[1]王凤敏,李涛,王运杰,等.影响农杆菌介导大豆子叶节遗传转化因素的研究[J].大豆科学,2011,30(04):557-562.[doi:10.11861/j.issn.1000-9841.2011.04.0557]
　WANG Feng-min,LI Tao,WANG Yun-jie,et al.Assessment of Factors Affecting Soybean Cotyledonary-node Agrobacterium-mediated Genetic Transformation[J].Soybean Science,2011,30(04):557-562.[doi:10.11861/j.issn.1000-9841.2011.04.0557]
点击复制

影响农杆菌介导大豆子叶节遗传转化因素的研究

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第30卷 期数: 2011年04期 页码: 557-562 栏目: 出版日期: 2011-08-25
Title:
Assessment of Factors Affecting Soybean Cotyledonary-node Agrobacterium-mediated Genetic Transformation
文章编号:
1000-9841（2011）04-0557-06
作者:
王凤敏; 李涛; 王运杰; 荆慧贤; 史晓蕾; 张孟臣; 邸锐; 蒋春志
河北省农林科学院 粮油作物研究所 国家大豆改良中心石家庄分中心，河北省作物遗传育种实验室，河北 石家庄050031
Author(s):
WANG Feng-min; LI Tao; WANG Yun-jie; JING Hui-xian; SHI Xiao-lei; ZHANG Meng-chen; DI Rui; JIANG Chun-zhi
Lab of Crops Genetic Breeding of Hebei,National Soybean Improvement Center Shijiazhuang Sub-Center,Institute of Food and Oil Crops,Hebei Academy of Agricultural and Forestry Sciences,Shijiazhuang 050031,Hebei,China
关键词:
大豆; GUS; 菌株; 基因型; 遗传转化
Keywords:
Soybean; GUS;  Agrobacterium tumefaciens strain; Genetypes; Genetic transformation
分类号:
S565.1
DOI:
10.11861/j.issn.1000-9841.2011.04.0557
文献标志码:
A
摘要:
利用GUS基因的稳定性和特异性及在植物体中瞬时表达的特点，对农杆菌介导大豆子叶节遗传转化过程中的一些影响因素（激素浓度、筛选剂浓度、基因型和农杆菌菌株）进行研究。结果表明：芽诱导阶段6-BA浓度为1.6 mg?L-1时诱导效率较高；适宜的草铵膦筛选浓度为3 mg?L-1；同时对24个大豆基因型的再生能力和对农杆菌菌株EHA101、LBA4404和EHA105的敏感性进行比较，再生能力最强的基因型为吉林小豆和Maverick,侵染能力最强的农杆菌菌株为EHA101,GUS阳性率为0～41.03%，其次是EHA105，LBA4404侵染效果最差；就受体基因型而言，吉林小豆和Maverick的GUS阳性率最高。EHA101侵染时，吉林小豆和Maverick的阳性率分别达40.25%和 29.89%；EHA105侵染时，吉林小豆和Maverick的阳性率分别达33.35%和 27.52%，而冀豆15、冀豆17和nf58的阳性率分别为12.47%、10.67%和9.11%。
Abstract:
Based on stability,specificity and transient expression of GUS gene in plant,we assessed factors affecting soybean Agrobacterium-mediated cotyledonary-node genetic transformation including hormone concentrations,selective agent concentration,genotypes,Agrobacterium tumefaciens strain.For shoot induction,a higher regeneration rate was obtained with 1.6 mg?L-16-BA;the suitable selective agent concentration of glufosinate was 3 mg?L-1.Based on their regeneration ability and sensitivity to Agrobacterium EHA101,EHA105 and LBA4404,‘Jilin xiaodou’ and ‘Maverick’ had higher regeneration capacity.The infection rate of ?Agrobacterium strain was EHA101(0-41.03%)better than EHA105(0-35%), LBA4404(0-18.27%)was the lowest.Based on the analysis of receptor genotypes,‘Jilinxiaodou’ and ‘Maverick’ was selected as the best receptor genotypes for the ?Agrobacterium-mediated cotyledonary-node transformation system.When infected by strain EHA101,GUS positive rate of ‘Jilinxiaodou’ and ‘Maverick’ reached 40.25% and 29.89%,respectively;while EHA105 infected soybean cotyledonary-node,percentage of GUS positive of ‘Jilinxiaodou’,‘Maverick’,‘Jidou15’,‘Jidou17’ and ‘nf58’ were 33.36%,27.52%,12.47%,10.67% and 9.11%,respectively.

