大豆ZIP基因家族鉴定及响应铝胁迫的表达分析

doi:10.13560/j.cnki.biotech.bull.1985.2025-0318

摘要/Abstract

摘要：

目的鉴定大豆锌铁转运蛋白（zinc-regulated transporter/iron-regulated transporter, ZIP）基因家族成员，分析GmZIP基因家族成员在铝胁迫下的表达模式，为阐明GmZIP家族的生物学功能奠定基础。方法通过生物信息学鉴定大豆基因组中GmZIP基因家族成员，根据鉴定结果构建其系统发育树，预测蛋白网络图，分析基因家族成员的共线性关系、基因结构及启动子顺式作用元件。经转录组分析GmZIP基因家族成员在大豆不同组织中及在铝胁迫下的表达模式。结果在大豆基因组中鉴定到26个GmZIP基因家族成员，分布在14条染色体上，分为Ⅰ-Ⅴ亚家族。GmZIP基因家族成员基因结构较为保守，均含有ZIP保守结构域，含有多个外显子，且不同成员之间外显子数量差异明显。转录组结果显示，大部分GmZIP成员在大豆根和根瘤组织均有较高表达水平；在铝胁迫下，有3个成员（GmZIP6、GmZIP12、GmZIP25）在耐铝大豆种质南农99-6中上调表达，在铝敏感大豆种质中豆32中2个上调表达（GmZIP20、GmZIP25）、1个下调表达（GmZIP23）。蛋白预测结果显示，26个GmZIP蛋白中有16个存在互作关系，其中铝胁迫下差异表达基因有GmZIP23、GmZIP25。荧光定量PCR检测发现，耐铝种质中GmZIP25在铝处理72 h后表达量最高，而铝敏感种质中GmZIP25在处理12 h时表达量达到最大值。结论推测GmZIP25在大豆响应铝胁迫中具有重要作用。

关键词: 大豆, GmZIP基因家族, 生物信息学, 铝胁迫

Abstract:

Objective Identifying and analyzing the expressions of GmZIP family members under aluminum stress in soybean lays the foundation for elucidating the biological function of GmZIP family. Method The members of GmZIP gene family were identified by bioinformatics method, which was used to establish the phylogenetic tree, predict protein network map and analyze the collinearity relationships, gene structure, and promoter cis-acting elements. Based on transcriptome sequencing, the expression patterns of GmZIP gene family in different tissues and under aluminum (Al) stress were analyzed. Result A total of 26 GmZIP were identified in the whole soybean genome, which were unevenly distributed on 14 chromosomes and divided into Ⅰ-Ⅴ subfamilies by phylogenetic analysis. The gene structure of GmZIP members were relatively conservative. All of GmZIP members contained ZIP conserved domain, multiple exons, and significantly different introns. Transcriptome analysis of different soybean tissues showed that most of GmZIP members had high expressions in the soybean root and nodule tissues. Three members of GmZIP (GmZIP6, GmZIP12, and GmZIP25) were up-regulated in Al-tolerant soybean germplasm Nannong 99-6, two members (GmZIP20 and GmZIP25) were up-regulated while one was down-regulated (GmZIP23) in Al-sensitive soybean germplasm soybean Zhongdou 32. The protein prediction results show that 16 of 26 GmZIP have interaction relationships, among which GmZIP23 and GmZIP25 were differentially expressed under Al toxicity stress. Based on fluorescence quantitative PCR detection, the expression of GmZIP25 in Al-resistant germplasm reached the highest at 72 h under aluminum stress treatment, while that in Al-sensitive germplasm reached the highest at 12 h. Conclusion These results suggest that GmZIP25 may play an important role in response to aluminum stress of soybean.

Key words: soybean, GmZIP gene family, bioinformatics, aluminum stress

黄国栋, 邓宇星, 程宏伟, 但焱南, 周会汶, 吴兰花. 大豆ZIP基因家族鉴定及响应铝胁迫的表达分析[J]. 生物技术通报, 2025, 41(9): 71-81.

HUANG Guo-dong, DENG Yu-xing, CHENG Hong-wei, DAN Yan-nan, ZHOU Hui-wen, WU Lan-hua. Genome-wide Identification and Expression Analysis of the ZIP Gene Family in Soybean[J]. Biotechnology Bulletin, 2025, 41(9): 71-81.

