摘要
以不同发育阶段的‘糯米糍’荔枝果皮和成熟时不同色泽表型的‘糯米糍’(红色)、‘乌皮荔’(紫色)和‘鸭姆笼’(绿色)荔枝果皮为试材,以甘油醛-3-磷酸脱氢酶(gapdh)基因做内标、类黄酮-3-O-葡糖基转移酶(ufgt)基因为目的基因,建立了基于SYBR Green I染料技术的Real-Time PCR检测体系,并分析ufgt基因在上述材料中的表达差异。结果表明:以cDNA为模板建立的标准曲线循环阈值(Ct)与标准cDNA模板在一定浓度范围内呈良好的线性关系,gapdh基因和ufgt基因标准曲线的相关系数分别为0.998和0.999,扩增效率分别为96.2%和95.3%,达到定量PCR分析的标准。利用该方法的分析结果表明,不同发育阶段的‘糯米糍’荔枝果皮和3个成熟时不同色泽品种荔枝果皮的ufgt基因表达有显著差异。
In this study,an efficient Real-Time PCR assay was developed for detection of UFGT gene expression difference using pericarp from different developmental stage of litchi cultivars ‘ Nuomici＇ and different colour phenotypes-‘Nuomici＇（red）,‘Wupili＇ （purple） and ‘Yamulong＇ （green） as materials and GAPDH gene as internal control gene based on SYBR Green I dying technique.The results showed that threshold cycle （Ct） of the standard curve displayed good linear relationship with the concentrations of standard cDNA in a certain range.The correlation coefficients of GAPDH and UFGT genes were 0.998 and 0.999 respectively,and amplification efficiency were 96.2% and 95.3% respectively.Significant difference were observed in UFGT expression in different developmental stages of ＇Nuomici＇ pericarp and three different litchi cultivars with different color phenotypes using this established Real-Time PCR protocol.The method could provide a sensitive and rapid technique to analyze anthocyanin biosynthetic genes in litchi.
出处
《热带作物学报》
CSCD
2010年第8期1253-1259,共7页
Chinese Journal of Tropical Crops
基金
国家自然科学基金项目"荔枝果皮花色素苷生物合成的分子机理及其调控"(No.30971985)
国家科技支撑计划项目(No.2006BAD01A1705)
现代农业产业技术体系建设专项资金(No.nycytx-32-01)资助
