Identification and Expression Analysis of the C3H Gene Family in Grona styracifolia across Different Varieties

doi:10.13560/j.cnki.biotech.bull.1985.2024-0883

Abstract

Abstract:

Objective The C3H family genes play a significant role in regulating plant growth and development. Investigating the characteristics and functions of the C3H gene family in Grona styracifolia will lay a foundation for studying its roles in the physiological and morphological formation of this species. Method Based on the comparison of transcriptome data between ‘Guangyao Da 1’ and wild G. styracifolia, the C3H family genes were identified. Bioinformatics techniques were utilized to analyze the physicochemical properties, chromosomal positions, phylogenetic relationships, conserved motifs, and domains of these genes. real-time quantitative PCR (RT-qPCR) was applied to examine the expression patterns of the C3H family genes in different tissues and varieties of G. styracifolia. The correlation between C3H genes and key genes involved in flavonoid biosynthesis was analyzed. Additionally, the total flavonoid content in the stem tips and bases of different varieties were measured. Result The G. styracifolia C3H gene family consists of 27 members distributed across 9 chromosomes, primarily encoding basic, unstable, and hydrophilic proteins. Phylogenetic analysis shows that the G. styracifoliaC3H family genes are divided into three groups, with members in the same group being relatively conserved. Analysis of conserved motifs and domains indicates that the most frequently occurring motif is related to auxin regulation, and there is cooperative interaction with domains such as ANKYR among the G. styracifolia C3H family members. Expression pattern analysis reveals that the G. styracifoliaC3H family genes mainly function in the stems of it and are highly expressed at the stem tips of the ‘Guangyao Da 1’ variety. Correlation analysis found that the expressions of G. styracifolia C3H family genes are often positively correlated with key genes in flavonoid biosynthesis. Total flavonoid content measurement showed that at the stem tip, the total flavonoid content of ‘Guangyao Da 1’ is significantly higher than that of wild creeping G. styracifolia, while the opposite is true at the stem base. Conclusion The GsC3H family genes may indirectly influence the agronomic traits of ‘Guangyao Da 1’ G. styracifolia by regulating the biosynthesis of flavonoids in the stem segments.

Key words: C3H family genes, Grona styracifolia, expression analysis, correlation analysis, total flavonoids, gene family identification, ‘Guangyao Da 1’

HUANG Jin-heng, HUANG Xi, ZHANG Jia-yan, ZHOU Xin-yu, LIAO Pei-ran, YANG Quan. Identification and Expression Analysis of the C3H Gene Family in Grona styracifolia across Different Varieties[J]. Biotechnology Bulletin, 2025, 41(4): 243-255.

