大葱成熟胚愈伤组织诱导及植株再生
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摘要
为了建立大葱组培高频再生体系,以‘铁杆大葱’的成熟胚为材料,探讨培养基种类、植物生长调节剂浓度等因素对大葱成熟胚愈伤组织诱导及再生的影响。结果表明,MS培养基适合大葱成熟胚愈伤组织诱导,大葱成熟胚愈伤组织诱导最适宜的培养基为MS+2,4-D (2～3 mg/L)。大葱成熟胚愈伤组织分化培养最适培养基为MS+6-BA (1.0 mg/L)+KT (1.0 mg/L)+NAA (0.1 mg/L),分化率达90%;对分化的再生苗移植于1/2 MS培养基上进行生根及壮苗培养,生根率达到100%,炼苗移栽后小苗的成活率达到90%以上。本研究为进一步优化大葱再生体系提供了依据,为后续大葱的遗传转化的研究起了推动作用。
To establish high frequ ency regeneration system of welsh onion tissue culture, mature embryos of welsh onion cultivar 'Tiegan onion' were selected as experimental materials, and the influence of basic medium and the concentration of plant growth regulator on callus induction and regeneration of mature embryos of welsh onion were studied. The results showed that the MS medium was suitable for callus induction of mature embryos of welsh onion, and the most suitable medium for callus induction of mature embryos of welsh onion was MS +2,4-D(2～3 mg/L). The optimum medium for callus differentiation and culture of mature embryos of welsh onion was MS+6-BA(1.0 mg/L)+KT(1.0 mg/L)+NAA(0.1 mg/L), with the differentiation rate of 90%. The differentiated regenerated seedlings were transplanted on 1/2 MS medium for rooting and strong seedling culture. The rooting rate of the regenerated plants was up to 100%, and the survival rate of seedlings after hardening and transplanting was more than 90%. This study would provide a basis for further optimization of the regeneration system of welsh onion, and could promote the subsequent research on genetic transformation of welsh onion.
To establish high frequ ency regeneration system of welsh onion tissue culture, mature embryos of welsh onion cultivar 'Tiegan onion' were selected as experimental materials, and the influence of basic medium and the concentration of plant growth regulator on callus induction and regeneration of mature embryos of welsh onion were studied. The results showed that the MS medium was suitable for callus induction of mature embryos of welsh onion, and the most suitable medium for callus induction of mature embryos of welsh onion was MS +2,4-D(2～3 mg/L). The optimum medium for callus differentiation and culture of mature embryos of welsh onion was MS+6-BA(1.0 mg/L)+KT(1.0 mg/L)+NAA(0.1 mg/L), with the differentiation rate of 90%. The differentiated regenerated seedlings were transplanted on 1/2 MS medium for rooting and strong seedling culture. The rooting rate of the regenerated plants was up to 100%, and the survival rate of seedlings after hardening and transplanting was more than 90%. This study would provide a basis for further optimization of the regeneration system of welsh onion, and could promote the subsequent research on genetic transformation of welsh onion.
引文
Huang X.M.,Li G.Y.,Liang Y.,and Chen D.,2005,Study on the variation of welsh onion chromosome ploidy in vitro culture,Zhongguo Nongxue Tongbao(Chinese Agricultural Science Bulletin),21(1):50-52(黄晓梅,李桂英,梁艳,陈典,2005,组织培养中大葱染色体倍性变异研究,中国农学通报,21(1):50-52)
Ibrahim A.M.,2016,Haploid induction of kenaf(Hibiscus Cannabinus L.),okra(Abelmoschus esculentus L.)and spring onion(Allium fistulosum L.)using anther,ovary and ovule cultures,Dissertation for Ph.D.,Faculty of Agro-Based Industry,University Malaysia Kelantan,Supervisor:Fatimah B.,pp.11-26
Ibrahim A.M.,Katat F.,Susanto D.,Kashiani P.,and Arifullah M.,2016,Haploid induction in spring onion(Allium fistulosum L.)via gynogenesis,Biotechnology,15(1):10-16
