DU Sheng-wei,LIU Ye-li,et al.Optimization of Soybean Transformation System and Transferring Dof 4 Gene into Soybean[J].Soybean Science,2010,29(03):398-402.[doi:10.11861/j.issn.1000-9841.2010.03.0398]
大豆转化体系的优化和Dof 4基因转入大豆的研究
- Title:
- Optimization of Soybean Transformation System and Transferring Dof 4 Gene into Soybean
- 文章编号:
- 1000-9841（2010）03-0398-05
- 分类号:
- S565.1
- 文献标志码:
- A
- 摘要:
- 研究优化了包括种子消毒方法、生长调节剂用量和抗生素种类在内的影响农杆菌介导大豆子叶节遗传转化效率的多个因素，并将Dof 4基因转入绥农14中。结果表明：氯气熏蒸消毒方法对大豆伤害小，子叶节丛生芽分化率高；菌液侵染时间和共培养时间控制直接影响长菌和丛生芽状态。芽诱导培养阶段，当6-BA 浓度为1.6 m·L-1，IBA浓度为0.1m·L-1，分化率较高，畸形率和愈伤状况都较轻。最优的抗生素组合为头孢噻肟钠（Cefotaxime Sodium）200 m·mL-1加羧苄青霉素(Carbenicillin) 250 m·mL-1。
- Abstract:
- In this paper, some major factors, which played important role in improving the transformation efficiency of soybean, such as the method of sterilizing the seeds, plant hormones, and antibiotics, were optimized. The Dof 4 was transferred into the soybean cultivar Suinong 14 by Agrobacterium-mediated cotyledonary node transformation. The result showed that, the chlorine suffocating was chosen to seeds sterilize, which did not harm to the seeds and have a higher differentiation rate for multiple shoot. However, duration of Agrobacterium infection and co-culture influenced the contamination and the state of multiple shoot. As for shoot induction, a higher regenerated rate, better callus induction rate, and lower deformity rate were obtained with 1.6 m·L－16-BA and 0.1m·mL-1IBA in this experiment. And the optimum combination of antibiotic and concentration in medium was 200 m·mL-1Cefotaxime Sodium and 250 mg·mL-1Carbenicillin.
参考文献/References:
[1]卜云萍, 李明春, 胡国武, 等. 大豆子叶节组培再生系统与农杆菌介导的基因转化系统的比较研究[J].南开大学学报(自然科学版), 2003, 36(1): 103-108. ( Bu Y P, Li C M, Hu G W, et al. The study of comparing the transformation system of Agrobacterium-mediated and regeneration system of cotyledon nod of soybean culture[J]. Acta Scientiarum Naturallum (Universitatis Nakaiensis), 2003, 36 (1): 103-108.) [2]Hinchee M A W, Connor Ward D V, Hewell C A, et al. Production of transgenic soybean plants using Agrobacterium mediated DNA transfer[J]. Biotechnology,1988, 19:91-99. [3]Parrott W A, Williams E G, Hilbrand D P, et al. Effect of genotypeon somatic embryogenesis form immature cotyledons of soybean[J]. Plant Cell Tissue and Organ Culture, 1989, 16 (1):15-21. [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 and Organ Culture, 1999, 56: 37-46. [5]李桂兰, 乔亚科, 杨少辉, 等. 农杆菌介导大豆子叶节遗传转化的研究[J]. 作物学报, 2005, 31(2): 170-176. ( Li G L, Qiao Y K, Yang S H, et al. Study of the Agrobacterium-mediated transformation systems of soybean cotyledonary node[J]. Acta Agronomica Sinica, 2005, 31(2): 170-176.) [6]张颖君, 高慧敏, 蒋春志, 等. 大豆种子脂肪酸含量的快速测定[J]. 大豆科学, 2008. 27(5): 859-862. ( Zhang Y J, Gao H M, Jiang C Z. et al. Fast analysis on fatty acids of soybean seed by gas chromatograph[J]. Soybean Science, 2008, 27(5): 859-862.) [7]徐杰, 胡国华, 张大勇. 大豆籽粒发育过程中脂肪酸组分的累积动态[J]. 作物学报, 2006, 32(11): 1759-1763. ( Xu J, Hu G H, Zhang D Y. Dynamic accumulation of fatty acids in grain maturing process of soybean[J]. Acta Agronomy Sinica, 2006, 32(11): 1759-1763.) [8]王萍, 高世庆, 郭永来, 等. 利用农杆菌介导将抗逆相关基因GmDREB导入大豆的研究[J]. 大豆科学, 2008, 27(1): 47-51. ( Wang P, Gao S Q, Guo Y L, et al. Transformation of stress resistance related gene GmDREB?into soybean via Agrobacterium-mediation[J]. Soybean Science, 2008, 27(1): 47-51.) [9]王萍, 吴颖, 季静, 等. 抗生素对大豆愈伤组织的诱导和生长的影响[J]. 遗传, 2001, 23(4 ): 321-324 ( Wang P, Wu Y, Ji J, et al. Effect of antibioties on induction of callus and callus growth in soybean[J]. Hereditas, 2001, 23(4):321-324.)
