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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

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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
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