芸薹属植物原生质体培养研究进展
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摘要
植物原生质体为遗传转化及遗传改良研究提供了有利的试验材料。采用原生质体技术可进行基因瞬时表达研究,可获得突变体材料,也可导入外源目的基因以改良植物种质资源,在芸薹属植物育种上具有重要的应用价值。本综述介绍了芸薹属植物原生质体培养及体细胞杂交的研究进展,主要从原生质体分离纯化、培养再生、原生质体融合及其在育种中的应用等方面进行了概述,详细阐述了影响芸薹属植物原生质体分离及培养的相关因素;分析了芸薹属植物原生质体研究中目前存在的主要问题;并对今后的研究方向和内容进行了展望,以期为芸薹属植物种质遗传改良工作提供参考。
Protoplast can be used as an excellent material in genetic transformation and improvement.It could be used for the research of gene transient expression,and obtain mutant materials,as well as import the exogenous genes for improving plant germplasm resources via the protoplast technology.Thus,it plays an important role in breeding of Brassica species.This paper reviewed advances in protoplast culture and fusion of Brassica species,which included the isolation,culture,regeneration and fusion of protoplast,as well as their application in Brassica species breeding.The relevant factors on protoplast culture and fusion were expounded in detail.The main problems in protoplast technology were analyzed,and the prospects of this technology in future work were also discussed,which could lay a foundation for genetic improvement in Brassica species.
Protoplast can be used as an excellent material in genetic transformation and improvement.It could be used for the research of gene transient expression,and obtain mutant materials,as well as import the exogenous genes for improving plant germplasm resources via the protoplast technology.Thus,it plays an important role in breeding of Brassica species.This paper reviewed advances in protoplast culture and fusion of Brassica species,which included the isolation,culture,regeneration and fusion of protoplast,as well as their application in Brassica species breeding.The relevant factors on protoplast culture and fusion were expounded in detail.The main problems in protoplast technology were analyzed,and the prospects of this technology in future work were also discussed,which could lay a foundation for genetic improvement in Brassica species.
引文
Cao X.,Da i Z.L.,Qin W.B.,and Pan Y.P.,2016,Advances in protoplast fusion technology,Zhongguo Nongxue Tongbao(Chinese Agricultural Science Bulletin),32(25):84-90(曹雪,戴忠良,秦文斌,潘跃平,2016,植物原生质体融合技术的研究进展,中国农学通报,32(25):84-90)
Cardi T.,and Earle E.D.,1997,Production of new CMS Brassica oleracea by transfer of'Anand'cytoplasm from Brassica rapa through protoplast fusion,Theor.Appl.Genet.,94(2):204-212
Chen L.P.,Zhang M.F.,Xiao Q.B.,Wu J.G.,and Hirata Y.,2004Plant regeneration from hypocotyl protoplasts of red cabbage(Brassica oleracea)by using nurse cultures,Plant Cell,Tissue and Organ Culture,77(2):133-138
Davey M.R.,Anthony P.,Power J.B.,and Lowe K.C.,2005Plant protoplasts:status and biotechnological perspectives Biotechnology Advances,23(2):131-171
