[1]杨秋姣,孙晓丽,孙明哲,等.转cry6Aa2m基因大豆遗传稳定性分析及农艺性状调查[J].大豆科学,2014,33(05):629-633.[doi:10.11867/j.issn.1000-9841.2014.05.0629]
　YANG Qiu-jiao,SUN Xiao-li,SUN Ming-zhe,et al.Genetic Stability Analysis and Agronomic Traits Investigation of cry6Aa2m Transgenic Glycine max[J].Soybean Science,2014,33(05):629-633.[doi:10.11867/j.issn.1000-9841.2014.05.0629]
点击复制

转cry6Aa2m基因大豆遗传稳定性分析及农艺性状调查

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第33卷 期数: 2014年05期 页码: 629-633 栏目: 出版日期: 2014-10-25
Title:
Genetic Stability Analysis and Agronomic Traits Investigation of cry6Aa2m Transgenic Glycine max
作者:
杨秋姣1; 2; 孙晓丽2; 孙明哲2; 陈超2; 赵超越2; 纪巍2; 朱延明1; 2
1.塔里木大学 生命科学学院，新疆 阿拉尔 843300；
2.东北农业大学 农业生物功能基因重点实验室，黑龙江 哈尔滨 150030
Author(s):
YANG Qiu-jiao1; 2; SUN Xiao-li2; SUN Ming-zhe2; CHEN Chao2; ZHAO Chao-yue2; JI Wei2; ZHU Yan-ming1; 2
1.School of Life Sciences,Tarim University,Alar 843300,China;
2.Key Laboratory of Agricultural Biological Functional Genes,Northeast Agricultural University,Harbin 150030,China
关键词:
大豆; 抗虫基因cry6Aa2m; 遗传稳定性; 农艺性状
Keywords:
Transgenic soybean; Cry6Aa2m gene; Genetic stability; Agronomic traits
分类号:
S565.1
DOI:
10.11867/j.issn.1000-9841.2014.05.0629
文献标志码:
A
摘要:
以前期获得的转抗虫基因cry6Aa2m大豆T4代植株4个株系为材料，采用PCR及RT-PCR检测方法进行了外源基因遗传稳定性及表达情况分析。并以受体品种(东农50）为对照，进行了产量性状、形态性状、品质性状及环境适应能力等农艺性状的调查。结果表明：cry6Aa2m基因能够在T4代转基因植株中遗传并表达；在产量性状上，除转基因株系DpC6b7单株结荚数显著低于对照外，其余3个株系的百粒重和单株结荚数与对照相比无显著差异；在形态性状上，除转基因株系DpC6b37株高显著低于对照外，其余株系的生育期、结荚习性、花色、叶形与对照也均无显著差异；在品质性状上，与对照相比，DpC6b37粗蛋白含量高2%，粗脂肪含量高5%。株系DpC6b7则粗蛋白含量高4%，粗脂肪含量低1%；在环境适应能力方面，转基因株系种子萌发率、落粒率、种子休眠期、杂草竞争性与对照均无显著差异。综合结果表明cry6Aa2m基因能够稳定遗传并表达，其中株系DpC6b4和DpC6b8所调查的农艺性状均不劣于受体品种。
Abstract:
In our previous studies,we had transformed the insect-resistant gene cry6Aa2m into Glycine max and obtained the T4 generation of the cry6Aa2m transgenic lines.In order to screen the transgenic soybean lines not only with stable expression of the cry6Aa2m gene but also sharing good agronomic traits,in the current study,we analyzed the genetic stability and investigated the agronomic traits of the transgenic soybean lines with gene cry6Aa2m.By using PCR and RT-PCR analysis,we verified the heredity and expression of the cry6Aa2m gene in the T4generation transgenic plants,which were investigated in this study.As expected,we showed over 85% PCR-positive plants of all the T4transgenic lines,and demonstrated the constitutive expression of the cry6Aa2m gene in the transgenic lines by using the semi-quantitative RT-PCR assays.In this aspect,we investigated and analyzed the agronomic traits of four transgenic lines(DpC6b4,DpC6b7,DpC6b8 and DpC6b37),including yield traits,morphological traits,quality traits and adaptive capacity to environment.Results of plant yield traits showed that all transgenic lines displayed no significant differences in the 100-seed weight and pods per plant,except for the significantly less pods per plant for line DpC6b7.As to the morphological trails,no significant differences were observed in growth period,pod bearing habit,flower color and leaf shape,except that the plant height of line DpC6b37 was significantly shorter than the non-transgenic soybean.Furthermore,we also found that the transgenic line DpC6b7 exhibited 4% higher in the protein content and 1% lower in the oil content than non-transgenic soybean,while line DpC6b37 displayed 2% and 5% higher,respectively.In addition,we also observed no obvious differences in germination rates,period of dormancy,weed species and quantity between transgenic and non-transgenic plants.Taken together,our results presented in this study suggested that the cry6Aa2m gene could be stably inherited and expressed in the transgenic Glycine max lines,and its overexpression did not affect plant agronomic traits.Among the four transgenic lines,DpC6b4 and DpC6b8,with relatively higher expression levels of the cry6Aa2m gene,displayed better agronomic traits than the non-transgenic plants,which made them as optimal candidates for further application in fields.

