Welcome to Acta Agronomica Sinica，

Acta Agron Sin ›› 2008, Vol. 34 ›› Issue (12): 2070-2076.doi: 10.3724/SP.J.1006.2008.02070

• CROP GENETICS & BREEDING · GERMPLASM RESOURCES · MOLECULAR GENETICS • Previous Articles     Next Articles

Expression of Protein Elicitor-Encoding Gene pemG1 in Tobacco (Nicotiana tobacum cv. Samsun NN) Plants and Enhancement of Resistance to TMV

MAO Jian-Jun,QIU De-Wen*,YANG Xiu-Feng,ZENG Hong-Mei,YUAN Jing-Jing   

  1. State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2008-04-21 Revised:2008-07-08 Online:2008-12-12 Published:2008-10-10
  • Contact: QIU De-Wen

PDF

2305

Cited

7

Abstract:

Protein elicitors are important signal molecules that trigger plants disease resistance. Defence responses will be induced once the elicitors are recognized by acceptors in plants. It is revealed that elicitor protein PemG1 from Magnaporthe grisea is able to increase hydrogen peroxide content of tobacco suspension cells. To study PemG1’s functions in plants, pemG1 gene was transferred into tobacco in the study. For this, plant expression vector pCAMBIA2300-Ubi-pemG1-Oc harboring elicitor-encoding gene pemG1 from Magnaporthe grisea was constructed. The maize ubiquitin promoter/octopine synthase terminator system and kanamycin-resistant gene npt II (neomycin phosphotransfers II) were used for constitutive expression systems. The vector was then introduced into Agrobacterium tumefaciens (strain AGL-1) by freeze-thaw method. Tobacco (Nicotiana tobacum cv. Samsun NN) primary transformants were produced by leaf disc transformation. The kanamycin-resistant regenerated plants were confirmed to be electropositive by PCR. Integration and expression of the pemG1 gene were further confirmed by Southern blotting and Western blotting, respectively. Then, transgenic tobacco plants of T2 generation were inoculated with Tobacco Mosaic Virus (TMV) at two different virus concentration. In comparison with TMV-infected wild-type SNN plants, PemG1-expressed plants displayed reduced hypersensitive-response lesions in both treatments. Furthermore, accumulation level of pemG1 steady-state transcripts was examined at 24 h after inoculation. The results indicated that the reduction of lesions corresponded to the accumulation of pemG1 steady-state transcripts as monitored by Northern analysis. All these indicated that the expression of pemG1 in tobacco plants improved the resistance to TMV.