El-Esawi M.A.,2016,Somatic hybridization and microspore culture in Brassica improvement,In:Anis M.,and Ahmad N(eds),Plant tissue culture:propagation,conservation and crop improvement,Springer,Singapore,pp.599-609
Fan C.L.,Tao L.,Lin N.,and Wu X.L.,2005,The protoplast isolation and purification of yellow-seed rape,Zhongguo Nongxue Tongbao(Chinese Agricultural Science Bulletin),21(2):43-44,68(范春丽,陶澜,林呐,吴晓亮,2005,甘蓝型黄籽油菜原生质体的游离和纯化,中国农学通报,21(2):43-44,68)
Fang Z.Y.,Sun P.T.,Liu Y.M.,Yang L.M,Wang X.W.,and Zhuang M.,2001,Investigation of different types of male sterility and application of dominant male sterility in cabbage,Zhongguo Shucai(China Vegetables),1(1):6-10(方智远,孙培田,刘玉梅,杨丽梅,王晓武,庄木,2001,几种类型甘蓝雄性不育的研究与显性不育系的利用,中国蔬菜,1(1):6-10)
Gaikwad K.,Kirti P.B.,Sharma A.,Prakash S.,and Chopra V.L.,1996,Cytogenetical and molecular investigations on somatic hybrids of Sinapis alba and Brassica juncea and their backcross progeny,Plant Breeding,115(6):480-483
Hagimori M.,Nagaoka M.,Kato N.,and Yoshikawa H.,1992,Production and characterization of somatic hybrids between the Japanese radish and caulflower,Theor.Appl.Genet.,84(7):819-824
Hou X.L.,Cao S.C.,She J.M.,Cai X.N.,and Lu W.Z.,2000,Plantlet regeneration from hypocotyl protoplast culture of Ogu CMS,non-heading Chinese cabbage,Yuanyi Xuebao(Acta Horticulturae Sinica),27(6):449-451(侯喜林,曹寿椿,佘建明,蔡小宁,陆维忠,2000,不结球白菜Ogu CMS下胚轴原生质体培养再生植株,园艺学报,27(6):449-451)
Hu Z.H.,Lang C.X.,Liu Z.H.,Huang R.Z.,Wu G.T.,Chen X.Y.,and Chen J.Q.,2002,Plant regeneration from hypocotyl protoplasts of Brassica napus,Zhejiang Nongye Xuebao(Acta Agriculturae Zhejiangensis),14(5):278-280(胡张华,郎春秀,刘智宏,黄锐之,吴关庭,陈笑芸,陈锦清,2002,甘蓝型油菜下胚轴原生质体培养再生植株,浙江农业学报,14(5):278-280)
Huang L.,2011,The Protoplast culture and plant regeneration of head cabbage,Thesis for M.S.,Zhejiang Normal University,Supervisor:Zhong X.M.,pp.4-5(黄玲,2011,结球甘蓝原生质体培养及植株再生,硕士学位论文,浙江师范大学,导师:钟新民,pp.4-5)
Li G.,Li B.Y.,Wang W.H.,Yue Z.C.,Zhong X.M.,and Hou X.L.,2012,Plant regeneration from hypocotyl protoplast culture of cabbage(Brassica oleracea L.var capitata),Xibei Zhiwu Xuebao(Acta Botanica Boreali-Occidentalia Sinica),32(12):2438-2443(李贵,李必元,王五宏,岳智臣,钟新民,侯喜林,2012,结球甘蓝下胚轴原生质体培养再生植株体系的优化研究,西北植物学报,32(12):2438-2443)
Lian Y.J.,Lin G.Z.,and Zhao X.M.,2011,Morphological and cytological characterization of somatic hybrids obtained by tri-parental protoplast fusion in Brassica species,Yuanyi Xuebao(Acta Horticulturae Sinica),38(11):2099-2110(廉玉姬,林光哲,赵小梅,2011,大白菜,青花菜和叶用芥菜体细胞杂交种形态学与细胞学特性鉴定,园艺学报,38(11)2099-2110)
Lian Y.J.,Lin G.Z.,Zhao X.M.,and Lim H.T.,2011,Production and genetic characterization of somatic hybrids between leaf mustard(Brassica juncea)and broccoli(Brassica oleracea)In Vitro Cellular&Developmental Biology,Plant,47(2)289-296
Lian Y.J.,Zhao X.M.,Lin G.Z.,and Lim H.T.,2012,Protoplas isolation and culture for somatic hybridisation of rapid cycling Brassica rapa with'Anand'CMS and Brassica juncea Plant Cell,Tissue and Organ Culture,109(3):565-572
