Welcome to Acta Agronomica Sinica,

Acta Agron Sin ›› 2010, Vol. 36 ›› Issue (12): 2028-2034.doi: 10.3724/SP.J.1006.2010.02028


Involvement of Heterotrimeric G Protein α and β Subunits in Defense Responses of Wheat to Puccinia triticina

YANG Jing-Jing,LI Ya-Ning*,LI Xing,LIU Da-Qun*   

  1. College of Plant Protection, Agricultural University of Hebei / Biological Control Center of Plant Diseases and Plant Pests of Hebei Province/ National Engineering Research Center for Agriculture in Northern Mountainous Areas, Baoding 071001, China
  • Received:2010-05-30 Revised:2010-08-03 Online:2010-12-12 Published:2010-10-09
  • Contact: LI Ya-Ning,E-mail:yaning22@yahoo.com.cn;LIU Da-Qun,E-mail:ldq@hebau.edu.cn

Abstract: Heterotrimeric GTP-binding protein (G protein), a kind of protein in living cells, plays an important role in signal transduction on plasma membrane. G protein is actively involved in many physiological processes in plant, such as seed germination, stem elongation, root growth, fruit development, and regulation of defense reaction system. To clarify the functions of G protein on wheat leaf rust resistance, we detected the roles of Gα and Gβ subunits at a gene expression level in the interaction between wheat (Triticum aestvum L.) and Puccinia triticina. The results may help to disclose the molecular mechanism of wheat leaf rust resistance and signal transduction. Typical compatible and incompatible interactions were constructed, from wheat leaf rust resistance near-isogenic lines TcLr1 and P. triticina 05-22-64/05-8-63①. Expression of gene encoding Gα or Gβ subunit was detected in these interactions by real-time fluorescence quantitative PCR. Using water as control, the activities of chitinase, β-1,3-glucanase, and the rate changes of reactive oxygen ions production were determined in the compatible and incompatible interactions of wheat and P. triticina, respectively. In contrast with inoculated by virulent or avirulent strain independently, wheat leaves were smeared in a thin layer by G protein inhibitor pertussis toxin and inoculated with avirulent strain after 24 h. The [1]expressions of gene for Gα and Gβ subunits were up-regulated when inoculated with the avirulent strain but down-regulated by the inoculation of the virulent strain. Different subunits of G protein showed different priorities in the process of disease resistance signal transduction. The β subunit gene expressed earlier and in a higher level than the α subunit gene. In addition, G protein increased the resistance to P. triticina through stimulating the defense enzyme activities and reactive oxygen ions. These results indicate that both Gα and Gβ subunits may be involved in the process of disease resistant signal transduction.

Key words: Wheat leaf rust, Heterotrimeric G protein &alpha, and &beta, subunits, Real-time quantitative PCR, Chitinase, Glucanase, Reactive oxygen ions

