[1]孙兴民,余智莹,张萌,等.葡萄未成熟胚诱导体细胞胚发生和植株再生与遗传鉴定[J].南京农业大学学报,2012,35(3):13-18.[doi:10.7685/j.issn.1000-2030.2012.03.003]
 SUN Xing-min,YU Zhi-ying,ZHANG Meng,et al.Somatic embryogenesis from immature zygotic embryos and genetic fidelity identificating of regenerated plants of grapevine[J].Journal of Nanjing Agricultural University,2012,35(3):13-18.[doi:10.7685/j.issn.1000-2030.2012.03.003]
点击复制

葡萄未成熟胚诱导体细胞胚发生和植株再生与遗传鉴定()
分享到:

《南京农业大学学报》[ISSN:1000-2030/CN:32-1148/S]

卷:
35卷
期数:
2012年3期
页码:
13-18
栏目:
出版日期:
2012-05-18

文章信息/Info

Title:
Somatic embryogenesis from immature zygotic embryos and genetic fidelity identificating of regenerated plants of grapevine
作者:
孙兴民 余智莹 张萌 董瑞奇 陶建敏
南京农业大学园艺学院, 江苏 南京 210095
Author(s):
SUN Xing-min YU Zhi-ying ZHANG Meng DONG Rui-qi TAO Jian-min
College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China
关键词:
葡萄未成熟合子胚体细胞胚发生流式细胞仪SSR
Keywords:
grapevineimmature zygotic embryossomatic embryogenesisflow cytometrySSR
分类号:
S663.1
DOI:
10.7685/j.issn.1000-2030.2012.03.003
文献标志码:
A
摘要:
以二倍体和四倍体葡萄杂交获得的未成熟合子胚为材料,诱导体细胞胚发生。采用的初生胚诱导培养基(SE培养基)为:NN69+1.80 mg·L-1 NAA+0.4 mg·L-1水解酪蛋白+2 g·L-1活性炭+30 g·L-1蔗糖+6 g·L-1琼脂;次生胚诱导培养基分别为:3/4MS+0.35 mg·L-1 IBA+30 g·L-1蔗糖+6 g·L-1琼脂(扩繁培养基)和MS+1.5 g·L-1 6-BA+0.2 g·L-1 IBA+30 g·L-1蔗糖+6 g·L-1琼脂(胚状体萌发培养基)。结果表明:初生胚诱导率为11.11%和16.67%,体细胞胚在扩繁培养基上再生植株。利用流式细胞仪和SSR标记检测源自同一未成熟合子胚(胚珠)的不同株系的遗传稳定性,结果显示,所测植株均为三倍体,且都来自杂交所得的未成熟合子胚。本研究所建立的体细胞胚诱导方法及遗传稳定性检测技术,能够为植物材料保存、扩繁,转基因应用和研究植物发育及极性建成等提供途径。
Abstract:
Somatic embryogenesis and plant regeneration were successfully established on Nitsch and Nitsch(NN69)medium from immature zygotic embryos(IZEs)of interploid crossing 2004-6-12 בAki Queen’.The optimum hormone combinations was NN69+1.80 mg·L-1 NAA+0.4 mg·L-1 casein hydroly sate+2 g·L-1 activated carbon+30 g·L-1 sucrose+6 g·L-1 agar(SE medium)for primary somatic embryogenesis directly from IZEs,with the frequency varying from 11.11% to 16.67% among IZEs of different sampling period after pollination.And importantly,somatic embryogenesis and normal germinating occurred synchronically.While the optimum medium for secondary somatic embryogenesis were 3/4MS+0.35 mg·L-1 IBA+30 g·L-1 sucrose+6 g·L-1 agar(multiplication medium)and MS+1.5 g·L-1 6-BA+0.2 g·L-1 IBA+30 g·L-1 sucrose+6 g·L-1 agar(somatic embryo germination medium).After subcultured on germination medium,somatic embryos converted into normal plantlets.The analysis of DNA content and genetic fidelity determined by flow cytometry and SSR of the regenerated plantlets clearly indicated that the four plantlets were triploid and the normal germinated plant and the somatic embryogenesis-derived plants came from the same immature zygotic embryos.We concluded that somaclonal variation was almost absent in our grapevine plant regeneration system,which was useful in research of transformation and embryo development and so on.

