Identification of the oat sHSP gene family and its transcript profiles in response to high temperature and aging

doi:10.11686/cyxb2023346

Abstract

Abstract:

Small heat shock proteins （sHSPs） are a type of protein that is encoded by nuclear genes in plants. The sHSPs are ubiquitous in plants， contain conserved ACD domains， and play important roles in aging as well as in responses to stresses such as high temperature and drought. In this study， 24 HSP20 （AsHSP20.1-AsHSP20.24） genes were identified in the oat genome using bioinformatics methods. The physical and chemical properties， protein structure， subcellular localization， phylogeny， conserved motifs， and conserved domains of the putative AsHSP20 family members were determined， and the chromosomal locations of their encoding genes and gene transcription profiles in response to high temperature and aging were systematically analyzed. The AsHSP20 genes were distributed on 17 chromosomes. The genes encoded polypeptides consisting of 136-529 amino acids， with molecular weights ranging from 14.9 to 58.1 kDa， and theoretical isoelectric points ranging from 5.30 to 8.79. Most HSP20 members were predicted to localize in the nucleus， cytoplasm， or chloroplast， and a few were predicted to localize in the plasma membrane， mitochondria， peroxisome， or extracellular space. Protein secondary and tertiary structure analyses showed that AsHSP20 members have β-folded structures. Based on analyses of conserved motif composition and phylogenetic relationships， the AsHSP20 gene family was classified into 11 subgroups. The conserved motifs were similar or identical among the members of the same subgroup， indicative of functional similarity among proteins in the same subgroup. Further analysis of AsHSP20 transcript profiles in seeds showed that AsHSP20.20 and AsHSP20.24 were down-regulated under both natural and artificial aging conditions. It is speculated that AsHSP20.20 and AsHSP20.24 are involved in regulating the reduction in oat seed viability during aging， and are candidate genes related to oat seed longevity. These genes have potential applications in anti-aging germplasm breeding. The results of this study provide valuable information on the regulatory mechanism of the AsHSP20 gene family in the aging process of oat， and also provide theoretical support for further research on oat HSP20 genes and the molecular mechanism of delayed aging in oat seeds.

Key words: oat, seed aging, HSP20 gene family, phyletic evolution, expression analysis

Yuan MA, Huan LIU, Gui-qin ZHAO, Jing-long WANG, Ran ZHANG, Rui-rui YAO. Identification of the oat sHSP gene family and its transcript profiles in response to high temperature and aging[J]. Acta Prataculturae Sinica, 2024, 33(8): 145-158.

Figures/Tables 8

References 67

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名称Name	正向引物Forward primer （5′-3′）	反向引物Reverse primer （5′-3′）
AsHSP20.1	GCCGCTCAGTATTATGCAAG	CCGATCAAGGGGTTCAAGTA
AsHSP20.2	TCAGCTGGAACGAATAGTCC	AGTAACGATGGAGAAGCCAG
AsHSP20.3	TAGGGGACCAATCTCTGTGT	TTGGTGCATTTCAGAAGGGT
AsHSP20.4	GAGCTCTTCTTCGGGAGC	ACCTTGACGTCGTCCTTG
AsHSP20.5	ATCTTCCAGGGGTGAAGAAG	CCACTTGTCGTTCTTGTCCT
AsHSP20.6	CATCTACTCTGCCGACCTC	TTCCTGGCGAAACTGCTC
AsHSP20.7	ATCTCTACTCTGCCGACCTC	GATGACGAGGTATCTCCCG
AsHSP20.8	TTTGTTGATCAGCAGAGGGT	TATGTATGGCATCCGAGACG
AsHSP20.9	CTCCGATAAGCCAACGGATA	TTGCTACTCGTTGCAGTTTG
AsHSP20.10	GAAGTACCTGCGGATGGA	GCTTCTCAACCAACACGGT
AsHSP20.11	AAGGAGGACGCCAAGTACC	CTTCTCCATGTCGGCGTT
AsHSP20.12	AAGTTCGTTAGGCGTTTTCG	GCTTCTTGACCTCGGTTTTG
AsHSP20.13	ACAAGAACGAGAAATGGCAC	ATATCTCGATGGCCTTCACC
AsHSP20.14	GAGCATCGACATCCTGGAG	CACCTTCACCTCCTCCTTG
AsHSP20.15	TGTTCAACACCCAGGGTTC	CAGGTCCTGCGAGAAGAAG
AsHSP20.16	CGCGTACAAGATGGACAAGA	CTCCTCCTTCTTGAGCTTGG
AsHSP20.17	GTTCAAGTTGCCAGACAATG	ATCTGGATGGATTTCACCTCG
AsHSP20.18	GTGTTTGACCCCTTCTCTCT	ACCTTCACCTCCTCCTTCTT
AsHSP20.19	GACGTACCTCGACTTCATGC	TCAGGTGCTCCTTCTTGAAC
AsHSP20.20	CTCTCTCGATCTCTTGGACC	TTCACCTCCTCCTTCTTGAG
AsHSP20.21	AGGGATGCCAATTTCTCGAT	TTCTTGTGTGAGGTGCTCTT
AsHSP20.22	GTGCATGGTTCTTCTTCACC	GAAAGCATCCATTGTGGCAT
AsHSP20.23	GACATTGTGGAGGACGAGAA	CCACCATCACGTTCACCT
AsHSP20.24	GTTTTTCTCGCGATGACCG	CCACCGCCTGAGAAGAGAT
AsActin（KP257585.1）	CATTGGTATGGAAGCTGCTG	CACTGAGCACAATGTTACCG

