高、低糖杧果品种MiFRK基因克隆、表达及生物信息学分析  

Cloning,expression and bioinformatic analysis of MiFRK genes in mango varieties with high or low sugar content

作　　者：郑斌[1] 王松标[1] 李新月 武红霞[1] 马小卫[1] 梁清志[1] 许文天[1] 李丽[1] ZHENG Bin;WANG Songbiao;LI Xinyue;WU Hongxia;MA Xiaowei;LIANG Qingzhi;XU Wentian;LI Li(Key Laboratory of Tropical Fruit Biology,Ministry of Agriculture and Rural Affairs/South Subtropical Crops Research Institute,Chinese Academy of Tropical Agricultural Sciences/Key Laboratory of Hainan Province for Postharvest Physiology and Technology of Tropical Horticulture Product,National Field Genebank for Tropical Fruit,Zhanjiang 524091,Guangdong,China;College of Tropical Crop Science,Yunnan Agricultural University,Puer 665099,Yunnan,China)

机构地区：[1]农业农村部热带果树生物学重点实验室·中国热带农业科学院南亚热带作物研究所·国家热带果树种质资源圃海南省热带园艺采后处理与保鲜重点实验室,广东湛江524091 [2]云南农业大学热带作物学院,云南普洱665099

出　　处：《果树学报》2022年第6期945-956,共12页Journal of Fruit Science

基　　金：国家自然科学基金青年科学基金项目(31901968);海南省自然科学基金项目(322MS117,2019CXTD410,319QN287);国家重点研发计划项目(2018YFD1000504,2019YFD1000504,2019YFD1000500);广东省现代农业产业技术体系创新团队建设专项资金(2019KJ108)。

摘　　要：【目的】了解MiFRK基因在高糖和低糖杧果品种中的差异。【方法】对在高糖和低糖杧果品种转录组数据中表达差异显著的MiFRK1和MiFRK2基因DNA序列和cDNA序列进行克隆,再进行一系列生物信息学分析,并利用qRT-PCR检测2个MiFRK基因在品种间及不同果实发育阶段的表达差异。【结果】MiFRK1基因序列在2个品种间无差异,MiFRK2差异位点为69个,所编码的氨基酸相似率为97.87%;MiFRK1和MiFRK2均含有PfkB家族保守结构域,均为稳定的、不含跨膜结构的疏水性蛋白;MiFRK1在低糖品种不同果实发育期相对表达量均高于高糖品种;MiFRK2在低糖品种幼果期的相对表达量显著高于高糖品种,且在后熟过程中降低,而在高糖品种后熟过程中升高。【结论】MiFRK1和MiFRK2均在2个杧果品种果糖含量差异调控中起一定作用。【Objective】Sweetness is one of the key factors for mango fruit quality.In our previous study,we found that the soluble sugar contents of Tainong No.1(high-sugar-content variety)and Renong No.1(low-sugar-content variety)had significant differences at different fruit development stages,in which the difference of fructose content was the largest.Moreover,it is well known that fructokinase(FRK),as a key gene of the sugar metabolism pathways,plays an important role in fructose phosphorylation.We also screened out four MiFRK genes from mango genomic data,and there was a significant difference in gene expression levels of the MiFRK1 and MiFRK2 between Tainong No.1 and Renong No.1 based on transcriptome data,suggesting that the MiFRK1 and MiFRK2 would be one of the key genes for mango sugar metabolism.This study intended to further understand the differences of the MiFRK1 and MiFRK2 and their encoded proteins between high-sugar-content mango variety Tainong No.1 and low-sugar-content mango variety Renong No.1.【Methods】Two mango varieties Tainong No.1 and Renong No.1 were selected as materials.Firstly,the DNA sequences of the MiFRK1 and MiFRK2 genes in two varieties were cloned respectively and their structural differences between the two varieties were analyzed.The cDNA sequences of the two MiFRK genes were further cloned from two varieties,and the physicochemical properties,structural domain,subcellular localization and phylogenetic trees of their encoded proteins were analyzed by bioinformatic software.Finally,the fruits of each variety in four stages were selected as samples,namely,young fruit stage(about 40 days after anthesis),fruit expansion stage,green maturity stage(harvest maturity stage)and full maturity stage(edible maturity stage,one week after harvest).The gene expression patterns of the two genes in fruits of two varieties at different fruit stages were detected by quantitative real-time PCR(qRT-PCR).【Results】The results showed that there was no difference in the DNA sequences of the Mi FRK1 bet

关 键 词：杧果 果糖激酶 糖代谢 生物信息学分析 

分 类 号：S667.7[农业科学—果树学]