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作物学报 ›› 2010, Vol. 36 ›› Issue (09): 1605-1609.doi: 10.3724/SP.J.1006.2010.01605

• 研究简报 • 上一篇    下一篇

玉米ZmPR4启子的克隆及其在小麦幼胚愈伤组织中的活性分析

王爱云1,2,庄洪涛2,3,**,张增艳2,*,张学文3,杜丽璞2,叶兴国2   

  1. 1 中南林业科技大学生命科学与技术学院, 湖南长沙 410004; 2 中国农业科学院作物科学研究所 / 农作物基因资源与基因改良国家重大科学工程 /农业部作物遗传育种重点开放实验室, 北京 100081; 3 湖南农业大学生物科学技术学院, 湖南长沙 410128
  • 收稿日期:2010-05-04 修回日期:2010-07-19 出版日期:2010-09-12 网络出版日期:2010-08-25
  • 通讯作者: 张增艳, E-mail: zhangzy@mail.caas.net.cn
  • 基金资助:

    本研究由国家重大科技专项（2008ZX08002-001）资助。

Cloning and Activity Analysis of Zea mays ZmPR4 Promoter in Wheat Immature Embryonic Cali

WANG Ai-Yun1,2, ZHUANG Hong-Tao2,3,**,ZHANG Zeng-Yan2,*,ZHANG Xue-Wen3,DU Li-Pu2,YE Xing-Guo2   

  1. 1 College of Life Science and Technology, Central South University of Forestry and Technology, Changsha 410004, China; 2 National Key Facility for Crop Gene Resources and Genetic Improvement, Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China;     3 College of Life Science and Technology, Hunan Agricultural University, Changsha 410128, China
  • Received:2010-05-04 Revised:2010-07-19 Published:2010-09-12 Published online:2010-08-25
  • Contact: ZHANG Zeng-Yan, E-mail: zhangzy@mail.caas.net.cn

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被引次数

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摘要： 以玉米B73基因组DNA为模板, 通过特异PCR扩增, 克隆出玉米启动子ZmPR4序列。序列分析表明, 该启动子与AJ969166序列同源性为100%。构建了ZmPR4或玉米泛素基因(Ubiquitin)启动子控制的报告基因GUS的表达载体。通过基因枪介导法转化小麦幼胚愈伤组织。瞬间表达实验表明, 在小麦幼胚愈伤组织中, 玉米ZmPR4启动子的本底表达活性明显比Ubi启动子的低, 但经纹枯病菌诱导后, ZmPR4 启动子控制的GUS基因的表达明显增强。PCR检测结果证实ZmPR4 启动子在小麦愈伤组织中具有表达活性, 能够驱动GUS基因的表达。因此, 玉米ZmPR4启动子在小麦抗病基因工程育种中具有潜在的应用价值。

关键词: 诱导型启动子, 克隆, 愈伤组织, 瞬间表达

Abstract: The ZmPR4 promoter was cloned from genomic DNA of maize inbred line B37 through specific PCR amplification. The promoter cloned had the same sequence as the fragment registered in GenBank under the accession number of AJ969166. Expression vectors driven by ZmPR4 promoter or Ubiquitin promoterwere constructed, which harbor GUS gene. The vectors were bombarded into wheat immature embryogenic calli. Histochemical assays of GUS activity, the ZmPR4 promoter was obviously expressed in wheat immature embryonic calli, but the expression activity was lower than that of Ubi Promoter. The ZmPR4 promoter could be induced by infection of Rhizoctonia cerealis. PCR assay confirmed the expression of GUS gene driven by ZmPR4 promoter. These results suggest that ZmPR4promoter has a use potential in molecular breeding of resistance in wheat.

