不同水稻品种OsVDAC3及3个互作蛋白基因OsV3IP1-3表达模式分析
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摘要
为研究OsVDAC3及互作蛋白基因OsV3IP1-3的表达模式,用生物信息学和定量PCR方法分析了水稻不育系‘粤泰A’(YTA)、保持系‘粤泰B’(YTB)和‘日本晴’(NIP)不同发育时期幼穗,四叶期根、鞘和叶中OsVDAC3和OsV3IP1-3的表达量。生物信息学分析表明在ATG上游含有21个与水稻生长发育及胁迫应答相关的顺式作用元件;RiceXPro和MSU Rice Genome Annotation Project水稻基因表达数据库分析结果表明在NIP中OsV3IP1只在幼苗表达,而OsVDAC3、OsV3IP2和OsV3IP3在整个生活周期都表达,但OsVDAC3在花药表达水平最高,OsV3IP2在生殖器官表达水平最高,OsV3IP3在嫩芽中表达量最高。定量PCR结果表明OsVDAC3和OsV3IP1-3基因在NIP、YTA和YTB 3个水稻品种中表达模式不同。OsVDAC3在YTB的1.5～4.0 cm幼穗表达水平非常高,OsV3IP1在3个品种的四叶期地上部都有非常高的表达,OsV3IP2和OsV3IP3在YTB和NIP中表达模式相似,而与YTA中差异明显。这些基因的表达模式与功能的关系有待进一步研究。
To study the expression patterns of OsVDAC3 and its interaction protein gene OsV3 IP1-3, the expression of OsVDAC3 and OsV3 IP1-3 in the inflorescences of different developmental stages and the root,sheath and leaf at the fourth-leaf stage from the rice sterile line 'Yuetai A'(YTA), the maintenance line'Yuetai B'(YTB) and 'Nipponbare'(NIP) were analyzed with bioinformatics and RT-PCR. The results of bioinformatics analysis showed that there were 21 Cis-action elements related with the growth and stress response of rice in the upstream of ATG. The analysis results of rice gene expression database from RiceXPro and MSU Rice Genome Annotation Project showed that OSV3 IP1 in Nipponbare was only expressed in the seedlings, but OsVDAC3, OSV3 IP2 and OSV3 IP3 were expressed throughout the life cycle, furthermore OsVDAC3 had the highest expression in anther, OSV3 IP2 had the highest expression in the reproductive organs, and OSV3 IP3 had the highest expression in the buds. The results of RT-PCR showed that the expression patterns of OsVDAC3 and OsV3 IP1-3 were different in NIP, YTA and YTB. The expression level of OsVDAC3 in young panicles(1.5-4.0 cm) of YTB was very high. OsV3 IP1 had very high expression at the fourth-leaf stage among the 3 cultivars,the expression patterns of OsV3 IP2 and OsV3 IP3 in YTB and NIP were similar, but were different from that in YTA. The relationship between the expression patterns and functions of these genes needs further study.
To study the expression patterns of OsVDAC3 and its interaction protein gene OsV3 IP1-3, the expression of OsVDAC3 and OsV3 IP1-3 in the inflorescences of different developmental stages and the root,sheath and leaf at the fourth-leaf stage from the rice sterile line 'Yuetai A'(YTA), the maintenance line'Yuetai B'(YTB) and 'Nipponbare'(NIP) were analyzed with bioinformatics and RT-PCR. The results of bioinformatics analysis showed that there were 21 Cis-action elements related with the growth and stress response of rice in the upstream of ATG. The analysis results of rice gene expression database from RiceXPro and MSU Rice Genome Annotation Project showed that OSV3 IP1 in Nipponbare was only expressed in the seedlings, but OsVDAC3, OSV3 IP2 and OSV3 IP3 were expressed throughout the life cycle, furthermore OsVDAC3 had the highest expression in anther, OSV3 IP2 had the highest expression in the reproductive organs, and OSV3 IP3 had the highest expression in the buds. The results of RT-PCR showed that the expression patterns of OsVDAC3 and OsV3 IP1-3 were different in NIP, YTA and YTB. The expression level of OsVDAC3 in young panicles(1.5-4.0 cm) of YTB was very high. OsV3 IP1 had very high expression at the fourth-leaf stage among the 3 cultivars,the expression patterns of OsV3 IP2 and OsV3 IP3 in YTB and NIP were similar, but were different from that in YTA. The relationship between the expression patterns and functions of these genes needs further study.
引文
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[18]胡颖.水稻基因OsVDAC3功能分析及互作蛋白的筛选[D].武汉:中南民族大学,2015.
[19]陈利维.水稻膜系统酵母双杂交cDNA文库及OsVDAC5诱饵蛋白载体的构建与鉴定[D].武汉:中南民族大学,2014.
