枣幼胚体细胞胚的发生及植株再生
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摘要
以小梨枣胚龄30~40d的幼胚为外植体,研究不同质量浓度6-苄基腺嘌呤(6-BA)、噻苯隆(TDZ)、吲哚丁酸(IBA)、萘乙酸(NAA)、脱落酸(ABA)及其组合对幼胚胚乳看护培养效果、体细胞胚发生、成熟及萌发的影响。结果表明,适宜小梨枣幼胚(30~40d)胚乳看护培养的培养基为MS+IBA0.2mg·L~(-1)+BA0.5mg·L~(-1)+NAA 0.1 mg·L~(-1),成胚率达62.07%;诱导胚性愈伤组织较适宜的培养基为MS+TDZ 0.6mg·L~(-1)+IBA1.5mg·L~(-1)+NAA 0.1mg·L~(-1),诱导率为53.32%;在MS+6-BA 0.3mg·L~(-1)+NAA0.02mg·L~(-1)组合下有利于体细胞胚的发生,发生率可达71.43%;ABA可促进体胚的正常发育,质量浓度为0.5mg·L~(-1)时,体细胞胚结构完整,体胚成熟率为21.83%;将成熟的体胚转接到含有IBA 0.03mg·L~(-1)+GA30.03mg·L~(-1)的培养基中,体细胞胚萌发形成完整植株。
The 30-40d-aged young embryos at globular stage of Ziziphus jujuba ‘Xiaolizao'were used as explants and cultured in MS basic medium with different kinds of plant hormones.The effects of different combinations of plant hormones,including 6-BA,TDZ,IBA,NAA and ABA,on endosperm nursing culture,somatic embryogenesis,somatic maturation and somatic germination were studied in vitro culture.The results showed that the suitable hormone combination for endosperm nursing culture of‘Xiaolizao'immature embryos(30-40d)was IBA0.2 mg·L~(-1)+BA0.5 mg·L~(-1)+ NAA0.1mg·L~(-1),with the embryo formation rate of 62.07%;the most suitable medium to induce embryogenic callus was TDZ0.6mg·L~(-1)+IBA1.5mg·L~(-1)+NAA0.1mg·L~(-1) with the induction rate of53.32%;the hormone combination of somatic embryogenesis was 6-BA0.3 mg·L~(-1)+NAA0.02mg·L~(-1) with the somatic embryogenesis rate of 71.43%;ABA can promote the normal development of somatic embryos,and when the mass concentration is 0.5mg·L~(-1),and the somatic embryo maturation rate is 21.83% with the complete somatic embryo structure;the mature somatic embryos were transferred to the culture medium contraining IBA 0.03mg·L~(-1)+GA30.03mg·L~(-1)to germinate and the complete plants were obtained.
The 30-40d-aged young embryos at globular stage of Ziziphus jujuba ‘Xiaolizao'were used as explants and cultured in MS basic medium with different kinds of plant hormones.The effects of different combinations of plant hormones,including 6-BA,TDZ,IBA,NAA and ABA,on endosperm nursing culture,somatic embryogenesis,somatic maturation and somatic germination were studied in vitro culture.The results showed that the suitable hormone combination for endosperm nursing culture of‘Xiaolizao'immature embryos(30-40d)was IBA0.2 mg·L~(-1)+BA0.5 mg·L~(-1)+ NAA0.1mg·L~(-1),with the embryo formation rate of 62.07%;the most suitable medium to induce embryogenic callus was TDZ0.6mg·L~(-1)+IBA1.5mg·L~(-1)+NAA0.1mg·L~(-1) with the induction rate of53.32%;the hormone combination of somatic embryogenesis was 6-BA0.3 mg·L~(-1)+NAA0.02mg·L~(-1) with the somatic embryogenesis rate of 71.43%;ABA can promote the normal development of somatic embryos,and when the mass concentration is 0.5mg·L~(-1),and the somatic embryo maturation rate is 21.83% with the complete somatic embryo structure;the mature somatic embryos were transferred to the culture medium contraining IBA 0.03mg·L~(-1)+GA30.03mg·L~(-1)to germinate and the complete plants were obtained.
引文
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[16]项伟波,赵金凯,吴家胜,等.香榧体细胞胚发生、发育的形态与细胞学观察[J].园艺学报,2015,42(8):1477-1486.XIANG W B,ZHAO J K,WU J SH,et al.Morphological and cytological observation of somatic embryogenesis and development in Torreyagrandis‘Merrillii’[J].Acta Horticulturae Sinica,2015,42(8):1477-1486(in Chinese with English abstract).
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[20]高述民,李凤兰.ABA对植物体细胞胚胎发育影响的研究进展[J].北京林业大学学报,2002,7(4):122-128.GAO SH M,LI F L.Advances in researches on effeches on effects of ABA on somatic embryogenesis[J].Journal of Beijing Forestry University,2002,7(4):122-128(in Chinese with English abstract).
[2]赵宁,冯建灿,叶霞,等.枣组织培养及相关生物技术研究进展[J].果树学报,2015,32(6):1241-1252.ZHAO N,FENG J C,YE X,et al.A review of tissue culyure and biotechnology in Chinese jujube[J].Journal of Fruit Science,2015,32(6):1241-1252(in Chinese with English abstract).
[3]陈维伦,郭东红.酸枣组织培养中胚状体的形成[J].植物生理学报,1981,7(1):83-84.CHEN W L,GUO D H.Embryoid formation of Zizipus jujuba var.spinosue Hu in tissue culture[J].Journal Plant Physiology,1981,7(1):83-84(in Chinese with English abstract).