Ma R.Q.,Song Z.X.,Huang Y.,and Yue W.H.,2009,Application progress of modern biotechnology on genetic research of welsh onion(Allium fistulosum L.),Shandong Nongye Kexue(Shandong Agricultural Sciences),(6):18-22(马荣群,宋正旭,黄粤,岳文辉,2009,现代生物技术在大葱遗传研究中的应用进展,山东农业科学,(6):18-22)
Mu C.H.,2002,Tissue culture of spring onion and the analysis of blood relationship among some Allium lines,Thesis for M.S.,Shandong Agriculture University,Supervisor:Yu Y.J.,pp.1-12(穆春华,2002,大葱组织培养与种质亲缘关系分析,硕士学位论文,山东农业大学,导师:于元杰,pp.1-12)
Sanglard N.A.,Amaral-silva P.M.,Sattler M.C.,Sattler M.C.,O-liveira S.C.D.,and Nunes A.C.P.,2017,From chromosome doubling to DNA sequence changes:outcomes of an improved in vitro procedure developed for allotriploid"Híbrido de Timor"(Coffea arabica L.×Coffea canephora Pierre ex A.Froehner),Plant Cell Tiss.Org.,131(2):223-231
Sheng Y.P.,Yang J.P.,Qi X.L.,Zhang S.,and Jiang L.Y.,2004,Plantlet regeneration from mature embryos of scallion in vitro,Shengwu Jishu(Biotechnology),14(3):54-55(盛艳萍,杨建平,齐宪磊,张松,姜璐琰,2004,大葱成熟胚离体培养植株再生,生物技术,14(3):54-55)
Sheng Y.P.,Yang J.P.,Zhang S.,Qi X.L.,and Li X.C.,2005,Studies on the changes of endogenous hormones during the course of regeneration from Allium fistulosum L.mature embryos culture,Yuanyi Xuebao(Acta Horticulturae Sinica),32(2):318-320(盛艳萍,杨建平,张松,齐宪磊,李新成,2005,大葱成熟胚离体再生过程中内源激素的变化,园艺学报,32(2):318-320)
Xie Z.X.,Zhang Y.X.,and Yu Y.J.,2004,Studies on vitrification characteristics of in vitro cultured Chinese onion(Allium fistulosum),Xibei Zhiwu Xuebo(Acta Botanica Boreali-occidentalia Sinica),24(11):2146-2149(谢芝馨,张玉喜,于元杰,2004,大葱组织培养中玻璃苗特性研究,西北植物学报,24(11):2146-2149)
Yang J.J.,Tan W.,Peng J.H.,and Yu Y.J.,2008a,The establishment and optimization of the regeneration system of welsh onion in vitro,Shandong Nongye Daxue Xuebao(Journal of Shandong Agricultural University(Natural Science Edition)),39(2):207-212(杨俊杰,谭伟,彭金环,于元杰,2008a,大葱组织培养及其再生体系的建立与优化,山东农业大学学报(自然科学版),39(2):207-212)
Yang J.J.,Tan W.,Peng J.H.,and Yu Y.J.,2008b,Establishment of the regeneration system of Zhangqiu welsh onion and tissue culture with different organs,Yunnan Nongye Daxue Xuebao(Journal of Yunnan Agricultural University),23(4):571-575(杨俊杰,谭伟,彭金环,于元杰,2008b,章丘大葱再生体系的建立及组培外植体的选择,云南农业大学学报,23(4):571-575)
Zhang S.,and Zhang Q.P.,1994,Study on regeneration plant via flower bud culture of welsh onion(Allium fistulosum L.),Shandong Nongye Daxue Xuebao(Journal of Shandong Agricultural University),25(3):277-282(张松,张启沛1994,大葱(Allium fistulosum L.)花蕾培养再生植株的研究,山东农业大学学报,25(3):277-282)
Zhang S.,and Zhang Q.P.,1995,Studies on propagation in vitro using immature leaf on welsh onion,Yuanyi Xuebao(Acta Horticulturae Sinica),22(2):161-165(张松,张启沛,1995,利用大葱幼叶进行组织培养微繁的研究,园艺学报,22(2):161-165)
Zong H.,2007,Tissue culture and primary studies on polyploid induction of wesh onion,Thesis for M.S.,Shandong Agriculture University,Supervisor:Chen Y.Q.,pp.50-72(宗红,2007,大葱组织培养及其多倍体诱导的初步研究,硕士学位论文,山东农业大学,导师:陈运起,pp.50-72)
Zong H.,Chen Y.Q.,Wang X.F.,and Gao L.M.,2006,Study on the hormone composition regeneration plant culture via mature embryo of welsh onion,Shandong Nongye Kexue(Shandong Agricultural Sciences),5:23-25(宗红,陈运起,王秀峰,高莉敏,2006,大葱成熟胚培养再生植株激素浓度及配比的研究,山东农业科学,5:23-25)
Ibrahim A.M.,2016,Haploid induction of kenaf(Hibiscus Cannabinus L.),okra(Abelmoschus esculentus L.)and spring onion(Allium fistulosum L.)using anther,ovary and ovule cultures,Dissertation for Ph.D.,Faculty of Agro-Based Industry,University Malaysia Kelantan,Supervisor:Fatimah B.,pp.11-26