[1]Gilman A G. G proteins: transducers of receptor-generated signals. Annu Rev Biochem, 1987, 56: 615-649
[2]Ma H, Yanofsky M F, Meyerowitz E M. Molecular cloning and characterization of GPA1, a G protein αlpha subunit gene from Arabidopsis thaliana. Proc Natl Acad Sci USA, 1990, 87: 3821-3825
[3]Ma H, Yanofsky M F, Huang H. Isolation and sequence analysis of TGA1 cDNAs encoding a tomato G protein α subunit. Gene, 1991, 107: 189-195
[4]Kim W Y, Cheong N E, Lee D C, Je D Y, Bahk J D, Cho M J, Lee S Y. Cloning and sequencing analysis of a full-length cDNA encoding a G protein α subunit, SGA1, from soybean. Plant Physiol, 1995, 108: 1315-1316
[5]Gotor C, Lam E, Cejudo F R, Romero L C. Isolation and analysis of the soybean SGA2 gene (cDNA) encoding a new member of the plant G-protein family of signal transducers. Plant Mol Biol, 1996, 32: 1227-1234
[6]Poulsen C, Mai X M, Borg S A. Lotus japonicus cDNA encoding an α subunit of a heterotrimeric G protein. Plant Physiol, 1994, 105: 1453-1454
[7]Seo H S, Kim H Y, Jeong J Y, Lee S Y, Bahk J D. Molecular cloning and characterization of RGA1 encoding a G protein α subunit from rice (Oryza sativa L. IR36). Plant Mol Biol, 1995, 27: 1119-1131
[8]Seo H, Choi C H, Lee S Y, Cho M J, Bahk J D. Biochemical characteristics of a rice (Oryza sativa L. IR36) G-protein α-subunit expressed in Escherichia coli. Biochem J, 1997, 324: 273-281
[9]Kang S G, Lee H J, Park E H, Suh S G. Molecular cloning characterization of cDNAs encoding heterotrimeric G protein α and β subunits from potato (Solanum tuberosum L.). Mol Cells, 2002, 13: 99-106
[10]Saalbach G, Natura B, Lein W, Buschmann P, Dahse I, Rohrbeck M, Nagy F. The α-subunit of a heterotrimeric G protein from tobacco, NtGPα1, functions in K+ channel regulation in mesophyll cells. J Exp Bot, 1999, 50: 53-61
[11]Ando S, Takumi S, Ueda Y, Ueda T, Mori N, Nakamura C. Nicotiana tabacum cDNAs encoding α and β subunits of a heterotrimeric GTP-binding protein isolated from hairy root tissues. Genes & Genet Syst, 2000, 75: 211-221
[12]Marsh J F, Kaufman L S. Cloning and characterization of PGA1 and PGA2: two G protein α-subunits from pea that promote growth in the yeast Saccharomyces cerevisiae. Plant J, 1999, 19: 237-247
[13]Kusnetsov V V, Oelmueller R. Isolation of cDNAs encoding the subunit α of heterotrimeric G proteins from Lupinus luteus (accession No. X99485). Plant Physiol, 1996, 112: 1399
[14]Pei Z M, Ghassemian M, Kwak C M, McCourt P, Schroeder J I. Role of farnesyltransferase in ABA regulation of guard cell anion channels and plant water loss. Science, 2000, 282: 287-290
[15]Hossain M S, Koba T, Harada K. Cloning and characterization of two full-length cDNAs, TaGA1 and TaGA2, encoding G-protein alpha subunits expressed differentially in wheat genome. Genes & Genet Syst, 2003, 78: 127-138
[16]Weiss C A, Garnaat C W, Mukai K, Hu Y, Ma H. Isolation of cDNAs encoding guanine nucleotide-binding protein beta-subunit homologues from maize (ZGB1) and Arabidopsis (AGB1). Proc Natl Acad Sci USA, 1994, 91: 9554-9558
[17]Ishikawa A, Isasaki Y, Asahi T. Molecular cloning and characterization of a cDNA for the beta subunit of a G protein from rice. Plant Cell Physiol, 1996, 37: 223-228
[18]Jones H D, Smith S J, Desikan R, Plakidou-Dymock S, Lovegrove A, Hooley R. Hetetotrimeric G proteins are implicated in gibberellin induction of α -amylase gene expression in wild oat aleurone. Plant Cell, 1998, 10: 245-254
[19]Kusnetsov V V, Oelmueller R. Isolation and characterization of cDNAs encoding the subunit β of heterotrimeric G proteins from N. tabacum (accession No. X98161). Plant Physiol, 1996, 111: 948
[20]Hossain M S, Koba T, Harada K. Cloning and characterization of a cDNA (TaGB1) encoding β subunit of heterotrimeric G-protein from common wheat cv. S615. Plant Biotechnol, 2003, 20: 153-158