参考文献/References:

[1] Steward F C,Mapes M O,Smith J.Growth and organized development of cultured cellsⅠ:Growth and division of freely suspended cells[J].American Journal of Botany,1958,45:693-703
[2] 潘晓,徐春香,陈厚彬.植物体细胞胚发生的研究进展[J].江西农业学报,2009,21(10):103-107
[3] Iocco P,Franks T,Thomas M R.Genetic transformation of major wine grape cultivars of Vitis vinifera L.[J].Transgenic Research,2001,10:105-112
[4] 陶建敏,庄智敏,章镇,等.葡萄基因转导研究进展[J].果树学报,2003,20(5):384-387
[5] 金万梅,董静,闫爱玲,等.葡萄器官离体再生和遗传转化体系的建立[J].园艺学报,2008,35(1):27-32
[6] Dhekney S A,Li Z T,Dutt M,et al.Agrobacterium-mediated transformation of embryogenic cultures and plant regeneration in Vitis rotundifolia Michx.(muscadine grape)[J].Plant Cell Report,2008,27:865-872
[7] Zimmerman J L.Somatic embryogenesis:a model for early development in higher plants[J].The Plant Cell,1993,5:1411-1423
[8] 闫爱玲,徐海英,张国军.葡萄体细胞胚发生研究进展[J].中外葡萄与葡萄酒,2008(1):30-33
[9] Das D K,Reddy M K,Upadhyaya K C,et al.An efficient leaf-disc culture method for the regeneration via somatic embryogenesis and transformation of grape(Vitis vinifera L.)[J].Plant Cell Report,2002,20:999-1005
[10] Martinelli L,Bragagna P,Poletti V,et al.Somatic embryogenesis from leaf- and petiole-derived callus of Vitis rupestris[J].Plant Cell Reports,1993,12:207-210
[11] Maillot P,Lebel S,Schellenbaum P,et al.Differential regulation of SERK,LEC1-Likev and Pathogenesis-Related genes during indirect secondary somatic embryogenesis in grapevine[J].Plant Physiology and Biochemistry,2009,47:743-752
[12] Gambino G,Ruffa P,Vallania R,et al.Somatic embryogenesis from whole flowers,anthers and ovaries of grapevine(Vitis spp.)[J].Plant Cell,Tissue and Organ Culture,2007,90:79-83
[13] 卢炳芝,于向荣,郭飞跃,等.葡萄花丝培养中的胚胎发生[J].果树科学,1995,12(增刊):37-40
[14] Kikkert J R,Striem M J,Vidal J R,et al.Long-term study of somatic embryogenesis from anthers and ovaries of 12 grapevine(Vitis sp.)genotypes[J].In Vitro Cell Developmental Biology-Plant,2005,41:232-239
[15] 李世诚,堀内昭作,望岡亮介.葡萄未熟胚的胚状体诱导和再生植株[J].上海农业学报,1992,8(2):27-32
[16] Dhekney S A,Li Z T,Gray D J.Factors influencing induction and maintenance of Vitis rotundifolia Michx.embryogenic cultures[J].Plant Cell,Tissue and Organ Culture,2011,105:175-180