[4]李登科,杜学梅,王永康,等.六月鲜枣愈伤组织诱导及胚状体发生[J].果树学报,2004,21(5):414-418.LI D K,DU X M,WANG Y K,et al.Callus induction and embryogenesis of Ziziphus jujube cv.Liuyuexian[J].Journal of Fruit Science,2004,21(5):414-418(in Chinese with English abstract).
[5]郝建平,金竹萍,王永康,等.枣合子胚和体细胞胚发育过程的观察与比较[J].分子细胞生物学报,2006,39(5):423-430.HAO J P,JIN ZH P,WANG Y K,et al.Study on embryo development and somatic embryogenesis of Jujube[J].Journal of Molecular Cell Biology,2006,39(5):423-430(in Chinese with English abstract).
[6]张存智.枣幼胚愈伤组织诱导和胚状体发生[J].北方园艺,2011(13):125-127.ZHANG C ZH.Callus induction and embryogenesis of Ziziphus jujuba[J].Norther Horticulture,2011(13):125-127(in Chinese with English abstract).
[7]王群瑛.枣胚与胚乳的发育[M].植物学报,1983,25(6):526-531.WANG Q Y.Study on development of embryo and endosperm in Chinese Jujube[J].Chinese Bulletin of Botany,1983,25(6):526-531(in Chinese with English abstract).
[8]VIKRANT R A.Somatic embryogenesis from immature and mature embryos of a minor millet Paspalum scrobiculatumt L[J].Plant Cell,Tissue and Organ Culture,2002,69(1):71-77.
[9]RUAN J Y,GEREND S J,HARDTER R,et al.Effect of nitrogen form and rootzone pH on growth and nitrogen uptake of tea(Ca-mellia sinensis)plants[J].Annals of Botany,2007,99(2):301-310.
[10]耿菲菲,肖丰坤,吴涛,等.思茅松成熟胚的胚性愈伤组织诱导与增殖[J].东北林业大学学报,2015,43(5):59-63.GENG F F,XIAO F K,WU T,et al.Induction and proliferation of embryonic calluses from mature zygotic embryos of Pinus kesiyavar.langbianensis[J].Journal of Northeast Forestry University,2015,43(5):59-63(in Chinese with English abstract).
[11]孙丹,李宏博,李千,等.北五味子体细胞胚发生过程中内源IAA、ABA和GA3含量的动态变化[J].植物生理学报,2013,49(1):70-74.SUN D,LI H B,LI Q,et al.Dynamic variation of endogenous hormone during somatic embryo development of Schisandra incarnata[J].Plant Physiology Journal,2013,49(1):70-74(in Chinese with English abstract).
[12]李玲,桂明春,管艳,等.激素对橡胶树次生体细胞胚成熟培养的影响[J].中国农学通报,2015,31(31):30-35.LI L,GUI M CH,GUAN Y,et al.Effects of plant hormones on maturation of secondary somatic embryos of Hevea brasilliensis[J].Chinese Agricultural Science Bulletin,2015,31(31):30-35(in Chinese with English abstract).
[13]齐力旺,韩一凡,韩素英,等.麦芽糖、NAA及ABA对华北落叶松体细胞胚成熟及生根的影响[J].林业科学,2004,40(1):52-57.QI L W,HAN Y F,HAN S Y,et al.Effects of Maltose,NAA and ABA on somatic maturation and radicle rooting of Larix principis-rupprechtii[J].Scientia Silvae Sinicae,2004,40(1):52-57(in Chinese with English abstract).
[14]STASOLLA C,KONG L,YEUNG E C,et al.Maturation of somatic embryos in conifers:morphogenesis,physiology,biochemistry and molecular biology[J].In Vitro Cell Dev-PL,2002,38(2):93-105.
[15]STASOLLA C,YEUNG E C.Recent advances in conifer somatic embryogenesis:Improving somatic embryo quality[J].Plant Cell,Tissue and Organ Culture,2003,74(1):15-35.
[16]项伟波,赵金凯,吴家胜,等.香榧体细胞胚发生、发育的形态与细胞学观察[J].园艺学报,2015,42(8):1477-1486.XIANG W B,ZHAO J K,WU J SH,et al.Morphological and cytological observation of somatic embryogenesis and development in Torreyagrandis‘Merrillii’[J].Acta Horticulturae Sinica,2015,42(8):1477-1486(in Chinese with English abstract).
[17]魏岳荣,杨护,黄秉智,等.Picloram,ABA和TDZ对香蕉体细胞胚胎发生的影响[J].园艺学报,2007,34(1):81-86.WEI Y R,YANG H,HUANG B ZH,et al.Effects of Picloram,ABA and TDZ on somatic embryogenesis of banana[J].Acta Horticulturace Sinica,2007,34(1):81-86(in Chinese with English abstract).
[18]ROHANI E R,ISMANIZAN I,NOOR N M.Somatic embryogenesis of Mangosteen[J].Plant Cell,Tissue and Culture,2012,110:251-259.
[19]于波,刘金梅,刘晓荣,等.白鹤芋胚性细胞悬浮培养和高效植株再生体系的建立[J].园艺学报,2015,42(4):721-730.YU B,LIU J M,LIU X R,et al.An efficient system of embryogenic cell suspension cultures and plant regeneration in Spathiphyllum cannifolium[J].Acta Horticulturae Scienca,2015,42(4):721-730(in Chinese with English abstract).
[20]高述民,李凤兰.ABA对植物体细胞胚胎发育影响的研究进展[J].北京林业大学学报,2002,7(4):122-128.GAO SH M,LI F L.Advances in researches on effeches on effects of ABA on somatic embryogenesis[J].Journal of Beijing Forestry University,2002,7(4):122-128(in Chinese with English abstract).