大蒜NAC基因家族的鉴定与低温表达分析

doi:10.13304/j.nykjdb.2021.0794

摘要/Abstract

摘要：

为挖掘大蒜NAC转录因子家族成员，研究其在低温胁迫下的表达特性，基于大蒜转录组的测序结果，对低温胁迫下大蒜NAC转录因子家族的响应进行分析。结果表明，共鉴定出49个大蒜NAC基因，根据系统发育特征将其分为10个亚族。大蒜NAC蛋白的序列长度为81~619 aa，分子量9 293.66~70 229.04 Da，且均为亲水蛋白。保守结构域分析发现，大部分NAC蛋白在N端具有保守性，其中18个NAC蛋白在N端均有motif 3、motif 4、motif 1、motif 2、motif 5保守域。从中随机挑选6个蛋白（AsNAC001、AsNAC010、AsNAC013、AsNAC014、AsNAC017、AsNAC047）对其进行蛋白二级及三级结构的预测，发现无规则卷曲是构成大蒜NAC蛋白的重要组成部分，其次是α-螺旋和延长链，且这6个NAC蛋白具有高度的相似性。实时荧光定量分析发现，检测的6个NAC基因中有4个（AsNAC001、AsNAC010、AsNAC013、AsNAC014）在低温胁迫12和24 h时显著上调表达；2个（AsNAC017、AsNAC047）在低温胁迫4 h时显著下调表达，其中AsNAC010基因的表达量显著高于其他基因，说明NAC转录因子在大蒜响应低温胁迫的过程中起重要调控作用，为大蒜NAC蛋白的生理调控功能的分析奠定了理论基础。

关键词: 大蒜, NAC基因家族, 低温胁迫, 基因表达

Abstract:

To dig up the members of NAC family of transcription factors in Allium sativum L. and discuss the different expression in low temperature stress，NAC transcription factors were identified based on the result of transcriptome sequencing， and the different expression was analyzed. The results showed that 49 members of NAC gene family were screened in Allium sativum L.， which were divided into 10 subfamilies. The length of sequences varied and ranged from 81 to 613 aa， and the molecular weights were 9 293.6~70 229.04 Da. The total average hydrophobic index were less than 0， which indicated all the NAC proteins were hydrophilic proteins. According to the analysis of conserved domain， most of them had conserved N-terminal， of which 18 NAC proteins had motif 3， motif 4， motif 1， motif 2， and motif 5 conserved domains at the N-terminu. AsNAC001， AsNAC010， AsNAC013， AsNAC014， AsNAC017 and AsNAC047 were randomly selected for predicting secondary and tertiary structures， and the results showed the random coil was the main component， and the alpha helix and extended strand were followed， which meant NAC proteins were highly similar to each other. The real-time fluorescence quantification showed 4 genes containing AsNAC001， AsNAC010， AsNAC013 and AsNAC014 were significantly upregulate expression under low temperature stress 12 and 24 h， while 2 genes containing AsNAC017 and AsNAC047 were significantly down-regulate expression under low temperature stress 4 h. Among them， the expression level of AsNAC010 gene was significantly higher than others. All the results indicated that the NAC transcription factors played an important role in responsing to low temperature stress of Allium sativum L.， which provided a theoretical foundation for further study on physiological regulation function of NAC genes in Allium sativum L..

Key words: Allium sativum L., NAC gene family, low temperature stress, gene expression

中图分类号:

S633.4

闫艺薇, 田洁. 大蒜NAC基因家族的鉴定与低温表达分析[J]. 中国农业科技导报, 2023, 25(4): 67-76.

Yiwei YAN, Jie TIAN. Identification and Expression Analysis of NAC Gene Family Under Low Temperature in Allium sativum L.[J]. Journal of Agricultural Science and Technology, 2023, 25(4): 67-76.

