西南农业学报2025，Vol.38Issue(8)：1637-1643,7.DOI:10.16213/j.cnki.scjas.2025.8.008
猕猴桃AcSPS1基因克隆及其在番茄中的过表达分析
Cloning of AcSPS1 gene from Actinidia chinensis and analysis of its over-expression in tomato
摘要
Abstract
[Objective]In the present study,the gene characteristics and role in fruit development of SPS in Actinidia chinensis were investiga-ted to provide a theoretical basis for improvement of kiwifruit quality.[Method]A.chinensis was used as the study material,and the highly ex-pressed AcSPS1 gene in kiwifruit was identified by bioinformatics methods.We conducted cloning,bioinformatics analysis,subcellular locali-zation and genetic transformation of AcSPS1 to obtain heterologous over-expression tomato lines.[Result]The full-length sequence of AcSPS1 was cloned from'Qihong'kiwifruit.The total length of the gene sequence was 3153 bp,encoding 1050 amino acids with a molecular weight of 117.98 kD and the Grand average of hydropathicity of-0.424.The analysis of protein secondary structure prediction showed that the pro-portion of Alpha helix was 41.52％,the proportion of Beta turn was 6.48％,the proportion of extended strand was 14.19％,and the propor-tion of random coil was 37.81％.The multiple alignment analysis of amino acid sequences revealed that the similarity between the amino acid sequences of AcSPS1 and CsSPS1 was the highest,reaching 90.39％.The phylogenetic tree analysis indicated that AcSPS1 and CsSPS1 had a close evolutionary relationship.Subcellular localization of Arabidopsis protoplasts showed that AcSPS1 was localized in the cytoplasm.Ac-SPS1 over-expression vector was constructed and heterogeneously expressed in Micro-Tom tomato,and 3 transgenic lines were obtained by PCR and western-blotting detection.Compared with wild-type tomatoes,the relative expression of AcSPS1 gene in the fruits of the three trans-genic lines was significantly increased.[Conclusion]The highly expressed AcSPS1 gene in kiwifruit is identified and cloned,with the protein located in the cytoplasm.Heterologous over-expressed tomato materials are successfully obtained,providing a reference for further exploration of its function in the biosynthesis of sucrose in kiwifruit fruits.关键词
猕猴桃/蔗糖磷酸合酶/基因克隆/番茄过表达分析Key words
Actinidia chinensis/Sucrose phosphate synthase/Gene cloning/Tomato over-expression analysis分类
农业科技引用本文复制引用
陈成,王依,刘凤丽,万春雁,钱亚明,狄华涛,霍恒志,阎永齐..猕猴桃AcSPS1基因克隆及其在番茄中的过表达分析[J].西南农业学报,2025,38(8):1637-1643,7.基金项目
江苏现代农业产业技术体系(JATS[2023]407) （JATS[2023]407）
江苏省农业科技自主创新项目[CX(23)2002] （23）
江苏省亚夫科技服务项目[KF(23)1203] （23）
