摘要
【目的】STOP1是植物耐铝毒的重要基因,为探明葡萄STOP1基因的结构特征及其表达量与其耐铝性的关系。【方法】以耐铝性弱的山葡萄(Vitis amurensis)和耐铝性强的小叶葡萄(V.sinocinerea)的叶片cDNA为模板,采用PCR和TA克隆技术获得VaSTOP1和VsSTOP1两基因序列,对其进行生物信息学分析和铝胁迫下的表达分析,并测定铝胁迫下的膜脂过氧化指标,对2个种STOP1基因的表达和膜脂过氧化物质的变化与耐铝性的关系进行比较。【结果】克隆获得的VaSTOP1和VsSTOP1基因序列长度分别为1782和1800 bp,分别编码594和600个氨基酸,相对分子量分别为67.03和67.48 kD,理论等电点为5.21和6.42;二者的二级结构中均有无规则卷曲>α-螺旋>折叠延伸链的规律,由α-螺旋、β-折叠片、β-转角和锌指结构等组成三级空间结构;进化树研究得出,VaSTOP1与VsSTOP1的同源性高达96%,二者与CcSTOP1、PnSTOP1、NtSTOP1、ErSTOP1的同源性均在80%以上。VaSTOP1基因表达量在铝胁迫28 d内呈先上升后下降的规律,较早出现峰值,而VsSTOP1基因表达量则稳定上升,这与二者的膜脂过氧化物质的变化规律基本一致。【结论】葡萄的耐铝性与其STOP1基因的表达量呈正相关,研究为进一步探明葡萄耐铝毒的分子调控机制提供了基因资源。
【Objective】STOP1 is an important gene for tolerance to aluminum(Al)-toxicity in plants.The purpose of the present study was to explore the relationship between the structural characteristics,expression level of STOP1 gene and its tolerance to Al in grape.【Method】Leaf cDNA of Vitis amurensis with weak tolerance to Al and V.sinocinerea with strong tolerance to Al were used as reference materials.Sequences of two genes VaSTOP1 and VsSTOP1 were obtained by PCR and TA cloning technology,and then bioinformatics analysis and expression analysis under Al stress were carried out.The membrane lipid peroxidation indices under Al stress were determined.Finally,the relationship between the expression of STOP1 genes,the changes of membrane lipid peroxidation of two species and tolerance to Al were compared.【Result】The sequences of VaSTOP1 and VsSTOP1 were 1782 and 1800 bp respectively,encoding 594 and 600 amino acids.Their relative molecular weights were 67.03 and 67.48 kD,and the theoretical isoelectric points were 5.21 and 6.42,respectively.The secondary structure in two species were the law of irregular curl>α-helix>folding extension chain,and the tertiary spatial structure were composed of zinc protein,α-helix,β-folded sheet andβ-rotation angle.The phylogenetic tree analysis found that the homology between VaSTOP1 and VsSTOP1 was as high as 96%,and the homology between them and CcSTOP1,PnSTOP1,NtSTOP1 and ErSTOP1 was more than 80%.The expression of VaSTOP1 gene increased first and then decreased within 28 days under Al stress,but the peak value appeared earlier,whereas the VsSTOP1 gene expression increased steadily,which was basically consistent with the change law of membrane lipid peroxidation.【Conclusion】The tolerance to Al of Vitis was positively correlated with the expression of STOP1 gene,which provided a reference for further exploring the molecular regulation mechanism of Vitis tolerance to Al.
作者
张永福
徐仕琴
陈姣
杨砚斌
任禛
ZHANG Yong-fu;XU Shi-qin;CHEN Jiao;YANG Yan-bin;REN Zhen(School of Agriculture and Life Sciences,Kunming University,Kunming 650214,China)
出处
《西南农业学报》
CSCD
北大核心
2022年第3期588-595,共8页
Southwest China Journal of Agricultural Sciences
基金
国家自然科学基金项目(32060645)
国家自然科学基金项目(31660559)
云南省高校高原特色果树优异种质资源挖掘与新品种选育科技创新团队建设项目
云南省地方本科高校基础研究联合专项资金重点项目(202101BA070001-036)。
关键词
葡萄
STOP1基因
序列分析
基因表达
铝毒
Vitis
STOP1 gene
Sequence analysis
Gene expression
Aluminum toxicity
