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浙江大学学报（农业与生命科学版）
浙江大学学报（农业与生命科学版）  2021, Vol. 47 Issue (3): 303-313    DOI: 10.3785/j.issn.1008-9209.2020.08.251
作物栽培与生理     
甘蓝型油菜钾离子转运载体HAK/KUP/KT家族的全基因组鉴定与分析
朱乐(),赵鑫泽,蒋立希()
浙江大学农业与生物技术学院作物科学研究所/浙江省作物种质资源重点实验室，杭州 310058
Genome-wide identification and analysis of potassium ion transporter HAK/KUP/KT family in rapeseed (Brassica napus L.)
Le ZHU(),Xinze ZHAO,Lixi JIANG()
Institute of Crop Science, College of Agriculture and Biotechnology, Zhejiang University/Zhejiang Key Laboratory of Crop Gene Resources, Hangzhou 310058, China
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摘要：

钾离子转运载体HAK/KUP/KT（high-affinity K＋/K＋ uptake permease/K＋ transporter）家族成员在植物对钾（K）的吸收和转运以及植物生长发育、耐盐性和渗透势的调节中起着重要作用。本研究以甘蓝型油菜（Brassica napus L.）为材料，共鉴定出45个HAK/KUP/KT基因，分别命名为BnHAK1～BnHAK45，并对油菜中的HAK/KUP/KT基因家族进行基因结构、进化和表达以及染色体定位分析。结果表明，甘蓝型油菜中的HAK/KUP/KT基因家族在染色体上呈现不均匀分布，其中在A01号染色体上分布最多，有5个BnHAKs基因。根据不同物种HAK/KUP/KT蛋白构建系统进化树，将其分为2个组群，其中双子叶植物中的拟南芥、甘蓝、白菜和甘蓝型油菜基因的保守性更强，亲缘关系更近。在表达量分析中，尽管HAK/KUP/KT基因家族的表达模式各不相同，但是总体而言，它们在根中的表达量最高，在角果中的表达量最低，这说明不同HAK/KUP/KT基因家族成员的分工是不同的。BnHAK4、BnHAK9基因在叶中特异性高表达，BnHAK11、BnHAK18和BnHAK23在根中特异性高表达，BnHAK7和BnHAK17在花中的表达量最高，说明该基因家族的表达具有组织特异性。本研究为进一步揭示甘蓝型油菜HAK/KUP/KT基因家族的功能机制提供了相应的参考依据。
关键词： 甘蓝型油菜;  钾离子转运载体HAK/KUP/KT;  基因家族分析    
Abstract:

The high-affinity K＋/K＋ uptake permease/K＋ transporter (HAK/KUP/KT) family members play important roles in the absorption and transport of potassium (K), regulations of plant growth and development, modulation/acquisition of salt tolerance, and osmotic potential regulation/control. However, this gene family has not been thoroughly studied in Brassica napus. In this study, forty-five BnHAK genes (BnHAK1-BnHAK45) were identified in the B. napus reference genome, and bioinformatic methods were used to resolve the gene structures, evolutionary analysis, expression patterns and chromosome position in different tissues of rape. The results showed that the members of the HAK/KUP/KT gene family were unevenly distributed on the chromosomes, with the most of them located on chromosome A01 which had five BnHAK genes. Phylogenetic analysis of HAK/KUP/KT proteins from multiple species showed that they could be divided into two groups. Among them, Arabidopsis, B. oleracea, B. rapa and B. napus had stronger gene conservation and closer genetic relationships. Although the expression patterns of the HAK/KUP/KT genes were different, in general, they exhibited highest and lowest expression levels in the roots and siliques, respectively, and this showed that there was a functional division of different HAK/KUP/KT gene family members. The expression levels of BnHAK4 and BnHAK9 were highest in the leaves, whilst the expression levels of BnHAK11, BnHAK18, and BnHAK23 were highest in the roots, and BnHAK7 and BnHAK17 were mostly expressed in the flowers, which indicated that the expression of this gene family had tissue specificity. This study provides a reference for further revealing the functional mechanism of HAK/KUP/KT gene family in B. napus.
Key words: Brassica napus    potassium ion transporter    HAK/KUP/KT    gene family analysis
收稿日期: 2020-08-25 出版日期: 2021-06-25
CLC:  S 565.4  
基金资助: 国家自然科学基金(31961143008)
通讯作者: 蒋立希     E-mail: 11616028@zju.edu.cn;jianglx@zju.edu.cn
作者简介: 朱乐（https://orcid.org/0000-0001-8410-3717），E-mail：11616028@zju.edu.cn
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引用本文:

朱乐,赵鑫泽,蒋立希. 甘蓝型油菜钾离子转运载体HAK/KUP/KT家族的全基因组鉴定与分析[J]. 浙江大学学报（农业与生命科学版）, 2021, 47(3): 303-313.

Le ZHU,Xinze ZHAO,Lixi JIANG. Genome-wide identification and analysis of potassium ion transporter HAK/KUP/KT family in rapeseed (Brassica napus L.). Journal of Zhejiang University (Agriculture and Life Sciences), 2021, 47(3): 303-313.

链接本文:

http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2020.08.251    或    http://www.zjujournals.com/agr/CN/Y2021/V47/I3/303

引物名称

Primer name

引物序列（5′→3′）

Primer sequence (5′→3′)

HAK1-FCAACAATCCAAAGATGAGCAGCGAC
HAK1-RGCTCCGATGAAGAGGAACCAAATG
HAK4-FCCTCCTCTTAGCTTACATGGGCCA
HAK4-RAACCTTGGGAAGCAGCCGAGA
HAK9-FGATGGGATTCTCACACCAGCTATGT
HAK9-RAGACGATGTAATACGGGTTCAGTGC
HAK13-FAGCCTCTTCCTTTTCATCCGCC
HAK13-RCTGGCTCGACCCAGTCACCG
HAK18-FGAAGCATGTGGGATTTGGACCA
HAK18-RGAGAGCTCCAATGATGTCTTCAGGG
HAK37-FAACAATCCAAAGATGAGCAGCGAC
HAK37-RGATAGGTGCAAAGAGCCAGCCTACT
ACTIN7-FGCTGACCGTATGAGCAAAG
ACTIN7-RAAGATGGATGGACCCGAC
表1  RT-qPCR引物序列
图1  甘蓝型油菜与其他植物HAK/KUP/KT蛋白的系统进化分析
图2  甘蓝型油菜HAK/KUP/KT基因家族的染色体定位
图3  甘蓝型油菜HAK/KUP/KT基因的系统发育、基因结构和蛋白结构域分析A. BnHAKs蛋白系统发育树分析；B. BnHAKs基因结构分析；C. BnHAKs蛋白的基序分析；D. 10种基序中的氨基酸保守性分析。UTR：非编码区；CDS：编码序列。
图4  甘蓝型油菜HAK/KUP/KT基因家族顺势作用元件分析
图5  甘蓝型油菜HAK/KUP/KT基因家族在不同组织中的表达量A. 转录表达分析；B. 荧光定量PCR表达分析。
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