‘砀山酥梨’及其褐皮芽变果皮中多胺类物质含量和相关基因表达分析
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国家现代农业产业技术体系建设专项(CARS-29-14);国家自然科学基金(31101519)


Polyamines Content,Cloning and Expression of the Relevant Genes in Exocarp of ‘Dangshansuli’ Pear and Its Russet Fruit Mutant
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    摘要:

    为研究多胺类物质在‘砀山酥梨’芽变品系‘锈酥’果皮褐色形成中的作用,以‘砀山酥梨’和‘锈酥’花后不同时期果皮为试材,采用HPLC方法测定果皮中多胺类物质的含量,利用RACE技术克隆多胺合成过程中的关键基因,通过Protparam网站与MEGA5.0软件分析相关基因蛋白的理化性质及其系统进化,用荧光定量qRT-PCR方法分析不同时期果皮中相关基因的相对表达量。结果表明:(1)除花后50 和150 d外,其它时期‘砀山酥梨’果皮中腐胺含量均显著高于‘锈酥’,花后50 d、75 d、150 d时,‘锈酥’果皮中的亚精胺、精胺含量显著高于‘砀山酥梨’果皮。(2)克隆出多胺合成的基因ADC、SPDS和SPMS(GenBank登录号分别为KM923903、KM923905和KM923906),预测ADC和SPMS均为水溶性蛋白、SPDS为疏水性蛋白,它们与苹果的亲缘关系最近。(3)荧光定量PCR结果显示,花后50 d,多胺合成中ADC、SPDS和SPMS在‘锈酥’果皮中表达量均显著高于‘砀山酥梨’。研究推测,‘锈酥’果皮中多胺代谢及相关基因的上调表达可能与‘锈酥’的果皮褐色形成有关。

    Abstract:

    To investigate the mechanism of russet skin formation in pear (Pyrus bretschneideri Rehd.),we used the exocarp of the wild type ‘Dangshansuli’ and its mutant type ‘Xiusu’ pear with russet fruits at different days after full bloom (DAFB) as material in this experiment.The content of polyamine was determined by High Performance Liquid Chromatography (HPLC).The key genes in polyamine biosynthesis were cloned by rapid amplification of cDNA ends (RACE),and their physicochemical properties were conducted by the protocol of Protparam website (http://web.expasy.org/protparam/) and the phylogenetic tree of putative proteins was constructed by MEGA5.0 software.The relative expressions of ADC,SPDS and SPMS genes at different stages were analyzed by using real-time quantitative reverse transcription-PCR (qRT-PCR).The results showed that:(1)The putrescine contents in the exocarp of ‘Dangshansuli’ was significantly higher than that in ‘Xiusu’ except for 50 and 150 DAFB.At 75 DAFB,the contents of spermidine and spermine in the exocarp of ‘Xiusu’ were higher than that in ‘Dangshansuli’.(2)The genes of arginine decarboxylase (ADC),spermidine synthase (SPDS) and spermine synthase (SPMS) were cloned,and were submitted to the database of the National Center for Biotechnology Information(NCBI) with the accession number KM923903,KM923905 and KM923906 ,respectively.The structure of putative proteins of ADC,SPDS and SPMS gene in pear was closer to that in apple.ADC and SPMS were all water-soluble by the prediction of their physicochemical properties.The SPDS was water-repellent by the prediction of their physicochemical properties.(3)The levels of relative expressions of ADC,SPDS and SPMS in the exocarp of ‘Xiusu’ were higher than that in ‘Dangshansuli’ at 50 DAFB.It was postulated that the metabolism of polyamine and up-regulation of relevant genes may be associated with the formation of russet fruit of ‘Xiusu’ pear.

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孙虹丽,王子腾,蒋向红,等.‘砀山酥梨’及其褐皮芽变果皮中多胺类物质含量和相关基因表达分析[J].西北植物学报,2015,35(8):1534-1540

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  • 在线发布日期: 2015-08-29
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