Key words: Inducible promoter, Cloning, Calli, Transient expression

[1] Chen Y-X(陈延熙), Tang W-H(唐文华), Zhang D-H(张敦华), Jian X-Y(简小鹰). A preliminary study on etiology of sharp eyespot of wheat in China. J Plant Prot (植物保护学报), 1986, 13(1): 39-44 (in Chinese with English abstract)
[2] Shi J-R(史建荣), Wang Y-Z(王裕中), Shen S-W(沈素文), Chen H-G(陈怀谷).Pathogenicity of Rhizoctonia cerealis to wheat in Jiangsu province. Jiangsu J Agric Sci (江苏农业学报), 1997, 13(3): 188-190 (in Chinese with English abstract)
[3] Wang Y-Z(王裕中). Occurrence of wheat sharp eyespot and its controlling. Plant Prot Technol Extension (植保技术与推广), 2001, 21(8): 39-41 (in Chinese)
[4] Mitsuhara I, Matsufuru H, Ohshima M, Kaku H, Nakajima Y, Murai N, Natori S, Ohashi Y. Induced expression of sarcotoxin IA enhanced host resistance against both bacterial and fungal patho- gens in transgenic tobacco
[J].Mol Plant-Microbe Interact
[5] Osusky M, Zhou G, Osuska L, Hancock R E, Kay W W, Misra A. Transgenic plants expressing cationic peptide chimeras exhibit broad-spectrum resistance to phytopathogens
[J].Nat Biotech
[6] Sharma A, Sharma R, Imamura M, Yamakawa M, Machii H. Transgenic expression of cecropin B, an antibacterial peptide from Bombyx mori, confers enhanced resistance to bacterial leaf blight in rice
[J].FEBS Lett
[7] Li Q, Lawrence C B, Xing H Y, Babbitt R A, Bass W T, Maiti I B, Everett N P. Enhanced disease resistance conferred by expression of an antimicrobial magainin analog in transgenic tobacco. Planta, 2001, 212: 635-639
[8] Carmona, M J, Molina A, Fernández, J A, López-Fando J J, García-Olmedo F. Expression of the a-thionin gene from barley in tobacco confers enhanced resistance to bacterial pathogens
[J].Plant J
[9] Mourgues F, Brisset M N, Chevreau E. Strategies to improve plant resistance to bacterial diseases through genetic engineering
[J].Trends Biotech
[10] Coca M, Bortolotti C, Rufat M, Peñas G, Eritja R, Tharreau D, Martinez del Pozo A, Messeguer J, San Segundo B. Transgenic rice plants expressing the antifungal AFP protein from Aspergillus giganteus show enhanced resistance to the rice blast fungus Magnaporthe grisea
[J].Plant Mol Biol
[11] Iwai T, Kaku H, Honkura R, Nakamura S, Ochiai H, Sasaki T, Ohashi Y. Enhanced resistance to seedtransmitted bacterial diseases in transgenic rice plants overproducing an oat cell-wall- bound thionin
[J].Mol Plant-Microbe Interact
[12] Kanzaki H, Nirasawa S, Saitoh H, Ito M, Nishihara M, Terauchi R, Nakamura I. Overexpression of the wasabi defensin gene confers enhanced resistance to blast fungus (Magnaporthe grisea) in transgenic rice
[J].Theor Appl Genet
[13] Jia S-R(贾士荣). Environment and food biosafety assessment of trangenic plants. Adv Bioeng (生物工程进展), 1997, 17(6): 37-42 (in Chinese with English abstract)
[14] Morris S H, Adley C C. Irish public perceptions and attitudes to modern biotechnology: an overview with a focus on GM foods
[J].Trends Biotechnol
[15] Simth N. Seeds of opportunity an assessment of the benefits, safety and oversight of plant genomics and agricultural biotechnology. Biotechnology Law Report. August 2000, 19(4): 449-536.
[16] Moreno A B, Peñas G, Rufat M, Bravo M J, Estopà M, Messeguer J, Segundo B S. Pathogen-induced production of the antifungal AFP protein from Aspergillus giganteus confers resistance to the blast fungus Magnaporthe grisea in transgenic rice
[J].Mol Plant- Microbe Interact
[17] Jefferson R A. Assaying chimeric genes in plants: the GUS gene fusion system
[J].Plant Mol Biol Rep
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