图/表 10

参考文献 40

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基因 Gene	上游引物 Forward primer （5'-3'）	下游引物 Reverse primer （5'-3'）	片段大小 Size （bp）
GmZIP6	GCGTGTTGTGTCCTTCTTTTAC	CTTAACAATGGGAGGCTCACC	167
GmZIP12	AGGAGGAATCAGGGGATGTG	GAAGCTCCCATAGAAATTCC	178
GmZIP20	TCCTTTTACCAACCATGGCCT	TCAACAATGGGAGGCTCACC	150
GmZIP23	TGACCAACCGCGTGATGCTG	GTGATGATGATCAGAGCTAG	127
GmZIP25	GAAGTGTAGGACTACTAGTG	GTCTCATGATGAGCATGGTG	126
Glyma.18G290800（Actin）	GCACCACCGGAGAGAAAATA	GTGCACAATTGATGGACCAG	123

基因 Gene	上游引物 Forward primer （5'-3'）	下游引物 Reverse primer （5'-3'）	片段大小 Size （bp）
GmZIP6	GCGTGTTGTGTCCTTCTTTTAC	CTTAACAATGGGAGGCTCACC	167
GmZIP12	AGGAGGAATCAGGGGATGTG	GAAGCTCCCATAGAAATTCC	178
GmZIP20	TCCTTTTACCAACCATGGCCT	TCAACAATGGGAGGCTCACC	150
GmZIP23	TGACCAACCGCGTGATGCTG	GTGATGATGATCAGAGCTAG	127
GmZIP25	GAAGTGTAGGACTACTAGTG	GTCTCATGATGAGCATGGTG	126
Glyma.18G290800（Actin）	GCACCACCGGAGAGAAAATA	GTGCACAATTGATGGACCAG	123

基因 Gene	基因ID Gene ID	氨基酸数量 Amino acid	相对分子质量 Mw（Da）	等电点 pI	不稳定系数 Instability index	脂肪系数 Aliphatic index	亲水性 Hydropathicity	亚细胞定位 Subcellular location
GmZIP1	Glyma.02G126000	360	38 468.30	7.62	26.61	104.06	0.551	细胞膜
GmZIP2	Glyma.05G137400	485	51 989.60	6.00	31.08	95.96	-0.033	细胞膜
GmZIP3	Glyma.06G052000	478	51 484.51	6.57	52.00	94.14	0.340	细胞膜
GmZIP4	Glyma.07G223200	356	38 455.58	8.61	33.40	108.54	0.569	细胞膜
GmZIP5	Glyma.08G092700	485	52 353.64	5.79	35.60	92.95	-0.139	细胞膜
GmZIP6	Glyma.08G164400	361	38 404.85	6.38	40.99	102.16	0.446	细胞膜
GmZIP7	Glyma.08G328000	349	37 559.76	6.06	32.77	107.65	0.668	细胞膜
GmZIP8	Glyma.09G271900	598	62 182.12	7.19	30.11	112.26	0.777	细胞膜
GmZIP9	Glyma.11G132500	272	28 959.34	9.10	47.79	114.38	0.693	细胞膜
GmZIP10	Glyma.11G169300	326	34 776.07	5.81	29.45	113.96	0.769	液泡膜
GmZIP11	Glyma.12G056900	256	27 593.55	9.19	45.62	114.26	0.586	细胞膜
GmZIP12	Glyma.13G004400	347	37 633.21	7.72	37.42	110.55	0.455	细胞膜
GmZIP13	Glyma.13G338300	350	37 989.51	8.43	30.78	105.63	0.549	细胞膜
GmZIP14	Glyma.13G340900	276	29 153.48	8.78	38.44	118.04	0.740	细胞膜
GmZIP15	Glyma.14G094902	135	14 389.27	6.50	43.26	120.00	1.076	液泡膜
GmZIP16	Glyma.14G196200	324	33 863.86	6.35	34.60	114.51	0.773	细胞膜
GmZIP17	Glyma.15G033500	276	29 241.54	7.95	37.60	117.68	0.717	细胞膜
GmZIP18	Glyma.15G036200	345	37 385.49	6.82	32.57	101.22	0.462	细胞膜
GmZIP19	Glyma.15G036300	342	36 681.97	8.18	31.29	106.08	0.617	细胞膜
GmZIP20	Glyma.15G262800	359	38 243.55	6.14	39.12	104.09	0.486	细胞膜
GmZIP21	Glyma.17G228600	393	43 130.94	5.73	40.52	98.52	0.475	细胞膜
GmZIP22	Glyma.18G060300	328	35 036.37	5.96	29.37	112.07	0.742	细胞膜
GmZIP23	Glyma.18G078600	360	38 721.95	6.08	35.02	108.97	0.577	细胞膜
GmZIP24	Glyma.18G217100	598	62 115.03	6.93	29.44	112.59	0.781	细胞膜
GmZIP25	Glyma.20G022500	358	38 705.73	6.86	34.80	105.20	0.548	细胞膜
GmZIP26	Glyma.20G063100	354	38 301.74	6.30	28.63	105.56	0.399	细胞膜