参考文献/References:

[1]Horsch R B,Fraley B T,Rogers S C.Inheritance of functional foreign genes in plants[J].Science,1984,223:496-498.

[2]Hinchee M A,Connor-Ward D V,Newell C,et al.Production of transgenic soybean plants using Agrobacterium-mediated DNA transfer[J]. Nature Biotechnology,1988,6:915-922.

[3]Cheng T Y,Saka H,Thanh H V.Plant regeneration from soybean cotyledon node segments in cultures[J].Plant Science Letters,1980,19:91-99.

[4]Zhang Z Y,Xing A Q.The use of glufosinate as a selective agent in Agrobacterium-mediated transformation of soybean[J]. Plant Cell Tissue Organ Culture,1999,56:37-46.

[5]陈秀华,柏锡,潘欣,等.转cry1 Lem基因大豆的培育及抗虫性检测[J].大豆科学,2009,28（6）:959-963.(Chen X H,Bai X,Pan X,et al.Cultivation of?cry1 Lem?gene transform soybean and insect resistant assay[J].Soybean Science,2009,28(6):959-963.)

[6]刘海坤,卫志明.利用根瘤农杆菌介导转化大豆成熟种胚尖获得转基因植株[J].植物生理与分子生物学学报,2004,30（6）:631-636.(Liu H K,Wei Z M.Transgenic soybean obtained withAgrobacterium tumefaciens-mediated transformation of Embryonic tip of soybean mature seeds[J].Journal of Plant Physiology and Molecular Biology,2004,30(6):631-636.)

[7]Dang W,Wei Z M.An optimized Agrobacterium-mediated transformation for soybean for expression of binary insect resistance genes[J].Plant Science,2007,173:381-389.

[8]刘金华,王丕武,武丽敏,等.脯氨酸、硝酸银对农杆菌转化大豆的影响[J].大豆科学,2003,12（2）:36-40.(Liu J H,Wang P W,Wu L M,et al.Effection of proline and AgNO3on?Agrobacterium-mediated transformation of soybean[J].Soybean Science,2003,12(2):36-40.)

[9]袁鹰,刘德璞,郑培和,等.大豆组织培养再生植株研究[J].大豆科学,2001,20（1）:9-13.(Yuan Y,Liu D P,Zheng P H,et al.Study on plant regeneration from soybean culture[J].Soybean Science,2001,20(1):9-13.)

[10]曲姗姗,刘丽君,寇坤,等.农杆菌介导法将Bt(cryIA)基因导入大豆的研究[J].大豆科学,2009,28（2）:186-194.(Qu S S,Liu L J,Kou K,et al.Transformation of?Bt(cryIA)?gene into soybean via Agrobacterium-mediated method[J].Soybean Science,2009,28(2):186-194.)

[11]许智宏.植物生物技术[M].上海:上海科学技术出版社,1998:321.(Xu Z H.Plant biotechnology[M].Shanghai:Shanghai Science and Technology Press,1998:321.)

[12]李文霞,宁海龙,吕文河,等.农杆菌介导大豆子叶节转化系统的优化[J].中国农业科学,2008,41（4）:971-977.（Li W X,Ning H L,Lu W H,et al.Optimization of the Agrobacterium-mediated transformation systems of soybean cotyledonary node[J].Scientia Agricultura Sinica,2008,41(4):971-977.）


相似文献/References:

