玉米大斑病菌MAPK基因StIME2的基因组定位蛋白质结构预测及表达分析

doi:10.3864/j.issn.0578-1752.2015.13.007

Abstract

Abstract: 【Objective】The objectives of this study are to determine the location in genome, investigate the protein structure of StIME2 of Setosphaeria turcica, and to compare the expression level of StIME2 at different developmental stages and under different stress conditions, such as temperature, oxygen stress, hyperosmolar stress, hence laying a foundation for future studying functions of StIME2.【Method】 Blast search tool was used to determine the exact location of StIME2 in genome of S. turcica. Physical and chemical properties of StIme2 was analyzed by employing online software ProtParam, and secondary structure of StIme2 was predicted by applying Software SOMPA. Three-dimensional structure of StIme2 was doped out using PHYRE2 server and the structure model was evaluated using online software SAVES and the aim was to obtain Laplace image (Ramachandran Plot). StIME2 expression level was detected at different developmental stages and stress conditions based on semi-quantitative RT-PCR analysis. 【Result】The identity of StIME2 was 98 105, located between 1 560 184 and 1 562 574 in the positive-strand of scaffold_7 in the genome of S. turcica. StIme2 shared characteristic conserved domains of MAPK-like protein kinase, α-helices and random coils, which were its main elements of secondary structure of StIme2. β strands were less and mainly located in N-terminal in this structure. Three-dimensional structure of the protein was composed of a smaller N-terminal and a bigger C-terminal. The expression level of StIME2 was the highest in conidium developmental periods, and was the lowest during appressorium development stage. Under different stress conditions, the expression level of StIME2 appeared the highest when the incubation temperature was at 28℃. In hyperosmotic stress conditions, the expression level of StIME2 increased with the increase of NaCl concentration, but the expression level of the gene was almost completely inhibited at higher stress conditions (0.8 mol·L^-1 NaCl). Under oxygen stress conditions, in H₂O₂ stress treatment, the expression level of StIME2 was heightened with the increase of the concentration of H₂O₂. And the expression level of the gene was the highest under 10 mmol·L^-1 H₂O₂. 【Conclusion】 StIME2 was located between 1 560 184 and 1 562 574 in the positive-strand of scaffold_7 in the genome of S. turcica. The characteristics of StIme2 were analyzed, and the result showed that StIme2 shared all conserved domains of MAPK kinase of plant pathogenic fungi. However, unlike three types of MAPK genes, which were widely reported in plant pathogens, StIme2 was rarely reported. The expression level of StIME2 in conidium developmental period in S. turcica was the highest. Thus, the authors speculated that StIME2 could play an important role in regulating the developmental processes in spore of the pathogen. Moreover, the expression level of StIME2 changed with the temperature and the highest expression level was at 28℃. The gene may be also involved in osmotic stress and oxygen stress reaction of the pathogen.

Key words: Setosphaeria turcica, StIME2, bioinformatics analysis, gene expression

GONG Xiao-dong, WANG Yue, ZHANG Pan, FAN Yong-shan, GU Shou-qin, HAN Jian-min, DONG Jin-gao. Analysis of the Genomic Location, Protein Structure Prediction and Expression of MAPK Gene StIME2 in Setosphaeria turcica[J].Scientia Agricultura Sinica, 2015, 48(13): 2549-2558.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://www.chinaagrisci.com/EN/10.3864/j.issn.0578-1752.2015.13.007

https://www.chinaagrisci.com/EN/Y2015/V48/I13/2549

References

[1] 孙淑琴, 温雷蕾, 董金皋. 玉米大斑病菌的生理小种及交配型测定. 玉米科学, 2005, 13(4): 112-113, 123.

Sun S Q, Wen L L, Dong J G. Identification of physiological races and mating type of Exserohilum turcicum. Journal of Maize Sciences, 2005, 13(4): 112-113, 123. (in Chinese)

[2] Degefu Y, Lohtander K, Paulin L. Expression patterns and phylogenetic analysis of two xylanase genes (htxyl1 and htxyl2) from Helminthosporium turcicum, the cause of northern leaf blight of maize. Biochimie, 2004, 86(2): 83-90.

[3] Zhao X H, Mehrabi R, Xu J R. Mitogen-activated protein kinase pathways and fungal pathogenesis. Eukaryotic Cell, 2007, 6(10): 1701-1714.

[4] Brefort T, Müller P, Kahmann R. The high-mobility-group domain transcription factor Rop1 is a direct regulator of prf1 in Ustilago maydis. Eukaryotic Cell, 2005, 4(2): 379-391.