基因 Gene	基因ID Gene ID	氨基酸数量 Amino acid	相对分子质量 Mw（Da）	等电点 pI	不稳定系数 Instability index	脂肪系数 Aliphatic index	亲水性 Hydropathicity	亚细胞定位 Subcellular location
GmZIP1	Glyma.02G126000	360	38 468.30	7.62	26.61	104.06	0.551	细胞膜
GmZIP2	Glyma.05G137400	485	51 989.60	6.00	31.08	95.96	-0.033	细胞膜
GmZIP3	Glyma.06G052000	478	51 484.51	6.57	52.00	94.14	0.340	细胞膜
GmZIP4	Glyma.07G223200	356	38 455.58	8.61	33.40	108.54	0.569	细胞膜
GmZIP5	Glyma.08G092700	485	52 353.64	5.79	35.60	92.95	-0.139	细胞膜
GmZIP6	Glyma.08G164400	361	38 404.85	6.38	40.99	102.16	0.446	细胞膜
GmZIP7	Glyma.08G328000	349	37 559.76	6.06	32.77	107.65	0.668	细胞膜
GmZIP8	Glyma.09G271900	598	62 182.12	7.19	30.11	112.26	0.777	细胞膜
GmZIP9	Glyma.11G132500	272	28 959.34	9.10	47.79	114.38	0.693	细胞膜
GmZIP10	Glyma.11G169300	326	34 776.07	5.81	29.45	113.96	0.769	液泡膜
GmZIP11	Glyma.12G056900	256	27 593.55	9.19	45.62	114.26	0.586	细胞膜
GmZIP12	Glyma.13G004400	347	37 633.21	7.72	37.42	110.55	0.455	细胞膜
GmZIP13	Glyma.13G338300	350	37 989.51	8.43	30.78	105.63	0.549	细胞膜
GmZIP14	Glyma.13G340900	276	29 153.48	8.78	38.44	118.04	0.740	细胞膜
GmZIP15	Glyma.14G094902	135	14 389.27	6.50	43.26	120.00	1.076	液泡膜
GmZIP16	Glyma.14G196200	324	33 863.86	6.35	34.60	114.51	0.773	细胞膜
GmZIP17	Glyma.15G033500	276	29 241.54	7.95	37.60	117.68	0.717	细胞膜
GmZIP18	Glyma.15G036200	345	37 385.49	6.82	32.57	101.22	0.462	细胞膜
GmZIP19	Glyma.15G036300	342	36 681.97	8.18	31.29	106.08	0.617	细胞膜
GmZIP20	Glyma.15G262800	359	38 243.55	6.14	39.12	104.09	0.486	细胞膜
GmZIP21	Glyma.17G228600	393	43 130.94	5.73	40.52	98.52	0.475	细胞膜
GmZIP22	Glyma.18G060300	328	35 036.37	5.96	29.37	112.07	0.742	细胞膜
GmZIP23	Glyma.18G078600	360	38 721.95	6.08	35.02	108.97	0.577	细胞膜
GmZIP24	Glyma.18G217100	598	62 115.03	6.93	29.44	112.59	0.781	细胞膜
GmZIP25	Glyma.20G022500	358	38 705.73	6.86	34.80	105.20	0.548	细胞膜
GmZIP26	Glyma.20G063100	354	38 301.74	6.30	28.63	105.56	0.399	细胞膜

Gene	TT_T_vs_TCK_T_Log₂FC	TT_T_vs_TCK_T_regulated	ST_T_vs_SCK_T_Log₂FC	ST_T_vs_SCK_T_regulated
GmZIP6	1.24	Up	0.39	False
GmZIP12	1.82	Up	-0.10	False
GmZIP20	0.898	False	1.14	Up
GmZIP23	-0.73	False	-1.01	Down
GmZIP25	4.25	Up	2.72	Up