金针菇PRX家族基因鉴定及其在子实体发育过程中的表达分析

doi:10.16420/j.issn.0513-353x.2023-0929

摘要/Abstract

摘要：

为了解金针菇中第Ⅲ类过氧化物酶（class Ⅲ peroxidases，PRX）家族基因的序列与表达特征，利用生物信息手段从金针菇全基因组中筛选鉴定PRX家族基因，对其基因结构、序列特征、染色体定位、顺式作用元件、表达模式等进行分析。结果显示，金针菇基因组中共有6个PRX编码基因，分布于6条不同染色体上，且定位于不同的亚细胞位置；进化分析表明，6个基因可以分到1CysPRX、2CysPRX、PRXQ、PRXⅡ和PRXⅤ等5个亚家族中；顺式作用元件分析表明，6个基因均含有与生长发育、非生物胁迫相关的顺式作用元件；表达模式分析显示，基因家族成员在金针菇不同发育时期、不同组织表达量存在差异，FfPRXQ1、FfPRXⅤ在成熟期菌盖中的表达量高于其他部位，Ff1CysPRX、FfPRXⅡ、FfPRXⅤ 在菌柄稳定区的表达量显著高于伸长区和过渡区，FfPRXQ2在菌柄中表达量显著高于菌盖，在菌盖发育不同时期表现为先上升后下降。上述研究结果表明，金针菇PRX家族基因参与了子实体发育调控过程，且表达模式存在明显差异。

关键词: 金针菇, 第Ⅲ类过氧化物酶, 子实体发育, 表达分析

Abstract:

To understand the sequence and expression characteristics of the PRX family genes in Flammulina filiformis，bioinformatics methods were used to screen and identify the PRX gene family sequences from the whole genome of F. filiformis. Subsequently，the members of the gene family were analyzed in terms of gene structure，sequence characterization，chromosomal localization，cis-acting elements，and expression patterns. The results showed that there were six PRX genes distributed on six different chromosomes and localized in different cellular locations. The evolutionary analysis showed that the six genes could be classified into five subfamilies，namely 1CysPRX，2CysPRX，PRXQ，PRXⅡ，and PRXⅤ. The analysis of the cis-acting elements showed that the six genes contained cis-acting elements related to the growth，development，abiotic stresses. Expression pattern analysis showed that members of the gene family had different expression in different developmental periods and tissues of F. filiformis. The expression of FfPRXQ1 and FfPRXⅤ were significantly higher in the pileus of maturation stage than that in other parts，and the expression of Ff1CysPRX，FfPRXⅡ，and FfPRXⅤ were higher in the stable region than the elongation region and transitional region of the stipe. The expression of FfPRXQ2 was significantly higher in the stipe than in the pileus，and the expression at different periods of cap development，showing a first increase and then a decrease. The above results indicated that the PRX family genes were involved in the developmental regulation of the fruiting-body of F. filiformis，and there were obvious differences in the expression patterns.

Key words: Flammulina filiformis, class Ⅲ peroxidase, fruiting-body development, expression analysis

仝宗军, 韩星, 段昕莲, 刘媛媛, 林俊彬, 甘颖, 陈杰, 谢宝贵, 甘炳成, 严俊杰. 金针菇PRX家族基因鉴定及其在子实体发育过程中的表达分析[J]. 园艺学报, 2025, 52(2): 337-348.

TONG Zongjun, HAN Xing, DUAN Xinlian, LIU Yuanyuan, LIN Junbin, GAN Ying, CHEN Jie, XIE Baogui, GAN Bingcheng, YAN Junjie. Sequence Characterization of PRX Family and Expression Analysis in the Process of Fruiting Body Development in Flammulina filiformis[J]. Acta Horticulturae Sinica, 2025, 52(2): 337-348.

