Please wait a minute...

中国生物工程杂志

China Biotechnology
China Biotechnology  2012, Vol. 32 Issue (02): 50-56    DOI:
    
Transformation and Identification of pta Gene into Wheat Cultivar Longchun22 Mediated by Agrobacterium tumefaciens
LI Shu-jie, LI Jin-wen, ZHANG Zheng-ying
Institute of Biotechnology, Gansu Academy of Agricultural Science, Lanzhou 730070,China
Download: HTML   PDF(1123KB) HTML
Export: BibTeX | EndNote (RIS)      

Abstract  

Agrobacterium tumefaciens-mediated wheat genetic transformation system was set up by using immature embryos of spring wheat cultivar Longchun 22 as explant. 4-day-preculture immature embryos were transformed with pta gene using Agrobacterium tumefaciens C58c1. Three transgenic plants were gained after G418 resistant screening and PCR analysis. Real-time PCR analysis showed that pta gene copy number in transgenic wheat plant is 2,1,3, respectively. The exogenous pta gene have integrated into three plants out of six T1 progeny plants confirmed by PCR analysis, and the transgenic wheat showed good resistance to aphids in greenhouse.



Key wordspta gene      Longchun 22 (Triticum aestivum L.)      Agrobacterium tumefaciens      Real-time      PCR analysis     
Received: 15 November 2011      Published: 25 February 2012
ZTFLH:  Q819  
Cite this article:

LI Shu-jie, LI Jin-wen, ZHANG Zheng-ying. Transformation and Identification of pta Gene into Wheat Cultivar Longchun22 Mediated by Agrobacterium tumefaciens. China Biotechnology, 2012, 32(02): 50-56.

URL:

https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/Y2012/V32/I02/50


[1] Cheng M, Fry J E, Pang S, et al. Genetic transformation of wheat mediated by Agrobacterium tumefaciens.Plant Physiol, 1997,115:971-980.

[2] Xia G M, Li Z Y, He C X, et al. Transgenic plant regeneration from wheat (Triticum aestivum L.) mediated by Agrobacterium tumefaciens. Acta Phytophysiol Sin,1999,25:22-28.

[3] Weir B, Gu X, Wang M B, et al. Agrobacterium tumefaciens-mediated transformation of wheat using suspension cells as a model system and green fluorescent protein as a visual marker. Aust J Plant Physiol, 2001, 28:807-818.

[4] Hu S, Metz C. Agrobacterium-mediated large-scale transformation of wheat (Triticum aestivum L.) using glyphosate selection. Plant Cell Reports,2003,21:1010-1019.

[5] Cheng M, Hu T C, Layton J, et al. Desiccation of plant tissues post-Agrobacterium infection enhances t-DNA delivery and increases stable transformation efficiency in wheat. In Vitro Cellular Developmental Biology, 2003, 39:595-604.

[6] 叶兴国,陈明,杜丽璞,等.小麦转基因方法及其评述.遗传,2011,33(5):422-430. Xe X G, Chen M, Du L P, et al.Description and evaluation of transformation approaches used in wheat.Hereditas, 2011, 33(5): 422-430.

[7] Khanna H K, Daggard G E. Agrobacterium tumefaciens-mediated transformation of wheat using a superbinary vector and a polyamine-supplemented regeneration medium. Plant Cell Reports, 2003, 21:429-436.

[8] Wu H, Sparks C, Amoah B, et al. Factors influencing successful Agrobacterium mediated genetic transformation of wheat. Plant Cell Reports, 2003, 21:659-668

[9] 叶兴国,Shirley S,徐惠君,等.小麦农杆菌介导转基因植株的稳定获得和检测.中国农业科学,2001,34(5):465-468. Ye X G,Shirley S,Xu H J,et al. Regular production of transgenic wheat mediated with Agrobacterium tumefaciens.Scientia Agricultura Sinica, 2001,34(5):465-468.

[10] 叶兴国,程红梅,徐惠君,等.转几丁质酶和p-1,3-葡聚糖酶双价基因小麦的获得和鉴定.作物学报,2005,31(5):583-586. Ye X G, Cheng H M, Xu H J, et al. Development of transgenic wheat plants with chitinase and β-1,3-glucosanase genes and their resistance to Fusarium Head Blight.Acta Agronomica Sinica, 2005,3 1(5):583-586.

[11] 张红宇,吴先军,唐克轩,等.半夏凝集素基因(pta)导入水稻及其表达的初步研究.遗传学报,2003,30(11):1013-1019. Zhang H Y, Wu X J,Tang K X, et al.A primary study of transferring the Pinellia tenata agglutinin (pta) gene into rice and expression. Acta Genetica Sinica, 2003, 30 (11): 1013-1019.

