野生蒙古韭组培快繁体系的建立
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摘要
蒙古韭属于葱科葱属,是一种具有营养价值、饲用价值、药用价值和生态价值的野生植物资源。用野生蒙古韭的鳞茎作为外植体接入诱导愈伤培养基(MS+1 mg/L NAA+2 mg/L 6-BA+30 g/L蔗糖+8 g/L琼脂,pH 5.2)中,经过40 d培养,可萌发愈伤组织和丛生芽;放入诱导分化培养基(MS+1 mg/L NAA+1 mg/L6-BA+30 g/L蔗糖+6 g/L琼脂,pH 5.2)中,增殖3~5代,陆续获得大量的不定芽;诱导生根培养基为1/2MS+0.5 mg/L NAA+0.2 mg/L 6-BA+30 g/L蔗糖+6 g/L琼脂(pH 5.2),约20 d左右可生根,每棵小苗3~4条根;幼苗移栽成活率约80%,可以短时间内获得大量的蒙古韭幼苗。蒙古韭组培快繁体系的建立为下一步蒙古韭重要基因的功能验证和蒙古韭转基因育种提供理论基础,也为蒙古韭分子生物学相关研究提供材料。
Alliu m mongolicum belongs to Allium,Alliaceae,which is a kind of wild plant resource with nutritional,feeding,medicinal and ecological value.The bulbus of wild A.mongolicum were used as explants and placed in callus induction(MS medium supplemented with 1 mg/L NAA and 2 mg/L 6-BA30 g/L Suc+8 g/L agar,pH 5.2).After about 40 days,the callus and adventitious shoot appeared.Then callus was placed in differentiation induction(MS medium supplemented with 1 mg/L NAA and 2 mg/L 6-BA30 g/L Suc+6 g/L agar,pH 5.2).A lot of adventitious shoot appeared after 3-5 generations.The rooting induction MS medium was 1/2 MS supplemented with 0.5 mg/L NAA and 0.2 mg/L 6-BA30 g/L Suc+6 g/L agar(pH 5.2).It takes root in about 20 days,and every seedling had 3-4 number roots.The survival rate of seedlings transplant reached about 80%.And researchers would get lots of seedlings in short time.The construction of plant regeneration system could lay the foundations for functional verification of key genes,and transgenetic breeding,also providing materials of molecular biology research in A.mongolicum.
Alliu m mongolicum belongs to Allium,Alliaceae,which is a kind of wild plant resource with nutritional,feeding,medicinal and ecological value.The bulbus of wild A.mongolicum were used as explants and placed in callus induction(MS medium supplemented with 1 mg/L NAA and 2 mg/L 6-BA30 g/L Suc+8 g/L agar,pH 5.2).After about 40 days,the callus and adventitious shoot appeared.Then callus was placed in differentiation induction(MS medium supplemented with 1 mg/L NAA and 2 mg/L 6-BA30 g/L Suc+6 g/L agar,pH 5.2).A lot of adventitious shoot appeared after 3-5 generations.The rooting induction MS medium was 1/2 MS supplemented with 0.5 mg/L NAA and 0.2 mg/L 6-BA30 g/L Suc+6 g/L agar(pH 5.2).It takes root in about 20 days,and every seedling had 3-4 number roots.The survival rate of seedlings transplant reached about 80%.And researchers would get lots of seedlings in short time.The construction of plant regeneration system could lay the foundations for functional verification of key genes,and transgenetic breeding,also providing materials of molecular biology research in A.mongolicum.
