大麦条纹病菌GH17基因家族的鉴定及表达分析

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

生物技术通报 ›› 2025, Vol. 41 ›› Issue (8): 146-154.doi: 10.13560/j.cnki.biotech.bull.1985.2025-0104

• 研究报告 • 上一篇

大麦条纹病菌GH17基因家族的鉴定及表达分析

任睿斌¹^,²(), 司二静¹^,², 万广有¹^,², 汪军成¹^,², 姚立蓉¹^,², 张宏¹^,², 马小乐¹^,², 李葆春¹^,³, 王化俊¹^,², 孟亚雄¹^,²()

^1.省部共建干旱生境作物学国家重点实验室甘肃省作物遗传改良与种质创新重点实验室，兰州 730070
^2.甘肃农业大学农学院，兰州 730070
^3.甘肃农业大学生命科学技术学院，兰州 730070

收稿日期:2025-01-24 出版日期:2025-08-26 发布日期:2025-08-14
通讯作者: 孟亚雄，男，博士，教授，研究方向：分子植物育种；E-mail: yxmeng1@163.com
作者简介:任睿斌，男，硕士研究生，研究方向：作物遗传育种；E-mail: 1318361099@qq.com
基金资助:
国家大麦青稞产业技术体系项目(CARS-05-03B-03);国家自然科学基金项目(31960426);国家自然科学基金项目(32160496);甘肃省自然科学基金重点项目(24JRRA637)

Identification and Expression Analysis of GH17 Gene Family of Pyrenophora graminea

REN Rui-bin¹^,²(), SI Er-jing¹^,², WAN Guang-you¹^,², WANG Jun-cheng¹^,², YAO Li-rong¹^,², ZHANG Hong¹^,², MA Xiao-le¹^,², LI Bao-chun¹^,³, WANG Hua-jun¹^,², MENG Ya-xiong¹^,²()

^1.State Key Laboratory of Arid Habitat Crop Science, Gansu Provincial Key Laboratory of Crop Genetic Improvement and Germplasm Innovation, Lanzhou 730070
^2.College of Agronomy, Gansu Agricultural University, Lanzhou 730070
^3.College of Life Science and Technology, Gansu Agricultural University, Lanzhou 730070

Received:2025-01-24 Published:2025-08-26 Online:2025-08-14

摘要/Abstract

摘要：

目的糖苷水解酶17（glycoside hydrolase 17, GH17）在植物、动物和真菌的多种生理及致病过程中发挥重要作用，对大麦条纹病菌PgGH17家族成员进行鉴定和分析，为后续深入研究GH17基因功能提供科学依据，同时为解析大麦条纹病菌的致病性基因功能和探究寄主‒病原互动机制奠定坚实的理论基础。方法以大麦条纹病菌为研究对象，通过生物信息学方法，对基因家族进行鉴定及分析，经实时荧光定量PCR（RT-qPCR）监测大麦条纹病菌感染过程中各基因不同时间点的表达变化。结果 PgGH17基因家族含有6个成员，氨基酸序列长度为305‒648 aa，均为亲水性蛋白，等电点为5.06‒8.66，脂肪指数为65.43‒82.00，信号肽长度为0‒22 aa，且大多定位在细胞外基质，只有1个基因具有跨膜结构。内含子数为1‒3个，进化分析表明，该基因家族成员可聚成7个亚族。RT-qPCR结果表明，除PgGH17-3外，其他PgGH17基因在侵染过程中的表达水平基本呈现先上升后下降的趋势。结论从大麦条纹病菌中鉴定出6个GH17家族成员，除GH17-3外，其他GH17基因在侵染过程中表达均增强，说明其参与了病原菌的致病性。

关键词: 大麦条纹病菌, 糖苷水解酶17（GH17）, 基因家族, 基因表达分析

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

任睿斌, 司二静, 万广有, 汪军成, 姚立蓉, 张宏, 马小乐, 李葆春, 王化俊, 孟亚雄. 大麦条纹病菌GH17基因家族的鉴定及表达分析[J]. 生物技术通报, 2025, 41(8): 146-154.

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.

图/表 7

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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

大麦条纹病菌GH17基因家族的鉴定及表达分析

Identification and Expression Analysis of GH17 Gene Family of Pyrenophora graminea

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