图/表 9

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基因名称 Gene name	氨基酸长度 Number of amino acid/aa	编码区长度 Length of cDNA/bp	等电点 Theoretical pI	分子量 Molecular weight/Da	不稳定指数 Instability index	脂溶系数 Aliphatic index	总平均疏水指数Grand average of hydropathicity
AsNAC001	317	966	5.48	36 739.14	40.50	60.60	-0.939
AsNAC002	227	692	9.18	25 415.27	48.38	56.65	-0.654
AsNAC003	314	966	6.22	36 414.20	35.74	67.36	-0.662
AsNAC004	299	911	6.34	34 235.70	29.90	57.06	-0.716
AsNAC005	297	905	4.98	34 391.81	44.56	64.31	-0.904
AsNAC006	181	555	9.86	20 697.99	25.75	77.07	-0.480
AsNAC007	284	866	5.93	32 876.85	38.61	63.84	-0.816
AsNAC008	174	533	9.83	19 776.87	33.91	74.08	-0.466
AsNAC009	295	899	8.61	33 523.98	39.65	65.15	-0.637
AsNAC010	220	670	7.17	25 613.94	35.54	66.45	-0.780
AsNAC011	81	247	9.79	9 293.66	29.75	53.09	-0.793
AsNAC012	350	1 067	4.95	40 488.80	43.51	69.63	-0.827
AsNAC013	299	914	8.07	34 120.71	37.43	63.24	-0.581
AsNAC014	311	948	6.83	35 302.75	45.96	74.63	-0.591
AsNAC015	153	466	9.60	17 763.48	36.73	72.61	-0.460
AsNAC016	142	433	9.54	16 467.63	30.22	68.59	-0.706
AsNAC017	241	735	6.17	27 870.55	40.27	59.83	-0.651
AsNAC018	234	713	5.74	27 139.67	41.73	55.77	-0.631
AsNAC019	196	597	8.62	22 554.44	21.80	61.68	-0.671
AsNAC020	256	780	6.34	28 965.66	40.18	63.52	-0.550
AsNAC021	205	625	8.75	23 361.40	38.21	60.78	-0.573
AsNAC022	185	567	9.45	21 445.02	20.73	72.16	-0.374
AsNAC023	159	484	9.47	18 236.02	23.81	61.95	-0.664
AsNAC024	151	460	9.40	17 500.04	36.29	57.48	-0.788
AsNAC025	147	448	9.70	16 631.15	31.41	69.12	-0.589
AsNAC026	322	985	8.94	36 669.70	40.31	68.17	-0.567
AsNAC027	256	780	8.57	29 357.41	30.19	61.37	-0.656
AsNAC028	144	439	9.10	16 917.46	34.03	56.25	-0.816
AsNAC029	156	475	9.67	17 769.38	43.29	73.78	-0.574
AsNAC030	198	603	5.25	22 305.54	44.75	62.47	-0.819
AsNAC031	619	1 887	4.78	70 229.04	46.44	70.39	-0.709
AsNAC032	176	542	9.19	20 419.75	38.67	73.64	-0.274
AsNAC033	238	728	9.17	27 182.77	34.76	60.21	-0.607
AsNAC034	272	835	7.80	31 081.00	43.57	64.89	-0.604
AsNAC035	209	637	7.00	23 813.71	30.07	73.68	-0.678
AsNAC036	276	841	4.95	31 051.17	32.52	67.50	-0.791

基因名称 Gene name	氨基酸长度 Number of amino acid/aa	编码区长度 Length of cDNA/bp	等电点 Theoretical pI	分子量 Molecular weight/Da	不稳定指数 Instability index	脂溶系数 Aliphatic index	总平均疏水指数Grand average of hydropathicity
AsNAC001	317	966	5.48	36 739.14	40.50	60.60	-0.939
AsNAC002	227	692	9.18	25 415.27	48.38	56.65	-0.654
AsNAC003	314	966	6.22	36 414.20	35.74	67.36	-0.662
AsNAC004	299	911	6.34	34 235.70	29.90	57.06	-0.716
AsNAC005	297	905	4.98	34 391.81	44.56	64.31	-0.904
AsNAC006	181	555	9.86	20 697.99	25.75	77.07	-0.480
AsNAC007	284	866	5.93	32 876.85	38.61	63.84	-0.816
AsNAC008	174	533	9.83	19 776.87	33.91	74.08	-0.466
AsNAC009	295	899	8.61	33 523.98	39.65	65.15	-0.637
AsNAC010	220	670	7.17	25 613.94	35.54	66.45	-0.780
AsNAC011	81	247	9.79	9 293.66	29.75	53.09	-0.793
AsNAC012	350	1 067	4.95	40 488.80	43.51	69.63	-0.827
AsNAC013	299	914	8.07	34 120.71	37.43	63.24	-0.581
AsNAC014	311	948	6.83	35 302.75	45.96	74.63	-0.591
AsNAC015	153	466	9.60	17 763.48	36.73	72.61	-0.460
AsNAC016	142	433	9.54	16 467.63	30.22	68.59	-0.706
AsNAC017	241	735	6.17	27 870.55	40.27	59.83	-0.651
AsNAC018	234	713	5.74	27 139.67	41.73	55.77	-0.631
AsNAC019	196	597	8.62	22 554.44	21.80	61.68	-0.671
AsNAC020	256	780	6.34	28 965.66	40.18	63.52	-0.550
AsNAC021	205	625	8.75	23 361.40	38.21	60.78	-0.573
AsNAC022	185	567	9.45	21 445.02	20.73	72.16	-0.374
AsNAC023	159	484	9.47	18 236.02	23.81	61.95	-0.664
AsNAC024	151	460	9.40	17 500.04	36.29	57.48	-0.788
AsNAC025	147	448	9.70	16 631.15	31.41	69.12	-0.589
AsNAC026	322	985	8.94	36 669.70	40.31	68.17	-0.567
AsNAC027	256	780	8.57	29 357.41	30.19	61.37	-0.656
AsNAC028	144	439	9.10	16 917.46	34.03	56.25	-0.816
AsNAC029	156	475	9.67	17 769.38	43.29	73.78	-0.574
AsNAC030	198	603	5.25	22 305.54	44.75	62.47	-0.819
AsNAC031	619	1 887	4.78	70 229.04	46.44	70.39	-0.709
AsNAC032	176	542	9.19	20 419.75	38.67	73.64	-0.274
AsNAC033	238	728	9.17	27 182.77	34.76	60.21	-0.607
AsNAC034	272	835	7.80	31 081.00	43.57	64.89	-0.604
AsNAC035	209	637	7.00	23 813.71	30.07	73.68	-0.678
AsNAC036	276	841	4.95	31 051.17	32.52	67.50	-0.791