Lin L.B.,Ma Z.Q.,Zhang C.L.,Liu Y.T.,Mao X.Q.,Xia G.Y.and Ma Z.Q.,2008,Studies on somatic hybridization between Brassica napus and Malthiola incana B.Br,Yunnan Nongye Daxue Xuebao(Journal of Yunnan Agricultural University),23(3):295-301(林良斌,马占强,张传利,刘雅婷毛孝强,厦国银,马忠琴,2008,甘蓝型油菜与紫罗兰体细胞杂交研究,云南农业大学学报,23(3):295-301)
Liu F.,Zhao H.,and Qin F.,2006,The protoplast culture of Brassica campestris ssp.pekinensis and plant regeneration via somatic embryogenesis,Zhiwuxue Tongbao(Chinese Bulletin of Botany),23(3):275-280(刘凡,赵泓,秦帆,2006结球白菜下胚轴原生质体培养及其体细胞胚植株再生植物学通报,23(3):275-280)
Radchuk V.V.,Ryschka U.,Schumann G.,and Klocke E.,2002Genetic transformation of cauliflower(Brassica oleracea var botrytis)by direct DNA uptake into mesophyll protoplasts Physiologia Plantarum,114(3):429-438
Sheng X.G.,Gu H.H.,Zhao Z.Q.,Yu H.F.,Wang J.S.,and Zhang X.H.,2011,Totipotency expression of plant protoplast and its application in Brassica oleracea L.vegetable breeding Zhongguo Shucai(China Vegetables),1(16):1-8(盛小光顾宏辉,赵振卿,虞慧芳,王建升,张晓辉,2011,植物原生质体全能性表达及其在甘蓝类蔬菜育种上的应用,中国蔬菜,1(16):1-8)
Sheng X.G.,Liu F.,Zhu Y.L.,Zhao H.,Zhang L.,and Chen B.,2008Production and analysis of intergeneric somatic hybrids between Brassica oleracea and Matthiola incana,Plant Cell Tissue and Organ Culture,92(1):55-62
Sigareva M.A.,and Earle E.D.,1997,Direct transfer of a cold-tolerant Ogura male sterile cytoplasm into cabbage(Brassica oleracea ssp.capitata)via protoplast fusion,Theor Appl.Genet.,94(2):213-220
Simmonds D.H.,Long N.E.,and Keller W.A.,1991,High plating efficiency and plant regeneration frequency in low density protoplast cultures derived from an embryogenic Brassica napus cell suspension,Plant Cell,Tissue&Organ Culture27(3):231-241
Sun B.,Fang L.,Liu N.,Yan H.Z.,Zhang Y.J.,Shi Q.Q.,and Wang Q.M.,2011,Studies on main nutritional components of Chinese kale among different organs,Yuanyi Xuebao(Acta Horticulturae Sinica),38(3):541-548(孙勃,方莉,刘娜,闫会转,张雅君,施倩倩,汪俏梅,2011,芥蓝不同器官主要营养成分分析,园艺学报,38(3):541-548)
Sun H.H.,2010,The study of somatic hybridization between Brassica napus and Sinapis arvensis by protoplast fusion,Thesis for M.S.,Henan Agricultural University,Supervisor:Ye Y.Z.,pp.8-9(孙慧慧,2010,甘蓝型油菜与新疆野生油菜原生质体融合培养研究,硕士学位论文,河南农业大学,导师:叶永忠,pp.8-9)
Swanson E.B.,Coumans M.P.,Brown G.L.,Patel J.D.,and Beversdorf W.D.,1988,The characterization of herbicide tolerant plants in Brassica napus after in vitro selection of microspores and protoplasts,Plant Cell Reports,7(2):83-87
Wang Q.B.,Zhang Y.Y.,Fang Z.Y.,Liu Y.M.,Yang L.M.,and Zhuang M.,2012,Chloroplast and mitochondrial SSR help to distinguish allo-cytoplasmic male sterile types in cabbage(Brassica oleracea L.var.capitata),Molecular Breeding,30(2):709-716
Wang X.J.,Chen S.R.,and Song M.,1993,Plant regeneration from protoplast of savoy cabbage(Brassica oleracea L.var.subauda),Zhiwu Xuebao(Acta Botanica Sinica),35(2):107-114(王晓佳,陈世儒,宋明,1993,皱叶甘蓝的原生质体培养与植株再生,植物学报,35(2):107-114)
