大白菜WSD基因家族鉴定及其在干旱胁迫下蜡粉近等基因系中的表达

doi:10.16420/j.issn.0513-353x.2025-0407

摘要/Abstract

摘要： 蜡酯合成酶（WSD）是催化长链脂肪酸与脂肪醇酯化反应合成蜡酯的关键酶。蜡酯含量增加有助于提高植物的耐旱性。基于生物信息学手段对大白菜WSD家族成员进行全基因组鉴定及染色体定位、蛋白质理化性质、基因结构、蛋白保守域结构、共线性关系、启动子顺式作用元件、组织特异性表达和在干旱胁迫下的表达模式等进行分析。结果鉴定到18个WSD基因成员，分成4个亚族，不均匀地分布在7条染色体上。WSD氨基酸长度和分子量的范围分别为415 ~ 554 和46.57 ~ 62.86 kD。基因结构分析发现WSD结构较为保守。WSD中存在3对串联复制基因和6对片段复制基因，共线性分析表明BrWSD1、BrWSD4、BrWSD8、BrWSD16、BrWSD17和BrWSD18共6个家族成员与拟南芥共线性的WSD基因发生三倍化，BrWSD3和BrWSD7共2个家族成员发生了两倍化。基于同源性比较和比较转录组分析，筛选到BrWSD1和BrWSD9两个候选基因可能参与大白菜干旱胁迫响应。

关键词: 大白菜, 蜡酯合成酶, 蜡质, 基因家族, 干旱

Abstract:

Wax synthase/diacylglycerol acyltransferase（WSD）is a key enzyme that catalyzes esterification reaction of long-chain fatty acids and fatty alcohols to synthesize wax esters. An increase in wax ester content helps to improve drought resistance in plant. Whole genome identification and chromosome localization，protein physicochemical properties，gene structure，protein conserved domain structure，collinear relationships，promoter cis-acting elements，tissue-specific expression and expression patterns under drought stress were analyzed based on bio-informatics methods. The results identified 18 WSD genes members classified into four subfamilies，which are unevenly distributed across seven chromosomes.The range of amino acids length and molecular weight for WSD is 415-554 and 46.57-62.86 kD，respectively. Gene structure analysis revealed that the WSD structure is relatively conserved. There are three pairs of tandem duplicated genes and six pairs of fragment duplicated genes in WSD. Collinearity analysis showed that six family members，including BrWSD1，BrWSD4，BrWSD8，BrWSD16，BrWSD17 and BrWSD18，exhibited tripling of the WSD gene in Arabidopsis thaliana，while two family members，BrWSD3 and BrWSD7，exhibited doubling. Based on homology comparison and comparative transcriptome analysis，two candidate genes BrWSD1 and BrWSD9 were identified which may be involved in the drought stress response of Chinese cabbage.

Key words: Chinese cabbage, wax ester synthase, wax, gene family, drought

王荣花, 刘栓桃, 赵智中, 李巧云, 许念芳, 张志刚, 王树彬. 大白菜WSD基因家族鉴定及其在干旱胁迫下蜡粉近等基因系中的表达[J]. 园艺学报, 2026, 53(4): 1143-1156.

WANG Ronghua, LIU Shuantao, ZHAO Zhizhong, LI Qiaoyun, XU Nianfang, ZHANG Zhigang, WANG Shubin. Identification of the WSD Family Gene in Chinese Cabbage and Expression Analysis Under Drought Stress in Waxy Near-Isogenic Lines[J]. Acta Horticulturae Sinica, 2026, 53(4): 1143-1156.

图/表 11

表1 用于qRT-PCR分析的引物序列

Table 1 Primer sequences for qRT-PCR analysis

基因名称Gene	正向引物（5′-3′）Forward primer sequence	反向引物（5′-3′）Reverse primer sequence
BrWSD1	GCGTAAAACATCGAACCCCG	CACCTCTTGAACCGGCCTTT
BrWSD6	TTGGTCGCTCAACAGTCGTA	ACAATGAGCGCCTGTGGAAA
BrWSD8	CACGGCTGTTCAATTCTCCG	TGCTGGAAAAGCGAGGATGA
BrWSD9	AGCTGTGGCTAGGTTTCACC	ATGCCTCGAGATCGCATGTT
BrWSD14	TTAGCGGTCGATGCAACTGT	TCGTAGAACATCTTGGCCGC
BrWSD15	TCATGAGCTCGACACACGAG	GGCTGAGTCTTCGGGCTATC
G6PD	GGGTATGCCAGGACTAAGCTC	GAATCATAAGGGCCACTCACAT

图1 大白菜WSD基因家族的染色体定位红色基因为串联基因簇

Fig. 1 Chromosomal localization of WSD gene family in Brassica rapa Red gene indicates the tandem gene cluster