基因名称 Gene name	氨基酸长度 Number of amino acid/aa	编码区长度 Length of cDNA/bp	等电点 Theoretical pI	分子量 Molecular weight/Da	不稳定指数 Instability index	脂溶系数 Aliphatic index	总平均疏水指数Grand average of hydropathicity
AsNAC037	290	884	6.45	33 210.39	45.73	63.59	-0.694
AsNAC038	176	548	9.74	20 481.63	39.16	79.77	-0.354
AsNAC039	147	448	9.72	16 705.32	31.72	70.41	-0.582
AsNAC040	179	555	9.74	21 143.43	51.78	68.60	-0.450
AsNAC041	152	463	7.78	17 151.33	42.10	56.51	-0.648
AsNAC042	145	442	9.46	16 896.43	38.54	62.48	-0.587
AsNAC043	149	454	9.46	17 444.08	36.77	63.42	-0.533
AsNAC044	130	396	9.80	15 114.11	16.76	50.23	-0.893
AsNAC045	149	454	9.39	17 409.75	35.18	66.04	-0.799
AsNAC046	140	426	9.43	16 452.80	46.91	63.36	-0.699
AsNAC047	251	765	5.24	28 836.27	31.96	54.74	-0.734
AsNAC048	260	792	5.04	29 891.58	47.35	94.42	-0.202
AsNAC049	261	796	8.63	29 843.57	37.33	67.59	-0.593

基因名称 Gene name	氨基酸长度 Number of amino acid/aa	编码区长度 Length of cDNA/bp	等电点 Theoretical pI	分子量 Molecular weight/Da	不稳定指数 Instability index	脂溶系数 Aliphatic index	总平均疏水指数Grand average of hydropathicity
AsNAC037	290	884	6.45	33 210.39	45.73	63.59	-0.694
AsNAC038	176	548	9.74	20 481.63	39.16	79.77	-0.354
AsNAC039	147	448	9.72	16 705.32	31.72	70.41	-0.582
AsNAC040	179	555	9.74	21 143.43	51.78	68.60	-0.450
AsNAC041	152	463	7.78	17 151.33	42.10	56.51	-0.648
AsNAC042	145	442	9.46	16 896.43	38.54	62.48	-0.587
AsNAC043	149	454	9.46	17 444.08	36.77	63.42	-0.533
AsNAC044	130	396	9.80	15 114.11	16.76	50.23	-0.893
AsNAC045	149	454	9.39	17 409.75	35.18	66.04	-0.799
AsNAC046	140	426	9.43	16 452.80	46.91	63.36	-0.699
AsNAC047	251	765	5.24	28 836.27	31.96	54.74	-0.734
AsNAC048	260	792	5.04	29 891.58	47.35	94.42	-0.202
AsNAC049	261	796	8.63	29 843.57	37.33	67.59	-0.593

编号 Code	α-螺旋 Alpha helix	延长链 Extended strand	无规卷曲 Random coil	β-转角 Beta turn
AsNAC001	31.55	9.46	54.89	4.10
AsNAC010	32.73	15.00	48.18	4.09
AsNAC013	16.05	15.38	64.88	3.68
AsNAC014	24.44	13.83	58.20	3.54
AsNAC017	13.69	20.33	62.24	3.73
AsNAC047	17.53	19.52	58.96	3.98