperialis Lem.,Zhiwu Shenglixue Tongxun(Plant Physiology Communications),43(1):117(司徒琳莉,张小军,宗宪春,任如意,2007,帝王秋海棠愈伤组织的诱导和植株再生,植物生理学通讯,43(1):117)
Tang F.L.,Huang N.Z.,Jiang N.,Xian K.H.,and Huang X.M.,2013,Tissue culture and rapid propagation of Begonia retinervia,Xinan Nongye Xuebao(Southwest China Journal of Agricultural Sciences),26(2):762-765(唐凤鸾,黄宁珍,蒋能,冼康华,黄雪梅,2013,突脉秋海棠组织培养及快速繁殖研究,西南农业学报,26(2):762-765)
Tian D.K.,Guan K.Y.,Guo R.X.,and Li J.X.,2001,Propagation and cultivation of Begonia versicolor,Guangxi Zhiwu(Guihaia),21(4):375-380(田代科,管开云,郭瑞贤,李景秀,2001,变色秋海棠的繁殖栽培,广西植物,21(4):375-380)
Tian D.K.,Li C.,Xiao Y.,Fu N.F.,Tong Y.,and Wu R.J.,2017,Occurrence and characteristics of natural hybridization in Begonia in China,Shengwu Duoyangxing(Biodiversity Science),25(6):654-674(田代科,李春,肖艳,付乃峰,童毅,吴瑞娟,2017,中国秋海棠属植物的自然杂交发生及其特点,生物多样性,25(6):654-674)
Xing G.M.,Wu H.H.,Li D.,and Xu X.W.,2013,Influencing factors of regeneration in vitro on Begonia tuberhybrida Voss cv Jinzhengri,Heilongjiang Nongye Kexue(Heilongjiang Agricultural Sciences),12(3):9-12(邢桂梅,吴海红,李丹,徐兴伟,2013,球根秋海棠金正日离体再生影响因素研究,黑龙江农业科学,12(3):9-12)
Xu Q.L.,Dong J.L.,Gao N.,Ruan M.Y.,Jia H.Y.,Zhang L.,and Wang C.Y.,2011,Transgenic lines of Begonia maculata generated by ectopic expression of PttKN1,Biologia,66(2):251-257
Xu Q.L.,Xie Y.H.,Dong J.L.,Ruan M.Y..,Hu X.,and Wang C.Y.,2011,Altered leaf morphology generated by overexpression of PttKN1 gene in Begonia maculata,Beifang Yuanyi(Northern Horticulture),5(21):98-102(徐全乐,谢亚红,董江陵,阮美煜,胡鑫,王崇英,2011,PttKN1基因过表达对斑叶竹节秋海棠叶型的影响,北方园艺,5(21):98-102)
Zheng R.X.,1985,Rapid propagation of Begonia rex by using the tissue culture of leaf blade,Yunan Zhiwu Yanjiu(Acta Botanica Yunnanica),7(1):125-127(郑若仙,1985,毛叶秋海棠叶片组织培养快速繁殖,云南植物研究,7(1):125-127)
Zong X.C.,and Zong C.H.,2010,Study on the rapid propagation with tissue culture of Begonia heracleifolia,Anhui Nongye Kexue(Journal of Anhui Agricultural Sciences),38(24):13561-13562(宗宪春,宗灿华,2010,枫叶秋海棠组织培养与快繁技术的研究,安徽农业科学,38(24):13561-13562)