[5] Yang S H, Sharrocks A D, Whitmarsh A J. Transcriptional regulation by the MAP kinase signaling cascades. Gene, 2003, 320(3): 3-21.

[6] Romeis T. Protein kinases in the plant defence response. Current Opinion in Plant Biology, 2001, 4: 407-414.

[7] 范永山, 刘颖超, 谷守芹, 桂秀梅, 董金皋. 植物病原真菌的MAPK基因及其功能. 微生物学报, 2004, 44(4): 547-551.

Fan Y S, Liu Y C, Gu S Q, Gui X M, Dong J G. Mitogen activated protein kinase genes and its functions in phytopathogenic fungus. Acta Microbiologica Sinica, 2004, 44(4): 547-551. (in Chinese)

[8] Idnurm A, Howlett B J. Pathogenicity genes of phytopathogenic fungi. Molecular Plant Pathology, 2001, 2(4): 241-255.

[9] Krens S F, Spaink H P, Snaar-Jagalska B E. Functions of the MAPK family in vertebrate-development. FEBS Letters, 2006, 580(21): 4984-4990.

[10] Chen R E, Thorner J. Function and regulation in MAPK signaling pathways: Lessons learned from the yeast Saccharomyces cerevisiae. Biochimica et Biophysica Acta, 2007, 1773(8): 1311-1340.

[11] Harel A, Bercovich S, Yarden O. Calcineurin is required for sclerotial development and pathogenicity of Sclerotinia sclerotiorum in an oxalic acid-independent manner. Molecular Plant-Microbe Interactions, 2006, 19(6): 682-693.

[12] Hamel L P, Nicole M C, Duplessis S, Ellis B E. Mitogen-activated protein kinase signaling in plant-interacting fungi: Distinct messages from conserved messengers. The Plant Cell, 2012, 24(4): 1327-1351.

[13] Xu J R. MAP kinases in fungal pathogens. Fungal Genetics and Biology, 2000, 31(3): 137-152.

[14] Garrido E, Voss U, Müller P, Castillo-Lluva S, Kahmann R, Pérez M J. The induction of sexual development and virulence in the smut fungus Ustilago maydis depends on Crk1, a novel MAPK protein. Genes & Development, 2004, 18(24): 3117-3130.

[15] 巩校东, 张晓玉, 田兰, 王星懿, 李坡, 张盼, 王玥, 范永山, 韩建民, 谷守芹, 董金皋. 玉米大斑病菌MAPK超家族的全基因组鉴定及途径模型建立. 中国农业科学, 2014, 47(9): 1715-1724.

Gong X D, Zhang X Y, Tian L, Wang X Y, Li P, Zhang P, Wang Y, Fan Y S, Han J M, Gu S Q, Dong J G. Genome-wide identification MAPK superfamily and establishment of the model of MAPK cascade pathway in Setosphaeria turcica. Scientia Agricultura Sinica, 2014, 47(9): 1715-1724. (in Chinese)

[16] 杨鹏雅, 李开绵, 周建国, 王文泉. 蓖麻WRKY基因的电子克隆及其生物信息学分析. 现代农业科学, 2009, 16(5): 22-25.

Yang P Y, Li K M, Zhou J G, Wang W Q. Identification and bioinformatical analysis of novel WRKY gene from Ricinus Communis. Modern Agricultural Sciences, 2009, 16(5): 22-25. (in Chinese)

[17] 武雪, 黄晓丽, 王喆之. 葡萄乙醇脱氢酶基因Ⅲ的电子克隆及生物信息学分析. 生物技术通报, 2009(5): 71-75.

Wu X, Huang X L, Wang Z Z. Electronic cloning and characterizations of ADHⅢ gene from vitisvinifera using bioinformatics tools. Biotechnology Bulletin, 2009(5): 71-75. (in Chinese)

[18] 张鑫, 曹志艳, 刘士伟, 郭丽媛, 董金皋. 玉米大斑病菌聚酮体合成酶基因StPKS功能分析. 中国农业科学, 2011, 44(8): 1603-1609.

Zhang X, Cao Z Y, Liu S W, Guo L Y, Dong J G. Functional analysis of polyketide synthase gene StPKS in Setosphaeria turcica. Scientia Agricultura Sinica, 2011, 44(8): 1603-1609. (in Chinese)

[19] Krylov D M,Nasmyth K,Koonin E V.Evolution of eukaryotic cell cycle regulation: stepwise addition of regulatory kinases and late advent of the CDKs.Current Biology, 2003, 13(2):173-177.

