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当前位置: 首页 > 文章 > 低温胁迫下西瓜同源二倍体和三倍体甲基化及基因表达的差异分析 园艺学报 2014,41 (11) 2313-2322
Position: Home > Articles > MSAP and Differential Expression of Homologous Diploid and Triploid Watermelon Under Cold Stress Acta Horticulturae Sinica 2014,41 (11) 2313-2322

低温胁迫下西瓜同源二倍体和三倍体甲基化及基因表达的差异分析

作  者：
杨炳艳;霍秀爱;刘云婷;段会军
单  位：
华北作物种质资源研究与利用教育部重点实验室;河北农业大学农学院
关键词：
西瓜;低温;MSAP;cDNA-AFLP;基因表达
摘  要：
为了揭示西瓜二倍体及三倍体在低温胁迫下的耐冷性分子机制,以西瓜二倍体自交系‘83166’(‘京欣1号’品种的母本)及其同源三倍体为材料,利用MSAP及c DNA-AFLP技术研究低温处理前后基因表达的差异,并对差异带进行克隆、测序和比对。28对MSAP引物扩增得到2 356个位点,其中二倍体经低温处理后总甲基化率下降8.5%,三倍体下降10.4%。将13条差异带与西瓜基因组数据库比对,有7条带确定是西瓜基因组序列。26对c DNA-AFLP引物组合扩增出1 267条带,其中二倍体上调表达占48.2%,下调表达占51.8%;三倍体上调表达占51.3%,下调表达占48.7%。21条差异带在NCBI中找到了同源序列基因,包括假定蛋白(38.1%)、能量与代谢(38.1%)、信号转导(9.5%)、物质运输(9.5%)以及DNA修饰(4.8%),另有9条无同源序列。低温胁迫后,三倍体去甲基化率与上调表达量均大于二倍体,而且三倍体诱导出更多的差异基因参与了能量代谢调控、信号转导、物质运输等过程,说明在抵御低温胁迫中三倍体比二倍体有更强的耐冷性。
译  名：
MSAP and Differential Expression of Homologous Diploid and Triploid Watermelon Under Cold Stress
作  者：
YANG Bing-yan;HUO Xiu-ai;LIU Yun-ting;DUAN Hui-jun;College of Agronomy,Agricultural University of Hebei,North China Key Laboratory for Crop Germplasm Resources of Education Ministry;
关键词：
watermelon;;cold stress;;MSAP;;cDNA-AFLP;;gene expression
摘  要：
The purpose of this study focused on molecular mechanism in low temperature of watermelon seeding,making homologous diploid and triploid of watermelon of female precocious Jingxin as materials,using MSAP and cDNA-AFLP analysis of genes differentially expressed before and after low temperature stress,and selected different bands to be cloned,sequenced and compared. A total of 2 356 locus were amplied after using 28 MSAP primer combinations. Levels of decline in the total methylation were 8.5% and 10.4% of the diploid and the triploid of watermelon,respectively. Took 13 different bands with watermelon genome database,identified 7 bands as watermelon genome sequence. After being screened with 26 cDNA-AFLP primer combinations,1 267 bands were amplied:48.2% with up-regulated expression,51.8% with down-regulated expression in the diploid of watermelon;51.3% with up-regulated expression,48.7% with down-regulated expression in the triploid of watermelon. NCBI Blast search showed that 21 bands were homologous to functional genes,including hypothetical protein(38.1%),energy and metabolism protein(38.1%),signal transduction protein(9.5%),material transportation protein(9.5%),and DNA modification gene(4.8%),the function of the other 9 bands remained unknown. Under low temperature stress,the demethylation rate and up-regulated genes of the triploid of watermelon were greater than the diploid of watermelon,the triploid of watermelon induced more genes involved in energy and metabolism,signal transduction and transportation process,illustrating that the triploid of watermelon had stronger resistance to low temperature than the diploid of watermelon.

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