表2 大白菜WSD蛋白理化性质及亚细胞定位预测

Table 2 Physicochemical properties and subcellular localization prediction of WSD proteins in Brassica rapa

基因 ID Gene ID	基因名称 Gene name	氨基酸数 No. of amino acids	分子量/kD Molecular weight	等电点 pI	不稳定系数 Instability index	脂肪族氨基酸指数 Aliphatic index	平均亲水系数 GRAVY	亚细胞定位 Subcellular localization
BraA01g024150	BrWSD1	511	57.93	9.15	43.60	93.48	-0.221	细胞质 Cytoplasm
BraA01g024160	BrWSD2	484	54.76	8.42	53.67	92.85	-0.165	细胞质 Cytoplasm
BraA02g004570	BrWSD3	467	52.32	9.04	44.20	97.69	-0.032	质膜 Plasma membrane
BraA02g015030	BrWSD4	482	54.48	9.17	37.41	98.03	-0.043	质膜 Plasma membrane
BraA02g021490	BrWSD5	477	53.47	8.71	43.47	101.13	0.105	细胞质 Cytoplasm
BraA02g021510	BrWSD6	478	53.73	8.33	45.92	105.75	0.110	细胞质 Cytoplasm
BraA03g005460	BrWSD7	474	53.30	8.96	37.91	89.45	-0.184	叶绿体 Chloroplast
BraA03g014460	BrWSD8	490	55.38	9.27	34.21	98.39	-0.009	质膜 Plasma membrane
BraA03g018490	BrWSD9	418	46.72	8.01	36.77	102.54	0.178	叶绿体 Chloroplast
BraA03g020550	BrWSD10	484	54.34	8.97	30.38	95.39	-0.068	叶绿体 Chloroplast
BraA03g020560	BrWSD11	476	53.94	9.08	30.01	97.00	-0.020	细胞质 Cytoplasm
BraA03g066590	BrWSD12	479	53.46	7.67	35.73	100.71	0.062	细胞质 Cytoplasm
BraA04g016040	BrWSD13	460	51.16	8.82	35.87	100.48	0.038	细胞质 Cytoplasm
BraA04g031560	BrWSD14	418	47.14	6.43	50.12	86.89	-0.261	叶绿体 Chloroplast
BraA05g015270	BrWSD15	543	61.54	9.46	37.74	92.49	-0.137	细胞质 Cytoplasm
BraA06g018340	BrWSD16	554	62.86	9.58	41.22	94.46	-0.225	细胞质 Cytoplasm
BraA06g025960	BrWSD17	415	46.57	6.74	42.54	90.70	-0.275	细胞核 Nuclear
BraA10g011970	BrWSD18	461	50.79	7.72	40.58	109.76	0.068	质膜 Plasma membrane

图2 大白菜WSD家族蛋白保守基序（A）和基因外显子/内含子结构的分布（B）

Fig. 2 Distribution of conserved motifs（A）and exon/intron structures in the WSD gene family of Brassica rapa（B）

图3 大白菜WSD蛋白Motif1氨基酸序列比对结果红色框为保守活性位点

Fig. 3 Motif1 amino acid sequence of WSD proteins in Brassica rapa The red box indicates the conserved active site

图4 大白菜WSD基因与拟南芥、甘蓝、油菜的蛋白系统进化树分析数字代表该分支在统计上的可信度

Fig. 4 Phylogenetic tree analysis of WSD proteins in Brassica rapa，Arabidopsis thaliana，Brassica oleracea and Brassica napus The numbers represent the statistical confidence of the branch

图5 大白菜WSD家族基因的共线性分析

Fig. 5 Collinearity analysis of Brassica rapa WSD family genes

图6 大白菜与拟南芥（A）和大白菜与甘蓝、油菜（B）WSD家族基因共线性分析

Fig. 6 Synteny analysis of WSD family genes across Brassica rapa and Arabidopsis thaliana（A），Brassica rapa and Brassica oleracea/Brassica napus（B）

图7 大白菜WSD家族基因启动子序列顺式作用元件分析

Fig. 7 Analysis of cis-acting elements in promoter sequence of WSD family genes in Brassica rapa

图8 大白菜WSD基因在不同组织/器官中的表达分析

Fig. 8 Expression analysis of WSD genes on different tissues/organs in Brassica rapa

图9 干旱胁迫下大白菜蜡质近等基因系WSD基因的表达分析不同小写字母表示处理间差异显著（P < 0.05，n = 3）

Fig. 9 WSD genes expression analysis in Brassica rapa waxy near isogenic lines under drought stress Different lowercase letters indicate significant differences among different treatments（P < 0.05，n = 3）

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