桉树焦枯病菌MDR基因生物信息学分析及功能解析
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摘要
通过生物信息软件及qPCR对桉树焦枯病菌MDR蛋白的序列特征、蛋白结构、理化性质及MDR基因在不同条件下的表达情况进行分析.结果表明,MDR转运蛋白均由α-折叠、无规则卷曲、β-转角,所占百分比α-折叠>β-转角>无规则卷曲.三级与二级结构预测均显示桉树焦枯病菌MDR具有典型的MDR蛋白结构,说明该蛋白家族的进化比较保守;CpABCB10基因参与调控分生孢子稳定性和萌发,CpABCB4和CpABCB9基因参与调控金属离子的转运,CpABCB7和CpABCB8基因参与调控桉树焦枯病菌的抗药机制.此外,CpABCB7基因还能参与调控桉树焦枯病菌的整个致病过程.
The sequence characteristics, protein structure, physicochemical properties of MDR protein and the expression of MDR gene under different conditions were analyzed. The results showed that the MDR transporters consits of α-helix, random coils, and β-sheet, and the proportion of α-helix, β-sheet, random coils were in a descending order. The predicted structure of tertiary structure and secondary structure are typical MDR protein structure, indicating that the evolution of this protein family is relatively conservative. CpABCB10 gene is involved in the regulation of conidial stability and germination while CpABCB4 and CpABCB9 genes are involved in the regulation of metal ion transport. CpABCB7 and CpABCB8 genes are involved in the regulation of the resistance mechanism of C.pseudoreteaudii. In addition, the CpABCB7 gene can also participate in the regulation of the pathogenic process of C.pseudoreteaudii.
The sequence characteristics, protein structure, physicochemical properties of MDR protein and the expression of MDR gene under different conditions were analyzed. The results showed that the MDR transporters consits of α-helix, random coils, and β-sheet, and the proportion of α-helix, β-sheet, random coils were in a descending order. The predicted structure of tertiary structure and secondary structure are typical MDR protein structure, indicating that the evolution of this protein family is relatively conservative. CpABCB10 gene is involved in the regulation of conidial stability and germination while CpABCB4 and CpABCB9 genes are involved in the regulation of metal ion transport. CpABCB7 and CpABCB8 genes are involved in the regulation of the resistance mechanism of C.pseudoreteaudii. In addition, the CpABCB7 gene can also participate in the regulation of the pathogenic process of C.pseudoreteaudii.
引文
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[2] ANDRADE A C, VAN NISTELROOY J G, PEERY R B, et al. The role of ABC transporters from Aspergillus nidulans in protection against cytotoxic agents and in antibiotic production[J]. Molecular & general genetics, 2000,263(6):966.
[3] NISHI K, YOSHIDA M, NISHIMURA M, et al. A leptomycin B resistance gene of Schizosaccharomyces pombe encodes a protein similar to the mammalian P-glycoproteins[J]. Molecular Microbiology, 1992,6(6):761-769.
[4] SUN C B, SURESH A, DENG Y Z, et al. A multidrug resistance transporter in magnaporthe is required for host penetration and for survival during oxidative stress[J]. Plant cell, 2006,18(12):3 686-3 705.
[5] 楼天灵.小麦赤霉病菌(Fusarium graminearum)ABC转运蛋白(ABCT)基因功能分析[D].杭州:浙江大学,2010.
[6] BOOTH T H, JOVANOVIC T, OLD K M, et al. Climatic mapping to identify high-risk areas for Cylindrocladium quinqueseptatum leaf blight on eucalypts in mainland South East Asia and around the world[J]. Environmental Pollution, 2000,108(3):365-372.
[7] 陈全助,陈慧洁,郭文硕,等. 桉树焦枯病菌(Calonectria pseudoreteaudii)生物学特性测定[J]. 福建林学院学报, 2014(4):328-332.
