石榴种皮总木质素含量及PgCOMT基因的克隆与表达.pdfVIP

热带亚热带植物学报　2015 ，23(1) ：65 ~ 73 Journal of Tropical and Subtropical Botany 石榴种皮总木质素含量及PgCOMT 基 因的克隆与 表达 * 张水明, 龚凌燕, 曹丹琴, 张永娟, 杨健 (安徽农业大学园艺学院，合肥 230036) 摘要： 为探讨石榴(Punica granatum L.)籽粒硬度与种皮总木质素含量 的相关性及种皮 COMT 基 因的表达方式 ，利用 Texture Analyser 质构仪和巯基乙酸法测定 6 个 品种 的成熟籽粒硬度及种皮总木质素含量 。结果表 明，6 个石榴品种 的籽粒硬度与种 皮总木质素含量呈正相关 ，相关系数为 0.9246。采用 RACE 技术从石榴种皮 中克隆得到 1 条长度为 1456 bp 的 PgCOMT 基 因 cDNA 序列(GenBank 登录号为 KJ713968) 。实时荧光定量 PCR 分析表 明， 在 ‘红玉石籽 ’、‘粉皮 ’、‘会理软籽 ’、‘蒙 PgCOMT 自甜 ’和 ‘突尼斯软籽 ’石榴种皮 中的相对表达量与石榴籽粒硬度较为一致 ；随着石榴种皮 的发育其表达量先下降后上升。这为 深入 了解石榴软籽性状 的产生机理奠定 了基础 。 关键词：石榴 ；籽粒硬度 ；木质素 ； 基 因；基 因表达 COMT doi: 10.11926/j .issn.1005–3395.2015.01.010 Total Lignin Content in Pomegranate Seed Coat and Cloning and Expression Analysis of PgCOMT Gene * ZHANG Shui-ming , GONG Ling-yan, CAO Dan-qin, ZHANG Yong-juan, YANG Jian (College of Horticulture, Anhui Agricultural University, Hefei 230036, China) Abstract: In order to understand the correlation between seed hardness and total lignin content in seed coat of pomegranate (Punica granatum), the seed hardness and total lignin content of six cultivars were measured by Texture Analyser and thioglycolic acid method. The results showed that the correlation coefficient was 0.9246 between seed hardness and total lignin content in seed coat. A full-length cDNA sequence of 1456 bp (GenBank accession No.: KJ713968) was cloned from pomegranate seed coat by RACE, named PgCOMT gene. Real- time quantitative PCR showed that the relative expression of PgCOMT in ‘Hongyushizi’, ‘Fenpi’, ‘Huiliruanzi’, ‘Mengzitian’ and ‘Tunisiruanzi’ seed coat had a similar trend with seed hardness. The relative expression of PgCOMT declined at first, and then rose with development stage of pomegranate seed coat the last stage. These lay a foundation for further study the mechanism of sof