LIU Zhengjie,LIU Yuntong,XU Shaozhong,et al.Cloning and Expression Analysis of Flavanone 3-hydroxylase SlF3H-like Gene From Purple Tomato[J].Northern Horticulture,2022,(17):26-33.[doi:10.11937/bfyy.20220892]
紫色番茄黄烷酮3-羟化酶基因SlF3H-like克隆与表达分析
- Title:
- Cloning and Expression Analysis of Flavanone 3-hydroxylase SlF3H-like Gene From Purple Tomato
- 关键词:
- 紫色番茄; SlF3H-like基因; 克隆; 花青素; 表达分析
- Keywords:
- purple tomato; SlF3H-like gene; cloning; anthocyanins; expression analysis
- 文献标志码:
- A
- 摘要:
- 以樱桃型紫色番茄‘砚紫1号’为试材,采用RT-PCR、生物信息学与qRT-PCR表达分析的方法,研究了该材料中黄烷酮3-羟化酶基因SlF3H-like的生物学信息,以期表征该基因序列特征和表达模式。结果表明:SlF3H-like基因开放阅读框(ORF)长度为1 089 bp,编码362个氨基酸;随机卷曲占据SlF3H-like蛋白结构元件的大多数,其次是α-螺旋和延伸链;‘砚紫1号’与其它物种的F3H氨基酸序列保守性非常高,SlF3H-like蛋白含有典型的非血红素双加氧酶和2OG-FeⅡ_Oxy加氧酶结构域;系统进化分析发现‘砚紫1号’与‘潘那利’番茄亲缘关系最近,其次是马铃薯。测定‘砚紫1号’发育过程中叶片和果皮中花青素含量,发现果皮中花青素含量随着果实直径的变大而逐渐积累,而叶片中花青素含量随着叶片直径的变大表现为先升后急剧下降。qRT-PCR分析SlF3H-like基因在‘砚紫1号’发育过程中果皮和叶片内的表达,发现该基因表达变化趋势与花青素含量的变化趋势类似,说明SlF3H-like基因在紫色番茄花青素积累的过程中起到重要的作用。
- Abstract:
- The cherry purple tomato ‘Yanzi No.1’ was used as test material.RT-PCR,bioinformatics and qRT-PCR expression analysis were used to study the biological information of flavanone 3-hydroxylase gene SlF3H-like,in order to characterize the sequence characteristics and expression pattern of the gene.The results showed that the open reading frame (ORF) of SlF3H-like gene was 1 089 bp,encoding 362 amino acids.The random coil was found to occupy the majority of structural elements of SlF3H-like protein,followed by α-helix and extended chain.The F3H amino acid sequence of ‘Yanzi No.1’ was highly conserved from other species.SlF3H-like protein contained typical DIOX-N superfamily and 2OG-FeⅡ_Oxy domains.Phylogenetic analysis showed that ‘Yanzi No.1’ was closely related to ‘Pannali’ tomato,followed by potato.The anthocyanin content in leaves and pericarp of ‘Yanzi No.1’ was measured during its development.It was found that the anthocyanin content in pericarp gradually accumulated with the increase of fruit diameter,while the anthocyanin content in leaves increased first and then decreased sharply with the increase of leaf diameter.qRT-PCR was used to analyze the expression of SlF3H-like gene in the pericarp and leaves of ‘Yanzi No.1’.Results showed that the change trend of this gene expression was similar to that of anthocyanin content,indicating that SlF3H-like gene played an important role in anthocyanin accumulation of purple tomato.