[12] 范媛媛,庞永珍,吴为胜,等.双价抗虫植物表达载体p3300-bt-pta的构建及转基因烟草的初步检测.上海交通大学学报,2004,22(1):1-6. Fan Y Y,Pang Y Z,Wu W S,et al.Construction of plant expression vectors containing binary insect resistance genes Bt CryIA(a)-pta or Bt CryIA(c)-pta and their expression in transgenic tobaccos. Jorunal of Shanghai Jiaotong University, 2004,22(1):1-6.

[13] 唐东芹,钱虹姝,唐克轩,等.根癌农杆菌介导半夏凝集素基因pBIXPTA对百合的转化.上海交通大学学报,2004,22(2):135-137. Tang D Q, Qian H S, Tang K X, et al. Genetic transformation of Lily with pBIXPTA gene by Agrobacterium tumefaciens. Jorunal of Shanghai Jiaotong University, 2004,22(2):135-137.

[14] 熊恒硕,康俊梅,杨青川,等.双价抗虫植物表达载体p3300-bt-pta转化苜蓿的初步研究.中国草地学报,2008,30(1):21-26. Xiong H S, Kang J M, Yang Q C, et al. A primary study on transferring plant expression vectors containing binary insect resistance genes Bt CryIA(a)-pta or Bt CryIA(c)-pta into alfalfa.Chinese Journal of Grassland, 2008,30(1):21-26.

[15] 李淑洁,王红梅,张正英.半夏凝集素基因对油菜的遗传转化及REAL-TIME PCR分析.作物杂志,2010,(4):52-55. Li S J, Wang H M,Zhang Z Y. Genetic transformation of Brassica napus with pta gene by Agrobacterium tumefaciens and real-time PCR analysis.Crops, 2010,(4):52-55.

[16] 张正英,令利军,王红梅,等.半夏凝集素基因的克隆及转基因烟草对蚜虫的抑制作用.植物保护,2010,36(6):21-25. Zhang Z Y, Ling L J, Wang H M.Cloning and transformation of pta gene and inhibitory effects of the transgenic tobacco plants on developments of aphid populations.Plant Protections, 2010,36(6):21-25.

[17] 李淑洁,张正英.REAL-TIME PCR方法测定转基因小麦中外源基因拷贝数.中国生物工程杂志,2010,30(3):90-94. Li S J, Zhang Z Y. Using real-time PCR to determine transgene copy number in wheat. China Biotechnology,2010,30(3):90-94.

[18] Weng H B, Pan A H,Yang L T, et al. Estimating number of transgene copies in transgenic rapeseed by real-time PCR assay with HMG I/Y as an endogenous reference gene.Plant Molecular Biology Reporter,2004,22:289-300.

[19] Ingham D J, Beer S,Miney S, et al. Quantitative real-time PCR assay for determining transgene copy number in transformed plants. BioTechniques,2001,31:132-140.

[20] Song P,Cai C,Skokut M, et al. Quantitative real-time PCR as a screening tool for estimating transgene copy number in WHISKERSTM-derived transgenic maize.Plant Cell Reports,2002,20:948-954.

[21] 杨立桃,赵志辉,丁嘉羽,等.利用实时荧光定量PCR法分析转基因水稻外源基因拷贝数. 中国食品卫生杂志,2005,17(2):140-144. Yang L T,Zhao Z H,Ding J Y,et al. Estimating copy number of transgenes in transformed rice by real-time quantitative PCR.Chinese Journal of Food Hygiene,2005,17(2):140-144.

[22] 王育花,赵 森,陈 芬,等.利用实时荧光定量PCR法检测转基因水稻外源基因拷贝数的研究.生命科学研究,2007,11(4):301-305. Wang Y H,Zhao S,Chen F,et al. Estimation of the copy number of exogenous gene in transgenic rice by real-time fluorescence quantitative PCR. Life Science Research,2007,11(4):301-305.

[23] Zhiwu Li, Jennifer L, Ying L,et al. Using real-time PCR to determine transgene copy number in wheat. Plant Molecular Biology Reporter, 2004,22:179-188.