Wang Y.P.,Sonntag K.,Rudloff E.,Groeneveld I.,Gramenz J.,and Chu C.C.,2006,Production and characterization of somatic hybrids between Brassica napus and Raphanus sativus,Plant Cell,Tissue Organ Culture,86(2):279-283
Wu Y.X.,2009,Protoplast preparation and transformation of bcplh RNAi in orange-heading Chinese cabbage,Thesis for M.S.,Northwest A&F University,Supervisor:Zhang L.G.,pp.20-24(武云霞,2009,橙色大白菜原生质体的制备及Bcp LHRNAi载体的遗传转化,硕士学位论文,西北农林科技大学,导师:张鲁刚,pp.20-24)
Xu Z.H.,Davey M.R.,and Cocking E.C.,1982,Plant regeneration from root protoplasts of Brassica,Plant Science Letters,24(1):117-121
Yao X.W.,Liu F.,Yun X.F.,Zhao H.,Ryschka U.,and Schumann G.,2005,Interspecific hybridization between Brassica oleracea L.var.botrytis and Brassica spinescens via asymmetric somatic protoplast fusion,Yuanyi Xuebao(Acta Horticulturae Sinica),32(6):1039-1044(姚星伟,刘凡,云兴福,赵泓,Ryschka U.,Schumann G.,2005,非对称体细胞融合获得花椰菜与Brassica spinescens的种间杂种,园艺学报,32(6):1039-1044)
Yu H.L.,Li Z.Y.,Yang L.M.,Liu Y.M.,Zhuang M.,Zhang L.G.,Lv H.H.,Li Z.S.,Han F.Q.,Liu X.P.,Fang Z.Y.,and Zhang Y.Y.,2017,Morphological and molecular characterization of the second backcross progenies of Ogu-CMS Chinese kale and rapeseed,Euphytica,213(2):55
Zhang Y.,Li C.Q.,Song H.Y.,and Qin J.S.,2007,The progress of protoplast culture research on Brassica vegetable crops,Shengwu Jishu Tongxun(Letters in Biotechnology),18(3):543-545(张艳,李成琼,宋洪元,秦家顺,2007,原生质体培养在芸薹属蔬菜作物中的研究进展,生物技术通讯,18(3):543-545)
Zhao J.L.,Lu B.D.,Liang A.H.,and Zhao M.H.,2005,Optimization of the system of protoplast culture and regeneration for Chinese cabbage,Xibei Zhiwu Xuebao(Acta Botanica Boreali-Occidentalia Sinica),25(3):546-551(赵军良,逯保德,梁爱华,赵美华,2005,大白菜原生质体培养再生体系的优化,西北植物学报,25(3):546-551)
Zhou B.,Nie Y.Z.,Zhao X.L.,and Li Y.H.,2008,Protoplast isolation of Brassica rapa'Tsuda'turnip and transient expression of green fluorescent protein,Shengwu Jishu Tongxun(Letters in Biotechnology),19(4):542-544(周波,聂玉哲,张晓磊,李玉花,2008,芜菁原生质体的分离及绿色荧光蛋白的瞬时表达,生物技术通讯,19(4):542-544)
Cardi T.,and Earle E.D.,1997,Production of new CMS Brassica oleracea by transfer of'Anand'cytoplasm from Brassica rapa through protoplast fusion,Theor.Appl.Genet.,94(2):204-212
Chen L.P.,Zhang M.F.,Xiao Q.B.,Wu J.G.,and Hirata Y.,2004Plant regeneration from hypocotyl protoplasts of red cabbage(Brassica oleracea)by using nurse cultures,Plant Cell,Tissue and Organ Culture,77(2):133-138
Davey M.R.,Anthony P.,Power J.B.,and Lowe K.C.,2005Plant protoplasts:status and biotechnological perspectives Biotechnology Advances,23(2):131-171
El-Esawi M.A.,2016,Somatic hybridization and microspore culture in Brassica improvement,In:Anis M.,and Ahmad N(eds),Plant tissue culture:propagation,conservation and crop improvement,Springer,Singapore,pp.599-609
Fan C.L.,Tao L.,Lin N.,and Wu X.L.,2005,The protoplast isolation and purification of yellow-seed rape,Zhongguo Nongxue Tongbao(Chinese Agricultural Science Bulletin),21(2):43-44,68(范春丽,陶澜,林呐,吴晓亮,2005,甘蓝型黄籽油菜原生质体的游离和纯化,中国农学通报,21(2):43-44,68)