[21]Mason M G,Botella J R.Completing the heterotrimer:Isolation and characterization of an Arabidopsis thaliana G protein γ-subunit cDNA. Proc Natl Acad Sci USA, 2000, 97: 14784-14788
[22]Mason M G, Botella J R. Isolation of a novel G-protein γ-subunit from Arabidopsis thaliana and its interaction with Gβ. Biochim Biophy Acta, 2001, 1520: 147-153
[23]Kato C, Mizutani T, Tamaki H, Kumagai H, Kamiya T, Hirobe A, Fujisawa Y, Kato H, Iwasaki Y. Characterization of heterotrimeric G protein complexes in rice plasma membrane. Plant J, 2004, 38: 320-331
[24]Zeng Z-L(曾志林), Dong H-T(董海涛), Li D-B(李德葆). Functions and related mechanisms of heterotrimeric G protein in plant. Acta Agric Zhejiangensis (浙江农业学报), 2007, 19(3): 248-252 (in Chinese with English abstract)
[25]Song S-S(宋水山), Yang W-X(杨文香), Ling Y-N(李亚宁), Liu D-Q(刘大群). Plant G protein and plant defense response. China Biotechnol (中国生物工程杂志), 2006, 26(3): 73-77(in Chinese with English abstract)
[26]Beffa R, Szell M, Meuwly P, Pay A, Vögeli-Lange R, Métraux J P, Neuhaus G, Meins F Jr, Nagy F. Cholera toxin elevates pathogen resistance and induces pathogenesis-related gene expression in tobacco. EMBO J, 1995, 14: 5753-5761
[27]Ortega X, Velásquez J C, Pérez L M. IP3 production in the hypersensitive response of lemon seeding against Alternaria alternata involves active protein tyrosine kinases but not a G protein. Biol Res, 2005, 38: 89-99
[28]Ortega X, Planco R, Castaneda P, Perez L M. Signal transduction in lemon seedling in the hypersensitive response against Alternaria alternata: participation calmodulin G-protein and protein kinases. Biol Res, 2002, 35: 373-383
[29]Komer J A. Genetics of resistance to wheat leaf rust. Annu Rev Phytopathol, 1996, 34: 435-455
[30]Yang J-J(杨静静), Li X(李星), Li Y-N(李亚宁), Liu D-Q(刘大群). Involvement of small GTP-binding protein gene Rab2 in resistance response of wheat to its leaf rust disease. Acta Agric Boreali-Sin (华北农学报), 2010, 25(6)(in press)
[31]Sambrook J, Russell D. Molecular Cloning: A Laboratory Manual, 3rd Edn. New York: Cold Spring Harbor Laboratory Press, 2001. pp 304-331
[32]Yue C-F(岳彩凤), Kang G-Z(康国章), Liu C(刘超), Guo T-C(郭天财), Zhu Y-J(朱云集), Shen B-Q(沈丙权). Cloning and sequencing of GAPDH gene in common wheat (Tritucum aestivum L.). Chin Agric Sci Bull (中国农学通报), 2008, 24(4): 94-98 (in Chinese with English abstract)
[33]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
[34]Zhang Z-Q(张占全), Song S-S(宋水山), Zhang Y-C(张英春), Yang W-X(杨文香), Liu D-Q(刘大群). Involvement of the heterotrimeric G protein in the defense responses of wheat to Puccinia triticina. Sci Agric Sin (中国农业科学), 2009, 42(1): 117-123 (in Chinese with English abstract)
[35]Zhou Z-F(周祖富), Li Z-A(黎兆安). Laboratory Guide for Plant Physiology (植物生理学实验指导). Nanning: Guangxi University Press, 2005. pp 99-101 (in Chinese)
[36]Shi Y-M(史益敏), Tang Z-C(汤章城), Wang G-Q(王国强). Laboratory Guide for Modern Plant Physiology (现代植物生理学实验指南). Beijing: Science Press, 1999. pp 128-129 (in Chinese)
[37]Boller T, Gehri A, Mauch F, Vögeli U. Chitinase in bean leaves: induction by ethylene, purification, properties, and possible function. Planta, 1983, 157: 22-31
[38]Michel L, Serge K, Pierrette G, Bernard F. Biological function of pathogenesis related proteins: four tobacco pathogenesis related proteins are chitinases. Proc Natl Acad Sci USA, 1987, 84: 6750-6754
[39]Serge K, Michel L, Pierrette G, Bernard F. Biological function of pathogenesis related proteins: four PR proteins of tobacco have β-1,3-glucanase activity. EMBO J, 1987, 6: 3209-3212
[40]Bradford M M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem, 1976, 72: 248-254
[41]Meng L(孟亮), Cui D-C(崔德才). Plant chitinases and its application in resistance to fungal disease. Lett Biotechnol (生物技术通讯), 2004, 15(4): 420-422 (in Chinese with English abstract)