名称Name	正向引物Forward primer （5′-3′）	反向引物Reverse primer （5′-3′）
AsHSP20.1	GCCGCTCAGTATTATGCAAG	CCGATCAAGGGGTTCAAGTA
AsHSP20.2	TCAGCTGGAACGAATAGTCC	AGTAACGATGGAGAAGCCAG
AsHSP20.3	TAGGGGACCAATCTCTGTGT	TTGGTGCATTTCAGAAGGGT
AsHSP20.4	GAGCTCTTCTTCGGGAGC	ACCTTGACGTCGTCCTTG
AsHSP20.5	ATCTTCCAGGGGTGAAGAAG	CCACTTGTCGTTCTTGTCCT
AsHSP20.6	CATCTACTCTGCCGACCTC	TTCCTGGCGAAACTGCTC
AsHSP20.7	ATCTCTACTCTGCCGACCTC	GATGACGAGGTATCTCCCG
AsHSP20.8	TTTGTTGATCAGCAGAGGGT	TATGTATGGCATCCGAGACG
AsHSP20.9	CTCCGATAAGCCAACGGATA	TTGCTACTCGTTGCAGTTTG
AsHSP20.10	GAAGTACCTGCGGATGGA	GCTTCTCAACCAACACGGT
AsHSP20.11	AAGGAGGACGCCAAGTACC	CTTCTCCATGTCGGCGTT
AsHSP20.12	AAGTTCGTTAGGCGTTTTCG	GCTTCTTGACCTCGGTTTTG
AsHSP20.13	ACAAGAACGAGAAATGGCAC	ATATCTCGATGGCCTTCACC
AsHSP20.14	GAGCATCGACATCCTGGAG	CACCTTCACCTCCTCCTTG
AsHSP20.15	TGTTCAACACCCAGGGTTC	CAGGTCCTGCGAGAAGAAG
AsHSP20.16	CGCGTACAAGATGGACAAGA	CTCCTCCTTCTTGAGCTTGG
AsHSP20.17	GTTCAAGTTGCCAGACAATG	ATCTGGATGGATTTCACCTCG
AsHSP20.18	GTGTTTGACCCCTTCTCTCT	ACCTTCACCTCCTCCTTCTT
AsHSP20.19	GACGTACCTCGACTTCATGC	TCAGGTGCTCCTTCTTGAAC
AsHSP20.20	CTCTCTCGATCTCTTGGACC	TTCACCTCCTCCTTCTTGAG
AsHSP20.21	AGGGATGCCAATTTCTCGAT	TTCTTGTGTGAGGTGCTCTT
AsHSP20.22	GTGCATGGTTCTTCTTCACC	GAAAGCATCCATTGTGGCAT
AsHSP20.23	GACATTGTGGAGGACGAGAA	CCACCATCACGTTCACCT
AsHSP20.24	GTTTTTCTCGCGATGACCG	CCACCGCCTGAGAAGAGAT
AsActin（KP257585.1）	CATTGGTATGGAAGCTGCTG	CACTGAGCACAATGTTACCG