Figures/Tables 11

References 31

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正向引物名称 Forward primer name	正向引物序列 Forward primer sequence （5′‒3′）	反向引物名称 Reverse primer name	反向引物序列 Reverse primer sequence （5′‒3′）
GsC3H1f	AACCAGAACCATTGGAGGAAA	GsC3H1r	TGATAGGCGACAGTTACTACCAAAT
GsC3H2f	CTTCCCTTGGAGCATACCCT	GsC3H2r	GAGGCACATAAGATTGAAGACCA
GsC3H3f	GCACTGTCTCCATCTCCGTTAC	GsC3H3r	TTGACCATCTGCTTTGACCC
GsC3H4f	GTCAAATGCCGTCAACAGGA	GsC3H4r	CGCAGATAGCGAAGGAAGGT
GsC3H5f	CGATTCCGCAGCACTTTCC	GsC3H5r	CACCCTTACCAGTTTCTTCGTCACA
GsC3H6f	CAACGCAGGTATGGAATCAAA	GsC3H6r	GGTAGACCCTTGTCACTGAGGTT
GsC3H7f	ATGGTGTAGTTACCTTATGTGGGATG	GsC3H7r	TGCTCGGACCTCTTGTTTCG
GsC3H8f	TACAAAGCGGAGCCAGAGC	GsC3H8r	GGTGGTAGCAGGTGTTAGAGGAGT
GsC3H9f	TCAAAGTCCGACCTTGTTCACG	GsC3H9r	TGCAGCCAGCACTCGAAAA
GsC3H10f	CCCCTTTGTTCACCCTGGAG	GsC3H10r	CACAGCTTGGTCCGATACTGG
GsC3H11f	GCTCAACTTACTAACCTGTCAGCCTC	GsC3H11r	CCGAAAGGCAGTCTTAGAGTCATC
GsC3H12f	GGATCTCCCAGCAAGATTGAC	GsC3H12r	GCTGCCGCTCTTTGAGTATCAT
GsC3H13f	CCCTCAAGATTCACTGTGGATG	GsC3H13r	TCTTTCTGGGAACTCGCCTAT
GsC3H14f	AGGCTGGGATTGCTGGAA	GsC3H14r	GAGATTGGACGGTAGGATAAACA
GsC3H15f	AAACGAAACGGACGCTGAC	GsC3H15r	GCATCGCCGAACTTTGAAC
GsC3H16f	TTCTCGCTCTGATCCGTCG	GsC3H16r	GCTCCAACAACCGTTCCAC
GsC3H17f	GCAAAGCGAAGCCAAAGTT	GsC3H17r	CTGAAAGGGCAGAGGACACC
GsC3H18f	CAATAGGTCGTCTTGGTGGTCG	GsC3H18r	CGCTGATCTTTGCCGTAGCA
GsC3H19f	CGAACACGCCGTCAACCTT	GsC3H19r	CCAAATGGGCAGCCAGAAG
GsC3H20f	TGAGTCTTACCCTGAGCGTCC	GsC3H20r	TGTAGCTCTTACTGCCGCATC
GsC3H21f	GAAGAGGAGCAGGGTAAGAAGC	GsC3H21r	CTGGCAAGTTGTGAGCAAGAGGT
GsC3H22f	GTCGTTTCAACCACCCTCG	GsC3H22r	TTTCTCACCCACTCGCAATG
GsC3H23f	ACAATGGTGACTCCGAGACAAC	GsC3H23r	GCATGTACCCGCCCTGAAT
GsC3H24f	AGGTGCTCCTTGTGACATTGTT	GsC3H24r	TGATTGAAAGCTCATCCCAGA
GsC3H25f	GGACGAAGATTTGTTGAAGCG	GsC3H25r	TCCTTCCAACGGAGGGTGT
GsC3H26f	TTTCTCCGATTCGCCACC	GsC3H26r	CGAGTTTAACATTCCGCATTG
GsC3H27f	CACCGAGCAAATCCAACCG	GsC3H27r	CACAGGAATAAGCATCGACCG
EF-1αf	GAGATGCCTGGTCTATTGTGGG	EF-1αr	GATTGGCTCAACTGTCAGATGCT

