马铃薯StSRP1的克隆、表达及生物信息学分析
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摘要
胁迫相关基因的表达导致代谢物的积累和生化与生理途径的改变,对于植物适应不利胁迫条件至关重要。通过研究马铃薯StSRP1的结构和在逆境胁迫中的表达情况,为验证马铃薯StSRP1在抵抗生物胁迫过程中的作用奠定了基础。运用电子克隆法获得StSRP1全长序列,利用生物信息学分析StSRP1的核酸序列、氨基酸序列、蛋白质结构和启动子,基于NCBI数据库,选取20条与StSRP1匹配度高的胁迫相关蛋白序列构建了系统进化树,对不同物种间的SRP蛋白进行比对。经ABA、MJ和病菌的诱导,对StSRP1进行qRT-PCR分析。克隆获得StSRP1全长867 bp,可编码288个氨基酸,无信号肽序列,无跨膜区,有34个可能的磷酸化位点,亲水性蛋白,定位于内质网。StSRP1的表达受ABA、MJ和病菌的调控。StSRP1可能参与马铃薯逆境胁迫应答过程
The expression of stress-related genes(StSRP1)leads to the accumulation of metabolites and changes in biochemical and physiological pathways,which are essential for plants to adapt to adverse stress conditions. By studying the structure of potato StSRP1 and its expression in stress,the foundation for verifying the role of potato StSRP1 in the process of resistance to biological stress is laid. Electronic cloning was used to clone the full-length sequence of StSRP1 gene,bioinformatics was to analyze the nucleic acid sequence,amino acid sequence,protein structure and promoter of StSRP1. Based on the NCBI database,20 strains of stress-related protein sequences highly matched with StSRP1 were selected to construct a phylogenetic tree,and SRP proteins among different species were compared. StSRP1 induced by ABA,MJ and pathogen was analyzed by qRT-PCR. The full length of the cloned sequence was 867 bp,encoding 288 amino acids,no signal peptide sequence,no transmembrane region,34 possible phosphorylation sites,hydrophilic protein,and localized to the endoplasmic reticulum. In sum,the expression of StSRP1 is regulated by ABA,MJ and pathogens. StSRP1 may be involved in the potato stress response process.
The expression of stress-related genes(StSRP1)leads to the accumulation of metabolites and changes in biochemical and physiological pathways,which are essential for plants to adapt to adverse stress conditions. By studying the structure of potato StSRP1 and its expression in stress,the foundation for verifying the role of potato StSRP1 in the process of resistance to biological stress is laid. Electronic cloning was used to clone the full-length sequence of StSRP1 gene,bioinformatics was to analyze the nucleic acid sequence,amino acid sequence,protein structure and promoter of StSRP1. Based on the NCBI database,20 strains of stress-related protein sequences highly matched with StSRP1 were selected to construct a phylogenetic tree,and SRP proteins among different species were compared. StSRP1 induced by ABA,MJ and pathogen was analyzed by qRT-PCR. The full length of the cloned sequence was 867 bp,encoding 288 amino acids,no signal peptide sequence,no transmembrane region,34 possible phosphorylation sites,hydrophilic protein,and localized to the endoplasmic reticulum. In sum,the expression of StSRP1 is regulated by ABA,MJ and pathogens. StSRP1 may be involved in the potato stress response process.
引文
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[8]Udawat P, Jha RK, Sinha D, et al. Overexpression of a cytosolic abiotic stress responsive universal stress protein(SbUSP)mitigates salt and osmotic stress in transgenic tobacco plants[J]. Front Plant Sci, 2016, 7(14):518.
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[12]Cuming AC. LEA proteins[J]. Seed Proteins, 1999:753-780.
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[14]He LY. Characteristics of strains of pseudomonas solanacearum from china[J]. Plant Disease, 1983, 67(12):1357-1361.
[15]Li Y, Jia F, Yu Y, et al. The SCF E3 ligase AtPP2-B11 plays a negative role in response to drought stress in Arabidopsis[J].Plant Molecular Biology Reporter, 2014, 32(5):943-956.
[16]ParkKY,KimWT,KimEY.Theproperlocalizationof RESPONSIVE TO DESICCATION 20 in lipid droplets depends on their biogenesis induced by STRESS-RELATED PROTEINS in vegetative tissues[J]. Biochemical and Biophysical Research Communications, 2017, 495(2):1885-1889.
[17]Krishnan HB, Natarajan SS, Oehrle NW, et al. Proteomic analysis of Pigeonpea(Cajanus cajan)seeds reveals the accumulation of numerous stress-related proteins[J]. Journal of Agricultural and Food Chemistry, 2017, 65(23):4572-4581.
[18]Cascardo JC, Almeida RS, Buzeli RA, et al. The phosphorylation state and expression of soybean BiP isoforms are differentially regulated following abiotic stresses[J]. J Biol Chem, 2000, 275(19):14494-14500.
[19]Brown D, Feeney M, Ahmadi M, et al. Subcellular localization and interactions among rubber particle proteins from Hevea brasiliensis[J]. Journal of Experimental Botany, 2017, 68(18):5045-5055.
[20]Udawat P, Jha RK, Mishra A, et al. Overexpression of a plasma membrane-localized SbSRP-like protein enhances salinity and osmotic stress tolerance in transgenic tobacco[J]. Front Plant Sci, 2017, 8:582.