[20] Schindler K,Benjamin K R,Martin A,Boglioli A,Herskowitz I, Winter E.The Cdk-activating kinase Cak1p promotes meiotic S phase through Ime2p.Molecular and Cellular Biology, 2003, 23(23):8718-8728.

[21] Schindler K, Winter E.Phosphorylation of Ime2 regulates meiotic progression inSaccharomyces cerevisiae. The Journal of Biological Chemistry, 2006, 281(27):18307-18316.

[22] McDonald C M, Wagner M, Dunham M J, Shin M E, Ahmed N T, Winter E. The Ras/cAMP pathway and the CDK-Like Kinase Ime2 regulate the MAPK Smk1 and spore morphogenesis in Saccharomyces cerevisiae. Genetics, 2009, 181(2): 511-523.

[23] Cereghino J L, Cregg J M. Heterologous protein expression in the methylotrophic yeast Pichia pastoris. FEMS Microbiology Reviews, 2000, 24(1): 45-66.

[24] Xia Z, Dickens M, Raingeaud J, Davis R J, Greenberg M E. Opposing effects of ERK and JNK-p38 MAP kinases on apoptosis. Science, 1995, 270(5240): 1326-1331.

[25] Johnson G L, Lapadat R L. Mitogen-activated protein kinase pathways mediated by ERK, JNK, and p38 protein kinases. Science, 2002, 298(5600): 1911-1912.

[26] Cook J G, Bardwell L, Thorner J. Inhibitory and activating functions for MAPK Kss1 in the S. cerevisiae filamentous-growth signalling pathway. Nature, 1997, 390: 85-88.

[27] Brückner S, Köhler T, Braus G H, Heise B, Bolte M, Mösch H U. Differential regulation of Tec1 by Fus3 and Kss1 confers signaling specificity in yeast development. Current Genetics, 2004, 46(6): 331-342.

[28] Yang H Y, Tatebayashi K, Yamamoto K, Saito H. Glycosylation defects activate filamentous growth Kss1 MAPK and inhibit osmoregulatory Hog1 MAPK. The EMBO Journal, 2009, 28(10): 1380-1391.

[29] Strudwick N, Brown M, Parmar V M, Schroder M. Ime1 and Ime2 are required for pseudohyphal growth of Saccharomyces cerevisiae on nonfermentable carbon sources. Molecular and Cellular Biology, 2010, 30(23): 5514-5530.

[30] Hutchison E A, Bueche J A, Glass N L. Diversification of a protein kinase cascade: IME-2 is involved in nonself recognition and programmed cell death in Neurospora crassa. Genetics, 2012, 192(2): 467-482.

[31] Garrido E, Pérez-Martín J. The crk1 gene encodes an Ime2-related protein that is required for morphogenesis in the plant pathogen Ustilago maydis. Molecular Microbiology, 2003, 47(3): 729-743.

