羽衣甘蓝β-胡萝卜素羟化酶基因的克隆及表达分析

doi:10.3969/j.issn.1004-1524.2019.01.11

浙江农业学报 ›› 2019, Vol. 31 ›› Issue (1): 80-85.DOI: 10.3969/j.issn.1004-1524.2019.01.11

羽衣甘蓝β-胡萝卜素羟化酶基因的克隆及表达分析

王玉书¹, 王欢², 郭宇¹, 周明慧¹, 陈璐¹, 陈阳¹

1.齐齐哈尔大学生命科学与农林学院,黑龙江省抗性基因工程与寒地生物多样性保护重点实验室,黑龙江齐齐哈尔 161006;
2.齐齐哈尔大学化学与化学工程学院,黑龙江齐齐哈尔 161006

收稿日期:2018-04-03 出版日期:2019-01-25 发布日期:2019-04-09
作者简介:王玉书(1985—),女,黑龙江富锦人,博士,副教授,主要从事园艺植物遗传与分子育种研究。E-mail:wangys1019@126.com
基金资助:
国家自然科学基金(31401908); 黑龙江省自然科学基金(C2016056); 黑龙江省普通高等学校青年创新人才培养计划(UNPYSCT-2017155); 黑龙江省省属高等学校基本科研业务费科研项目(135209313)

Cloning and expression analysis of β-carotene hydroxylase gene from kale (Brassica oleracea L. var. acephala)

WANG Yushu¹, WANG Huan², GUO Yu¹, ZHOU Minghui¹, CHEN Lu¹, CHEN Yang¹

1.Key Laboratory of Resistance Gene Engineering and Preservation of Biodiversity in Cold Areas, College of Life Sciences, Agriculture and Forestry, Qiqihar University, Qiqihar 161006, China;
2. College of Chemistry and Chemical Engineering, Qiqihar University, Qiqihar 161006, China

Received:2018-04-03 Online:2019-01-25 Published:2019-04-09

摘要/Abstract

摘要： 以羽衣甘蓝(Brassica oleracea L. var. acephala)为试验材料,采用同源克隆和RT-PCR技术,克隆得到羽衣甘蓝β-胡萝卜素羟化酶的cDNA全长,命名为BoBCH(GenBank登录号为MH016242)。序列分析表明,该cDNA序列长906 bp,编码301个氨基酸,分子量33.8 ku,理论等电点为9.67。保守结构域分析表明,BoBCH属于FA_hydroxylase蛋白超家族。系统发育分析结果表明,羽衣甘蓝与结球甘蓝处于同一分支,其亲缘关系最近。TMHMM和Wolf-Psort进行跨膜区分析及亚细胞定位,结果表明BoBCH蛋白有4个跨膜区域,可能定位于叶绿体中发挥作用。qRT-PCR检测结果表明,BoBCH在紫叶羽衣甘蓝DH系D07的根、茎、叶中均有表达,在叶片中表达量最高,茎次之,根中表达量最低;不同发育时期的检测结果表明,BoBCH在观赏期叶片中表达最高,在幼苗期和莲座期表达水平较低。

关键词: 羽衣甘蓝, β-胡萝卜素羟化酶基因, 克隆, 表达分析

Abstract: The β-carotene hydroxylase gene from kale (Brassica oleracea L. var. acephala) was cloned by homology-based cloning and RT-PCR strategies, named as BoBCH(GenBank accession number: MH016242). Sequence analysis showed that the full-length of BoBCH cDNA was 906 bp, which encoded a protein with 301 amino acids, an estimated molecular weight was 33.8 ku and an isoelectric point was 9.67. The results of conservative structural domain analysis showed that BoBCH belonged to the FA_hydroxylase protein superfamily. Phylogenetic tree analysis revealed that the kale had the closest evolutionary relationship with Brassica oleracea. Subcellular localization analysis of TMHMM and Wolf-Psort showed that it might be targeted to the chloroplast with four transmembrane regions. The qRT-PCR test results indicated that the highest was leaf, the next was stem, and the lowest level was in root; in the different periods of leaves, the expression level in the ornamental period was high, and the expression level was lower in seedling stage and rosette stage.

Key words: kale, β-carotene hydroxylase gene, cloning, expression analysis

中图分类号:

S635.9

王玉书, 王欢, 郭宇, 周明慧, 陈璐, 陈阳. 羽衣甘蓝β-胡萝卜素羟化酶基因的克隆及表达分析[J]. 浙江农业学报, 2019, 31(1): 80-85.

WANG Yushu, WANG Huan, GUO Yu, ZHOU Minghui, CHEN Lu, CHEN Yang. Cloning and expression analysis of β-carotene hydroxylase gene from kale (Brassica oleracea L. var. acephala)[J]. , 2019, 31(1): 80-85.

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羽衣甘蓝β-胡萝卜素羟化酶基因的克隆及表达分析

Cloning and expression analysis of β-carotene hydroxylase gene from kale (Brassica oleracea L. var. acephala)

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