[1] MENG Yu-Yu, WEI Chun-Ru, FAN Run-Qiao, YU Xiu-Mei, WANG Xiao-Dong, ZHAO Wei-Quan, WEI Xin-Yan, KANG Zhen-Sheng, LIU Da-Qun. TaPP2-A13 gene shows induced expression pattern in wheat responses to stresses and interacts with adaptor protein SKP1 from SCF complex [J]. Acta Agronomica Sinica, 2021, 47(2): 224-236.
[2] Fang-Meng DUAN, Qiu-Lan LUO, Xue-Li LU, Na-Wei QI, Xian-Shun LIU, Wen-Wen SONG. Cloning of the Key Gene ZmCYP90B1 in Brassinosteroids Biosynthesis from Zea mays and Its Response to Adversity Stresses [J]. Acta Agronomica Sinica, 2018, 44(03): 343-356.
[3] LIU Hui-Yun,WANG Wan-Qing,LI Xin, WANG Ke,WANG Long,DU Li-Pu,YANG Yue-Ming,YE Xing-Guo. Glu-B1 Silencing Influences Protein Body Formation and Expression of Genes Regulating Synthesis and Processing of Seed-Storage Protein in Somatic mutant Wheat AS208 [J]. Acta Agron Sin, 2017, 43(05): 691-700.
[4] ZHAI Yu-Shan, DENG Yu-Qing, DONG Meng, XU Qian, CHENG Guang-Yuan, PENG Lei, LIN Yan-Quan*,XU Jing-Sheng*. Cloning and Characterization of Light Harvesting Chlorophyll a/b-Binding Protein Coding Gene (ScLhca3) in Sugarcane [J]. Acta Agron Sin, 2016, 42(09): 1332-1341.
[5] SU Wei-Hua**,LIU Feng**,HUANG Long,SU Ya-Chun,HUANG Ning,LING Hui,WU Qi-Bin,ZHANG Hua,QUE You-Xiong*. Cloning and Expression Analysis of a Ca2+/H+ Antiporter Gene from Sugarcane [J]. Acta Agron Sin, 2016, 42(07): 1074-1082.
[6] LIU Feng**,SU Wei-Hua**,HUANG Long,XIAO Xin-Huan,HUANG Ning,LING Hui,SU Ya-Chun,ZHANG Hua,QUE You-Xiong. Isolation and Characterization of a Na+/H+Antiporter Gene from Sugarcane [J]. Acta Agron Sin, 2016, 42(04): 501-512.
[7] FAN Wei,LI Xue-Jiao,GUAN Ming-Li,MIAO Liu-Yang,SHI Jia-Nan,DOU Shi-Juan,LIU Li-Juan,LI Li-Yun,LIU Guo-Zhen. Transcriptional and Translational Characterization of Rice Chitinase Genes [J]. Acta Agron Sin, 2014, 40(04): 571-580.
[8] YANG Hua,GAO Xiang,CHEN Qi-Jiao,ZHAO Wan-Chun,DONG Jian,LI Xiao-Yan. Isolation, Characterization and Farinograph Analysis of Novel HMW-GSs from Dasypyrum villosum [J]. Acta Agron Sin, 2014, 40(04): 600-610.
[9] HAO Jun-Jie,HU Yu-Wei,GUO Xiao-Qin,ZhAO Fu-An,JIA Xin-He,GUO Li-Juan,ZHANG Zhi-Xing,WANG Qing-Dong. Analysis of Resistance to Verticillium Wilt in Cotton by Reciprocal Grafting and Real-time Quantitative PCR [J]. Acta Agron Sin, 2013, 39(07): 1179-1186.
[10] HU E-E, ZHANG Na, LI Lin-Mao, YANG Wen-Xiang, LIU Da-Qun. Analysis of Wheat Leaf Rust Resistance Genes in 14 Wheat Cultivars or Lines [J]. Acta Agron Sin, 2011, 37(12): 2158-2166.
[11] CAO Li, WANG Zhen-Lin, DAI Zhong-Min, YIN Yan-Ping, DI Hua-Xu, NI Yang-Li, CA Tie, LI Yong, WANG Beng, CHEN Er-Ying, GUO Dun-Xiang, CHEN Xiao-Guang. Effect of Nitrogen Fertilization Timing on HMW-GS Accumulation and GMP Size Distribution in Wheat Grains [J]. Acta Agron Sin, 2011, 37(12): 2241-2250.
[12] XU Tian-Tian, CA Jian, HONG Bei, QI Ceng-Jun, DAI Ting-Bei, CAO Wei-Xing, JIANG Dong. T6VS?6AL Chromosome Translocation Does Not Alter Accumulations of High Molecular Weight Glutenin Subunits and Glutenin Macropolymer in Wheat Grain [J]. Acta Agronomica Sinica, 2011, 37(11): 2059-2065.
[13] ZHANG Chang-Wei, LING Yang-Hua, SANG Xian-Chun, LI Bing, ZHAO Fang-Meng, YANG Zheng-Lin, LI Yun-Feng, FANG Li-Kuai, HE Guang-Hua. Transgenic Rice Lines Harboring McCHIT1 Gene from Balsam Pear (Momordica charantia L.) and Their Blast Resistance [J]. Acta Agron Sin, 2011, 37(11): 1991-2000.
[14] MA Xiong-Feng,YU Chun-Ming,TANG Shou-Wei,ZHU Ai-Guo,WANG Yan-Zhou,ZHU Si-Yuan,. Cloning and Tissue Expression of Acting1 Gene in Different Fiber Development Phases of Ramie [Boehmeria nivea (Linn.) Gaud] [J]. Acta Agron Sin, 2010, 36(1): 101-108.
[15] ZHOU Yue, WANG Long, ZHANG Ye, LI Zai-Feng, LIU Da-Qun. Molecular Mapping of Leaf Rust Resistance Genes in Wheat Line Tian 95HF2 [J]. Acta Agron Sin, 2010, 36(08): 1265-1269.
Full text



No Suggested Reading articles found!