基因名Gene name	重命名Rename	ORF （bp）	aa	Mw （Da）	IP	II	PH	SLP
AVESA.00010b.r2.6AG1028780.1	AsHSP20.1	1281	426	46386.23	8.51	46.41	-0.123	Chl
AVESA.00010b.r2.6AG1028780.2	AsHSP20.2	1035	344	37976.51	5.76	41.26	-0.162	Chl
AVESA.00010b.r2.6DG1171000.1	AsHSP20.3	573	190	21219.33	6.59	47.41	-0.153	Chl
AVESA.00010b.r2.7AG1212080.1	AsHSP20.4	411	136	14949.18	8.01	41.96	-0.302	Per
AVESA.00010b.r2.3AG0406840.2	AsHSP20.5	456	151	16900.14	6.20	44.96	-0.597	Cyt
AVESA.00010b.r2.2DG0350420.1	AsHSP20.6	468	155	17251.45	6.19	52.50	-0.408	Cyt
AVESA.00010b.r2.2CG0275380.1	AsHSP20.7	468	155	17304.55	6.51	49.79	-0.434	Cyt
AVESA.00010b.r2.4AG0636730.1	AsHSP20.8	1557	518	57622.58	6.63	50.48	-0.459	Pl
AVESA.00010b.r2.3AG0433390.1	AsHSP20.9	828	275	29972.66	8.79	42.43	-0.759	Ex
AVESA.00010b.r2.7CG0689040.1	AsHSP20.10	480	159	17528.28	5.37	36.19	-0.382	Cyt
AVESA.00010b.r2.3DG0518610.1	AsHSP20.11	480	159	17508.25	5.71	36.38	-0.458	Cyt
AVESA.00010b.r2.3AG0406840.1	AsHSP20.12	456	151	17034.29	5.83	45.89	-0.647	Cyt
AVESA.00010b.r2.3CG0454310.1	AsHSP20.13	456	151	16926.12	6.00	56.17	-0.570	Cyt
AVESA.00010b.r2.6CG1118770.1	AsHSP20.14	534	177	19298.95	6.76	48.07	-0.473	Nuc
AVESA.00010b.r2.7DG1398280.1	AsHSP20.15	687	228	24631.75	5.30	41.45	-0.420	Mit
AVESA.00010b.r2.1CG0084310.1	AsHSP20.16	678	225	24304.46	5.61	45.15	-0.498	Mit
AVESA.00010b.r2.4CG1325780.1	AsHSP20.17	474	157	17411.57	5.80	54.23	-0.628	Nuc
AVESA.00010b.r2.1DG0138620.1	AsHSP20.18	450	149	16732.84	5.36	48.94	-0.714	Nuc
AVESA.00010b.r2.4AG0623840.1	AsHSP20.19	663	220	23889.56	5.41	47.70	-0.556	Ex
AVESA.00010b.r2.4CG1325690.1	AsHSP20.20	465	154	17198.33	5.68	44.63	-0.633	Nuc
AVESA.00010b.r2.5DG1005630.2	AsHSP20.21	1590	529	58060.51	6.29	48.53	-0.474	Pl
AVESA.00010b.r2.4CG1324540.2	AsHSP20.22	1590	529	58067.52	6.21	50.25	-0.465	Pl
AVESA.00010b.r2.5AG0799690.1	AsHSP20.23	738	245	26922.35	6.77	53.50	-0.642	Chl
AVESA.00010b.r2.7CG0688840.1	AsHSP20.24	618	205	22139.24	7.91	39.45	-0.279	Nuc