正向引物名称 Forward primer name	正向引物序列 Forward primer sequence （5′‒3′）	反向引物名称 Reverse primer name	反向引物序列 Reverse primer sequence （5′‒3′）
GsC3H1f	AACCAGAACCATTGGAGGAAA	GsC3H1r	TGATAGGCGACAGTTACTACCAAAT
GsC3H2f	CTTCCCTTGGAGCATACCCT	GsC3H2r	GAGGCACATAAGATTGAAGACCA
GsC3H3f	GCACTGTCTCCATCTCCGTTAC	GsC3H3r	TTGACCATCTGCTTTGACCC
GsC3H4f	GTCAAATGCCGTCAACAGGA	GsC3H4r	CGCAGATAGCGAAGGAAGGT
GsC3H5f	CGATTCCGCAGCACTTTCC	GsC3H5r	CACCCTTACCAGTTTCTTCGTCACA
GsC3H6f	CAACGCAGGTATGGAATCAAA	GsC3H6r	GGTAGACCCTTGTCACTGAGGTT
GsC3H7f	ATGGTGTAGTTACCTTATGTGGGATG	GsC3H7r	TGCTCGGACCTCTTGTTTCG
GsC3H8f	TACAAAGCGGAGCCAGAGC	GsC3H8r	GGTGGTAGCAGGTGTTAGAGGAGT
GsC3H9f	TCAAAGTCCGACCTTGTTCACG	GsC3H9r	TGCAGCCAGCACTCGAAAA
GsC3H10f	CCCCTTTGTTCACCCTGGAG	GsC3H10r	CACAGCTTGGTCCGATACTGG
GsC3H11f	GCTCAACTTACTAACCTGTCAGCCTC	GsC3H11r	CCGAAAGGCAGTCTTAGAGTCATC
GsC3H12f	GGATCTCCCAGCAAGATTGAC	GsC3H12r	GCTGCCGCTCTTTGAGTATCAT
GsC3H13f	CCCTCAAGATTCACTGTGGATG	GsC3H13r	TCTTTCTGGGAACTCGCCTAT
GsC3H14f	AGGCTGGGATTGCTGGAA	GsC3H14r	GAGATTGGACGGTAGGATAAACA
GsC3H15f	AAACGAAACGGACGCTGAC	GsC3H15r	GCATCGCCGAACTTTGAAC
GsC3H16f	TTCTCGCTCTGATCCGTCG	GsC3H16r	GCTCCAACAACCGTTCCAC
GsC3H17f	GCAAAGCGAAGCCAAAGTT	GsC3H17r	CTGAAAGGGCAGAGGACACC
GsC3H18f	CAATAGGTCGTCTTGGTGGTCG	GsC3H18r	CGCTGATCTTTGCCGTAGCA
GsC3H19f	CGAACACGCCGTCAACCTT	GsC3H19r	CCAAATGGGCAGCCAGAAG
GsC3H20f	TGAGTCTTACCCTGAGCGTCC	GsC3H20r	TGTAGCTCTTACTGCCGCATC
GsC3H21f	GAAGAGGAGCAGGGTAAGAAGC	GsC3H21r	CTGGCAAGTTGTGAGCAAGAGGT
GsC3H22f	GTCGTTTCAACCACCCTCG	GsC3H22r	TTTCTCACCCACTCGCAATG
GsC3H23f	ACAATGGTGACTCCGAGACAAC	GsC3H23r	GCATGTACCCGCCCTGAAT
GsC3H24f	AGGTGCTCCTTGTGACATTGTT	GsC3H24r	TGATTGAAAGCTCATCCCAGA
GsC3H25f	GGACGAAGATTTGTTGAAGCG	GsC3H25r	TCCTTCCAACGGAGGGTGT
GsC3H26f	TTTCTCCGATTCGCCACC	GsC3H26r	CGAGTTTAACATTCCGCATTG
GsC3H27f	CACCGAGCAAATCCAACCG	GsC3H27r	CACAGGAATAAGCATCGACCG
EF-1αf	GAGATGCCTGGTCTATTGTGGG	EF-1αr	GATTGGCTCAACTGTCAGATGCT

基因名 Gene name	氨基酸数量 Amino acid count/aa	相对分子质量 Relative molecular mass/Da	等电点 Isoelectric point	不稳定系数 Instability index	脂肪系数 Aliphatic index	亲水性 Hydrophilicity
GsC3H1	495	55 557.14	5.54	49.44	70.95	-0.761
GsC3H2	424	46 321.03	8.10	58.80	54.81	-0.586
GsC3H3	730	79 921.63	5.79	60.57	69.51	-0.460
GsC3H4	371	40 788.29	8.00	49.81	62.53	-0.705
GsC3H5	480	52 287.69	5.29	59.91	51.79	-0.785
GsC3H6	493	54 600.68	4.87	60.30	49.80	-0.936
GsC3H7	433	46 906.96	7.49	24.93	75.59	-0.212
GsC3H8	746	81 503.49	5.64	51.67	73.35	-0.422
GsC3H9	658	72 045.97	8.36	63.21	68.50	-0.440
GsC3H10	724	77 943.18	5.58	59.10	71.93	-0.346
GsC3H11	724	81 391.28	8.48	47.52	57.36	-0.991
GsC3H12	703	76 020.01	6.18	55.96	66.93	-0.437
GsC3H13	442	48 626.94	8.41	60.92	57.87	-0.514
GsC3H14	426	46 874.77	8.52	64.47	52.72	-0.587
GsC3H15	358	39 956.98	6.83	69.76	51.70	-0.649
GsC3H16	362	38 873.85	8.43	61.71	55.47	-0.520
GsC3H17	677	74 159.42	6.28	59.02	67.19	-0.501
GsC3H18	295	31 251.62	9.39	40.62	57.32	-0.466
GsC3H19	296	31 324.95	9.43	33.71	63.38	-0.409
GsC3H20	469	50 124.30	8.92	55.90	61.11	-0.418
GsC3H21	350	388 98.46	7.26	50.05	60.00	-0.722
GsC3H22	487	508 96.03	8.63	66.72	53.72	-0.384
GsC3H23	323	35 624.70	8.42	56.44	46.87	-0.724
GsC3H24	366	40 294.76	8.06	47.42	53.03	-0.700
GsC3H25	522	59 915.16	6.69	62.08	63.14	-0.912
GsC3H26	255	28 686.03	8.25	59.92	49.33	-0.696
GsC3H27	369	41 084.06	6.94	77.35	46.75	-0.696

