桂花类胡萝卜素异构酶基因的克隆及表达分析

doi:10.13560/j.cnki.biotech.bull.1985.2016-1163

生物技术通报 ›› 2017, Vol. 33 ›› Issue (6): 89-96.doi: 10.13560/j.cnki.biotech.bull.1985.2016-1163

桂花类胡萝卜素异构酶基因的克隆及表达分析

张超, 刘玉成, 王艺光, 付建新, 赵宏波

浙江农林大学风景园林与建筑学院,临安 311300

收稿日期:2017-01-05 出版日期:2017-06-26 发布日期:2017-06-19
作者简介:张超,男,博士,研究方向：观赏植物分子生物学;E-mail：zhangchao24804@gmail.com
基金资助:
国家自然科学基金项目（31501790）,浙江省自然科学基金（LQ15C160004和LQ16C160003）,浙江农林大学科研发展基金人才启动项目（2016FR031）

Cloning and Expression Analysis of Carotenoid Isomerase Gene in Osmanthus fragrans

ZHANG Chao, LIU Yu-cheng, WANG Yi-guang, FU Jian-xin, ZHAO Hong-bo

School of Landscape Architecture,Zhejiang Agriculture and Forestry University,Lin’an 311300

Received:2017-01-05 Published:2017-06-26 Online:2017-06-19

摘要/Abstract

摘要： 利用已构建的桂花‘堰虹桂’花瓣转录组数据库中Unigene序列信息,结合RT-PCR技术对1个桂花类胡萝卜素异构酶基因（OfCRTISO）的最大阅读框进行扩增,并测序验证。生物信息学分析表明OfCRTISO基因含有长为1 842 bp的开放读码框,编码613个氨基酸残基。桂花OfCRTISO与已知胡麻科和茄科植物的CRTISO序列相似性最高,系统进化树分析发现OfCRTISO与芝麻CRTISO（XP_011077785）亲缘关系最近,聚为同一小支。表达分析发现,OfCRTISO基因在桂花‘堰虹桂’花蕾前期花序中的表达量极低,花蕾后期其表达量逐渐增加,初开期达到高峰,而盛开期时,其表达量显著下降。对桂花不同花色品种花瓣中OfCRTISO基因的表达分析表明,桂花初开期OfCRTISO基因的表达量可能影响盛开期各品种花瓣中类胡萝卜素的积累。

关键词: 桂花, 花色, 类胡萝卜素, 类胡萝卜素异构酶基因

Abstract: Based on the sequence information of Unigene of Osmanthus fragrans ‘Yanhong Gui’ from the petal transcriptome database,sequence of open reading frame in one carotenoid isomerase gene（OfCRTISO）from Osmanthus fragrans was amplified using RT-PCR technology and confirmed by sequencing. Bioinformatics analysis indicated that OfCRTISO contained a 1 842 bp open reading frame,encoding 613 amino acid residues. OfCRTISO from Osmanthus fragrans shared high sequence similarity with CRTISOs from Pedaliaceae and Solanaceae plants. Phylogenetic analysis showed that OfCRTISO and Sesamum indicum CRTISO（XP_011077785）gathered into one branch,revealing that these two proteins showed the closest genetic relationship. Gene expression analysis implicated that the expression level of OfCRTISO was extremely low at bud former stage,gradually increased at bud later stage,and reached the peak at initial flowering stage;however,it significantly decreased at full flowering stage. According to the expression analysis of OfCRTISO in the petals of sweet osmanthus cultivars with different flower color,it was demonstrated that the expression level of OfCRTISO at initial flowering stage might have an impact on the carotenoid accumulation in the petals of sweet osmanthus cultivars at full flowering stage.

Key words: Osmanthus fragransl, petal color, carotenoids, carotenoid isomerase gene

张超, 刘玉成, 王艺光, 付建新, 赵宏波. 桂花类胡萝卜素异构酶基因的克隆及表达分析[J]. 生物技术通报, 2017, 33(6): 89-96.

ZHANG Chao, LIU Yu-cheng, WANG Yi-guang, FU Jian-xin, ZHAO Hong-bo. Cloning and Expression Analysis of Carotenoid Isomerase Gene in Osmanthus fragrans[J]. Biotechnology Bulletin, 2017, 33(6): 89-96.

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桂花类胡萝卜素异构酶基因的克隆及表达分析

Cloning and Expression Analysis of Carotenoid Isomerase Gene in Osmanthus fragrans

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