《植物生理学报》 2017, 53(8): 1507-1514

红肉苹果果实发育过程中花青苷含量变化及其合成相关基因表达分析

孙晓红^1,2,3, 刘源霞², 孙欣^2,4, 柏素花^2,3, 侯鸿敏², 张玉刚², 祝军², 戴洪义^2,*

¹湖南农业大学园艺园林学院, 长沙410128; ²青岛农业大学园艺学院, 青岛市园艺植物遗传改良与育种重点实验室, 山东青岛266109; ³青岛农业大学生命科学学院, 山东省高校植物生物技术重点实验室, 山东青岛266109; ⁴南京农业大学园艺学院, 南京210095

通信作者：戴洪义；E-mail: hydai@qau.edu.cn

摘　要：

本试验以白肉苹果(Malus pumila) ‘澳洲青苹’为对照, 研究了4个红肉苹果(M. sieversii f. neidzwetzkyana)品种(系)贵妃、‘红勋1号’、红勋5号、新疆1号在果实发育5个时期果肉花青苷含量, 利用实时荧光定量PCR检测了花青苷合成相关结构基因和转录因子的表达水平。结果表明: (1)红肉苹果果肉中的花青苷含量在花后30 d最高, 随后下降, 红勋5号和新疆1号在花后120和150 d又有明显提高。(2)苹果花青苷合成的酶基因CHS、F3H和DFR在所有品种(系)果实发育的各个时期相对表达量相差不大; ANS基因在新疆1号和红勋5号的前3个发育时期表达量是‘澳洲青苹’的约9倍至约18倍, 在后两个发育时期表达量降低; UFGT基因在‘红勋1号’、新疆1号和红勋5号果肉发育的各个时期中表达量均较高; MYB10转录因子在新疆1号和红勋5号盛花后30和60 d时的表达量为‘澳洲青苹’的约70倍至约90倍; TTG1、bHLH33在新疆1号和红勋5号中相对表达量高于bHLH3。(3)贵妃和‘红勋1号’ MYB10启动子基因型为R1R6, 红勋5号和新疆1号启动子基因型为R6R6, ‘澳洲青苹’启动子基因型为R1R1。研究结果为红肉苹果分子育种提供理论参考。

关键词：红肉苹果; 花青苷; 基因表达

收稿：2017-03-10   修定：2017-06-14

资助：山东省良种产业化工程项目(620902)、青岛市民生科技计划项目(15-9-2-99-nsh)、国家现代苹果产业技术体系项目(CARS-28)、国家自然科学基金(31372032)和山东省优秀中青年科学家科研奖励基金(BS2014SW027)。

Changes in anthocyanin content and analysis of relative gene expression during fruit development of Malus sieversii f. neidzwetzkyana

SUN Xiao-Hong^1,2,3, LIU Yuan-Xia², SUN Xin^2,4, BAI Su-Hua^2,3, HOU Hong-Min², ZHANG Yu-Gang², ZHU Jun², DAI Hong-Yi^2,*

¹Horticulture & Landscape College, Hunan Agricultural University, Changsha 410128, China; ²College of Horticulture / Key Laboratory for Genetic Improvement and Breeding of Horticultural Plant of Qingdao City, Qingdao Agricultural University, Qingdao, Shandong 266109, China; ³College of Life Sciences / Key Laboratory of Plant Biotechnology of Shandong Province, Qingdao Agricultural University, Qingdao, Shandong 266109, China; ⁴ College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China

Corresponding author: DAI Hong-Yi; E-mail: hydai@qau.edu.cn

Abstract:

In order to understand the mechanism of anthocyanin biosynthesis and speed up the breeding of red flesh apple (Malus sieversii f. neidzwetzkyana), changes in anthocyanin content during the development of fruits of red flesh apples, Guifei, ‘Hongxun No. 1’, Hongxun No. 5, Xinjiang No. 1, and white flesh apple (M. pumila), ‘Granny Smith’, were studied. Expressions of genes related to anthocyanin synthesis were analyzed by quantitative real time PCR. The result show (1) the highest anthocyanin contents of red flesh apples were found on 30 d after full bloom (DAFB), then the contents decreased, and the contents of Hongxun No. 5 and Xinjiang No. 1 increased on 120 and 150 DAFB. (2) The relative expression levels of CHS, F3H and DFR in different varieties (strains) during different developmental periods were not significantly different; the ANS in Hongxun No. 5 and Xinjiang No. 1 had maximal values during the first three developmental periods, which was around 9~18 times as high as that in ‘Granny Smith’. The expression level of ANS decreased rapidly during latter periods. The expressions of UFGT in ‘Hongxun No. 1’, Xinjiang No. 1 and Hongxun No. 5 were higher. In Xinjiang No. 1 and Hongxun No. 5, MYB10 transcription factor showed higher expression on 30 and 60 DAFB, respectively, which was around 70~90 times as high as that in ‘Granny Smith’. The expressions of TTG1 and bHLH33 were higher than that of bHLH3 in Xinjiang No. 1 and Hongxun No. 5. (3) The genotype of MYB10 promoter of Guifei and ‘Hongxun No. 1’ was R1R6, and that of Hongxun No. 5 and Xinjiang No. 1 was R6R6. The genotype of promoter in ‘Granny Smith’ was R1R1. These results would provide a theoretical basis for the breeding of red flesh apple.

Key words: Malus sieversii f. neidzwetzkyana; anthocyanin; gene expression