[17] Yang X M,An L Z,Xiong Y C,et al.Somatic embryogenesis from immature zygotic embryos and monitoring the genetic fidelity of regenerated plants in grapevine[J].Biologia Plantarum,2008,52(2):209-214
[18] 刘倩,周蓓蓓,张其林,等.葡萄雄性不育种质特性及其花粉败育的细胞学研究[J].果树学报,2010,27(4):514-520
[19] Gaspero G D,Peterlunger E,Testolin R,et al.Conservation of microsatellite loci within the genus Vitis[J].Theoretical Applied Genetics,2000,101:301-308
[20] Leal F,Loureiro J,Rodriguez E,et al.Nuclear DNA content of Vitis vinifera cultivars and ploidy level analyses of somatic embryo-derived plants obtained from anther culture[J].Plant Cell Reports,2006,25:978-985
[21] Bassam B J,Caetano-Anolles G,Gresshoff P M.Fast and sensitive silver staining of DNA in polyacrylamide gels[J].Analytical Biochemistry,1991,196:80-83
[22] 何荣荣,张雅丽,崔邵艳,等.不同因素对葡萄次生胚诱导的影响[J].农业生物技术学报,2009,17(6):1123-1124
[23] Emershad R L,Ramming D W.In-ovulo embryo culture of Vitis vinifera L.cv.’Thompson seedless’[J].American Journal of Botany,1984,71:873-877
[24] Park S M,Hiramatsu M,Wakana A.Aneuploid plants derived from crosses with triploid grapes through immature seed culture and subsequent embryo culture[J].Plant Cell,Tissue and Organ Culture,1999,59:125-133
[25] Perrin M,Gertz C,Masson J Z.High efficiency initiation of regenerable embryogenic callus from anther filaments of 19-grapevine genotypes grown worldwide[J].Plant Science,2004,167:1343-1349
[26] 杨晓明,安黎哲,王雅梅,等.酿酒葡萄’神索’体胚发生及再生体系遗传稳定性分析[J].园艺学报,2006,33(6):1317-1320
[27] Prado M J,Eleazar R,Laura R,et al.Detection of somaclonal variants in somatic embryogenesis-regenerated plants of Vitis vinifera by flow cytometry and microsatellite markers[J].Plant Cell,Tissue and Organ Culture,2010,103:49-59
[28] Pinto D L P,de Almeida Barros B,Viccini L F,et al.Ploidy stability of somatic embryogenesis-derived Passiflora cincinnata Mast.plants as assessed by flow cytometry[J].Plant Cell,Tissue and Organ Culture,2010,103:71-79
[29] Loureiro J,Pinto G,Lopes T,et al.Assessment of ploidy stability of somatic embryogenesis process in Quercus suber L.using flow cytometry[J].Planta,2005,221:815-822
[30] Stamp J A,Meredith C P.Proliferative somatic embryogenesis from zygotic embryos of grapevine[J].Journal of American Society for Horticultural Science,1988,113:941-945