Fang Z.Y.,Sun P.T.,Liu Y.M.,Yang L.M,Wang X.W.,and Zhuang M.,2001,Investigation of different types of male sterility and application of dominant male sterility in cabbage,Zhongguo Shucai(China Vegetables),1(1):6-10(方智远,孙培田,刘玉梅,杨丽梅,王晓武,庄木,2001,几种类型甘蓝雄性不育的研究与显性不育系的利用,中国蔬菜,1(1):6-10)
Gaikwad K.,Kirti P.B.,Sharma A.,Prakash S.,and Chopra V.L.,1996,Cytogenetical and molecular investigations on somatic hybrids of Sinapis alba and Brassica juncea and their backcross progeny,Plant Breeding,115(6):480-483
Hagimori M.,Nagaoka M.,Kato N.,and Yoshikawa H.,1992,Production and characterization of somatic hybrids between the Japanese radish and caulflower,Theor.Appl.Genet.,84(7):819-824
Hou X.L.,Cao S.C.,She J.M.,Cai X.N.,and Lu W.Z.,2000,Plantlet regeneration from hypocotyl protoplast culture of Ogu CMS,non-heading Chinese cabbage,Yuanyi Xuebao(Acta Horticulturae Sinica),27(6):449-451(侯喜林,曹寿椿,佘建明,蔡小宁,陆维忠,2000,不结球白菜Ogu CMS下胚轴原生质体培养再生植株,园艺学报,27(6):449-451)
Hu Z.H.,Lang C.X.,Liu Z.H.,Huang R.Z.,Wu G.T.,Chen X.Y.,and Chen J.Q.,2002,Plant regeneration from hypocotyl protoplasts of Brassica napus,Zhejiang Nongye Xuebao(Acta Agriculturae Zhejiangensis),14(5):278-280(胡张华,郎春秀,刘智宏,黄锐之,吴关庭,陈笑芸,陈锦清,2002,甘蓝型油菜下胚轴原生质体培养再生植株,浙江农业学报,14(5):278-280)
Huang L.,2011,The Protoplast culture and plant regeneration of head cabbage,Thesis for M.S.,Zhejiang Normal University,Supervisor:Zhong X.M.,pp.4-5(黄玲,2011,结球甘蓝原生质体培养及植株再生,硕士学位论文,浙江师范大学,导师:钟新民,pp.4-5)
Li G.,Li B.Y.,Wang W.H.,Yue Z.C.,Zhong X.M.,and Hou X.L.,2012,Plant regeneration from hypocotyl protoplast culture of cabbage(Brassica oleracea L.var capitata),Xibei Zhiwu Xuebao(Acta Botanica Boreali-Occidentalia Sinica),32(12):2438-2443(李贵,李必元,王五宏,岳智臣,钟新民,侯喜林,2012,结球甘蓝下胚轴原生质体培养再生植株体系的优化研究,西北植物学报,32(12):2438-2443)
Lian Y.J.,Lin G.Z.,and Zhao X.M.,2011,Morphological and cytological characterization of somatic hybrids obtained by tri-parental protoplast fusion in Brassica species,Yuanyi Xuebao(Acta Horticulturae Sinica),38(11):2099-2110(廉玉姬,林光哲,赵小梅,2011,大白菜,青花菜和叶用芥菜体细胞杂交种形态学与细胞学特性鉴定,园艺学报,38(11)2099-2110)
Lian Y.J.,Lin G.Z.,Zhao X.M.,and Lim H.T.,2011,Production and genetic characterization of somatic hybrids between leaf mustard(Brassica juncea)and broccoli(Brassica oleracea)In Vitro Cellular&Developmental Biology,Plant,47(2)289-296
Lian Y.J.,Zhao X.M.,Lin G.Z.,and Lim H.T.,2012,Protoplas isolation and culture for somatic hybridisation of rapid cycling Brassica rapa with'Anand'CMS and Brassica juncea Plant Cell,Tissue and Organ Culture,109(3):565-572
Lin L.B.,Ma Z.Q.,Zhang C.L.,Liu Y.T.,Mao X.Q.,Xia G.Y.and Ma Z.Q.,2008,Studies on somatic hybridization between Brassica napus and Malthiola incana B.Br,Yunnan Nongye Daxue Xuebao(Journal of Yunnan Agricultural University),23(3):295-301(林良斌,马占强,张传利,刘雅婷毛孝强,厦国银,马忠琴,2008,甘蓝型油菜与紫罗兰体细胞杂交研究,云南农业大学学报,23(3):295-301)
