结球甘蓝PRX基因家族全基因组鉴定与逆境条件下的表达分析

doi:10.13560/j.cnki.biotech.bull.1985.2023-0073

摘要/Abstract

摘要：

III类过氧化物酶（class III peroxidases, PRX）在植物生长发育和胁迫响应中发挥重要的作用，本研究利用生物信息学方法对结球甘蓝PRX基因家族进行鉴定，预测其结构和功能，并分析PRX基因在逆境条件下的表达模式，旨在明确甘蓝PRX基因家族的进化关系和功能。结果表明，结球甘蓝基因组中共鉴定出125个BoPRX基因家族成员，不均匀的分布在9条染色体上；编码氨基酸173-488 aa，大部分为亲水蛋白；亚细胞定位预测BoPRX基因大部分定位在叶绿体上。系统进化分析将甘蓝PRX蛋白分为5个亚族，每个亚族的成员都含有相似的外显子/内含子结构和蛋白质保守基序；启动子区域分析发现，BoPRX启动子序列中含有多种与激素和逆境等胁迫响应相关的顺式调控元件；基于转录组数据的热图分析显示，BoPRXs表达在叶绿体中最多。荧光定量PCR分析显示，6个BoPRX基因均受NaCl和PEG诱导上调；在ABA处理下，除BoPRX100外其余基因的表达在大部分时间被抑制，这些基因均受到ABA的差异调控，说明这些BoPRX基因都参与了ABA信号通路。综上所述，本研究揭示了PRX的复杂调控依赖于PRX的类型和信号分子，为进一步分析甘蓝PRX家族关键成员的功能提供了有价值的信息。

关键词: 甘蓝, PRX基因, 逆境胁迫, 表达分析

Abstract:

Class III peroxidases（PRX）play an important role in plant growth and development and stress response. In this study, the class III PRX gene family of cabbage was identified by bioinformatics, its function and structure were predicted, and the expression patterns of PRX gene under different abiotic stresses were analyzed, aiming to clarify the functional and evolutionary relationship of PRX gene family in cabbage genome. The results showed that a total of 125 BoPRX gene family members were identified in the cabbage genome, and were unevenly distributed on 9 chromosomes, the encoded 173-488 amino acids, and most were hydrophilic. Subcellular localization predicted that most BoPRX genes were located in chloroplasts. Phylogenetic analysis showed that the members of PRX family genes in the cabbage were classified into 5 groups, and members of each group had similar exon/intron structures and protein-conserved motifs. Promoter region analysis revealed that the BoPRX promoter sequence contained a variety of cis-regulatory elements related to abiotic stress responses such as hormones and stress. Heat map analysis of transcriptome data demonstrated that BoPRXs expression was the highest in chloroplasts. Fluorescence quantitative PCR analysis showed that 6 BoPRX genes were induced to be up-regulated by NaCl and PEG. Under ABA stress, except for BoPRX100, the expressions of the remaining genes were inhibited at most time, the expressions of the BoPRXs were all differentially regulated by ABA, indicating that they were all involved in the ABA signaling pathway. These findings reveal a complex regulation of PRXs that is dependent on the type of PRX, and the signaling molecule. It provides valuable information for further analysis of the functions of key members of the PRX family of cabbage.

Key words: cabbage, PRX gene, abiotic stress, expression analysis

葛雯冬, 王腾辉, 马天意, 范震宇, 王玉书. 结球甘蓝PRX基因家族全基因组鉴定与逆境条件下的表达分析[J]. 生物技术通报, 2023, 39(11): 252-260.

GE Wen-dong, WANG Teng-hui, MA Tian-yi, FAN Zhen-yu, WANG Yu-shu. Genome-wide Identification of the PRX Gene Family in Cabbage（Brassica oleracea L. var. capitata）and Expression Analysis Under Abiotic Stress[J]. Biotechnology Bulletin, 2023, 39(11): 252-260.

图/表 7

表1 荧光定量PCR引物

Table 1 Primer information for qRT-PCR

基因名称Gene name	正向引物 Forward primer（5'-3'）	反向引物Reverse primer（5'-3'）
BoActin	CGTGACCTTACTGACTACC	CTCCATCTCCTGCTCGTA
BoPRX12	TCTACGACAGTTCATGCCCAAACG	GTGGAAGTGAAGACGAAGGATGCTC
BoPRX44	CCAGAGCGACCAAGAGTTGTTCTC	TCCTTGTCATTGCGTCCACGAAC
BoPRX99	TCCAACGCCACTGACACAATTCC	ATCCTATTCATCGCCTCCACAAACG
BoPRX100	GTCCGATGCTCAACTTACACCTACC	GTGGAAGTGAAGACGAAGGATGCTC
BoPRX113	TCAGTCGAGTTGGCAGGAGGTC	AAGCGTGAAGAATGGAGCAGGAAG
BoPRX116	CCAGAGCGACCAAGAGTTGTTCTC	TTCCCATCCTTTTCATTGCCTCCAC

图1 BoPRX基因家族成员染色体定位

Fig. 1 Chromosome mapping of BoPRX gene family members

图2 拟南芥与甘蓝PRX蛋白系统发育进化树

Fig. 2 Phylogenetic tree of PRX protein of Arabidopsis and cabbage

图3 BoPRX蛋白基序组成（A）和BoPRX基因外显子-内含子结构（B）

Fig. 3 Motif composition of BoPRX proteins（A）and exon-intron structure of BoPRX genes（B）

图4 BoPRX顺式作用元件分析分布

Fig. 4 Cis-elements of BoPRX gene family

图5 BoPRX基因在不同部位的转录组表达谱

Fig. 5 Expression profiles of BoPRX genes in different tissues

图6 BoPRX基因在NaCl、PEG和ABA胁迫下表达模式

Fig. 6 Expression patterns of BoPRX genes under NaCl，PEG and ABA stresses

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