参考文献/References:

［1］刁琢,许艳丽.中国大豆胞囊线虫抗源筛选及抗病育种研究进展[J].大豆科技,2008(5):14-16.(Diao Z,Xu Y L.Research progress of the screening and utilization of resistant soybean sources and resistance breeding against soybean cyst nematode in China[J].Soybean Science & Technology,2008(5):14-16.)

[2]Hartwig E E.Breeding productive soybean cultivars resistant to soybean cyst nematode for the southern United State[J].Plant Disease,1981,65:303-307.
[3]Li X Q,Wei J Z,Tan A,et al.Resistance to root-knot nematode in tolnato roots expressing a nematieidal Bacillus thuringiensis crystal protein[J].Plant Biotechnology,2007,5(4):455-464.
[4]Li X Q,Tan A,Michael V.Expression of Cry5B protein from Bacillus thuringiensis in plant roots confers resistance to root-knot nematode[J].Biological Control,2008,47(1):97-102.
[5]Guo S,Liu M,Peng D,et al.New strategy for isolating novel nematicidal crystal protein genes from Bacillus thuringiensis strain YBT-1518[J].Applied and Environmental Microbiology,2008,11:6997-7001.
[6]Yu Z,Bai P,Ye W,et al.A novel negative regulatory factor for nematicidal cry protein gene expression in Bacillus Thuringiensis[J].Microbiology Biotechnology,2008,18(6):1033-1039.
[7]纪巍,杨丽坤,柏锡,等.Cry6Aa2m基因植物表达载体构建及大豆的遗传转化[J].东北农业大学学报,2014,45(1):59-64.(Ji W,Yang L K,Bai X,et al.Construction of plant expression vector of cry6Aa2m gene and transformation into Glycine max[J].Journal of Northeast Agricultural University,2014,45(1):59-64.)
[8]朱延明，张凤，柏锡,等.转SCMRP基因大豆“中间试验”农艺性状调查[J].东北农业大学学报,2013,44(10):11-13.(Zhu Y M,Zhang F,Bai X,et al.Agronomic performance investigation of field trial of transgenic SCMRP soybean[J].Journal of Northeast Agricultural University,2013,44(10):11-13.)
[9]赵彦平,赵春海.植物转基因育种的分析与研究[J].生物技术通报,2011(3):72-77.(Zhao Y P,Zhao C H.Research and analysis of transgenic breeding in plants[J].Biotechnology Bulletin,2011(3):72-77.)
[10]岳同卿,郎志宏,王延峰,等.转BtCryAh基因抗虫玉米的获得及其遗传稳定性分析[J].农业生物技术学报,2010,18(4):638-644.(Yue T Q,Lang Z H,Wang Y F,et al.Acquirement of the transgenic maize harboring BtCry1Ah gene and analysis of its inheritable stability[J].Journal of Agricultural Biotechnology,2010,18(4):638-644.)
[11]林凡敏,柏锡,樊超,等.转GsGST14耐盐碱基因大豆的农艺性状调查[J].大豆科学,2013,32(1):56-58.(Li F M,Bai X,Fan C,et al.Investigation and analysis of the main agronomic traits of different transgenic soybean line with GsGST14 gene[J].Soybean Science,2013,32(1):56-58.)
[12]赵超越,柏锡,樊超,等.转GsCa2+-ATPase基因大豆主要农艺性状调查与分析[J].大豆科学,2013,31(6):903-906.(Zhao C Y,Bai X,Fan C,et al.Investigation and analysis of the main agronomic traits of different transgenic soybean line with GsCa2+-ATPase gene[J].Soybean Science,2013,31(6):903-906.)
[13]樊超,柏锡,赵超越,等.转GsSAMS基因大豆主要农艺性状调查与分析[J].大豆科学,2013,31(2):252254.(Fan C,Bai X,Zhao C Y,et al.Investigation and analysis of the main agronomic traits of different transgenic soybean line with GsSAMS gene[J].Soybean Science,2013,31(2):252-254.)
[14]马炳田,李平,朱桢,等.转抗虫基因优良籼型恢复系的获得及外源基因的遗传稳定性研究[J].中国水稻科学,2002,16(3):211-215.(Ma B T,Li P,Zhu Z,et al.Turn insect-resistant gene fine indica restorer for exogenous gene and its genetic stability study[J].Chinese Journal of Rice Science,2002,16(3):211-215.)

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(05):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(05):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(05):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(05):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(05):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(05):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(05):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(05):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(05):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(05):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(2011ZX08004-002)；国家自然科学基金(31171578)；黑龙江省高校科技创新团队建设计划(2011TD055)；国家高技术研究发展计划“863计划”（2008AA-10Z153）；国家基础科学人才培养基金(J1210069)。

第一作者简介：杨秋姣(1988-)，女，硕士，主要从事植物基因工程与分子生物学研究。E-mail:yangqiujiao1@126.com。
通讯作者：朱延明(1955-)，男，教授，博导，主要从事植物基因工程与分子生物学研究。E-mail:ymzhu2001@aliyun.com。

更新日期/Last Update: 2014-12-24