基因名Gene name	重命名Rename	ORF （bp）	aa	Mw （Da）	IP	II	PH	SLP
AVESA.00010b.r2.6AG1028780.1	AsHSP20.1	1281	426	46386.23	8.51	46.41	-0.123	Chl
AVESA.00010b.r2.6AG1028780.2	AsHSP20.2	1035	344	37976.51	5.76	41.26	-0.162	Chl
AVESA.00010b.r2.6DG1171000.1	AsHSP20.3	573	190	21219.33	6.59	47.41	-0.153	Chl
AVESA.00010b.r2.7AG1212080.1	AsHSP20.4	411	136	14949.18	8.01	41.96	-0.302	Per
AVESA.00010b.r2.3AG0406840.2	AsHSP20.5	456	151	16900.14	6.20	44.96	-0.597	Cyt
AVESA.00010b.r2.2DG0350420.1	AsHSP20.6	468	155	17251.45	6.19	52.50	-0.408	Cyt
AVESA.00010b.r2.2CG0275380.1	AsHSP20.7	468	155	17304.55	6.51	49.79	-0.434	Cyt
AVESA.00010b.r2.4AG0636730.1	AsHSP20.8	1557	518	57622.58	6.63	50.48	-0.459	Pl
AVESA.00010b.r2.3AG0433390.1	AsHSP20.9	828	275	29972.66	8.79	42.43	-0.759	Ex
AVESA.00010b.r2.7CG0689040.1	AsHSP20.10	480	159	17528.28	5.37	36.19	-0.382	Cyt
AVESA.00010b.r2.3DG0518610.1	AsHSP20.11	480	159	17508.25	5.71	36.38	-0.458	Cyt
AVESA.00010b.r2.3AG0406840.1	AsHSP20.12	456	151	17034.29	5.83	45.89	-0.647	Cyt
AVESA.00010b.r2.3CG0454310.1	AsHSP20.13	456	151	16926.12	6.00	56.17	-0.570	Cyt
AVESA.00010b.r2.6CG1118770.1	AsHSP20.14	534	177	19298.95	6.76	48.07	-0.473	Nuc
AVESA.00010b.r2.7DG1398280.1	AsHSP20.15	687	228	24631.75	5.30	41.45	-0.420	Mit
AVESA.00010b.r2.1CG0084310.1	AsHSP20.16	678	225	24304.46	5.61	45.15	-0.498	Mit
AVESA.00010b.r2.4CG1325780.1	AsHSP20.17	474	157	17411.57	5.80	54.23	-0.628	Nuc
AVESA.00010b.r2.1DG0138620.1	AsHSP20.18	450	149	16732.84	5.36	48.94	-0.714	Nuc
AVESA.00010b.r2.4AG0623840.1	AsHSP20.19	663	220	23889.56	5.41	47.70	-0.556	Ex
AVESA.00010b.r2.4CG1325690.1	AsHSP20.20	465	154	17198.33	5.68	44.63	-0.633	Nuc
AVESA.00010b.r2.5DG1005630.2	AsHSP20.21	1590	529	58060.51	6.29	48.53	-0.474	Pl
AVESA.00010b.r2.4CG1324540.2	AsHSP20.22	1590	529	58067.52	6.21	50.25	-0.465	Pl
AVESA.00010b.r2.5AG0799690.1	AsHSP20.23	738	245	26922.35	6.77	53.50	-0.642	Chl
AVESA.00010b.r2.7CG0688840.1	AsHSP20.24	618	205	22139.24	7.91	39.45	-0.279	Nuc

名称 Name	α-螺旋 Alpha-helix	延伸链 Extended strand	无规则卷曲 Random coil	β-转角 Beta turn	β-折叠 Beta sheet	名称 Name	α-螺旋 Alpha-helix	延伸链 Extended strand	无规则卷曲 Random coil	β-转角 Beta turn	β-折叠 Beta sheet
AsHSP20.1	40.38	17.84	37.09	4.69	17	AsHSP20.13	17.22	19.21	58.28	5.30	40
AsHSP20.2	43.90	15.12	35.76	5.23	20	AsHSP20.14	23.16	20.90	48.59	7.34	36
AsHSP20.3	30.00	23.68	39.47	6.84	32	AsHSP20.15	28.95	21.93	43.42	5.70	22
AsHSP20.4	23.53	22.79	47.06	6.62	48	AsHSP20.16	21.33	16.89	57.78	4.00	24
AsHSP20.5	23.18	20.53	50.33	5.96	38	AsHSP20.17	15.29	19.75	57.32	7.64	38
AsHSP20.6	17.42	24.52	49.68	8.39	30	AsHSP20.18	30.20	18.79	44.30	6.71	39
AsHSP20.7	18.06	25.16	48.39	8.39	32	AsHSP20.19	43.18	16.82	29.55	10.45	30
AsHSP20.8	19.88	18.53	54.63	6.95	27	AsHSP20.20	17.53	22.08	55.19	5.19	38
AsHSP20.9	42.55	12.00	40.00	5.45	16	AsHSP20.21	18.15	20.23	56.14	5.48	23
AsHSP20.10	37.74	20.75	37.11	4.40	36	AsHSP20.22	16.45	18.34	59.55	5.67	24
AsHSP20.11	33.96	20.13	40.88	5.03	36	AsHSP20.23	20.82	15.10	61.22	2.86	24
AsHSP20.12	25.17	19.21	48.34	7.28	38	AsHSP20.24	42.93	16.59	34.63	5.85	30