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Acta Agron Sin ›› 2011, Vol. 37 ›› Issue (10): 1897-1903.doi: 10.3724/SP.J.1006.2011.01897

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Utilization of Tissue Specific Expressing Promoter RSS1P in TiERF1 Transgenic Wheat

1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实验室，北京 100081; 2山东农业大学农学院，山东泰安 2701082; 3江苏省农业科学院，江苏南京 210014   

  1. 1 National Key Facility for Crop Gene Resources and Genetic Improvement / Key Laboratory of Crop Genetic and Breeding, Ministry of Agriculture / Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China; 2 College of Agronomy, Shandong Agricultural University, Tai’an 271018, China; 3 Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China
  • Received:2010-11-05 Revised:2011-06-25 Online:2011-10-12 Published:2011-07-28
  • Contact: 张增艳, E-mail: zhangzy@mail.caas.net.cn, Tel: 010-82108781; 李斯深, E-mail: ssli@sdau.edu.cn E-mail:lizhao0504@163.com

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Abstract: A sucrose synthase-1 promoter (RSS1P) was cloned from rice leaf, and subcloned in-fused with 5′-terminate of the gene of ethylene responsive factor of Thinopyrum intermedium (TiERF1) to form the RSS1P::TiERF1 expressing cassette. The expressing cassette replaced the Ubi::bar gene expression cassette of pAHC20 to produce the phloem-specific expressing vector pA20-RSS1P::TiERF1. The selective mark bar was deleted in the pA20-RSS1P::TiERF1 vector, which is safe for the ecosystem. Using the method of bombardment, the vector DNAs of pA20-RSS1P::TiERF1 andpAHC20 with bar gene were co-transformed to young embryo callus of wheat cultivar Yangmai 12. The RSS1P::TiERF1 transgenic plants in T0 and T1 generations were subjected to PCR, PCR-Southern, RT-PCR, and Q-RT-PCR analyses, and Rhizoctonia cerealis resistance tests. The molecular detection results showed that the transgene RSS1P::TiERF1 was introduced into Yangmai 12 and was inheritable. In the RSS1P::TiERF1 transgenic wheat plants, expression of TiERF1 was detected in root, stem, and leaf except seed. The highest expression level was observed in root. Compared to the untransformated Yangmai 12, resistance to R. cerealis in the RSS1P::TiERF1 transgenic wheat plants was obviously improved, which was similar to that in the Ubi::TiERF1 transgenic wheat plants. The agronomic traits of RSS1P::TiERF1 transgenic plants had no obvious changes compared to untransformated Yangmai 12. These results suggest that RSS1P promoter is feasible to be used for developing new transgenic wheat germplasm.

Key words: Rice sucrose synthase-1 promoter, Phloem-specific expression, Transgenic wheat, Ethylene responsive factor of Thinopyrum intermedium