Key words: Protein elicitor, pemG1, Tobacco, Expression, TMV

[1]Michael G H. Microbial elicitors and their receptors in plants. Ann Rev Phytopathol, 1996, 34: 387-412
[2]Darvill A G, Albersheim P. Phytoalexins and their elicitors-a defense against microbial infection in plants. Annu Rev Plant Physiol, 1984, 35: 243-293
[3]Templeton M D, Lamb C J. Elicitors and defense gene activa-tion. Plant Cell Environ, 1988, 11: 395-401
[4]Wei Z M, Laby R J, Zumoff C H, Bauer D W, He S Y, Collmer A, Beer S V. Harpin, elicitor of the hypersensitive response produced by the plant pathogen Erwinia amylovora. Science, 1992, 257: 85-88
[5]Baker C J, Orlandi E W, Mock N M. Harpin, an elicitor of the hypersensitive response in tobacco caused by Erwinia amy-lovora, elicit active oxygen production in suspension cells. Plant Physiol, 1993, 102: 1341-1344
[6]Ricci P, Bonnet P, Huet J C, Sallantin M, Beauvais C F, Bru-neteau M, Billard V, Michel G, Pernollet J C. Structure and activity of proteins from pathogenic fungi Phytophthora eli- citing necrosis and acquired resistance in tobacco. Eur J Bio-chem, 1989, 183: 555-563
[7]Baillieul F, Genetat I, Kopp M, Saindrenan P, Fritig B, Kauffrnann S. A new elicitor of the hypersensitive response in tobacco: A fungal glycoprotein elicits cell death, expression of defense genes, production of salicylic acid, and induction of systemic acquired resistance. Plant J, 1995, 8: 551-560
[8]Nespoulous C, Huet J C, Pernollet J C. Structure-function re-lationships of α and β elicitins signal proteins involved in the plant—Phytophthora interaction. Planta, 1992, 186: 551-557
[9]Huet J C, Nespoulous C, Pernollet J C. Structure of elicitin isoforms secreted by Phytophthora dreschleri. Phytochemis-try, 1992, 31: 1471-1476
[10]Boissy G, De L F, Kahn R, Huet J C, Bridogne G, Pernollet J C, Brunie S. Crytal structure of a fungal elicitor secreted by Phytophthora cryptogea, a member of a novel class of plant necrotic protein. Structure, 1996, 4: 1429-1432
[11]Ricci P, Trentin F, Bonnet P, Vemond P, Mouton-Perronet F, Bnuneteau M. Differential production of parasiticein, an elicitor of necrosis an d resistance in tobacco, by isolates of Phytophthora parasitica. Plant Pathol, 1992, 41: 298-307
[12]Zhang H-M(张宏明), Cai Y-Y(蔡以滢), Chen J(陈珈). Elici-tation of the hypersensitive responses in tabacco by a 10.6 kD proteinaceous elicitor from Phytophthora palmi. Acta Bot Sin (植物学报), 1999, 41(11): 1183-1186(in Chinese with Eng-lish abstract)
[13]Zhang Z-G(张正光), Wang Y-C(王源超), Zheng X-B(郑小波). Bioactivity and stability of 90 kD extracellular protein elicitor from Phytophthora boehmeriae. Acta Phytopathol Sin (植物病理学报), 2001, 31(3): 213-218(in Chinese with Eng-lish abstract)
[14]Qiu D-W(邱德文). Microbe protein pesticide and its prospect. Chin J Biol Control (中国生物防治), 2004, 20(2): 91-94(in Chinese with English abstract)
[15]Qiu D-W(邱德文), Xiao Y-L(肖友伦), Yao Q(姚庆), Li L(李丽), Liu Z(刘峥). Effect of activator protein on cucumber growth and the activities of dehydrogenase, peroxidase and phenylalanine ammonia lyase. Chin J Biol Control (中国生物防治), 2005, 21(1): 41-44(in Chinese with English abstract)
[16]Xu F(徐锋), Yang Y(杨勇), Xie F-J(谢馥交), Liu Z(刘铮), Qiu D-W(邱德文), Yang X-F(杨秀芬). Effect of the activator protein from magnaporthe grisea on plant growth and physiological activities. Acta Agric Boreali-Sin (华北农学报), 2006, 21(5): 1-5(in Chinese with English abstract)
[17]Shao M(邵敏), Wang J-S(王金生). Transformation of rice with hrfA gene and resistance of the transgenic plant to bacte-rial leaf blight. J Nanjing Agric Univ (南京农业大学学报), 2004, 27(4): 36-40 (in Chinese with English abstract)
[18]Meng F-H(孟凡宏), Song C-F(宋从凤), Ji Z-L(纪兆林), Wang J-S(王金生). Effect of transgenic tobacco expressing HarpinXoo and its N-terminal sequence on TMV resistance. J Nanjing Agric Univ (南京农业大学学报), 2007, 30(3): 47-52(in Chinese with English abstract)
[19]Horsch R B, Fry J E, Fraley R T. A simple and general method for transferring genes into plants. Science, 1985, 227: 1229-1231
[20]Del P O, Lam E. Expression of the baculovirus p35 protein in tobacco affects cell death progression and compromises N gene-mediated disease resistance response to Tobacco mosaic virus. Mol Plant-Microbe Interact, 2003, 16: 485-494
[21]Fischer U, Droge-Laser W. Overexpression of NtERF5, a new member of the tobacco ethylene response transcription factor family enhances resistance to tobacco mosaic virus. Mol Plant-Microbe Interact, 2004, 17: 1162-1171
[22]Sinisterra X H, Polston J E, Abouzid A M, Hiebert E. To-bacco plants transformed with a modified coat protein of to-mato mottle Begomovirus show resistance to virus infection. Phytopathology, 1999, 89: 701-706
[23]Kobayashi K, Cabral S, Calamante G, Maldonado S, Mentaberry A. Transgenic tobacco plants expressing the po-tato virus X open reading frame 3 gene develop specific re-sistance and necrotic ring symptoms after infection with the homologous virus. Mol Plant-Microbe Interact, 2001, 14: 1274-1285
