不结球白菜花青苷合成负调控基因BrcLBD39的克隆和表达分析及其对外源6-BA的响应

[目的]本文旨在探究LBD39基因在不结球白菜紫色和绿色材料中的特性及其对外源6-BA的响应,为花青苷调控机制奠定理论基础。[方法]以不结球白菜紫色自交系NJZX1-3及其绿色突变体NJZX1-0为材料,同源克隆LBD39基因的全长,应用生物信息学方法分析其核酸和蛋白序列及其进化关系,并在线预测蛋白质二级和三级结构。采用外源6-BA处理后测定叶片中花青苷含量,并通过实时荧光定量PCR技术测定了LBD39基因在外源6-BA处理后的表达水平。[结果]在2个不同材料中克隆获得的LBD39基因完全相同,序列长为876 bp,包含编码区（长为699 bp）及非编码区（长为177 bp）。其编码区序列含有...

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Bibliographic Details
Published in南京农业大学学报 Vol. 41; no. 1; pp. 49 - 56
Main Author 董慧杰;侯喜林;韩克;张志硕;胡春梅
Format Journal Article
LanguageChinese
Published 南京农业大学园艺学院/作物遗传与种质创新国家重点实验室,江苏 南京,210095 2018
Subjects
LBD39
不结球白菜
同源克隆
基因表达
序列分析
同源克隆
不结球白菜
序列分析
homology-based cloning
基因表达
LBD39
sequence analysis
non-heading Chinese cabbage
gene expression
Online AccessGet full text
ISSN1000-2030
DOI10.7685/jnau.201704035

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Summary:[目的]本文旨在探究LBD39基因在不结球白菜紫色和绿色材料中的特性及其对外源6-BA的响应,为花青苷调控机制奠定理论基础。[方法]以不结球白菜紫色自交系NJZX1-3及其绿色突变体NJZX1-0为材料,同源克隆LBD39基因的全长,应用生物信息学方法分析其核酸和蛋白序列及其进化关系,并在线预测蛋白质二级和三级结构。采用外源6-BA处理后测定叶片中花青苷含量,并通过实时荧光定量PCR技术测定了LBD39基因在外源6-BA处理后的表达水平。[结果]在2个不同材料中克隆获得的LBD39基因完全相同,序列长为876 bp,包含编码区（长为699 bp）及非编码区（长为177 bp）。其编码区序列含有1个长为696 bp的开放阅读框（ORF）,编码232个氨基酸,将该基因命名为Brc LBD39。亚细胞定位预测该蛋白分布在细胞核中,蛋白相对分子质量为25.30×103,等电点为8.85。进化树分析表明,Brc LBD39蛋白序列与大白菜Bra LBD39相似性为100%,其次与油菜和甘蓝关系最近,相似性分别为99%和93%。外源6-BA处理24 h以后紫色材料NJZX1-3叶片中花青苷含量均高于对照,而绿色突变体叶片基本不含花青苷;Brc LBD39基因在2个材料中具有相似的表达模式,即先下降后上升而后又下降,其中在处理后48 h时Brc LBD39基因表达量急剧增加,在紫色和绿色材料中增量分别为74.84%和49.87%。此外,Brc LBD39表达量在绿色材料中明显高于紫色材料。[结论]不结球白菜中花青苷负调控基因Brc LBD39既响应外源6-BA,又与叶片中花青苷的积累密切相关,在花青苷的生物合成过程中起着重要的调控作用。
Bibliography:DONG Huijie, HOU Xilin, HAN Ke,ZHANG Zhishuo, HU Chunmei （ College of Horticulture/State Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095, China）
[ Objectives ] The aim of the study is to investigate the characteristic of LBD39 gene responsive to 6-BA in non-heading Chinese cabbage,providing the theoretical basis of the regulation of anthocyanins. [Methods] In this experiment, the full length of LBD39 gene was cloned from two materials of non-heading Chinese cabbage ,that was the purple self-line NJZX1-3 and its green mutant line NJZX1-0. The gene sequences and protein sequences were analyzed by hioinformatics, and the econdary structure and tertiary structure of BrcLBD39 protein were predicted online. The materials were treated by 6-BA, and the expression level of LBD39 was determined by quantitative real-time PCR and the total anthocyanins content was also measured. [ Results ] The study indicated that the sequences of the two cDNA clones were coincid
ISSN:1000-2030
DOI:10.7685/jnau.201704035