Welcome to Acta Agronomica Sinica,

Acta Agron Sin ›› 2009, Vol. 35 ›› Issue (2): 370-374.doi: 10.3724/SP.J.1006.2009.00370

• RESEARCH NOTES • Previous Articles     Next Articles

Cloning and Analyzing of G-protein Beta-Subunit Gene in Rhizoctonia solani Causing Soybean Sharp Eyespot

MA Bing-Tian1,2,QU Guang-Lin1,HUANG Wen-Juan1,LIN Yu-Fan1,LI Shi-Gui1,2,*   

  1. 1Rice Research Institute, Sichuan Agricultural University, Chengdu 611130,China;2 Key Laboratory of Crop Genetic Resoures and Improvement, Ministry of Education, Sichuan Agricultural University,Ya'an 625014,China
  • Received:2008-05-18 Revised:2008-09-10 Online:2009-02-12 Published:2008-12-12
  • Contact: LI Shi-Gui


Soybean sharp eyespotis one of the most serious diseases in world. The protein encoded by G-protein β-subunit (Guanine nucleotide binding protein beta-subunit) gene plays an important role in pathogenesis mechanism. In this paper, the G-protein β-subunit from Rhizoctonia solani (Teleomorph: Thanatephorus cucumeris)causing soybean sharp eyespot was identified. The genome of 1 864 and 1 047 bp open reading frame (ORF) were amplified by PCR and RT-PCR. The gene included 4 introns and 5 exons. Introns ranged in size from 54 to 65 bp, and their sequences complied with the rule of “5'-gt” and “ag-3'” (GenBank Accession No. EU663628). The ORF encoded 348-amino acid polypeptide with 38.24 kD of calculated molecular weight and 6.31 of pI. There were two alpha-helixes and seven beta-sheets including four beta-strands each in its amino acid secondary structure. Two alpha-helixes in its N-terminal and seven beta-sheets formed barrel structure by non-regular curl in the tertiary structure. The deduced amino acid sequence of β-subunit was identical to that from Rhizoctonia solani (GenBank Accession No. EU267677, AY884129), Lentinula edodes (GenBank Accession No. AAT74567), Coprinopsis cinerea (GenBank Accession Number EAU92269), Ustilago maydis (GenBank Accession Number AAN33051) and Filobasidiella neoformans (GenBank Accession No. AAD03596) with 99.43%, 89.19%, 87.97%, 83.66%, 80.23%, and 79.72%, respectively. The amplified ORF was ligated into the prokaryotic fusion expression vector pGEX-4T-2. E. coli BL21 was transformed with this recombinant vector and induced by IPTG for expression. The result indicated that the protein size of ORF matched the prediction. This cloning of this gene provides the evidence for controlling hyphal growth, development and virulence in R. solani.

Key words: Soybean, Rhizoctonia solani, Pathology, G-protein beta-subunit, Signal transduction