Ibrahim A.M.,Katat F.,Susanto D.,Kashiani P.,and Arifullah M.,2016,Haploid induction in spring onion(Allium fistulosum L.)via gynogenesis,Biotechnology,15(1):10-16
Ma R.Q.,Song Z.X.,Huang Y.,and Yue W.H.,2009,Application progress of modern biotechnology on genetic research of welsh onion(Allium fistulosum L.),Shandong Nongye Kexue(Shandong Agricultural Sciences),(6):18-22(马荣群,宋正旭,黄粤,岳文辉,2009,现代生物技术在大葱遗传研究中的应用进展,山东农业科学,(6):18-22)
Mu C.H.,2002,Tissue culture of spring onion and the analysis of blood relationship among some Allium lines,Thesis for M.S.,Shandong Agriculture University,Supervisor:Yu Y.J.,pp.1-12(穆春华,2002,大葱组织培养与种质亲缘关系分析,硕士学位论文,山东农业大学,导师:于元杰,pp.1-12)
Sanglard N.A.,Amaral-silva P.M.,Sattler M.C.,Sattler M.C.,O-liveira S.C.D.,and Nunes A.C.P.,2017,From chromosome doubling to DNA sequence changes:outcomes of an improved in vitro procedure developed for allotriploid"Híbrido de Timor"(Coffea arabica L.×Coffea canephora Pierre ex A.Froehner),Plant Cell Tiss.Org.,131(2):223-231
Sheng Y.P.,Yang J.P.,Qi X.L.,Zhang S.,and Jiang L.Y.,2004,Plantlet regeneration from mature embryos of scallion in vitro,Shengwu Jishu(Biotechnology),14(3):54-55(盛艳萍,杨建平,齐宪磊,张松,姜璐琰,2004,大葱成熟胚离体培养植株再生,生物技术,14(3):54-55)
Sheng Y.P.,Yang J.P.,Zhang S.,Qi X.L.,and Li X.C.,2005,Studies on the changes of endogenous hormones during the course of regeneration from Allium fistulosum L.mature embryos culture,Yuanyi Xuebao(Acta Horticulturae Sinica),32(2):318-320(盛艳萍,杨建平,张松,齐宪磊,李新成,2005,大葱成熟胚离体再生过程中内源激素的变化,园艺学报,32(2):318-320)
Xie Z.X.,Zhang Y.X.,and Yu Y.J.,2004,Studies on vitrification characteristics of in vitro cultured Chinese onion(Allium fistulosum),Xibei Zhiwu Xuebo(Acta Botanica Boreali-occidentalia Sinica),24(11):2146-2149(谢芝馨,张玉喜,于元杰,2004,大葱组织培养中玻璃苗特性研究,西北植物学报,24(11):2146-2149)
Yang J.J.,Tan W.,Peng J.H.,and Yu Y.J.,2008a,The establishment and optimization of the regeneration system of welsh onion in vitro,Shandong Nongye Daxue Xuebao(Journal of Shandong Agricultural University(Natural Science Edition)),39(2):207-212(杨俊杰,谭伟,彭金环,于元杰,2008a,大葱组织培养及其再生体系的建立与优化,山东农业大学学报(自然科学版),39(2):207-212)
Yang J.J.,Tan W.,Peng J.H.,and Yu Y.J.,2008b,Establishment of the regeneration system of Zhangqiu welsh onion and tissue culture with different organs,Yunnan Nongye Daxue Xuebao(Journal of Yunnan Agricultural University),23(4):571-575(杨俊杰,谭伟,彭金环,于元杰,2008b,章丘大葱再生体系的建立及组培外植体的选择,云南农业大学学报,23(4):571-575)
Zhang S.,and Zhang Q.P.,1994,Study on regeneration plant via flower bud culture of welsh onion(Allium fistulosum L.),Shandong Nongye Daxue Xuebao(Journal of Shandong Agricultural University),25(3):277-282(张松,张启沛1994,大葱(Allium fistulosum L.)花蕾培养再生植株的研究,山东农业大学学报,25(3):277-282)
Zhang S.,and Zhang Q.P.,1995,Studies on propagation in vitro using immature leaf on welsh onion,Yuanyi Xuebao(Acta Horticulturae Sinica),22(2):161-165(张松,张启沛,1995,利用大葱幼叶进行组织培养微繁的研究,园艺学报,22(2):161-165)
Zong H.,2007,Tissue culture and primary studies on polyploid induction of wesh onion,Thesis for M.S.,Shandong Agriculture University,Supervisor:Chen Y.Q.,pp.50-72(宗红,2007,大葱组织培养及其多倍体诱导的初步研究,硕士学位论文,山东农业大学,导师:陈运起,pp.50-72)
Zong H.,Chen Y.Q.,Wang X.F.,and Gao L.M.,2006,Study on the hormone composition regeneration plant culture via mature embryo of welsh onion,Shandong Nongye Kexue(Shandong Agricultural Sciences),5:23-25(宗红,陈运起,王秀峰,高莉敏,2006,大葱成熟胚培养再生植株激素浓度及配比的研究,山东农业科学,5:23-25)