[1] CHEN Cui-xia,WANG Xiao-long,JIANG Tai-jiao,CAO Zong-fu,LI Tian-jun,YU Lei,YU Yu-fei,CAI Rui-kun,GAO Hua-fang,Ma Xu. Platform Construction for the Early-Warning Forecast in Prevention and Control of Influenza Based on Multi-Source Heterogeneous Big-Data Mining[J]. China Biotechnology, 2020, 40(1-2): 109-115.
[2] LIU Li-yan,LIU Qi-qi,ZHANG Ying,WANG Sheng-qi. The Study of a Novel Nucleic Acid Detection Technology by Double-stranded Probe Real-time PCR[J]. China Biotechnology, 2020, 40(11): 28-34.
[3] Qi ZHANG,Lin YAO,Yan-hua JIANG,Feng-ling LI,Yuan ZHANG,Dong-qin XU,Wen-jia ZHU,Ying-ying GUO,Lian-zhu WANG,Yu-xiu ZHAI. Development of Armored RNA Reference Material of Norovirus Based on Qbeta Bacteriophage[J]. China Biotechnology, 2018, 38(1): 42-50.
[4] FU Li-wen, ZHANG Yu, YI Han, LI Xue, ZHU Nai-shuo. Establishment and Application of Multiplex Fluorescent Real-time PCR for Detecting Six Kinds of Animal Derived Materials[J]. China Biotechnology, 2017, 37(9): 48-59.
[5] Yan-yan LIU,Hui-rong LI,Yue HU,Yang-yang FAN,Xiang-ming LI,Qing-qing TAN,Jia-qiang WU,Xun BU. Multiplex Fluorescent Real-time PCR Detection of Fox, Mink, Raccoon and Dog Derived Materials in Feedstuff[J]. China Biotechnology, 2017, 37(12): 67-76.
[6] ZHANG Li-li, XU Bi-yu, LIU Ju-hua, JIA Cai-hong, ZHANG Jian-bin, JIN Zhi-qiang. Analysis of Banana MaASR1 Gene Expression Profiles in Arabidopsis Under Drought Stress[J]. China Biotechnology, 2017, 37(11): 59-73.
[7] REN Shuang, ZHU Hong-liang. Establishment of Taqman Quantitative PCR System to Estimate Copy Numbers of Exogenous Transgene in Genome Edited Tomato[J]. China Biotechnology, 2017, 37(10): 72-80.
[8] DONG Juan, LI Fo-sheng, LUO Feng-Xue, XIA Fang, ZHU Shu-hua, TANG Lin. Cloning and Expression Analysis of Rice miRNA3026 Promoter and Thioredoxin OsTxnDC9[J]. China Biotechnology, 2016, 36(1): 29-37.
[9] XU Deng-an, ZHAO Chun-qin, ZHANG Chi-hong, CHEN Jing. Expression Patterns of a Root-specific Barley Aquaporin Gene HvTIP2;1 and Promoter[J]. China Biotechnology, 2015, 35(7): 15-21.
[10] ZHANG Yu, BAI Dou, ZHU Nai-shuo. Multiplex Taqman Real-time PCR for Detection of Malignant Tumor Metastasis Rate in Cancer Metastasis Model in Nude Mice[J]. China Biotechnology, 2015, 35(4): 66-73.
[11] ZHOU Yu-cong, XIE Qiu-jin, SONG Kai, YANG Zhao-hui, CHEN Jie, LI Ya-qian. Optimal Agrobacterium tumefaciens-Mediated Transformation Technology Applied in Gene Knockout for Trichoderma atroviride[J]. China Biotechnology, 2015, 35(12): 58-64.
[12] HE Ya-nan, CHEN Xiao-li, REN Xiao-xia, HAO Hai-sheng, QIN Tong, ZHAO Xue-ming, LU Yong-qiang, WANG Dong. Research on Rurification of Mouse Spermatogonial Stem Cells Using Magnetic Microbeads[J]. China Biotechnology, 2014, 34(7): 38-43.
[13] ZHU Cai-hong, LI Shui-gen, QI Li-wang, HAN Su-ying. Agrobacterium tumefaciens-mediated Transformation of Larix leptolepis Embryogenic Tissue[J]. China Biotechnology, 2013, 33(5): 75-80.
[14] LI Fu-wei, ZHANG Shu-ya, REN Shuo, ZHANG Rui-hao, JIANG Yu, GU Shun-zhang, GE Xin, ZHANG Lei, BAO Jian-qiang. Detection of Gadiformes by Real-time PCR Assay[J]. China Biotechnology, 2012, 32(12): 80-85.
[15] LI Ming, LIU Meng, HUANG Yun-yan, ZHOU Li-ying, SUN Xin, LU Fu-ping. Establishment and Optimization of Agrobacterium tumefaciens-mediated Transformation System of Aspergillus niger[J]. China Biotechnology, 2012, 32(01): 56-63.