[1]Zhai Z-H(翟中和), Wang X-Z(王喜忠), Ding M-X(丁明孝). Cell Biology (细胞生物学). Beijing: Higher Education Press, 2000. pp 124–157(in Chinese)
[2]Ford C E, Skiba N P, Bae H, Daaka Y, Reuveny E, Shekter L R, Rosal R, Weng G, Yang C S, Iyengar R, Miller R J, Jan L Y, Lefkowitz R J, Hamm H E. Molecular basis for interations of G protein βγ subunits with effectors. Science, 1998, 280: 1271–1274
[3]Chen J-L(陈巨莲), Ni H-X(倪汉祥), Sun J-R(孙京瑞), Weng G. G protein β1γ2 subunits purification and their interaction with adenylyl cyclase. Sci China (Ser C) (中国科学?C辑), 2003, 33(1): 56–64 (in Chinese)
[4]Hou Y M, Chang V, Capper A B, Taussig R, Gautam N. G protein β subunit types differentially interact with a muscarinic receptor but not adenylyl cyclase type II or phospholipase C-β2/3. J Biol Chem, 2001, 276: 19982–19988
[5]Kasahara S, Nuss D L. Targeted disruption of a fungal G-protein β subunit gene results in increased vegetative growth but reduced viru-lence. Mol Plant Microbe Int, 1997, 10: 984–993
[6]Latijnhouwers M, Govers F. A Phytophthora infestans G-Protein β subunit is involved in sporangium formation. Eukaryot Cell, 2003, 2: 971–977
[7]Zeller C E, Parnell S C, Dohlman H G. The RACK1 ortholog Asc1 functions as a G-protein β-Subunit coupled to glucose responsiveness in yeast. J Biol Chem, 2007, 282: 25168–25176
[8]Delgado-Jarana J, Martínez-Rocha A L, Roldán-Rodriguez R, Ron-cero M I, Di Pietro A. Fusarium oxysporum G-protein beta subunit Fgb1 regulates hyphal growth, development, and virulence through multiple signalling pathways. Fungal Genet Biol, 2005, 42: 61–72
[9]Chen J-L(陈巨莲), Weng G-Z, Ni H-X(倪汉祥). The advancement of G protein and coupled signal transduction pathways. Chin J Biotech-nol (生物工程学报), 2001, 17(2): 113–117(in Chinese with English abstract)
[10]Ruiz-Velasco V, Ikeda S R, Puhl H L. Cloning, tissue distribution and functional expression of the human G protein β4-subunit. Physiol Genomics, 2002, 8: 41–50
[11]Lupas A N, Lupas J M, Stock J B. Do G protein subunits associate via a three-stranded coiled coil? FEBS Lett, 1992, 314: 105–108
[12]Claphan D E, Neer E J. New roles for G-protein βγ-dimers in trans-membrane signaling. Nature, 1993, 365: 403–406
[13]Wang D S, Shaw R, Winkelmann J C, Shaw G. Binding of PH do-mains of β-adrenergic receptor kinase and β-spectrin to WD40/ β-transducin repeat containing regions of the β-subunit of trimeric G-proteins. Biochem Biophys Res Commun, 1994, 203: 29–35
[14]Weiss C A, Garnaat C W, Mukai K, Hu Y, Ma H. Isolation of cDNAs encoding GTP-binding protein β-subunit homologues from maize (ZGB1) and Arabidopsis (AGB1). Proc Natl Acad Sci USA, 1994, 91: 9554–9558
[15]Ishikawa A, Iwasaki Y, Asahi T. Molecular cloning and characteriza-tion of a cDNA for the β-subunit of a G protein from rice. Plant Cell Physiol, 1996, 37: 223–228
[16]Kaydamov C, Tewes A, Adler K, Manteuffel R. Molecular charac-terization of cDNAs encoding G protein α and β subunits and study of their temporal and spatial expression patterns in Nicotiana plum-baginifolia Viv. Biochim Biophys Acta, 2000, 149: 143–160
[17]Wang P, Perfect J R, Heitman J. The G-protein β subunit GPB1 is re-quired for mating and haploid fruiting in Cryptococcus neoformans. Mol Cell Biol, 2000, 20: 352–362
[1] CHEN Ling-Ling, LI Zhan, LIU Ting-Xuan, GU Yong-Zhe, SONG Jian, WANG Jun, QIU Li-Juan. Genome wide association analysis of petiole angle based on 783 soybean resources (Glycine max L.) [J]. Acta Agronomica Sinica, 2022, 48(6): 1333-1345.
[2] YANG Huan, ZHOU Ying, CHEN Ping, DU Qing, ZHENG Ben-Chuan, PU Tian, WEN Jing, YANG Wen-Yu, YONG Tai-Wen. Effects of nutrient uptake and utilization on yield of maize-legume strip intercropping system [J]. Acta Agronomica Sinica, 2022, 48(6): 1476-1487.
