Welcome to Acta Agronomica Sinica,

Acta Agron Sin ›› 2017, Vol. 43 ›› Issue (08): 1254-1258.doi: 10.3724/SP.J.1006.2017.01254

• RESEARCH NOTES • Previous Articles    

Application of Pisum sativum Terminator rbc-T in Wheat Transformation

WANG Xiao-Ting1,2,HUANG Suo2,XU Zhao-Shi2,LI Lian-Cheng2,MA You-Zhi2,CHEN-Ming2,*,MIN Dong-Hong1,*   

  1. 1College of Agronomy, Northwest A&F University / State Key Laboratory of Crop Stress Biology for Arid Areas, Yangling 712100, China; 2 National Key Facility for Crop Gene Resource and Genetic Improvement / Key Laboratory of Biology and Genetic Improvement of Triticeae Crops, Ministry of Agriculture / Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2017-01-20 Revised:2017-04-20 Online:2017-08-12 Published:2017-04-27
  • Contact: Min Donghong, E-mail: mdh2493@126.com, Tel: 13609123593; Chen Ming, E-mail: chenming02@caas.cn, Tel: 010-82108750 E-mail:daisy19920829@foxmail.com
  • Supported by:

    This study was supported by the National Major Project for Developing New GM Crops (2014ZX08002-003B, 2016ZX08002002, 2016ZX08002005).


The fungal- or bacterial-origin vector elements are commonly used in transgenic crops, which raise great concerns about the safety of GM foods. In the study, we isolated the pea (Pisum sativum) terminator, rbc-T, and constructed a transformation vector by replacing the Agrobacterium terminator nos-T with rbc-T. The minimum transformation fragment consisting of Ubiquitin promoter, GUS reporter gene and terminator rbc-T was transformed into common wheat by particle bombardment method. Meanwhile a minimum transformation fragment with ?the Agrobacterium terminator nos-T was used as the control. The T2 transgenic lines were regenerated after PCR validation of the transformation. In the transgenic lines carrying both kinds of terminators, GUS gene expression was detected by GUS histochemical staining and enzyme activity analysis, indicating that the pea terminator rbc-T is applicable as an alternative of the Agrobacterium terminator nos-T in transgenic wheat research.

Key words: Nopaline synthase terminator (nos-T), Pea terminator (rbc-T), Transgenic wheat, GUS reporter gene

