Identification and Expression Analysis of GH17 Gene Family of Pyrenophora graminea

doi:10.13560/j.cnki.biotech.bull.1985.2025-0104

Abstract

Abstract:

Objective Glycoside hydrolase 17 (GH17) plays a significant role in various physiological and pathogenic processes in plants, animals and fungi. The identification and preliminary analysis of the PgGH17 family members in the Pyrenophora graminea provide a scientific basis for further in-depth research on the function of GH17 genes, and concurrently lay a solid theoretical foundation for understanding the function of pathogenicity genes in the P. graminea and exploring the host-pathogen interaction mechanism. Method Using P. graminea as the research subject, we identified and analyzed gene families through bioinformatics approaches. Subsequently, real-time quantitative PCR (RT-qPCR) was employed to monitor the temporal expression patterns of these genes during the infection process. Result The PgGH17 gene family comprises six members, with amino acid sequence lengths ranging from 305 to 648 aa. All proteins are hydrophilic, exhibiting isoelectric points (pI) of 5.06–8.66 and aliphatic indices of 65.43–82.00. Signal peptide lengths vary from 0 to 22 aa, with most members localized to the extracellular matrix; only one gene harbors a transmembrane domain. Intron numbers range from 1 to 3 per gene. Phylogenetic analysis revealed that these family members cluster into seven distinct subclades. Quantitative reverse transcription PCR (RT-qPCR) results demonstrated that, with the exception of PgGH17-3, the expressions of other PgGH17 genes during infection generally present first an initial upregulation followed by subsequent downregulation. Conclusion Six GH17 family members are identified in P. graminea, and with the exception of GH17-3, all show upregulated expressions during infection, suggesting their functional involvement in the pathogenicity of this fungal pathogen.

Key words: Pyrenophora graminea, glycoside hydrolase 17 (GH17), gene family, gene expression analysis

REN Rui-bin, SI Er-jing, WAN Guang-you, WANG Jun-cheng, YAO Li-rong, ZHANG Hong, MA Xiao-le, LI Bao-chun, WANG Hua-jun, MENG Ya-xiong. Identification and Expression Analysis of GH17 Gene Family of Pyrenophora graminea[J]. Biotechnology Bulletin, 2025, 41(8): 146-154.

Figures/Tables 7

References 53

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基因ID Gene ID	基因 Gene	正向引物 Forward primer （5′‒3′）	反向引物 Reverse primer （5′‒3′）
P. graminea_GLEAN_10001959	PgGH17-1	ACCGTCTACCACACCGTCTA	GGGTAGTTAGGAGGAGCGGA
P. graminea_GLEAN_10003434	PgGH17-2	CCTACGCCTACACCATCACC	CACGGGTCTTTGGGTTGTTG
P. graminea_GLEAN_10004508	PgGH17-3	AAATGCTTGTCTGAGGCCGA	TGTGGGATCCACGTATGCAC
P. graminea_GLEAN_10006809	PgGH17-4	GCAATGAACCAGTACCCCGA	CTGAGTCGGCCATGTTGTCT
P. graminea_GLEAN_10007745	PgGH17-5	TGGACCGTCTACGTCAATGC	AACAGTTTGGTGGTAGGCGT
P. graminea_GLEAN_10008820	PgGH17-6	AGGCACCGACTATGAAGCTG	TTGAAGCCGTAGTCGAGCAG
Pg-Actin	Pg-Actin	GCGGTTACACCTCTCTACCACC	AGTCTGGATCTCCTGCTCAAAG

基因ID Gene ID	基因 Gene	正向引物 Forward primer （5′‒3′）	反向引物 Reverse primer （5′‒3′）
P. graminea_GLEAN_10001959	PgGH17-1	ACCGTCTACCACACCGTCTA	GGGTAGTTAGGAGGAGCGGA
P. graminea_GLEAN_10003434	PgGH17-2	CCTACGCCTACACCATCACC	CACGGGTCTTTGGGTTGTTG
P. graminea_GLEAN_10004508	PgGH17-3	AAATGCTTGTCTGAGGCCGA	TGTGGGATCCACGTATGCAC
P. graminea_GLEAN_10006809	PgGH17-4	GCAATGAACCAGTACCCCGA	CTGAGTCGGCCATGTTGTCT
P. graminea_GLEAN_10007745	PgGH17-5	TGGACCGTCTACGTCAATGC	AACAGTTTGGTGGTAGGCGT
P. graminea_GLEAN_10008820	PgGH17-6	AGGCACCGACTATGAAGCTG	TTGAAGCCGTAGTCGAGCAG
Pg-Actin	Pg-Actin	GCGGTTACACCTCTCTACCACC	AGTCTGGATCTCCTGCTCAAAG

基因 Gene	氨基酸数 Number of amino acids （aa）	分子质量 Molecular weight （Da）	等电点 Isoelectric point	亲水性平均值 Grand average of hydropathicity （GRAVY）	不稳定系数 Instability index	跨膜结构数 Number of transmembrane structures	脂肪指数 Aliphatic index	信号肽长度 Length of signal peptide	亚细胞定位 Subcellular localization
PgGH17-1	530	55 200.02	5.09	-0.165	48.99	0	65.96	16	Extracellular matrix
PgGH17-2	454	48 488.13	8.66	-0.105	33.78	0	75.86	17	Extracellular matrix
PgGH17-3	443	46 301.96	5.29	-0.185	41.99	0	75.12	22	Extracellular matrix
PgGH17-4	648	70 125.37	5.67	-0.507	40.27	1	65.43	0	Plasma membrane
PgGH17-5	320	34 680.98	5.18	-0.164	14.52	0	82.00	19	Extracellular matrix
PgGH17-6	305	32 198.21	5.06	-0.125	19.54	0	68.33	19	Extracellular matrix

基因 Gene	氨基酸数 Number of amino acids （aa）	分子质量 Molecular weight （Da）	等电点 Isoelectric point	亲水性平均值 Grand average of hydropathicity （GRAVY）	不稳定系数 Instability index	跨膜结构数 Number of transmembrane structures	脂肪指数 Aliphatic index	信号肽长度 Length of signal peptide	亚细胞定位 Subcellular localization
PgGH17-1	530	55 200.02	5.09	-0.165	48.99	0	65.96	16	Extracellular matrix
PgGH17-2	454	48 488.13	8.66	-0.105	33.78	0	75.86	17	Extracellular matrix
PgGH17-3	443	46 301.96	5.29	-0.185	41.99	0	75.12	22	Extracellular matrix
PgGH17-4	648	70 125.37	5.67	-0.507	40.27	1	65.43	0	Plasma membrane
PgGH17-5	320	34 680.98	5.18	-0.164	14.52	0	82.00	19	Extracellular matrix
PgGH17-6	305	32 198.21	5.06	-0.125	19.54	0	68.33	19	Extracellular matrix

保守基序 Conserved motifs	氨基酸序列 Sequence of amino acids	氨基酸数目 Number of amino acids
Motif 1	DJIVGISVGNEDLYR	15
Motif 2	GKPVWVAETGWP	12
Motif 3	RJYGTDCNQTQNVIQSAIKHAIK	23
Motif 4	NFYQDWFCPRLDYGGDYFWFSAFDEPWK	28
Motif 5	DIIGANVHPFF	11
Motif 6	GFNYGAKWG	9
Motif 7	LLLGJWLD	8
Motif 8	QANRGMGFPM	10
Motif 9	RNQFRG	6
Motif 10	RFWDEVGCW	9