[8] CHEN S F, LOMBARD L, ROUX J, et al. Novel species of Calonectria associated with Eucalyptus leaf blight in Southeast China[J]. Persoonia, 2011,26:1-12.
[9] LOMBARD L, CROUS P W, WINGFIELD B D, et al. Phylogeny and systematics of the genus Calonectria[J]. Studies in Mycology, 2010,66:31-69.
[10] CHEN Q Z, GUO W S, FENG L Z, et al. Transcriptome and proteome analysis of Eucalyptus infected with Calonectria pseudoreteaudii[J]. Journal of Proteomics, 2015,115:117-131.
[11] 刘宏毅,陈全助,叶小真,等.桉树焦枯病菌ABC转运蛋白的鉴定与分析[J].林业科学研究,2017(4):685-692.
[12] YE X, LIU H, JIN Y, et al. Transcriptomic analysis of Calonectria pseudoreteaudii during various stages of Eucalyptus infection[J]. PLoS One, 2017,12(1)[2017-01-10].https://doi.org/10.1371/journal.pone.0169598
[13] 陈英.桉树焦枯病的研究[D].福州:福建农林大学,2004.
[14] 郑美珠.桉树焦枯病发病规律的研究[J].福建林学院学报,2006,26(4):339-343.
[15] 冯丽贞,刘玉宝,郭素枝,等.桉树叶片的解剖结构与其对焦枯病抗性的关系[J].电子显微学报,2008,27(3):229-234.
[16] JIN M S, OLDHAM M L, ZHANG Q, et al. Crystal structure of the multidrug transporter P-glycoprotein from Caenorhabditis elegans[J]. Nature, 2012,490,7421:566-569.
[17] LI J, JAIMES K F, ALLER S G. Refined structures of mouse P-glycoprotein[J]. Protein Science, 2014,23(1):34-46.
[18] ESSER L, ZHOU F, PLUCHINO K M, et al. Structures of the multidrug transporter P-glycoprotein reveal asymmetric ATP binding and the mechanism of polyspecificity[J]. Journal of Biological Chemistry, 2017,292(2):446-461.
[19] ALLER S G. Structure of P-Glycoprotein reveals a molecular basis for poly-specific drug binding[J]. Biophysical Journal, 2010,98(3):755.
[20] GUPTA A, CHATTOO B B. Functional analysis of a novel ABC transporter ABC4 from Magnaporthe grisea[J]. FEMS Microbiology Letter, 2008,278(1):22-28.
[21] KIM Y, PARK S, KIM D, et al. Genome-scale analysis of ABC transporter genes and characterization of the ABCC type transporter genes in Magnaporthe oryzae[J]. Genomics, 2013,101(6):354-361.
[22] ZWIERS L, ROOHPARVAR R, de WAARD M A. MgAtr7, a new type of ABC transporter from Mycosphaerella graminicola involved in iron homeostasis[J]. Fungal Genetics & Biology, 2007,44(9):853-863.
[23] 王金辉.粉红粘帚菌ABC转运蛋白超家族耐绿针假单孢杆菌代谢产物的基因表达模式[D].郑州:河南农业大学,2012.
[24] HULVEY J, JTP J, SANG H, et al. Overexpression of ShCYP51B and ShatrD in Sclerotinia homoeocarpa isolates exhibiting practical field resistance to a demethylation inhibitor fungicide[J]. Applied & Environmental Microbiology, 2012,78(18):6 674-6 682.
[25] STEFANATO F L, ABOU-MANSOUR E, BUCHALA A, et al. The ABC transporter BcatrB from Botrytis cinerea exports camalexin and is a virulence factor on Arabidopsis thaliana[J]. Plant journal, 2009,58(3):499-510.
[26] FLEISSNER A, SOPALLA C, WELTRING K. An ATP-binding cassette multidrug-resistance transporter is necessary for tolerance of Gibberella pulicaris to phytoalexins and virulence on potato tubers[J]. Molecular plant-microbe interactions, 2002,15(2):102.