苹果梨果实着色相关酶基因片段克隆及表达分析 - 吉林农业大学学报.pdfVIP

吉林农业大学学报 2012，05 Journal of Jilin Agricultural University E-mail:jlndxb@ 苹果梨果实着色相关酶基因片段克隆及表达分析* ** 孙百灵，曲柏宏 ，李 伟，王 旭 延边大学农学院，延吉 133002 摘 要：通过 RT–PCR 技术对苹果梨果实着色相关酶（PAL 、CHI 、UFGT ）基因片段进行了克隆，并且采用半定量 RT–PCR 技术进行了 3 种酶基因在不同发育阶段的表达分析。结果表明：苹果梨与其他植物 PAL 、CHI 、UFGT 的 相同氨基酸和相似氨基酸的百分比都约为 90% ，分别与鸭梨、砂梨、西洋梨氨基酸序列聚类关系最近；套袋抑制 PAL 、CHI 、UFGT 3 种酶基因的表达，去袋后，3 种酶基因的表达量明显增加。 关键词：苹果梨；PAL 基因；CHI 基因；UFGT 基因；半定量 RT–PCR ；表达分析 中图分类号：S661.2 文献标识码：A 文章编号：1000-5684(2012) 网络出版地址： /kcms/detail/22.1100.S1547.004.html Cloning and Expression Analysis of Gene Fragments of Related Fruit-Coloring Enzymes in Pingguoli SUN Bai-ling, QU Bai-hong, LI Wei, WANG Xu College of Agriculture, Yanbian University, Yanji 133002,China Abstract: In the study, fragments of genes coding related fruit-coloring enzymes(PAL, CHI and UFGT) in Pingguoli were cloned through RT–PCR, and their expressions in different developmental stages were analyzed using Semi-Quantitative RT–PCR. The results suggested that the percentages of the same amino acids (including the similar ones) that compose PAL, CHI and UFGT in Pingguoli and other plants were all about 90%, and amino acid sequence of Pingguoli showed closer clustering relationships with those of Pyrus bretschneideri Rehd, P. Pyrifolia Nakai and Pyrus communis respectively. Besides, bagging inhibited the expressions of PAL, CHI and UFGT genes, while the expression levels of these genes increased after the bag was s taken off. Keywords: Pingguoli; PAL gene; CHI gene; UFGT gene; semi-quantitative RT–PCR; expression analysis 果实的红色或紫色是花青素的颜色，通常以花 苹果梨是吉林省特产，因其抗寒，适应性强， 青苷的形式存在。花青苷在形成过程中伴随着多种 肉质酥脆多汁，个大，石细胞少，特别是阳面着红 酶活性的变化，其中苯丙氨酸解氨酶（phenylalanine 色，深受人们青睐。目前，除了杨长红等[5]在基于 ammonia lyase ，PAL ）、查尔酮异构酶（chalc