Expression Analysis of the Catalase Genes FfCAT1 and FfCAT2 in Flammulina filiformis

doi:10.16420/j.issn.0513-353x.2024-0744

Acta Horticulturae Sinica ›› 2025, Vol. 52 ›› Issue (7): 1769-1779.doi: 10.16420/j.issn.0513-353x.2024-0744

• Genetic & Breeding·Germplasm Resources·Molecular Biology • Previous Articles Next Articles

Expression Analysis of the Catalase Genes FfCAT1 and FfCAT2 in Flammulina filiformis

ZHANG Tuo², NIE Menghan², LIU Yuanyuan²^,³, DUAN Xinlian¹, HAN Xing¹, XIE Baogui²^,^**(), and YAN Junjie¹^,^**()

¹ Chengdu Agricultural Science and Technology Center，Institute of Urban Agriculture，Chinese Academy of Agricultural Sciences，Chengdu 610213，China
² Mycological Research Center，College of Life Sciences，Fujian Agriculture and Forestry University，Fuzhou 350002，China
³ Institute of Plant Protection，Beijing Academy of Agriculture and Forestry Sciences，Beijing 100080，China

Received:2025-04-11 Revised:2025-06-10 Online:2025-07-23 Published:2025-07-23
Contact: XIE Baogui, and YAN Junjie

Abstract

Abstract:

In order to explore the role of catalase gene（CAT）in the development of fruit body in Flammulina filiformis，two catalase genes were identified based on the whole-genome of F. filiformis，named FfCAT1 and FfCAT2，respectively. The length open reading frame of the FfCAT1 gene was 1 557 bp and encoded 518 amino acids. The length open reading frame of the FfCAT2 gene was 1 692 bp and encoded 563 amino acids. The proteins encoded by both genes were located in the cytoplasm，contained no transmembrane domains. However，FfCAT1 did not have a signal peptide，while FfCAT2 had signal peptide between amino acid residues 18-19. Conserved domain and phylogenetic tree analysis showed that FfCAT1 and FfCAT2 belonged to monofunctional catalases. The qRT-PCR results suggested that FfCAT1 was highly expressed in the pileus of maturation stage and the expression level was significantly up-regulated in the ejection spore stage of pileus. The expression level in the lamella was significantly higher than that in the context. FfCAT2 was highly expressed in the pileus of elongation stage，but the expression of FfCAT2 gene was less different in different stages of development of the pileus，and it was significantly down-regulated when the pileus diameter was 3.3 cm. There were no significant differences in the lamella and context of 0.9 cm diameter pileus. These results indicated that two FfCAT genes played different roles in the growth and development of the pileus. FfCAT1 was the major gene involved in the regulation of the development of the pileus and was related to the ejection of spores on the lamella.

Key words: Flammulina filiformis, catalase, fruit body development, expression analysis, sexual reproduction

ZHANG Tuo, NIE Menghan, LIU Yuanyuan, DUAN Xinlian, HAN Xing, XIE Baogui, and YAN Junjie. Expression Analysis of the Catalase Genes FfCAT1 and FfCAT2 in Flammulina filiformis[J]. Acta Horticulturae Sinica, 2025, 52(7): 1769-1779.

Figures/Tables 8

Table 1 Primers sequence in this study

用途Function	引物名称Primer name	上游引物（5′-3′）Forward primer	下游引物（5′-3′）Reverse primer
PCR验证 PCR validation	FfCAT1	TCTTGACACCTTTCCTCGTAG	ACATTGCTTGCTTGTTTATTT
PCR验证 PCR validation	FfCAT2	GGTCCTCTCGTCGGTCGGCAATC	TTCCCACAGCATCCTCCGTCCTG
荧光定量PCR验证 qRT-PCR	QFfCAT1	CTCACTTTGACCGAGAGCGTAT	TGTTGGAGGTAATGTCGGAGAG
	QFfCAT2	AGCGCCCATATGTTGACGAA	TTTCCGAGATGACCAGCGAC
	QFfGAPDH	CCTCTGCTCACTTGAAGGGT	GCGTTGGAGATGACTTTGAA

Fig. 1 Schematic map of the FfCAT1 and FfCAT2 genes of Flammulina filiformis

Fig. 2 Tertiary structures of Flammulina filiformis FfCAT proteins

Fig. 3 Phylogenetic tree of three types of catalase constructed based on the neighbor-joining method（NJ） The numbers at the branches represent the self-check confidence（Bootstrap = 1000）

Fig. 4 Multiple sequence alignment and conserved domain analysis of monofunctional catalases amino acid sequences between Flammulina filiformis and other basidiomycetes Underlines denote conserved domains，and inverted triangles mark highly conserved proximal and distal amino acid residues

Fig. 5 The gene expression patterns of FfCAT1 and FfCAT2 during fruit body developments of Flammulina filiformis The different lowercase letters indicate significant differences at the 0.05 level

Fig. 6 Morphological characteristics of Fv01 pileus during development（A）and the gene expression patterns of FfCAT1 and FfCAT2（B）during pileus developments of Flammulina filiformis Fv01 The different lowercase letters indicate significant differences at the 0.05 level，n = 3

Fig. 7 The gene expression patterns of FfCAT1 and FfCAT2 in pileus of Flammulina filiformis ** P < 0.01，n = 3

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Expression Analysis of the Catalase Genes FfCAT1 and FfCAT2 in Flammulina filiformis

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