大白菜DABB基因家族的全基因组鉴定及盐碱胁迫下的表达分析

doi:10.13560/j.cnki.biotech.bull.1985.2024-0817

摘要/Abstract

摘要：

目的 DABB（dimeric α+β barrel domain protein, Dabb）是含1‒2个二聚化α+β桶状结构域的蛋白质，在多种植物逆境胁迫响应过程中发挥作用。预测大白菜DABB蛋白质的特性，并探究这些基因在盐碱胁迫耐性不同的大白菜品种中响应盐碱胁迫的表达变化，为研究大白菜DABB基因家族的抗逆功能提供理论依据。方法通过生物信息学方法对大白菜DABB家族进行全基因组范围鉴定，分析其亲缘关系、所编码蛋白质理化性质、染色体定位情况和基因结构；利用实时荧光定量PCR技术分析这些基因在不同大白菜品种中盐碱胁迫下的表达模式。结果在大白菜中共鉴定出7个DABB基因，分布在5条染色体上，亲缘关系较近的DABB基因结构也较为相似；DABB基因编码蛋白质氨基酸数为79-495 aa，相对分子质量为8.94-54.19 kD，大部分为稳定蛋白质，亚细胞定位预测显示，多数DABB蛋白质定位于细胞质膜。盐碱胁迫处理下，6个大白菜DABB基因具有差异表达响应，但在耐盐碱性不同的大白菜品种中表达模式不同，其中，BrcDABB1-1和BrcDABB2表达变化较为剧烈。结论在大白菜基因组中共鉴定出7个含有DABB结构域蛋白质的编码基因，其中6个基因在盐碱胁迫处理下具有差异表达响应，这些基因不同的表达模式可能与大白菜的盐碱胁迫耐性相关。

关键词: 大白菜, DABB基因, 盐碱胁迫, 全基因组鉴定, 表达分析

Abstract:

Objective DABB (dimeric α+β barrel domain protein) is a kind of protein containing 1‒2 dimerized α+β barrel domains, which plays roles in the responses of various plants to stress. This work aims to predict the properties of DABB proteins in Chinese cabbage (Brassica rapa ssp. pekinensis), investigate the expression changes of these genes in response to salt-alkali stress in Chinese cabbage varieties with different salt-alkali stress tolerances, and provide theoretical evidence for studying the resistant functions of DABB proteins in Chinese cabbage. Method The DABB gene family of Chinese cabbage was identified genome-widely by bioinformatics methods, and the genetic relationship, physical and chemical properties of the encoded proteins, chromosome localizations and gene structures were analyzed, real-time fluorescence quantitative PCR was used to analyze the expression patterns of these genes in different Chinese cabbage varieties under salt-alkali stress. Result A total of 7 DABB genes were identified in the genome of Chinese cabbage, distributed in 5 chromosomes, the DABBs with close relatives were similar in structure; the amino acid number of the encoded protein was 79-495 aa, the relative molecular weight was 8.94-54.19 kD, most of them were stable proteins, and subcellular localization prediction showed that most of the DABB proteins were located in the plasma membrane. Under salt-alkali stress treatments, 6 DABB genes of Chinese cabbage performed differentially expression response, but the expression patterns were different in the Chinese cabbage cultivars with different salt-alkali stress tolerance. Conclusion Totally 7 genes encoding the protein containing DABB domains are identified in the Chinese cabbage genome, 6 of them show differentially expression response under salt-alkali stress treatments, the different expression patterns of these genes may be related to the salt-alkali stress tolerance of Chinese cabbage.

Key words: Chinese cabbage, DABB gene, salt-alkali stress, genome-wide identification, expression analysis

孙天国, 衣兰, 秦旭洋, 乔梦雪, 谷新颖, 韩艺, 沙伟, 张梅娟, 马天意. 大白菜DABB基因家族的全基因组鉴定及盐碱胁迫下的表达分析[J]. 生物技术通报, 2025, 41(4): 156-165.

SUN Tian-guo, YI Lan, QIN Xu-yang, QIAO Meng-xue, GU Xin-ying, HAN Yi, SHA Wei, ZHANG Mei-juan, MA Tian-yi. Genome-wide Identification of the DABB Gene Family in Brassica rapa ssp. pekinensis and Expression Analysis under Saline and Alkali Stress[J]. Biotechnology Bulletin, 2025, 41(4): 156-165.

图/表 9

表1 实时荧光定量PCR引物

Table 1 Primers information for RT-qPCR

引物名称 Primer name	引物序列 Primer sequence （5′-3′）
BrcActin-F	GGAGCTGAGAGATTCCGTTG
BrcActin-R	GAACCACCACTGAGGACGAT
BrcDABB1-1-F	CCGACGAGACCAAGAAGGAGATTATG
BrcDABB1-1-R	CTACGAAGGAGGAGGAACCAAGAAC
BrcDABB1-2-F	GGATGCTAAGTCGGAGATTGTGGATGTG
BrcDABB1-2-R	ACCCAGAAACCAAACCAGAAGGAACC
BrcDABB1-3-F	ATGGATGTGGTTAAGGAGAAGTCTGC
BrcDABB1-3-R	AAGGTACAACGAACTCAACAACAATGG
BrcDABB2-F	TCACGAGATGCTTAGACAAGGATTCAC
BrcDABB2-R	GAGAACTCAACGTGAAGAGGATGGCC
BrcDABB3-F	CTAAGGCTGATGAGATCTTGAAGGGCTC
BrcDABB3-R	GGCGACGGTGAAGTCAAGAATCACG
BrcHS1-1-F	GGAAGCAAAGGGACCAGTGAAGC
BrcHS1-1-R	AGTCTATGACCAGCACTTTATCCAAGC
BrcHS1-2-F	GGAAGCAAAGGGACCAGTGAAGC
BrcHS1-2-R	TGTAGTCTATAACCAACACTTTATCCAAGC