[3] YU Chun-Miao, ZHANG Yong, WANG Hao-Rang, YANG Xing-Yong, DONG Quan-Zhong, XUE Hong, ZHANG Ming-Ming, LI Wei-Wei, WANG Lei, HU Kai-Feng, GU Yong-Zhe, QIU Li-Juan. Construction of a high density genetic map between cultivated and semi-wild soybeans and identification of QTLs for plant height [J]. Acta Agronomica Sinica, 2022, 48(5): 1091-1102.
[4] LI A-Li, FENG Ya-Nan, LI Ping, ZHANG Dong-Sheng, ZONG Yu-Zheng, LIN Wen, HAO Xing-Yu. Transcriptome analysis of leaves responses to elevated CO2 concentration, drought and interaction conditions in soybean [Glycine max (Linn.) Merr.] [J]. Acta Agronomica Sinica, 2022, 48(5): 1103-1118.
[5] PENG Xi-Hong, CHEN Ping, DU Qing, YANG Xue-Li, REN Jun-Bo, ZHENG Ben-Chuan, LUO Kai, XIE Chen, LEI Lu, YONG Tai-Wen, YANG Wen-Yu. Effects of reduced nitrogen application on soil aeration and root nodule growth of relay strip intercropping soybean [J]. Acta Agronomica Sinica, 2022, 48(5): 1199-1209.
[6] WANG Hao-Rang, ZHANG Yong, YU Chun-Miao, DONG Quan-Zhong, LI Wei-Wei, HU Kai-Feng, ZHANG Ming-Ming, XUE Hong, YANG Meng-Ping, SONG Ji-Ling, WANG Lei, YANG Xing-Yong, QIU Li-Juan. Fine mapping of yellow-green leaf gene (ygl2) in soybean (Glycine max L.) [J]. Acta Agronomica Sinica, 2022, 48(4): 791-800.
[7] LI Rui-Dong, YIN Yang-Yang, SONG Wen-Wen, WU Ting-Ting, SUN Shi, HAN Tian-Fu, XU Cai-Long, WU Cun-Xiang, HU Shui-Xiu. Effects of close planting densities on assimilate accumulation and yield of soybean with different plant branching types [J]. Acta Agronomica Sinica, 2022, 48(4): 942-951.
[8] DU Hao, CHENG Yu-Han, LI Tai, HOU Zhi-Hong, LI Yong-Li, NAN Hai-Yang, DONG Li-Dong, LIU Bao-Hui, CHENG Qun. Improving seed number per pod of soybean by molecular breeding based on Ln locus [J]. Acta Agronomica Sinica, 2022, 48(3): 565-571.
[9] ZHOU Yue, ZHAO Zhi-Hua, ZHANG Hong-Ning, KONG You-Bin. Cloning and functional analysis of the promoter of purple acid phosphatase gene GmPAP14 in soybean [J]. Acta Agronomica Sinica, 2022, 48(3): 590-596.
[10] WANG Juan, ZHANG Yan-Wei, JIAO Zhu-Jin, LIU Pan-Pan, CHANG Wei. Identification of QTLs and candidate genes for 100-seed weight trait using PyBSASeq algorithm in soybean [J]. Acta Agronomica Sinica, 2022, 48(3): 635-643.
[11] ZHANG Guo-Wei, LI Kai, LI Si-Jia, WANG Xiao-Jing, YANG Chang-Qin, LIU Rui-Xian. Effects of sink-limiting treatments on leaf carbon metabolism in soybean [J]. Acta Agronomica Sinica, 2022, 48(2): 529-537.
[12] YU Tao-Bing, SHI Qi-Han, NIAN-Hai , LIAN Teng-Xiang. Effects of waterlogging on rhizosphere microorganisms communities of different soybean varieties [J]. Acta Agronomica Sinica, 2021, 47(9): 1690-1702.
[13] SONG Li-Jun, NIE Xiao-Yu, HE Lei-Lei, KUAI Jie, YANG Hua, GUO An-Guo, HUANG Jun-Sheng, FU Ting-Dong, WANG Bo, ZHOU Guang-Sheng. Screening and comprehensive evaluation of shade tolerance of forage soybean varieties [J]. Acta Agronomica Sinica, 2021, 47(9): 1741-1752.
[14] CAO Liang, DU Xin, YU Gao-Bo, JIN Xi-Jun, ZHANG Ming-Cong, REN Chun-Yuan, WANG Meng-Xue, ZHANG Yu-Xian. Regulation of carbon and nitrogen metabolism in leaf of soybean cultivar Suinong 26 at seed-filling stage under drought stress by exogenous melatonin [J]. Acta Agronomica Sinica, 2021, 47(9): 1779-1790.
[15] ZHANG Ming-Cong, HE Song-Yu, QIN Bin, WANG Meng-Xue, JIN Xi-Jun, REN Chun-Yuan, WU Yao-Kun, ZHANG Yu-Xian. Effects of exogenous melatonin on morphology, photosynthetic physiology, and yield of spring soybean variety Suinong 26 under drought stress [J]. Acta Agronomica Sinica, 2021, 47(9): 1791-1805.
Full text



No Suggested Reading articles found!