[24]Southern E M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol, 1975, 98: 503
[25]Sambrook J, Russell D. Molecular Cloning: A Laboratory Manual, 3rd edn. New York: Cold Spring Harbor Laboratory Press, 2001. pp 304-331
[26]Amero S A, James T C, Elgin S C R. Production of antibodies using proteins in gel bands. In: Walker J M ed. The Protein Protocols Handbook, 2nd edn. New Jersey: Humana Press, 2002. pp 941-944
[27]Wu Y-B(吴艳兵), Xie L-Y(谢荔岩), Xie L-H(谢联辉), Lin Q-Y(林奇英), Lin S-F(林诗发). A preliminary study on anti-TMV activity of polysaccharide from coprinus comatus. Chin Agric Sci Bull (中国农学通报), 2007, 23(5): 338-341 (in Chinese with English abstract)
[28]Strathmann A, Kuhlmann M, Heinekamp T, Dr?ge-Laser W. BZI-1 specifically heterodimerises with the tobacco bZIP transcription factors BZI-2, BZI-3/TBZF and BZI-4, and is functionally involved in flower development. Plant J, 2001, 28: 397-408
[1] Zuo Tong-Hong, Zhang He-Cui, Zeng Jing, Zhu Li-Quan. Molecular cloning and expression analysis of BoPUB3L associated with self-incompatibility in Brasscia oleracea [J]. Acta Agronomica Sinica, 2026, 52(6): 1698-1710.
[2] Wang Wen-Yuan, Yan Xue-Jia, Liu Yu-Lin, Sun Xiao-Tong, Li Ya-Nan, Tang Xin-Hua, Shi Ying. Screening of low-light-tolerant potato varieties and cloning and functional analysis of the transcription factor gene StPIF3 [J]. Acta Agronomica Sinica, 2026, 52(6): 1631-1645.
[3] Hu Zhao, Qian Run, Xie Feng-Pu, Ying Su-Ping. Genome-wide identification and expression analysis of the SPX gene family in rice under phosphorus treatment [J]. Acta Agronomica Sinica, 2026, 52(6): 1902-1912.
[4] Sun Shu-Feng, Xu Zhen-Nan, Huang Jia-Xin, Weng Jian-Feng, Li Xin-Hai. Genome-wide identification of the maize MAPK gene family and its response to Fusarium verticillioides infection [J]. Acta Agronomica Sinica, 2026, 52(5): 1291-1308.
[5] Han Ya-Xin, He Guan-Hua, Zhang Xiao-Qiong, Zhang Deng-Feng, Li Yong-Xiang, Liu Xu-Yang, Wang Tian-Yu, Li Yu, Zou Hua-Wen, Li Chun-Hui. Identification of maize lateral root density genes resources through integrated RNA-seq and BSA-seq analyses [J]. Acta Agronomica Sinica, 2026, 52(5): 1341-1352.
[6] Tian Li-Tao, Ding Ning, Wang Shu-Lin, Qi En-Fang, Zhang Rong, Wang Rui-Rui, Ma Li-Wen, Li Jian-Wu, Yang Jiang-Wei. Genome-wide identification of the Argonaute gene family and its induction by late blight in potato (Solanum tuberosum L.) [J]. Acta Agronomica Sinica, 2026, 52(4): 1116-1126.
[7] Wang Ling, Hu Hao, Song Jia-Feng, Cheng Jie-Lan, Chen Ying, Zheng Ting-Ting, Lyu Zhao-Yan, Zhu Xiao-Biao, Hou Hua-Lan. Cloning and functional validation of UDP-glycosyltransferase gene StUGT52 in potato [J]. Acta Agronomica Sinica, 2026, 52(3): 665-676.
[8] Yang Zong-Tao, Yang Ting, Wang Yu-Tong, Ai Jing, Li Yan-Ye, Liu Jia-Yong, Deng Jun, Zhao Yong, Zhang Yue-Bin. Identification and expression analysis of the CLC gene family in sugarcane [J]. Acta Agronomica Sinica, 2026, 52(3): 722-734.
[9] Meng Cheng, Wang Zhe. Genome-wide identification and expression analysis of the ZmPFK gene family under biotic and abiotic stresses in maize [J]. Acta Agronomica Sinica, 2026, 52(3): 764-779.
[10] Yang Yue, Zhang Xin-Xin, He Zeng-Hui, Li Rui-Dong, Pan Yu-Jie, Li Jia-Kang, Du Wei, Xu Da-Yong, Du Jin-Song. Non-destructive prediction and visualization of major chemical components in tobacco leaves using hyperspectral imaging [J]. Acta Agronomica Sinica, 2026, 52(3): 922-935.
[11] Yang Biao, Du Shuai-Kang, Zhang Ji-Wang, Shi Ying, Zhang Li-Li. Genome-wide identification of class III POD gene family in potato and its expression profile analysis [J]. Acta Agronomica Sinica, 2026, 52(2): 405-420.
[12] Yu Kai-Hang, Zhou Hong-Bin, Luo Liang-Zha, Wang Mei-Li, Jiang Rui-Mei, Dong-Chen Wen-Hua, Li Shi-Jin, Mao Xiao-Qiang, Chen Sheng-Wei. Cloning and expression analysis of the HvLRR-RLK-510 gene encoding a leucine-rich repeat receptor-like kinase in barley [J]. Acta Agronomica Sinica, 2026, 52(2): 421-432.
[13] Zhan Ge-Rui, Yu Wen, Li Feng, Wu Ming-Zhu, Xu Xin, Luo Zhao-Peng, Wu Sheng-Xin, Yang Jun, Zhang Zhi-Qiang, Wang Zhong. Functional study of NtWRKY6 in response to ABA expression and regulation of polyphenol synthesis [J]. Acta Agronomica Sinica, 2026, 52(2): 446-458.
[14] Zhang Li-Lan, Yang Jun, Wang Rang-Jian. Identification of candidate genes related to glycoside aroma precursor content in tea plant using WGCNA [J]. Acta Agronomica Sinica, 2026, 52(2): 494-513.
[15] Liu Di, Li Rui-Yuan, Shi Mao-Zhu, Li Hong-You, Chen Qing-Fu, Shi Tao-Xiong. Phenotypic characterization and transcriptomic analysis of the semi-dwarf mutant sd3 in Tartary buckwheat [J]. Acta Agronomica Sinica, 2026, 52(1): 316-328.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
Add: No. 80 Xueyuan South Rd, Beijing 100081, P. R. China E-mail: zwxb301@caas.cn
Supported by Beijing Magtech Co.Ltd