图1 大白菜和拟南芥DABB蛋白系统进化分析

Fig. 1 Phylogenetic analysis of Chinese cabbage (Brassica rapa ssp. pekinensis) and Arabidopsis DABB proteins

表2 大白菜DABB蛋白质家族成员信息及特性预测分析

Table 2 Membership information and property prediction of Chinese cabbage DABB protein family

蛋白质 Protein	基因座位号 Gene locus number	氨基酸数量 Number of amino acids/aa	相对分子质量 Relative molecular mass/kD	理论等电点 Theoretical isoelectric point	不稳定系数 Instability coefficient	平均疏水系数 Average hydrophobicity	亚细胞定位预测 Prediction of subcellular localizations
BrcDABB1-1	BraA08g002970.4C	201	22.328 56	5.27	30.79	-0.125	叶绿体 Chloroplast
BrcDABB1-2	BraA01g031610.4C	495	54.185 79	9.01	59.22	-0.159	细胞核 Nucleus
BrcDABB1-3	BraA05g019150.4C	187	20.350 27	4.87	37.96	-0.034	细胞质膜 Cell membrane
BrcDABB2	BraA10g019230.4C	111	12.321 08	5.22	22.95	0.109	细胞质膜 Cell membrane
BrcDABB3	BraA02g010040.4C	111	12.429 29	5.92	35.71	-0.009	细胞质膜 Cell membrane
BrcHS1-1	BraA01g037190.4C	107	12.111 95	5.41	27.24	-0.085	细胞质基质 Cytoplasm
BrcHS1-2	BraA01g037170.4C	79	8.936 29	5.21	19.30	-0.059	叶绿体 Chloroplast

图2 大白菜DABB蛋白质保守基序（A）及结构域（B）分析

Fig. 2 Analysis of conserved motifs (A) and structural domains (B) of Chinese cabbage DABB proteins

图3 大白菜DABB基因结构

Fig. 3 Structure of Chinese cabbage DABB genes

图4 大白菜DABB基因启动子顺式作用元件预测分布TC-rich repeats为逆境应答元件；TATC-box、GARE-motif、P-box为赤霉素响应元件；LTR为冷害胁迫应答元件；TCA-element、SARE为水杨酸响应元件；ABRE为脱落酸响应元件；ARE为抗氧化反应元件；AuxRR-core为生长素响应元件；G-Box、GT1-motif为光响应元件；RY-element为参与种子特异性调控的顺式作用调控元件；CGTCA-motif、TGACG motif为茉莉酸甲酯响应元件；CAT-box为与分生组织表达有关的元件；GCN4-motif为参与胚乳表达的元件；MBS为干旱胁迫应答元件；CCAAT-box、MRE、MBSI分别为MYBHv1结合位点、参与光响应的MYB结合位点以及参与黄酮类生物合成基因调控的MYB结合位点

Fig. 4 Distribution of cis-acting elements predicted in the promoters of Chinese cabbage DABB genesTC-rich repeats is a stress response element; TATC-box, GARE-motif, P-box are gibberellin response elements; LTR is a cold stress response element; TCA-element and SARE are salicylic acid response elements; ABRE is an abscisic acid response element; ARE is an antioxidant response element; AuxRR-core is a growth hormone response element; G-Box, GT1-motif are light response elements; RY-element is a cis-acting regulatory element involved in seed-specific regulation; CGTCA-motif, TGACG motif are methyl jasmonate response elements; CAT-box is an element related to the expression of meristematic tissue; GCN4-motif is a element involved in endosperm expression; MBS is a drought stress response element; CCAAT-box, MRE and MBSI are MYBHv1 binding sites, MYB binding sites are involved in light response and MYB binding sites are involved in gene regulation of flavonoid biosynthesis

图5 大白菜DABB基因在不同组织的表达分析不同颜色代表相对表达量

Fig. 5 Expression analysis of Chinese cabbage DABB genes in different tissuesDifferent colors indicate relative expression

图6 大白菜DABB基因在不同浓度NaCl处理下的表达分析不同小写字母表示差异显著（P<0.05）；JT为‘金小童’大白菜品种，RC为‘日本春秋’大白菜品种。下同

Fig. 6 Expression analysis of Chinese cabbage DABB genes under NaCl treatment of different concentrationsDifferent lowercase letters indicate significant differences (P<0.05); JT is ‘Jinxiaotong’ cultivar, and RC is ‘Ribenchunqiu’ cultivar. The same below

图7 大白菜DABB基因在不同浓度Na2CO3处理下的表达分析

Fig. 7 Expression analysis of Chinese cabbage DABB genes under Na2CO3 treatment of different concentrations

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