Liu F.,Zhao H.,and Qin F.,2006,The protoplast culture of Brassica campestris ssp.pekinensis and plant regeneration via somatic embryogenesis,Zhiwuxue Tongbao(Chinese Bulletin of Botany),23(3):275-280(刘凡,赵泓,秦帆,2006结球白菜下胚轴原生质体培养及其体细胞胚植株再生植物学通报,23(3):275-280)
Radchuk V.V.,Ryschka U.,Schumann G.,and Klocke E.,2002Genetic transformation of cauliflower(Brassica oleracea var botrytis)by direct DNA uptake into mesophyll protoplasts Physiologia Plantarum,114(3):429-438
Sheng X.G.,Gu H.H.,Zhao Z.Q.,Yu H.F.,Wang J.S.,and Zhang X.H.,2011,Totipotency expression of plant protoplast and its application in Brassica oleracea L.vegetable breeding Zhongguo Shucai(China Vegetables),1(16):1-8(盛小光顾宏辉,赵振卿,虞慧芳,王建升,张晓辉,2011,植物原生质体全能性表达及其在甘蓝类蔬菜育种上的应用,中国蔬菜,1(16):1-8)
Sheng X.G.,Liu F.,Zhu Y.L.,Zhao H.,Zhang L.,and Chen B.,2008Production and analysis of intergeneric somatic hybrids between Brassica oleracea and Matthiola incana,Plant Cell Tissue and Organ Culture,92(1):55-62
Sigareva M.A.,and Earle E.D.,1997,Direct transfer of a cold-tolerant Ogura male sterile cytoplasm into cabbage(Brassica oleracea ssp.capitata)via protoplast fusion,Theor Appl.Genet.,94(2):213-220
Simmonds D.H.,Long N.E.,and Keller W.A.,1991,High plating efficiency and plant regeneration frequency in low density protoplast cultures derived from an embryogenic Brassica napus cell suspension,Plant Cell,Tissue&Organ Culture27(3):231-241
Sun B.,Fang L.,Liu N.,Yan H.Z.,Zhang Y.J.,Shi Q.Q.,and Wang Q.M.,2011,Studies on main nutritional components of Chinese kale among different organs,Yuanyi Xuebao(Acta Horticulturae Sinica),38(3):541-548(孙勃,方莉,刘娜,闫会转,张雅君,施倩倩,汪俏梅,2011,芥蓝不同器官主要营养成分分析,园艺学报,38(3):541-548)
Sun H.H.,2010,The study of somatic hybridization between Brassica napus and Sinapis arvensis by protoplast fusion,Thesis for M.S.,Henan Agricultural University,Supervisor:Ye Y.Z.,pp.8-9(孙慧慧,2010,甘蓝型油菜与新疆野生油菜原生质体融合培养研究,硕士学位论文,河南农业大学,导师:叶永忠,pp.8-9)
Swanson E.B.,Coumans M.P.,Brown G.L.,Patel J.D.,and Beversdorf W.D.,1988,The characterization of herbicide tolerant plants in Brassica napus after in vitro selection of microspores and protoplasts,Plant Cell Reports,7(2):83-87
Wang Q.B.,Zhang Y.Y.,Fang Z.Y.,Liu Y.M.,Yang L.M.,and Zhuang M.,2012,Chloroplast and mitochondrial SSR help to distinguish allo-cytoplasmic male sterile types in cabbage(Brassica oleracea L.var.capitata),Molecular Breeding,30(2):709-716
Wang X.J.,Chen S.R.,and Song M.,1993,Plant regeneration from protoplast of savoy cabbage(Brassica oleracea L.var.subauda),Zhiwu Xuebao(Acta Botanica Sinica),35(2):107-114(王晓佳,陈世儒,宋明,1993,皱叶甘蓝的原生质体培养与植株再生,植物学报,35(2):107-114)
Wang Y.P.,Sonntag K.,Rudloff E.,Groeneveld I.,Gramenz J.,and Chu C.C.,2006,Production and characterization of somatic hybrids between Brassica napus and Raphanus sativus,Plant Cell,Tissue Organ Culture,86(2):279-283
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