[1]李军, 瞿绍洪. 用无终止子的RFP报告基因捕获水稻终止子研究. 浙江农业学报, 2012, 24: 758–763 Li J, Qu S H. PolyA trapping by using terminatorless RFP reporter gene in rice. Acta Agric Zhejiangensis, 2012, 24: 758–763 (in Chinese with English abstract) [2]陈贞. 转基因作物的多重PCR检测体系的研究. 暨南大学硕士学位论文, 广东广州, 2011 Chen Z. Multiplex PCR for Detection of Genetically Modified Crops. MS Thesis of Jinan University, Guangzhou, China, 2011 (in Chinese with English abstract) [3]焦改丽, 孟钊红, 聂安全, 南芝润, 张换样, 李俊峰, 王娇娟, 赵俊侠, 李燕娥, 郭三堆. 花椰菜花叶病毒(CaMV)35S启动子在转基因棉花中的表达. 作物学报, 2004, 30: 1135–1139 Jiao G L, Meng Z H, Nie A Q, Nan Z R, Zhang H Y, Li J F, Wang J J, Zhao J X, Li Y E, Guo S D. Expression of CaMV35S promoter in transgenic cotton. Acta Agron Sin, 2004, 30: 1135–1139 (in Chinese with English abstract) [4]牛东东, 郝育杰, 荣瑞娟, 韦汉福, 兰金苹, 史佳楠, 魏健, 李雪姣, 杨烁, 奚文辉, 武鹏程, 刘丽娟, 吴琳, 刘斯奇, 尹长城, 刘国振. 转基因水稻中GUS蛋白质的检测及其表达特征. 中国农业科学, 2014, 47: 2715–2722 Niu D D, Hao Y J, Rong R J, Wei H F, Lan J P, Shi J N, Wei J, Li X J, Yang S, Xi W H, Wu P C, Liu L J, Wu L, Liu S Q, Yin C C, Liu G Z. Detection of GUS protein and its expression pattern in transgenic rice plants. Sci Agric Sin, 2014, 47: 2715–2722 (in Chinese with English abstract) [5]Dipak K S, Nrisingha D, Indu B M. pSiM24 Is a novel versatile gene expression vector for transient assays as well as stable expression of foreign genes in plants. PLoS One, 2014, 9: e98988. [6]徐琼芳, 李连城, 陈孝, 马有志, 叶兴国, 张增艳, 徐惠君, 辛志勇. 基因枪法获得GNA转基因小麦植株的研究. 中国农业科学, 2001, 34: 5–8 Xu Q F, Li L C, Chen X, Ma Y Z, Ye X G, Zhang Z Y, Xu H J, Xin Z Y. Study on the obtaining of transgenic wheats with GNA alien gene by biolistic particle. Sci Agric Sin, 2001, 34: 5–8 (in Chinese with English abstract) [7]Lipp M, Brodmann P, Pietsch K, Pauwels J, Anklam E. IUPAC collaborative trial study of a method to detect genetically modified soybeans and maize in dried powder. JAOAC Int, 1999, 82: 923–928 [8]Jefferson R A, Kavanagh T A, Bevan M W. GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J, 1987, 6: 3901–3907 [9]Kevan M A G, Angela T M, Stanislav V, Karel B, Richard J I, Jill S G. Analysis of genetically modified plant gene expression using GUS fluorimetry. Mol Biotechnol, 2000, 14: 235–239 [10]Chen Y J, Liu P, Alec A K N, Jennifer A N B, Kevin C, Todd P, Christopher A V. Characterization of 582 natural and synthetic terminators and quantification of their design constraints. Nat Methods, 2013, 10: 659–664 [11]张发云, 尹维波, 胡赞民. 毛白杨叶绿体基因启动子和终止子的克隆及其功能鉴定. 农业生物技术学报, 2007, 15: 439–445 Zhang F Y, Yin W B, Hu Z M. Cloning and function identification of the promoters and terminator of Populus tomentosa chloroplast genes. J Agric Biotechnol, 2007, 15: 439–445 (in Chinese with English abstract) [12]金双侠, 韩杰, 刘小云, 刘冠泽, 王一娴, 唐文鑫, 张献龙. SDS-蛋白酶法分离棉花cpDNA及psbA基因启动子、终止子克隆. 分子植物育种, 2007, 5: 683–689 Jin S X, Han J, Liu X Y, Liu G Z, Wang Y X, Tang W X, Zhang X L. Chroloplast DNA extraction in cotton by SDS-proteinase method and cloning of the promoter and terminator of psbA gene. Mol Plant Breed, 2007, 5: 683–689 (in Chinese with English abstract) [13]钟山, 罗大极, 吴刚, 徐婧, 汪亚平, 朱作言. 两种不同终止子在转基因鲤鱼中的促生长效应. 遗传, 2009, 31: 831–836 Zhong S, Luo D J, Wu G, Xu J, Wang Y P, Zhu Z Y. The growth promoting activity of two different kinds of reconstructed terminators in transgenic common carp. Hereditas (Beijing), 2009, 31: 831–836 (in Chinese with English abstract)