[20]Sato Y,Takehisa H,Kamatsuki K,et al.RiceXPro version 3.0:expanding the informatics resource for rice transcriptome[J].Nucleic Acids Research,2013,41:1206-1213.
[21]Eisen M B,Spellman P T,Brown P O,et al.Cluster analysis and display of genome-wide expression patterns[J].Proc.Natl.Acad.Sci.U.S.A.,1998,95:4863-14868.
[2]McCommis K S,Baines C P.The Role of VDAC in Cell Death:Friend or Foe?[J].Biochimica et Biophysica Acta.,2012,1818(6):1444-1450.
[3]Shoshan-Barmatz V,De Pinto V,Zweckstetter M,et al.VDAC,a multi-functional mitochondrial protein regulating cell life and death[J].Molecular Aspects of Medicine,2010,31(3):227-285.
[4]Lee AC,Xu X,Blachly-Dyson E,et al.The role of yeast VDACgenes on the permeability of the mitochondrial outer membrane[J].Journal Membrane Biology,1998,161:173-181.
[5]Menard G,Evrard B,Bureau M,et al.Cerebral distribution of the B-36 VDAC protein in rat,cow and man brain:immunocytochemical study[J].Cell Molecular Biollogy,1994,40(3):295-300.
[6]Kusano T,Tateda C,Berberich T,et al.Voltage-dependent anion channels:their roles in plant defense and cell death[J].Plant Cell Rep.,2009,28:1301-1308.
[7]Tateda C,Watanabe K,Kusano T,et al.Molecular and genetic characterization of the gene family encoding the voltage-dependent anion channel in Arabidopsis[J].Journal Experimental Botany.2011,62:4773-4785.
[8]Al Bitar F,Roosens N,Smeyers M,et al.Sequence analysis,transcriptional and posttranscriptional regulation of the rice vdac family[J].Biochimica Biophysica Acta.2003,1625:43-51.
[9]Asakura T,Hirose S,Katamine H,et al.Isolation and proteomic analysis of rice Golgi membranes:cis-Golgi membranes labeled with GFP-SYP31[J].Plant Biotechnol,2006,23:475-485
[10]Godbole A,Mitra R,Dubey A K,et al.Bacterial expression,purifcation and characterization of a rice voltagedependent,anionselective channel isoform,OsVDAC4[J].Journal Membrane Biology,2011,244:67-80.
[11]Xu X,Tan Y P,Cheng G,et al.Genomic survey and gene expression analysis of the VDAC gene family in rice[J].Genetics and Molecular Research,2015,14(4):15683-15696.
[12]钟英健.4个水稻OsVDAC蛋白的亚细胞定位[D].武汉:中南民族大学,2015.
[13]Wandrey M,Trevaskis B,Brewin N,et al.Molecular and cell biology of a family of voltage-dependent anion channel porins in Lotus japonicus[J].Plant Physiology,2004,134:182-193.
[14]Wang J,Zhang L D,Zuo K J,et al.Cloning and expressional studies of the voltage-dependent anion channel gene from Brassica rapa L.[J].Journal of Integrative Plant Biology,2006,48:197-203.
[15]Desai M K,Mishra R N,Verma D,et al.Structural and functional analysis of a salt stress inducible gene encoding voltage dependent anion channel(VDAC)from pearl millet(Pennisetum glaucum)[J].Plant Physiology Biochemtry,2006,44:483-493.
[16]Tateda C,Yamashita K,Takahashi F,et al.Plant voltage-dependent anion channels are involved in host defense against Pseudomonas cichorii and in Bax-induced cell death[J].Plant Cell Report,2009,28:41-51.
[17]Elkeles A,Devos KM,Graur D,et al.Multiple c DNAs of wheat voltage-dependent anion channels(VDAC):isolation,differential expression,mapping and evolution[J].Plant Molecular Biollogy,1995,29:109-124.
[18]胡颖.水稻基因OsVDAC3功能分析及互作蛋白的筛选[D].武汉:中南民族大学,2015.
[19]陈利维.水稻膜系统酵母双杂交cDNA文库及OsVDAC5诱饵蛋白载体的构建与鉴定[D].武汉:中南民族大学,2014.
[20]Sato Y,Takehisa H,Kamatsuki K,et al.RiceXPro version 3.0:expanding the informatics resource for rice transcriptome[J].Nucleic Acids Research,2013,41:1206-1213.
[21]Eisen M B,Spellman P T,Brown P O,et al.Cluster analysis and display of genome-wide expression patterns[J].Proc.Natl.Acad.Sci.U.S.A.,1998,95:4863-14868.