桂花β-胡萝卜素羟化酶基因的克隆与序列分析  

Cloning and Sequence Analysis of β-carotene Hydroxylase Genes in Osmanthus fragrans

作　　者：程楠 项黛徽 沈雅园 付建新[1] 张超[1] CHENG Nan;XIANG Daihui;SHEN Yayuan;FU Jianxin;ZHANG Chao(School of Landscape Architecture,Zhejiang Agriculture and Forestry University,Hangzhou 311300,China)

机构地区：[1]浙江农林大学风景园林与建筑学院,浙江杭州311300

出　　处：《河南农业科学》2019年第1期99-104,共6页Journal of Henan Agricultural Sciences

基　　金：国家自然科学基金项目(31501790);浙江省自然科学基金项目(LY19C160002);浙江农林大学科研发展基金人才启动项目(2016FR031);浙江农林大学大学生科研训练项目(2013200053)

摘　　要：为揭示桂花不同品种β-胡萝卜素羟化酶(β-carotene hydroxylase,HYB)基因的序列差异,利用已构建的桂花花瓣转录组数据库中Unigene序列信息,结合RT-PCR技术对桂花品种玉玲珑、金球桂、堰虹桂和日香桂中2个β-胡萝卜素羟化酶基因(HYB1和HYB2)的最大阅读框进行扩增,并测序验证。生物信息学分析结果表明,4个桂花品种HYB1基因均含有长为909 bp的开放阅读框,编码302个氨基酸残基; HYB2基因均含有长为915 bp的开放阅读框,编码304个氨基酸残基。4个桂花品种HYB1和HYB2推导的氨基酸序列均具备保守的组氨酸结构域。HYB1基因核苷酸序列存在27个变异位点,其氨基酸序列存在8个变异位点; HYB2基因核苷酸序列存在12个变异位点,其氨基酸序列存在3个变异位点。进化树分析结果发现,4个桂花品种的HYB1和HYB2蛋白与茄科植物番茄和辣椒HYB蛋白亲缘性最高。To reveal the sequence difference ofβ-carotene hydroxylase(HYB)genes in different cultivars of Osmanthus fragrans,based on the Unigene sequence information from petal transcriptome database of O.fragrans,the open reading frames(ORFs)of twoβ-carotene hydroxylase genes HYB 1 and HYB 2 in four cultivars Yulinglong,Jinqiugui,Yanhonggui and Rixianggui,were amplified with RT-PCR technique and verificated by sequencing method in this study.Bioinformatics analysis revealed that ORF sequences of all HYB 1 genes in four cultivars were 909 bp,encoding 302 amino acid residues,while ORF sequences of all HYB 2 genes in four cultivars were 915 bp,encoding 304 amino acid residues.Conserved histidine structure domains were found present in the predicted amino acid residues of HYB 1 and HYB 2 in four cultivars.There were 27 nucleotide mutation sites in the HYB 1 gene sequences and 8 amino acid residues mutation sites in its predicted protein sequences in four cultivars.Accordingly,there were 12 nucleotide mutation sites in the HYB 2 gene sequences and 3 amino acid residues mutation sites in its predicted protein sequences.Phylogenetic analysis indicated that HYB1 and HYB2 were much closer to HYB proteins from Lycopersicon esculentum and Capsicum annuum of Solanaceae plants.

关 键 词：桂花 花色 Β-胡萝卜素羟化酶 克隆 序列分析 

分 类 号：S685.13[农业科学—观赏园艺]