基因名 Gene name	氨基酸数量 Amino acid count/aa	相对分子质量 Relative molecular mass/Da	等电点 Isoelectric point	不稳定系数 Instability index	脂肪系数 Aliphatic index	亲水性 Hydrophilicity
GsC3H1	495	55 557.14	5.54	49.44	70.95	-0.761
GsC3H2	424	46 321.03	8.10	58.80	54.81	-0.586
GsC3H3	730	79 921.63	5.79	60.57	69.51	-0.460
GsC3H4	371	40 788.29	8.00	49.81	62.53	-0.705
GsC3H5	480	52 287.69	5.29	59.91	51.79	-0.785
GsC3H6	493	54 600.68	4.87	60.30	49.80	-0.936
GsC3H7	433	46 906.96	7.49	24.93	75.59	-0.212
GsC3H8	746	81 503.49	5.64	51.67	73.35	-0.422
GsC3H9	658	72 045.97	8.36	63.21	68.50	-0.440
GsC3H10	724	77 943.18	5.58	59.10	71.93	-0.346
GsC3H11	724	81 391.28	8.48	47.52	57.36	-0.991
GsC3H12	703	76 020.01	6.18	55.96	66.93	-0.437
GsC3H13	442	48 626.94	8.41	60.92	57.87	-0.514
GsC3H14	426	46 874.77	8.52	64.47	52.72	-0.587
GsC3H15	358	39 956.98	6.83	69.76	51.70	-0.649
GsC3H16	362	38 873.85	8.43	61.71	55.47	-0.520
GsC3H17	677	74 159.42	6.28	59.02	67.19	-0.501
GsC3H18	295	31 251.62	9.39	40.62	57.32	-0.466
GsC3H19	296	31 324.95	9.43	33.71	63.38	-0.409
GsC3H20	469	50 124.30	8.92	55.90	61.11	-0.418
GsC3H21	350	388 98.46	7.26	50.05	60.00	-0.722
GsC3H22	487	508 96.03	8.63	66.72	53.72	-0.384
GsC3H23	323	35 624.70	8.42	56.44	46.87	-0.724
GsC3H24	366	40 294.76	8.06	47.42	53.03	-0.700
GsC3H25	522	59 915.16	6.69	62.08	63.14	-0.912
GsC3H26	255	28 686.03	8.25	59.92	49.33	-0.696
GsC3H27	369	41 084.06	6.94	77.35	46.75	-0.696

基因名 Gene name	α-螺旋 Alpha-helix/%	延伸链 Extended strand/%	无规则卷曲 Random coil/%	亚细胞定位 Subcellular localization
GsC3H1	50.71	6.26	43.03	细胞核
GsC3H2	0.00	5.19	94.81	细胞核
GsC3H3	26.85	3.70	69.45	细胞核
GsC3H4	25.34	1.89	72.78	细胞核
GsC3H5	3.12	4.17	92.71	细胞核
GsC3H6	8.52	4.06	87.42	细胞核
GsC3H7	2.31	40.18	57.51	叶绿体
GsC3H8	33.91	2.41	63.67	细胞核
GsC3H9	28.88	4.71	66.41	细胞核
GsC3H10	24.59	3.59	71.82	细胞核
GsC3H11	9.67	1.10	89.23	细胞核
GsC3H12	17.21	6.12	76.67	细胞核
GsC3H13	0.00	4.75	95.25	细胞核
GsC3H14	1.64	4.23	94.13	细胞核
GsC3H15	8.10	5.87	86.03	细胞核
GsC3H16	0.00	6.35	93.65	细胞核
GsC3H17	24.67	4.28	71.05	细胞核
GsC3H18	10.17	5.42	84.41	过氧化物酶体
GsC3H19	6.76	6.42	86.82	细胞质
GsC3H20	0.00	6.18	93.82	细胞核
GsC3H21	7.43	2.29	90.29	线粒体
GsC3H22	0.00	3.90	96.10	细胞核
GsC3H23	7.43	3.10	89.47	细胞核
GsC3H24	17.49	3.01	79.51	细胞核
GsC3H25	10.15	4.02	85.82	细胞核
GsC3H26	9.41	8.24	82.35	细胞核
GsC3H27	13.28	6.78	79.95	细胞核