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(03):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(03):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(03):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(03):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(03):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(03):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(03):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(03):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(03):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(03):46.[doi:10.3969/j.issn.1000-9841.2013.01.011]
[11]郝荣华,邵 群,杨素欣,等.根癌农杆菌介导的大豆子叶节转化体系的优化[J].大豆科学,2012,31(02):167.[doi:10.3969/j.issn.1000-9841.2012.02.002]
HAO Rong-hua,SHAO Qun,YANG Su-xin,et al.Optimization of Agrobacterium-mediated Soybean Transformation using the Cotyledonary Node[J].Soybean Science,2012,31(03):167.[doi:10.3969/j.issn.1000-9841.2012.02.002]
[12]郑丽红,季 静,王 罡,等.适于子叶节和胚尖再生体系的大豆基因型筛选[J].大豆科学,2012,31(02):212.[doi:10.3969/j.issn.1000-9841.2012.02.011]
ZHENG Li-hong,JI Jing,WANG Gang,et al.Selection of Suitable Soybean Genotype Based on Cotyledon Node and Embryonic Tip Regeneration Systems[J].Soybean Science,2012,31(03):212.[doi:10.3969/j.issn.1000-9841.2012.02.011]
[13]王 伟,王 罡,季 静,等.大豆胚尖再生体系的优化及与子叶节再生体系的比较[J].大豆科学,2012,31(03):353.[doi:10.3969/j.issn.1000-9841.2012.03.004]
WANG Wei,WANG Gang,JI Jing,et al.Optimization of Embryonic Tip Regeneration System and Comparison with Cotyledonary Node Regeneration System in Soybean[J].Soybean Science,2012,31(03):353.[doi:10.3969/j.issn.1000-9841.2012.03.004]
[14]姚丙晨,沈艳茹,韩 雪,等.大豆子叶节和胚尖再生体系的比较及大豆SR1基因的遗传转化[J].大豆科学,2012,31(03):364.[doi:10.3969/j.issn.1000-9841.2012.03.006]
YAO Bing-chen,SHEN Yan-ru,HAN Xue,et al.Comparison with Cotyledonary Node and Embryonic Tip Regeneration System in Soybean\[Glycine max(L.)Merrill\] and Genetic Transformation of SR1 [J].Soybean Science,2012,31(03):364.[doi:10.3969/j.issn.1000-9841.2012.03.006]
[15]陈秀华,柏锡,潘欣,等.基因大豆的培育及抗虫性检测[J].大豆科学,2009,28(06):959.[doi:10.11861/j.issn.1000-9841.2009.06.959]
CHEN Xiu-hua,BAI Xi,PAN Xin,et al.Cultivation of cry 1Iem Gene Transformed Soybean and Insect Resistant Assay[J].Soybean Science,2009,28(03):959.[doi:10.11861/j.issn.1000-9841.2009.06.959]
[16]赖冰冰,韩阳,李春风,等.大豆子叶节植株再生体系的研究[J].大豆科学,2011,30(02):303.[doi:10.11861/j.issn.1000-9841.2011.02.0303]
LAI Bing-bing,HAN Yang,LI Chun-feng,et al.Research on Cotyledonary Nodes Regeneration System of Soybean[J].Soybean Science,2011,30(03):303.[doi:10.11861/j.issn.1000-9841.2011.02.0303]
[17]赵晓雯,吴芳芳,狄少康,等.农杆菌介导的大豆子叶节遗传转化技术流程及操作要点[J].大豆科学,2011,30(03):362.[doi:10.11861/j.issn.1000-9841.2011.03.0362]
ZHAO Xiao-wen,WU Fang-fang,DI Shao-kang,et al.Technique flow and Key Operation Points of Agrobacterium-mediated Genetic Transformation of Soybean Cotyledonary Node[J].Soybean Science,2011,30(03):362.[doi:10.11861/j.issn.1000-9841.2011.03.0362]
[18]朱红林,沙爱华,符秀梅,等.转录调控基因GmLEC1转化大豆及转化方法的比较[J].大豆科学,2010,29(01):7.[doi:10.11861/j.issn.1000-9841.2010.01.0007]
ZHU Hong-lin,SHA Ai-hua,FU Xiu-mei,et al.Cloning and Transformation Study of Transcription Factor GmLEC1 in Soybean[J].Soybean Science,2010,29(03):7.[doi:10.11861/j.issn.1000-9841.2010.01.0007]
[19]寇坤,刘丽君,曲姗姗,等.大豆新品系黑农56子叶节再生体系的优化[J].大豆科学,2009,28(03):400.[doi:10.11861/j.issn.1000-9841.2009.03.0400]
KOU Kun,LIU Li-jun,QU Shan-shan,et al.Improvement of Regeneration System in New Soybean Line Heinong 56[J].Soybean Science,2009,28(03):400.[doi:10.11861/j.issn.1000-9841.2009.03.0400]
[20]姬月梅,陈受宜,李英慧,等.农杆菌介导大豆子叶节遗传转化体系的优化研究[J].大豆科学,2008,27(01):26.[doi:10.11861/j.issn.1000-9841.2008.01.0026]
JI Yue-mei,CHEN Shou-yi,LI Ying-hui,et al.Optimization of Genetic Transformation System from Soybean Cotyledon Mediated by Agrobacterium[J].Soybean Science,2008,27(03):26.[doi:10.11861/j.issn.1000-9841.2008.01.0026]
备注/Memo
基金项目：国家高技术研究发展计划资助项目（2006AA100104-3）；国家转基因专项资助项目（2008ZX08004-001-3）。