[21]Qi XT, Cui QH, Luo Y, et al. Zn stress-induced inhibition of bean PvSR2-GUS fusion gene splicing is gene-specific in transgenic tobacco[J]. Journal of Plant Physiology, 2009, 11(166):1223-1227.
[22]Takashi H, Kazuo S. Research on plant abiotic stress responses in the post-genome era:past, present and future[J]. Plant J, 2010,61(6):1041-1052.
[2]Dennis MS, Light DR. Rubber elongation factor from Hevea brasiliensis. identification, characterization, and role in rubber biosynthesis[J]. J Biol Chem, 1989, 264(31):18608-18617.
[3]Kim EY, Park KY, Seo YS, et al. Arabidopsis small rubber particle protein homolog SRPs play dual roles as positive factors for tissue growth and development and in drought stress responses[J]. Plant Physiology, 2016, 170(4):2494-2510.
[4]Jing D, Zhang J, Xia Y, et al. Proteomic analysis of stress-related proteins and metabolic pathways in Picea asperata somatic embryos during partial desiccation[J]. Plant Biotechnology Journal, 2017,15(1):27-38.
[5]Sano N, Masaki S, Tanabata T, et al. Proteomic analysis of stressrelated proteins in rice seeds during the desiccation phase of grain filling[J]. Plant Biotechnology, 2013, 30(2):147-156.
[6]Hong JP, Kim WT. Isolation and functional characterization of the Ca-DREBLP1 gene encoding a dehydration-responsive element binding-factor-like protein 1 in hot pepper(Capsicum annuum L.cv. Pukang)[J]. Planta, 2005, 220(6):875-888.
[7]Eun Yu K, Young Sam S, Hanna L, et al. Constitutive expression of CaSRP1, a hot pepper small rubber particle protein homolog,resulted in fast growth and improved drought tolerance in transgenic Arabidopsis plants[J]. Planta, 2010, 232(1):71-83.
[8]Udawat P, Jha RK, Sinha D, et al. Overexpression of a cytosolic abiotic stress responsive universal stress protein(SbUSP)mitigates salt and osmotic stress in transgenic tobacco plants[J]. Front Plant Sci, 2016, 7(14):518.
[9]Sarkar NK, Kim YK, Grover A. Rice sHsp genes:genomic organization and expression profiling under stress and development[J].BMC Genomics, 2009, 10(1):393-411.
[10]Gary F, Janae W, Williams JJ, et al. Effect of mutation of the tetratricopeptide repeat and asparatate-proline 2 domains of Sti1 on Hsp90 signaling and interaction in Saccharomyces cerevisiae[J].Genetics, 2006, 172(1):41-51.
[11]Mittler R. Oxidative stress, antioxidants and stress tolerance[J].Trends Plant Sci, 2002, 7(9):405-410.
[12]Cuming AC. LEA proteins[J]. Seed Proteins, 1999:753-780.
[13]Ni XM, Tian ZD, Liu J, et al. StPUB17, a novel potato UND/PUB/ARM repeat type gene, is associated with late blight resistance and NaCl stress[J]. Plant Science, 2010, 178(2):158-169.
[14]He LY. Characteristics of strains of pseudomonas solanacearum from china[J]. Plant Disease, 1983, 67(12):1357-1361.
[15]Li Y, Jia F, Yu Y, et al. The SCF E3 ligase AtPP2-B11 plays a negative role in response to drought stress in Arabidopsis[J].Plant Molecular Biology Reporter, 2014, 32(5):943-956.
[16]ParkKY,KimWT,KimEY.Theproperlocalizationof RESPONSIVE TO DESICCATION 20 in lipid droplets depends on their biogenesis induced by STRESS-RELATED PROTEINS in vegetative tissues[J]. Biochemical and Biophysical Research Communications, 2017, 495(2):1885-1889.
[17]Krishnan HB, Natarajan SS, Oehrle NW, et al. Proteomic analysis of Pigeonpea(Cajanus cajan)seeds reveals the accumulation of numerous stress-related proteins[J]. Journal of Agricultural and Food Chemistry, 2017, 65(23):4572-4581.
[18]Cascardo JC, Almeida RS, Buzeli RA, et al. The phosphorylation state and expression of soybean BiP isoforms are differentially regulated following abiotic stresses[J]. J Biol Chem, 2000, 275(19):14494-14500.
[19]Brown D, Feeney M, Ahmadi M, et al. Subcellular localization and interactions among rubber particle proteins from Hevea brasiliensis[J]. Journal of Experimental Botany, 2017, 68(18):5045-5055.
[20]Udawat P, Jha RK, Mishra A, et al. Overexpression of a plasma membrane-localized SbSRP-like protein enhances salinity and osmotic stress tolerance in transgenic tobacco[J]. Front Plant Sci, 2017, 8:582.
[21]Qi XT, Cui QH, Luo Y, et al. Zn stress-induced inhibition of bean PvSR2-GUS fusion gene splicing is gene-specific in transgenic tobacco[J]. Journal of Plant Physiology, 2009, 11(166):1223-1227.
[22]Takashi H, Kazuo S. Research on plant abiotic stress responses in the post-genome era:past, present and future[J]. Plant J, 2010,61(6):1041-1052.