相似文献/References:

[1]宗成文,章镇,房经贵,等.葡萄LEAFY基因启动子的克隆与序列分析[J].南京农业大学学报,2007,30(4):20.[doi:10.7685/j.issn.1000-2030.2007.04.005]
 ZONG Cheng-wen,ZHANG Zhen,FANG Jing-gui,et al.Cloning and sequence analysis of LEAFY gene promoter from grape(Vitis vinifera×V.labrusca)[J].Journal of Nanjing Agricultural University,2007,30(3):20.[doi:10.7685/j.issn.1000-2030.2007.04.005]
[2]黄非,盛炳成.宿晓红葡萄与葡萄属有关种亲缘关系初探[J].南京农业大学学报,1993,16(04):49.[doi:10.7685/j.issn.1000-2030.1993.04.009]
 Huang Fei Sheng Bingcheng.STUDY ON THE RELATIONSHIP BETWEEN SUXIAOHONG GRAPE AND SOME SPECIES OF VITIS[J].Journal of Nanjing Agricultural University,1993,16(3):49.[doi:10.7685/j.issn.1000-2030.1993.04.009]
[3]周培根,戚晓玉,P.A.Braell,等.葡萄中β-damascenone前体的初步纯化[J].南京农业大学学报,1989,12(03):79.[doi:10.7685/j.issn.1000-2030.1989.03.017]
[4]韩浩章,姜卫兵,费宪进,等.葡萄和油桃自然休眠解除过程中H2O2含量和抗氧化酶活性的变化[J].南京农业大学学报,2007,30(1):50.[doi:10.7685/j.issn.1000-2030.2007.01.010]
 HAN Hao-zhang,JIANG Wei-bing,FEI Xianjin,et al.Changes in H_2O_2 content and activities of antioxidant enzymes of grape and nectarine during natural dormancy-release[J].Journal of Nanjing Agricultural University,2007,30(3):50.[doi:10.7685/j.issn.1000-2030.2007.01.010]
[5]郭磊,上官凌飞,房经贵*,等.葡萄EST鄄SSR 标记的开发及其应用[J].南京农业大学学报,2011,34(4):23.[doi:10.7685/j.issn.1000-2030.2011.04.005]
 GUO Lei,SHANGGUAN Ling-fei,FANG Jing-gui *,et al.Development of SSR markers from grape ESTs and its application[J].Journal of Nanjing Agricultural University,2011,34(3):23.[doi:10.7685/j.issn.1000-2030.2011.04.005]
[6]王晨,张演义,房经贵,等.葡萄microRNA156b和microRNA172c及其靶基因在冬芽二次成花过程中的表达特性研究[J].南京农业大学学报,2012,35(4):59.[doi:10.7685/j.issn.1000-2030.2012.04.011]
 WANG Chen,ZHANG Yan-yi,FANG Jing-gui,et al.Spatiotemporal expression of microRNA156b and microRNA172c and their target genes during flower development of winter buds growing on cut-back treated shoots of grapevine[J].Journal of Nanjing Agricultural University,2012,35(3):59.[doi:10.7685/j.issn.1000-2030.2012.04.011]
[7]王西成,王晨,房经贵,等.葡萄VvGA2ox1基因克隆、亚细胞定位及时空表达分析[J].南京农业大学学报,2013,36(1):29.[doi:10.7685/j.issn.1000-2030.2013.01.006]
 WANG Xicheng,WANG Chen,FANG Jinggui,et al.Cloning,subcellular localization and spatiotemporal expression of VvGA2ox1 gene from grapevine[J].Journal of Nanjing Agricultural University,2013,36(3):29.[doi:10.7685/j.issn.1000-2030.2013.01.006]
[8]谢荔,成学慧,冯新新,等.氨基酸肥料对‘夏黑’葡萄叶片光合特性与果实品质的影响[J].南京农业大学学报,2013,36(2):31.[doi:10.7685/j.issn.1000-2030.2013.02.006]
 XIE Li,CHENG Xuehui,FENG Xinxin,et al.Effects of an amino acid fertilizer on the leaf photosynthesis and fruit quality of ‘Summer Black’grape[J].Journal of Nanjing Agricultural University,2013,36(3):31.[doi:10.7685/j.issn.1000-2030.2013.02.006]
[9]任国慧,上官凌飞,房经贵,等.葡萄DELLA家族成员VvRGA和VvRGL1的预测、验证及生物信息学分析[J].南京农业大学学报,2013,36(3):15.[doi:10.7685/j.issn.1000-2030.2013.03.003]
 REN Guohui,SHANGGUAN Lingfei,FANG Jinggui,et al.The prediction,validation and bioinformatics analysis of the grape DELLA family members about VvRGA and VvRGL1[J].Journal of Nanjing Agricultural University,2013,36(3):15.[doi:10.7685/j.issn.1000-2030.2013.03.003]
[10]任国慧,陶然,王晨,等.葡萄浆果着色与UFGT和MYBA基因表达量的关系研究[J].南京农业大学学报,2013,36(4):30.[doi:10.7685/j.issn.1000-2030.2013.04.006]
 REN Guohui,TAO Ran,WANG Chen,et al.The research of the relationship between coloring and UFGT and MYBA gene expression level of the grape berry[J].Journal of Nanjing Agricultural University,2013,36(3):30.[doi:10.7685/j.issn.1000-2030.2013.04.006]

备注/Memo

备注/Memo:
收稿日期:2011-06-20
基金项目:国家948项目(2011-G28);江苏省农业科技自主创新资金项目(cx(10)110);江苏省农业资源开发局项目(2012-kj59)
作者简介:孙兴民,硕士研究生。
通讯作者:陶建敏,教授,主要从事果树遗传育种和生物技术研究,E-mail:taojianmin@njau.edu.cn。
更新日期/Last Update: 1900-01-01