葡萄MAPK基因家族鉴定及逆境胁迫下的表达分析

Identification of Grape MAPK Gene Family and Expression Analysis under Adversity Stress

探究葡萄MAPK家族成员的特性及潜在功能,为葡萄抗逆基因资源的挖掘和利用提供理论依据。本研究克隆获得葡萄MAPK基因家族成员并对其进行生物信息学分析,根据基因芯片数据分析各成员的组织表达特征,结合qRT-PCR明确其对非生物胁迫的响应情况。结果表明,葡萄中共获得43个MAPK成员,分别命名为VvMAPK1~VvMAPK43,分布于15条染色体上,其中有6个成员集中分布于18号染色体上,5个分布于5号染色体上。氨基酸大小为114~1520aa,VvMAPK15编码的氨基酸序列最长,为1520个氨基酸残基,VvMAPK23编码的氨基酸序列最短,为114个。相对分子量集中为12.26~16.70ku,等电点为4.94~10.01。芯片数据分析发现,经ABA处理后,VvMAPK5、VvMAPK7和VvAMAPK13的表达量明显下调,在盐、PEG和低温处理下,VvMAPK6、VvMAPK9和VvMAPK13的表达量明显上调;不同成员在不同组织以及同一组织的不同生长阶段表达水平差异显著。qRT-PCR分析发现,大多数VvMAPK家族成员受到200mmol·L^(-1) NaCl的诱导,表达水平较对照上调显著,VvMAPK9、VvMAPK27、VvMAPK29和VvMAPK38在500μmol·L^(-1) ABA、200mmol·L^(-1) NaCl和10%PEG处理后均上调表达显著,对3种非生物胁迫的响应强烈。研究结果表明,葡萄MAPK家族成员在不同非生物胁迫下存在不同的潜在功能,为葡萄基因改良和遗传育种提供理论基础。

The function and characteristics of grapevine MAPK transcription factor family genes were explored to provide theoretical basis for utilizing grapevine stress resistance gene resources.Bioinformatics was used to analyze the physical and chemical properties,system evolution,gene chip expression,cis-acting elements,motif and expression level under stress of grapevine MAPK gene family.The results showed that 43 MAPK family genes,named as VvMAPK1-VvMAPK43,distributed on 15 chromosomes,among which 6 members were concentrated on the 18th chromosome and 5 on the 5th chromosome.The amino acid size of grapevine MAPK transcription factor family was between 114-1520 aa,and VvMAPK15 had the longest amino acid sequence with 1520 amino acid residues while VvMAPK23 had the shortest amino acid sequence,only 114 amino acid residues.The relative molecular mass concentrated in 12.26-16.70 ku.Isoelectric point was concentrated between 4.94-10.01.qRT-PCR analysis showed that VvMAPK2,VvMAPK3,VvMAPK6 and VvMAPK7 showed a downward trend compared with the control.VvMAPK1,VvMAPK2,VvMAPK3,VvMAPK5,VvMAPK6 and VvMAPK7 were all down regulated after being treated at 500μmol·L^(-1) ABA,200 mmol·L^(-1) NaCl and 4℃.The microarray data showed that after ABA treatment,the expression of VvMAPK5,VvMAPK7 and VvMAPK13 genes were significantly down regulated,while under treatments of salt,PEG and low temperature,the expression of VvMAPK6,VvMAPK9 and VvMAPK13 was significantly up regulated.