参考文献/References:
[1]陈丽,王定林,王跃云,等.番茄新品种砚紫1号选育[J].辣椒杂志,2018(3):35-39.[2]张园,张婉荣,林春,等.富花青素紫色番茄组织培养再生体系的建立[J].北方园艺,2018(18):41-47.[3]张园,徐绍忠,毛自朝,等.紫色番茄SlWD40-like基因克隆与表达分析[J].西北农业学报,2019,28(4):586-593.[4]YAN S,CHEN N,HUANG Z,et al.Anthocyanin fruit encodes an R2R3-MYB transcription factor,SlAN2-like,activating the transcription of SlMYBATV to fine-tune anthocyanin content in tomato fruit[J].New Phytologist,2019,225:2048-2063.[5]许志茹,崔国新,李春雷,等.芜菁黄烷酮3-羟化酶基因的克隆、序列分析及表达[J].分子植物育种,2008,6(4):787-792.[6]FERREYRA M F,RIUS S P,CASATI P.Flavonoids:Biosynthesis,biological functions,and biotechnological applications[J].Frontiers in Plant Science,2012,3(222):1-15.[7]王海竹,闫海芳,徐启江.红穗和白穗醋栗F3H的克隆及其在果实成熟过程中的表达分析[J].园艺学报,2016,43(10):2003-2011.[8]MARTIN C,PRESCOTT A,MACKAY S.Control of anthocyanin biosynthesis in flowers of Antirrhinum majus[J].Plant Journal,1991(1):37-49.[9]BRITSCH L,RUHNAU-BRICH B,FORKMANN G.Molecular cloning,sequence analysis,and in vitro expression of flavanone 3 beta-hydroxylase from Petunia hybrid[J].Journal of Biological Chemistry,1992,267(8):5380-5387.[10]DAVIES K M.A cDNA clone for flavanone 3-hydroxylase from Malus[J].Plant Physiology,1993,103(1):291.[11]DEBOO G B,ALBERTSEN M C,TAYLOR L P.Flavanone-3-hydroxylase transcripts and flavonol accumulation are temporally coordinate in maize anthers[J].Plant Journal,1995,7(5):703-713.[12]OWENS D K,CROSBY K C,RUNAC J,et al.Biochemical and genetic characterization of Arabidopsis flavanone-3-β-hydroxylase[J].Plant Physiology and Biochemistry,2008,46(10):833-843.[13]SHEN X,MARTENS S,CHEN M,et al.Cloning and characterization of a functional flavanone-3β-hydroxylase gene from Medicago truncatula[J].Molecular Biology Reports,2010,37 (7):3283-3289.[14]LIU M,LI X,LIU Y,et al.Regulation of flavanone 3-hydroxylase gene involved in the flavonoid biosynthesis pathway in response to UV-B radiation and drought stress in the desert plant,Reaumuria soongorica[J].Plant Physiology and Biochemistry,2013,73:161-167.[15]张华玲,黄元射,杨春贤,等.苦荞黄烷酮3-羟化酶基因F3H的克隆及序列分析[J].西北植物学报,2010,30(3):447-452.[16]ZHANG Y,HU Z,CHU G,et al.Anthocyanin accumulation and molecular analysis of anthocyanin biosynthesis-associated genes in eggplant (Solanum melongena L.)[J].Journal of Agricultural and Food Chemistry,2014,62(13):2906-2912.[17]赵乐,马利刚,张金燕,等.独行菜LaF3H基因克隆、序列分析及原核表达[J].中草药,2018,49(23):5626-5632.[18]SONG X,DIAO J,JI J,et al.Molecular cloning and identification of a flavanone 3-hydroxylase gene from Lycium chinense,and its overexpression enhances drought stress in tobacco[J].Plant Physiology and Biochemistry,2016,98:89-100.[19]袁岐,张春利,赵婷婷,等.番茄GRF 转录因子家族的生物信息学分析[J].分子植物育种,2017,15(8):2949-2956.[20]孙蓉,何周凤,陈惠,等.角蛋白酶kerC基因的克隆及序列分析[J].基因组学与应用生物学,2017,36(5):1965-1970.[21]LIU W,DONG D,YANG Z,et al.Polymerase Spiral Reaction (PSR):A novel isothermal nucleic acid amplification method[J].Scientific Reports,2015(5):12723.[22]马小磊,高霞莉,叶雯青,等.甘薯茎叶中花青素的定位及含量测定[J].植物学研究,2017,6(2):31-38.[23]黄春红,高燕会,朱玉球,等.石蒜黄烷酮3-羟化酶基因LrF3H的克隆及表达分析[J].园艺学报,2013,40(5):960-970.[24]宋祥忠,黄克江,王瑞珩.白芨(Bletilla striata)黄烷酮3-羟化酶基因的克隆和表达分析[J].分子植物育种,2019,17(14):4586-4591.[25]许明,伊恒杰,郭佳鑫,等.藤茶黄烷酮3-羟化酶基因AgF3H的克隆及表达分析[J].西北植物学报,2020,40(2):185-192.[26]张佩佩,张亮,郑凤霞,等.植物叶片中花青素的积累规律及生物学作用[J].北方园艺,2014(20):188-192.
相似文献/References:
[1]张园,张婉荣,林春,等.富花青素紫色番茄组织培养再生体系的建立[J].北方园艺,2018,42(18):47.[doi:10.11937/bfyy.20180312]
ZHANG Yuan,ZHANG Wanrong,LIN Chun,et al.Establishment of Tissue Culture and Regeneration System of Purple Tomato With Rich Anthocyanidin[J].Northern Horticulture,2018,42(17):47.[doi:10.11937/bfyy.20180312]
备注/Memo
第一作者简介:刘正杰(1984-),男,博士,讲师,现主要从事植物分子生物学等研究工作。E-mail:lzj1022@163.com.责任作者:林春(1971-),女,博士,副教授,现主要从事蔬菜功能基因组学等研究工作。E-mail:357315093@qq.com.基金项目:云南省农业基础研究联合专项面上资助项目(2018FG001(-019));云南省基础研究计划面上资助项目(202101AT070714)。收稿日期:2022-03-11