引文
Hao J.G.,and Jia J.F.,2009,Establishment of plant regeneration system from protoplasts of leaf base-derived calli in Allium mongolicum Regel,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),28(5):998-1001(郝建国,贾敬芬,2009,沙葱叶基愈伤组织原生质体再生体系的建立,基因组学与应用生物学,28(5):998-1001)
Hao J.P.,Zhou X.M.,and Li S.Q.,1995,Tissue culture and plant regeneration of Chinese chive,Shanxi Daxue Xuebao(Journal of Shanxi University(Natural Science Edition)),18(1):59-62(郝建平,周小梅,李绍清,1995,韭菜花序培养与植株再生,山西大学学报(自然科学版),18(1):59-62)
Nair A.S.,and Seo B.B.,1993,Plantlet regeneration from callus initiated from flower buds in the wild species Allium senescens var.minor,Plant Cell Tissue and Organ Culture,34:205-207
Shuto H.,Abe T.,and Sasahara T.,1993,In vitro propagation of plants from root apex-derived calli in Chinese chive(Allium tuberosumRottle)and garlic(Allium satirum L.),Japanese Journal of Breeding,43:349-355
Tang S.M.,Zhang Y.P.,Gao J.,Cui G.Y.,and Ao Y.,2010,Content changes on of endogenous hormones of Allium mongolicum Regel leaf during growth and development,XinjiangNongye Kexue(Xinjiang Agricultural Sciences),47(S2):34-37(唐式敏,张亚平,高杰,崔国盈,敖英,2010,蒙古韭生长发育过程中叶片的内源激素含量变化,新疆农业科学,47(S2):34-37)
Wang G.Y.,2007,Root tip tissue culture of Chinese chive and plant regeneration,Beifang Yuanyi(Northern Horticulture),(12):199-200(王桂英,2007,韭菜根尖培养及植株再生,北方园艺,(12):199-200)
Wang M.Y.,Wang G.Y.,Tian J.Y.,Fu G.S.,and Zhang Y.P.,2005,The tissue culture of flower organ in Chinese chive Hebei Nongye Daxue Xuebao(Journal of Agricultural University ofHebei),28(2):40-43(王明耀,王桂英,田金玉,付国森,张云平,2005,韭菜花器官的组织培养,河北农业大学学报,28(2):40-43)
Wang Q.L.,Xu Y.Z.,Wang P.T.,and Li X.,2010,Analysis on nutritional quality of leeks in Hexi region,Caoye Kexue(Pratacultural Science),27(10):170-172(王勤礼,许耀照,王佩汤,李霞,张文斌,王泽浩,殷学贵,2010,河西地区韭菜营养品质分析,草业科学,27(10):170-172)
Wang X.J.,Zhang F.L.,Yang Z.R.,and Hao L.Z.,2011,Relationship between characteristic of Allium mongolicum seed coat,embryo,seed leach liquor and seed dormancy,Zhiwu Shengli Xuebao(Plant Physiology Journal),47(6):589-594(王晓娟,张凤兰,杨忠仁,郝丽珍,2011,沙葱种皮特性,种胚及种子浸提液与种子休眠的关系,植物生理学报,47(6):589-594)
Zee S.Y.,19 97,Tissue culture and differentiation of Chinese chive,Hort Science,12(3):264
Zhang M.L.,Gao J.L.,and Sui Y.J.,1998,Primary study on germination of Allium mongolicum Regel seeds,Neimenggu Nongmuxueyuan Xuebao(Journal of Inner Mongolia Institute of Agriculture&Animal Husbandry),19(1):115-117(张美莉,高聚林,隋艳杰,1998,蒙古韭种子发芽特性初报,内蒙古农牧学院学报,19(1):115-117)
Zhang S.,Da K.D.,Cao C.X.,Cao S.R.,and Huang J.L.,2002,Efficient plant regeneration via root tip culture of Allium tuberosum Rottl.ex Spreng,Yuanyi Xuebao(Acta Horticulturae Sinica),29(2):141-144(张松,达克东,曹辰兴,曹淑荣,黄金丽,2002,韭菜组织培养高频植株再生体系的研究,园艺学报,29(2):41-144)
Zhang T.Y.,Chen H.K.,and Zhang C.R.,2014,Polymorphic microsatellite markers for Allium mongolicum Regel(Amaryllidaceae),Genes Genet.Syst.,89:133-136
Zhang Y.H.,Liu S.Z.,Yu Q.S.,He F.L.,and Zhang J.H.,2014,Geographical distribution and seed characteristics of Allium mongolicum in China,Zhongguo Sahmo(Journal of Desert Research),34(2):391-395(张莹花,刘世增,尉秋实,何芳兰,张进虎,2014,蒙古韭(Allium mongolicum)的分布和结实特性,中国沙漠,34(2):391-395)
Zhang Y.H.,Yu Q.S.,Zhang Q.,Hu X.K.,Hu J.,and Fan B.L.,2017,Regional-scale differentiation and phylogeography of a desert plant Allium mongolicum(Liliaceae)inferred from chloroplast DNA sequence variation,Plant Syst.Evol.,303:451-466