[1]刘章雄,李卫东,孙石,等.1983～2010年北京大豆育成品种的亲本地理来源及其遗传贡献[J].大豆科学,2013,32(01):1.[doi:10.3969/j.issn.1000-9841.2013.01.002]
　LIU Zhang-xiong,LI Wei-dong,SUN Shi,et al.Geographical Sources of Germplasm and Their Nuclear Contribution to Soybean Cultivars Released during 1983 to 2010 in Beijing[J].Soybean Science,2013,32(04):1.[doi:10.3969/j.issn.1000-9841.2013.01.002]
[2]李彩云,余永亮,杨红旗,等.大豆脂质转运蛋白基因GmLTP3的特征分析[J].大豆科学,2013,32(01):8.[doi:10.3969/j.issn.1000-9841.2013.01.003]
　LI Cai-yun,YU Yong-liang,YANG Hong-qi,et al.Characteristics of a Lipid-transfer Protein Gene GmLTP3 in Glycine max[J].Soybean Science,2013,32(04):8.[doi:10.3969/j.issn.1000-9841.2013.01.003]
[3]王明霞,崔晓霞,薛晨晨,等.大豆耐盐基因GmHAL3a的克隆及RNAi载体的构建[J].大豆科学,2013,32(01):12.[doi:10.3969/j.issn.1000-9841.2013.01.004]
　WANG Ming-xia,CUI Xiao-xia,XUE Chen-chen,et al.Cloning of Halotolerance 3 Gene and Construction of Its RNAi Vector in Soybean (Glycine max)[J].Soybean Science,2013,32(04):12.[doi:10.3969/j.issn.1000-9841.2013.01.004]
[4]张春宝,李玉秋,彭宝,等.线粒体ISSR与SCAR标记鉴定大豆细胞质雄性不育系与保持系[J].大豆科学,2013,32(01):19.[doi:10.3969/j.issn.1000-9841.2013.01.005]
　ZHANG Chun-bao,LI Yu-qiu,PENG Bao,et al.Identification of Soybean Cytoplasmic Male Sterile Line and Maintainer Line with Mitochondrial ISSR and SCAR Markers[J].Soybean Science,2013,32(04):19.[doi:10.3969/j.issn.1000-9841.2013.01.005]
[5]卢清瑶,赵琳,李冬梅,等.RAV基因对拟南芥和大豆不定芽再生的影响[J].大豆科学,2013,32(01):23.[doi:10.3969/j.issn.1000-9841.2013.01.006]
　LU Qing-yao,ZHAO Lin,LI Dong-mei,et al.Effects of RAV gene on Shoot Regeneration of Arabidopsis and Soybean[J].Soybean Science,2013,32(04):23.[doi:10.3969/j.issn.1000-9841.2013.01.006]
[6]杜景红,刘丽君.大豆fad3c基因沉默载体的构建[J].大豆科学,2013,32(01):28.[doi:10.3969/j.issn.1000-9841.2013.01.007]
　DU Jing-hong,LIU Li-jun.Construction of fad3c Gene Silencing Vector in Soybean[J].Soybean Science,2013,32(04):28.[doi:10.3969/j.issn.1000-9841.2013.01.007]
[7]张力伟,樊颖伦,牛腾飞,等.大豆“冀黄13”突变体筛选及突变体库的建立[J].大豆科学,2013,32(01):33.[doi:10.3969/j.issn.1000-9841.2013.01.008]
　ZHANG Li-wei,FAN Ying-lun,NIU Teng-fei?,et al.Screening of Mutants and Construction of Mutant Population for Soybean Cultivar "Jihuang13”[J].Soybean Science,2013,32(04):33.[doi:10.3969/j.issn.1000-9841.2013.01.008]
[8]盖江南,张彬彬,吴瑶,等.大豆不定胚悬浮培养基因型筛选及基因枪遗传转化的研究[J].大豆科学,2013,32(01):38.[doi:10.3969/j.issn.1000-9841.2013.01.009]
　GAI Jiang-nan,ZHANG Bin-bin,WU Yao,et al.Screening of Soybean Genotypes Suitable for Suspension Culture with Adventitious Embryos and Genetic Transformation by Particle Bombardment[J].Soybean Science,2013,32(04):38.[doi:10.3969/j.issn.1000-9841.2013.01.009]
[9]王鹏飞,刘丽君,唐晓飞,等.适于体细胞胚发生的大豆基因型筛选[J].大豆科学,2013,32(01):43.[doi:10.3969/j.issn.1000-9841.2013.01.010]
　WANG Peng-fei,LIU Li-jun,TANG Xiao-fei,et al.Screening of Soybean Genotypes Suitable for Somatic Embryogenesis[J].Soybean Science,2013,32(04):43.[doi:10.3969/j.issn.1000-9841.2013.01.010]
[10]刘德兴,年海,杨存义,等.耐酸铝大豆品种资源的筛选与鉴定[J].大豆科学,2013,32(01):46.[doi:10.3969/j.issn.1000-9841.2013.01.011]
　LIU De-xing,NIAN Hai,YANG Cun-yi,et al.Screening and Identifying Soybean Germplasm Tolerant to Acid Aluminum[J].Soybean Science,2013,32(04):46.[doi:10.3969/j.issn.1000-9841.2013.01.011]

备注/Memo

基金项目：转基因生物新品种培育重大专项资助项目（2009ZX08004-001B）。

第一作者简介：王凤敏（1981-），女，硕士，主要从事大豆遗传转化研究。E-mail:wangfm89@163.com。
通讯作者：蒋春志（1964-），女，硕士，研究员，从事大豆遗传育种工作。E-mail:jchzh64@sohu.com。

更新日期/Last Update: 2014-08-15