[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 eye-spot of wheat in China. Acta Phytophyl Sin (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, l3(3): 188?190 (in Chinese with English abstract)
[3]Dong N(董娜), Zhang Z-Y(张增艳), Xin Z-Y(辛志勇). Isolation and expression analysis of a pathogen-induced ERF gene in Triticum aestivum L. Sci Agric Sin (中国农业科学), 2008, 41(4): 946?953 (in Chinese with English abstract)
[4]Shi J-R(史建荣), Wang Y-Z(王裕中), Chen H-G(陈怀谷), Shen S-W(沈素文). Screening techniques and evaluation of wheat resistance to sharp eyespot caused by Rhizoctonia cerealis. Acta Phytophyl Sin (植物保护学报), 2000, 27(2): 107?112 (in Chinese with English abstract)
[5]Yang L-J(杨立军), Yang X-J(杨小军), Yu D-Z(喻大昭), Wang S-N(王绍南). Resistance evaluation of wheat cultivars (lines) to Rhizoctonia cerealis Varder Hoeven and screening of its resistance resources. Plant Prot (植物保护), 2001, 27(2): 4?7 (in Chinese with English abstract)
[6]Wan Y-X(万映秀), Wang W-X(王文相), Zhang P-Z(张平治), Cao W-X(曹文昕), Zhao L(赵莉). Identification techniques and screening of sharp eyespot resistance of wheat. Chin Agric Sci Bull (中国农学通报), 2009, 25(7): 223?226 (in Chinese with English abstract)
[7]Zhang X-C(张小村), Li S-S(李斯深), Zhao X-H(赵新华), Li R-J(李瑞军). Genetic analysis on resistance to sharp eyespot by using fifteen populations of recombinant inbred lines in wheat. J Triticeae Crops (麦类作物学报), 2004, 24(3): 13?16 (in Chinese with English abstract)
[8]Tang T(汤颋), Ren L-J(任丽娟), Cai S-B(蔡士宾), Wu J-Z(吴纪中), Lu W-Z(陆维忠), Chen J-M(陈建民), Ma H-X(马鸿翔). Study on QTL mapping of sharp eyespot resistance (Rhizoctonia cerealis) in wheat ARz. J Triticeae Crops (麦类作物学报), 2004, 24(4): 11?16 (in Chinese with English abstract)
[9]Cai S-B(蔡士宾), Ren L-J(任丽娟), Yan W(颜伟), Wu J-Z(吴纪中), Chen H-G(陈怀谷), Wu X-Y(吴小有), Zhang X-Y(张仙义). Germplasm development and mapping of resistance to sharp eyespot (Rhizoctonia cerealis) in wheat. Sci Agric Sin (中国农业科学), 2006, 39(5): 928?934 (in Chinese with English abstract)
[10]Liang H X, Lu Y, Liu H X, Wang F D, Xin Z Y, Zhang Z Y. A novel activator-type ERF of Thinopyrum intermedium, TiERF1, positively regulates defence responses. J Exp Bot, 2008, 59: 3111?3120
[11]Lu Y(路妍), Zhang Z-Y(张增艳), Ren L-J(任丽娟), Liu B-Y(刘宝业), Liao Y(廖勇), Xu H-J(徐惠君), Du L-P(杜丽璞), Ma H-X(马鸿翔), Ren Z-L(任正隆), Jing J-X(井金学), Xin Z-Y(辛志勇). Molecular analyses on Rs-AFP2 transgenic wheat plants and their resistance to Rhizoctonia cerealis. Acta Agron Sin (作物学报), 2009, 35(4): 640?646 (in Chinese with English abstract)
[12]Chen L, Zhang Z Y, Liang H X, Liu H X, Du L P, Xu H J, Xin Z Y. Overexpression of TiERF1 enhances resistance to sharp eyespot in transgenic wheat. J Exp Bot, 2008, 59: 4195?4204
[13]Oñate-Sánchez L, Anderson J P, Young J, Singh K B. AtERF14, a member of the ERF family of transcription factors, plays a nonredundant role in plant defense. Plant Physiol, 2007, 143: 400?409
[14]Shi Y, Wang M B, Powell K S, Van Damme E, Hilder V A, Gatehouse A M R, Boulter D, Gatehouse J A. Use of the rice sucrose synthase-1 promoter to direct phloem-specific expression of β-glucuronidase and snowdrop lectin genes in transgenic tobacco plants. J Exp Bot, 1994, 45: 623?631
[15]Li Y-C(李永春), Zhang X-Y(张宪银), Xue Q-Z(薛庆中). Cloning and specific expression of rice sucrose synthase gene promoter in transgenic rice plants. Acta Agron Sin (作物学报), 2002, 28(5): 586?590 (in Chinese with English abstract)
[16]Murray M G, Thompson W F. Rapid isolation of high molecular weight plant DNA. Nucl Acids Res, 1980, 8: 4321?4325
[17]Wang M B, Boulter D, Gatehouse J A. A complete sequence of the rice sucrose synthase-1 (RSs1) gene. Plant Mol Biol, 1992, 19: 881?885
[18]Christensen A H, Quail P H. Ubiquitin promoter-based vectors for high-level expression of selectable and/or screenable marker genes in monocotyledonous plants. Transgenic Res, 1996, 5: 213?218
[19]Xu H-J(徐惠君), Pang J-L(庞俊兰), Ye X-G(叶兴国), Du L-P(杜丽璞), Li L-C(李连城), Xin Z-Y(辛志勇), Ma Y-Z(马有志), Chen J-P(陈剑平), Chen J(陈炯), Cheng S-H(程顺和), Wu Y-H(吴宏亚). Study on the gene transferring of Nib8 into wheat for its resistance to the yellow mosaic virus by bombardment. Acta Agron Sin (作物学报), 2001, 27(6): 684?689 (in Chinese with English abstract)
[20]Sharp P J, Kries M, Shewry P R, Gale M D. Location of β-amylase sequences in wheat and its relatives. Theor Appl Genet, 1988, 75: 286?290
[21]Zhang Z-Y(张增艳), Xu J-S(许景升), Liu Y-G(刘耀光), Wang X-P(王晓萍), Lin Z-S(林志珊), Xin Z-Y(辛志勇). Isolation of resistance gene candidates by a resistance gene analog of Thinopyrum intermedium and pooled-PCR. Acta Agron Sin (作物学报), 2004, 30(3): 189?195 (in Chinese with English abstract)
[22]Livak K J, Schmittgen T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2–ΔΔCT method. Methods, 2001, 25: 402?408
[23]Higo K, Ugawa Y, Iwamoto M, Korenaga T. Plant cis-acting regulatory DNA elements (PLACE) database. Nucl Acids Res, 1999, 27: 297?300
[24]Prestridge D S. SIGNAL SCAN: a computer program that scans DNA sequences for eukaryotic transcriptional elements. Comput Appl Biosci, 1991, 7: 203?206
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