Zhao J.H.,Li Q.F.,Na R.T.Y.,and Si Q.,2010,Study on pollen viability and stigma receptivity of three species of Allium,Caoye Kexue(Journal of Inner Mongolia University),27(4):93-96(赵金花,李青丰,那仁图雅,斯琴,2010,3种野生葱属植物花粉活力和柱头可授性研究,草业科学,27(4):93-96)
Hao J.P.,Zhou X.M.,and Li S.Q.,1995,Tissue culture and plant regeneration of Chinese chive,Shanxi Daxue Xuebao(Journal of Shanxi University(Natural Science Edition)),18(1):59-62(郝建平,周小梅,李绍清,1995,韭菜花序培养与植株再生,山西大学学报(自然科学版),18(1):59-62)
Nair A.S.,and Seo B.B.,1993,Plantlet regeneration from callus initiated from flower buds in the wild species Allium senescens var.minor,Plant Cell Tissue and Organ Culture,34:205-207
Shuto H.,Abe T.,and Sasahara T.,1993,In vitro propagation of plants from root apex-derived calli in Chinese chive(Allium tuberosumRottle)and garlic(Allium satirum L.),Japanese Journal of Breeding,43:349-355
Tang S.M.,Zhang Y.P.,Gao J.,Cui G.Y.,and Ao Y.,2010,Content changes on of endogenous hormones of Allium mongolicum Regel leaf during growth and development,XinjiangNongye Kexue(Xinjiang Agricultural Sciences),47(S2):34-37(唐式敏,张亚平,高杰,崔国盈,敖英,2010,蒙古韭生长发育过程中叶片的内源激素含量变化,新疆农业科学,47(S2):34-37)
Wang G.Y.,2007,Root tip tissue culture of Chinese chive and plant regeneration,Beifang Yuanyi(Northern Horticulture),(12):199-200(王桂英,2007,韭菜根尖培养及植株再生,北方园艺,(12):199-200)
Wang M.Y.,Wang G.Y.,Tian J.Y.,Fu G.S.,and Zhang Y.P.,2005,The tissue culture of flower organ in Chinese chive Hebei Nongye Daxue Xuebao(Journal of Agricultural University ofHebei),28(2):40-43(王明耀,王桂英,田金玉,付国森,张云平,2005,韭菜花器官的组织培养,河北农业大学学报,28(2):40-43)
Wang Q.L.,Xu Y.Z.,Wang P.T.,and Li X.,2010,Analysis on nutritional quality of leeks in Hexi region,Caoye Kexue(Pratacultural Science),27(10):170-172(王勤礼,许耀照,王佩汤,李霞,张文斌,王泽浩,殷学贵,2010,河西地区韭菜营养品质分析,草业科学,27(10):170-172)
Wang X.J.,Zhang F.L.,Yang Z.R.,and Hao L.Z.,2011,Relationship between characteristic of Allium mongolicum seed coat,embryo,seed leach liquor and seed dormancy,Zhiwu Shengli Xuebao(Plant Physiology Journal),47(6):589-594(王晓娟,张凤兰,杨忠仁,郝丽珍,2011,沙葱种皮特性,种胚及种子浸提液与种子休眠的关系,植物生理学报,47(6):589-594)
Zee S.Y.,19 97,Tissue culture and differentiation of Chinese chive,Hort Science,12(3):264
Zhang M.L.,Gao J.L.,and Sui Y.J.,1998,Primary study on germination of Allium mongolicum Regel seeds,Neimenggu Nongmuxueyuan Xuebao(Journal of Inner Mongolia Institute of Agriculture&Animal Husbandry),19(1):115-117(张美莉,高聚林,隋艳杰,1998,蒙古韭种子发芽特性初报,内蒙古农牧学院学报,19(1):115-117)
Zhang S.,Da K.D.,Cao C.X.,Cao S.R.,and Huang J.L.,2002,Efficient plant regeneration via root tip culture of Allium tuberosum Rottl.ex Spreng,Yuanyi Xuebao(Acta Horticulturae Sinica),29(2):141-144(张松,达克东,曹辰兴,曹淑荣,黄金丽,2002,韭菜组织培养高频植株再生体系的研究,园艺学报,29(2):41-144)
Zhang T.Y.,Chen H.K.,and Zhang C.R.,2014,Polymorphic microsatellite markers for Allium mongolicum Regel(Amaryllidaceae),Genes Genet.Syst.,89:133-136
Zhang Y.H.,Liu S.Z.,Yu Q.S.,He F.L.,and Zhang J.H.,2014,Geographical distribution and seed characteristics of Allium mongolicum in China,Zhongguo Sahmo(Journal of Desert Research),34(2):391-395(张莹花,刘世增,尉秋实,何芳兰,张进虎,2014,蒙古韭(Allium mongolicum)的分布和结实特性,中国沙漠,34(2):391-395)
Zhang Y.H.,Yu Q.S.,Zhang Q.,Hu X.K.,Hu J.,and Fan B.L.,2017,Regional-scale differentiation and phylogeography of a desert plant Allium mongolicum(Liliaceae)inferred from chloroplast DNA sequence variation,Plant Syst.Evol.,303:451-466
Zhao J.H.,Li Q.F.,Na R.T.Y.,and Si Q.,2010,Study on pollen viability and stigma receptivity of three species of Allium,Caoye Kexue(Journal of Inner Mongolia University),27(4):93-96(赵金花,李青丰,那仁图雅,斯琴,2010,3种野生葱属植物花粉活力和柱头可授性研究,草业科学,27(4):93-96)