[1] SHI Lei, MIAO Li-Juan, HUANG Bing-Yan, GAO Wei, ZHANG Zong-Xin, QI Fei-Yan, LIU Juan, DONG Wen-Zhao, ZHANG Xin-You. Characterization of the promoter and 5'-UTR intron in AhFAD2-1 genes from peanut and their responses to cold stress [J]. Acta Agronomica Sinica, 2021, 47(9): 1703-1711.
[2] WANG Rui,ZHU Meng-Lin,GAO Fang-Yuan,REN Juan-Sheng,LU Xian-Jun,REN Guang-Jun,LIN Yong-Jun. Designing, Construction and Functional Characterization of Tissue-specific Synthetic Promoter in Rice [J]. Acta Agron Sin, 2017, 43(06): 789-794.
[3] WANG Yong-Xia,DU Xin-Hua,XU Wei-Gang,QI Xue-Li,LI Yan,WANG Hui-Wei,HU Lin. Photosynthetic Characteristics of Transgenic Wheat Expressing Maize C4-Type NADP-ME Gene [J]. Acta Agron Sin, 2016, 42(04): 600-608.
[4] SHEN Fang-Di,HONG Yan-Tao,DU Li- Pu,XU Hui-Jun,MA Ling-Jian,ZHANG Zeng-Yan. Expression of Apoptosis Inhibitor Genes OpIAP and p35 Enhances Resistance to Rhizoctonia cerealis in Transgenic Wheat [J]. Acta Agron Sin, 2015, 41(10): 1490-1499.
[5] YANG Kun,LIU Xin,DU Li-Pu,YE Xing-Guo,ZHANG Zeng-Yan. Development and Characterization of AcAMP-sn Transgenic Wheat with Enhanced Resistance to Wheat Take-all [J]. Acta Agron Sin, 2014, 40(01): 22-28.
[6] LIU Fei,YANG Li-Hua,WANG Ai-Yun,MA Xiao-Fei,DU Li-Pu,LIU Xin,LI Pan-Song,ZHANG Zeng-Yan,MA Ling-Jian. Molecular Detection and Take-all Response Assays of TiERF1-RC7 Transgenic Wheat [J]. Acta Agron Sin, 2013, 39(11): 2094-2098.
[7] YANG Li-Hua,WANG Jin-Feng,DU Li-Pu,XU Hui-Jun,WEI Xue-Ning,LI Zhao,MA Ling-Jian,ZHANG Zeng-Yan. Generation and Characterization of PgPGIP1 Transgenic Wheat Plants with Enhanced Resistance to Take-All and Common Root Rot [J]. Acta Agron Sin, 2013, 39(09): 1576-1581.
[8] DANG Liang,SU Zhen-Qi,YE Xing-Guo,XU Hui-Jun,LI Zhao,SHAO Yan-Jun,ZHANG Zeng-Yan. Overexpression of BvGLP1 in Transgenic Wheat Enhances Resistance to Common Root Rot [J]. Acta Agron Sin, 2013, 39(02): 368-372.
[9] ZHU Xiu-Liang,LI Zhao,DU Li-Pu,XU Hui-Jun,YANG Li-Hua,ZHUANG Hong-Tao,MA Ling-Jian,ZHANG Zeng-Yan. Development and Characterization of Wheat Lines with Resistance to Take-All and Powdery Mildew Diseases [J]. Acta Agron Sin, 2012, 38(12): 2178-2184.
[10] DANG Liang,WANG Ai-Yun,XU Hui-Jun,ZHU Xiu-Liang,DU Li-Pu,SHAO Yan-Jun,ZHANG Zeng-Yan. Development and Characterization of GmPGIP3 Transgenic Wheat Yangmai 18 with Enhanced Resistance to Common Root Rot [J]. Acta Agron Sin, 2012, 38(10): 1833-1838.
[11] WANG Jin-Feng,DU Li-Pu,LI Zhao,HUANG Su-Ping,YE Xing-Guo,FENG Dou,ZHANG Zeng-Yan. Development and Characterization of SN1 Transgenic Wheat Plants with Enhanced Resistance to Rhizoctonia cerealis and Bipolaris sorokiniana [J]. Acta Agron Sin, 2012, 38(05): 773-779.
[12] TUN Qiong, HU Wei-Gang, LI Yan, JI Hua-Li, HU Lin, ZHANG Lei, HAN Lin-Lin. Physiological Characteristics of Photosynthesis in Transgenic Wheat with Maize C4-PEPC Gene under Field Conditions [J]. Acta Agron Sin, 2011, 37(11): 2046-2052.
[13] SUN Yong-Wei, NIE Li-Na, MA Wei-Zhi, XU Zhao-Shi, JIA Lan-Qin. Cloning and Functional Analysis of Viviparous-1 Promoter in Wheat [J]. Acta Agron Sin, 2011, 37(10): 1743-1751.
[14] LI Zhao, PENG Hong-Chao, DU Li-Pu, ZHOU Miao-Beng, CA Shi-Bin, XU Hui-Jun, LI Shi-Shen, ZHANG Ceng-Yan. Utilization of Tissue Specific Expressing Promoter RSS1P in TiERF1 Transgenic Wheat [J]. Acta Agron Sin, 2011, 37(10): 1897-1903.
[15] FANG Xiao-Liang, LIU Wei, AN Jing, WANG Qiang-Guo. Isolation and Characterization of an Embryo-specific Promoter OsESP1 from Rice [J]. Acta Agron Sin, 2011, 37(10): 1904-1909.
Full text



No Suggested Reading articles found!