图/表 10

表1 qRT-PCR引物

Table 1 Primers used in real-time quantitative PCR

基因 Gene	正向引物（5′-3′） Forward sequence	反向引物（5′-3′） Reverse sequence
Ff1CysPRX	CTCCCCTTCACCATCCGAAC	TTCCAATTGACGGGGGTAGC
Ff2CysPRX	TCTTGGCCCCGATCTCAAAC	ACTGTGATTTGGCGAAGGGT
FfPRXQ1	GCGAGAGTTGATTTCCGCAC	CGAGCTTTGGACTATCGGCT
FfPRXQ2	AGAGGGCGACGTTTGTGATT	ACGTCAAAGTCTCCGCAACA
FfPRXⅡ	CCTACATTCCCTACGCACCC	TAGGGCGGGAGATGGTTTTG
FfPRXⅤ	TACCCGACGTCAAGTTGCTC	GCACGCCGATAATGACGAAC
FfGAPDH	CCTCTGCTCACTTGAAGGGT	GCGTTGGAGATGACTTTGAA
FfRas	TCAATGCGACGAGTAAAGAGAGG	CATAGGTCCCACATCTACATTTCG

表2 金针菇PRX家族6个成员氨基酸序列特征

Table 2 Characteristic analysis of amino acid sequences of six members of the PRX family of Flammulina filiformis

蛋白名称 Protein name	氨基酸长度/aa Protein length	分子量/kD Molecular weight	等电点 Theoretical pI	跨膜数量 Transmembr- anehelices number	信号肽 Signal peptide	亚细胞定位（得分） Sub-cellular localization（score）
Ff1CysPRX	239	26.79	6.39	0	无No	线粒体Mitochondrial（4.0）
Ff2CysPRX	206	22.72	5.08	1	无No	细胞质Cytoplasmic（5.6）
FfPRXⅡ	171	18.32	5.24	1	无No	细胞质Cytoplasmic（3.5）
FfPRXⅤ	193	20.63	5.69	2	无No	膜结合线粒体Membrane-bound Peroxisomal（8.8）
FfPRXQ1	225	24.45	7.61	0	无No	细胞核Nuclear（2.4）
FfPRXQ2	175	19.19	7.92	1	无No	膜结合线粒体 Membrane-bound Mitochondrial（8.0）

图1 金针菇PRX家族基因系统发育分析

Fig. 1 Phylogenetic tree of PRX family genes of Flammulina filiformis and its orthologous in multiple species

图2 金针菇PRX家族蛋白氨基酸序列保守结构域括号内为CDD数据库蛋白质保守结构域编号

Fig. 2 Conserved domain analysis of amino acid sequences of the PRX family of Flammulina filiformis Protein conserved domain numbers in the CDD database are in parentheses

图3 金针菇PRX家族基因染色体定位

Fig. 3 Chromosome location of FfPRX family genes in the Flammulina filiformis

图4 金针菇PRX家族基因结构

Fig. 4 Gene structure of FfPRX family genes in the Flammulina filiformis

图5 金针菇PRX家族基因启动子序列顺式作用元件分析

Fig. 5 Analysis of cis-acting elements in promoter sequence of FfPRX family genes

图6 金针菇PRX家族基因在子实体不同发育时期的相对表达量利用单因素方差分析进行显著性分析，不同小写字母代表不同时期差异显著，P < 0.05，n = 3。下同

Fig. 6 Relative expression levels of FfPRX family genes at developmental periods of Flammulina filiformis One-way ANOVA was used for significance analysis. The different lowercase letters over the columns within the same graph denote significant differences. P < 0.05，n = 3. The same below

图7 金针菇PRX家族在菌柄不同区段的相对表达量

Fig. 7 Relative expression levels of FfPRX family genes in different stipe regions of Flammulina filiformis

图8 金针菇PRX家族基因在菌盖开伞过程中的相对表达量

Fig. 8 Relative expression levels of FfPRX family genes in different pileus of Flammulina filiformis

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