Related Articles 15

[1]	ZHANG KeKun,CHEN KeQin,LI WanPing,QIAO HaoRong,ZHANG JunXia,LIU FengZhi,FANG YuLin,WANG HaiBo. Effects of Irrigation Amount on Berry Development and Aroma Components Accumulation of Shine Muscat Grape in Root-Restricted Cultivation [J]. Scientia Agricultura Sinica, 2023, 56(1): 129-143.
[2]	GU LiDan,LIU Yang,LI FangXiang,CHENG WeiNing. Cloning of Small Heat Shock Protein Gene Hsp21.9 in Sitodiplosis mosellana and Its Expression Characteristics During Diapause and Under Temperature Stresses [J]. Scientia Agricultura Sinica, 2023, 56(1): 79-89.
[3]	LI ShiJia,LÜ ZiJing,ZHAO Jin. Identification of R2R3-MYB Subfamily in Chinese Jujube and Their Expression Pattern During the Fruit Development [J]. Scientia Agricultura Sinica, 2022, 55(6): 1199-1212.
[4]	LAI ChunWang, ZHOU XiaoJuan, CHEN Yan, LIU MengYu, XUE XiaoDong, XIAO XueChen, LIN WenZhong, LAI ZhongXiong, LIN YuLing. Identification of Ethylene Synthesis Pathway Genes in Longan and Its Response to ACC Treatment [J]. Scientia Agricultura Sinica, 2022, 55(3): 558-574.
[5]	SHU JingTing,SHAN YanJu,JI GaiGe,ZHANG Ming,TU YunJie,LIU YiFan,JU XiaoJun,SHENG ZhongWei,TANG YanFei,LI Hua,ZOU JianMin. Relationship Between Expression Levels of Guangxi Partridge Chicken m6A Methyltransferase Genes, Myofiber Types and Myogenic Differentiation [J]. Scientia Agricultura Sinica, 2022, 55(3): 589-601.
[6]	GUO ShaoLei,XU JianLan,WANG XiaoJun,SU ZiWen,ZHANG BinBin,MA RuiJuan,YU MingLiang. Genome-Wide Identification and Expression Analysis of XTH Gene Family in Peach Fruit During Storage [J]. Scientia Agricultura Sinica, 2022, 55(23): 4702-4716.
[7]	KANG Chen,ZHAO XueFang,LI YaDong,TIAN ZheJuan,WANG Peng,WU ZhiMing. Genome-Wide Identification and Analysis of CC-NBS-LRR Family in Response to Downy Mildew and Powdery Mildew in Cucumis sativus [J]. Scientia Agricultura Sinica, 2022, 55(19): 3751-3766.
[8]	YuXia WEN,Jian ZHANG,Qin WANG,Jing WANG,YueHong PEI,ShaoRui TIAN,GuangJin FAN,XiaoZhou MA,XianChao SUN. Cloning, Expression and Anti-TMV Function Analysis of Nicotiana benthamiana NbMBF1c [J]. Scientia Agricultura Sinica, 2022, 55(18): 3543-3555.
[9]	JIN MengJiao,LIU Bo,WANG KangKang,ZHANG GuangZhong,QIAN WanQiang,WAN FangHao. Light Energy Utilization and Response of Chlorophyll Synthesis Under Different Light Intensities in Mikania micrantha [J]. Scientia Agricultura Sinica, 2022, 55(12): 2347-2359.
[10]	YUAN JingLi,ZHENG HongLi,LIANG XianLi,MEI Jun,YU DongLiang,SUN YuQiang,KE LiPing. Influence of Anthocyanin Biosynthesis on Leaf and Fiber Color of Gossypium hirsutum L. [J]. Scientia Agricultura Sinica, 2021, 54(9): 1846-1855.
[11]	SHU JingTing,JI GaiGe,SHAN YanJu,ZHANG Ming,JU XiaoJun,LIU YiFan,TU YunJie,SHENG ZhongWei,TANG YanFei,JIANG HuaLian,ZOU JianMin. Expression Analysis of IGF1-PI3K-Akt-Dependent Pathway Genes in Skeletal Muscle and Liver Tissue of Yellow Feather Broilers [J]. Scientia Agricultura Sinica, 2021, 54(9): 2027-2038.
[12]	ZHAO Ke,ZHENG Lin,DU MeiXia,LONG JunHong,HE YongRui,CHEN ShanChun,ZOU XiuPing. Response Characteristics of Plant SAR and Its Signaling Gene CsSABP2 to Huanglongbing Infection in Citrus [J]. Scientia Agricultura Sinica, 2021, 54(8): 1638-1652.
[13]	ZHAO Le,YANG HaiLi,LI JiaLu,YANG YongHeng,ZHANG Rong,CHENG WenQiang,CHENG Lei,ZHAO YongJu. Expression Patterns of TETs and Programmed Cell Death Related Genes in Oviduct and Uterus of Early Pregnancy Goats [J]. Scientia Agricultura Sinica, 2021, 54(4): 845-854.
[14]	ZHU FangFang,DONG YaHui,REN ZhenZhen,WANG ZhiYong,SU HuiHui,KU LiXia,CHEN YanHui. Over-expression of ZmIBH1-1 to Improve Drought Resistance in Maize Seedlings [J]. Scientia Agricultura Sinica, 2021, 54(21): 4500-4513.
[15]	YUE YingXiao,HE JinGang,ZHAO JiangLi,YAN ZiRu,CHENG YuDou,WU XiaoQi,WANG YongXia,GUAN JunFeng. Comparison Analysis on Volatile Compound and Related Gene Expression in Yali Pear During Cellar and Cold Storage Condition [J]. Scientia Agricultura Sinica, 2021, 54(21): 4635-4649.

Metrics

Viewed

Full text

Abstract

Cited

Shared

Discussed

Comments

Recommended 0

No Suggested Reading articles found!

Analysis of the Genomic Location, Protein Structure Prediction and Expression of MAPK Gene StIME2 in Setosphaeria turcica

RichHTML

PDF