工业大麻HD-Zip基因家族鉴定与干旱胁迫下的表达分析

doi:10.7668/hbnxb.20194303

摘要/Abstract

摘要：

HD-Zip在植物抗逆过程中起着重要的调节作用,为了解大麻基因组中HD-Zip基因家族的特征和潜在功能,并为进一步研究CsHDZ基因对干旱胁迫下的调控提供重要线索,根据大麻全基因组数据库中的参考序列,利用生物信息学的方法对大麻HD-Zip基因家族进行全基因组鉴定,并系统分析其理化特性、进化发育、基因结构、启动子顺式结合元件并测定干旱胁迫下的表达模式。结果表明,大麻HD-Zip基因家族共筛选出20个CsHDZs基因,分别定位在7条染色体上,属于4个亚家族,编码氨基酸204 ~ 837 个,理论等电点4.54~9.04,属不稳定蛋白,亚细胞定位预测全部定位于细胞核;蛋白保守基序分析显示,CsHDZs共有10个保守基序,其中motif 1和motif 9为各成员共有;启动子调控元件分析发现,CsHDZs富含多种逆境胁迫应答相关元件。实时荧光定量分析结果显示,CsHDZ1/8/10基因转录水平在干旱前期无显著差异,而后期显著上调。研究表明CsHDZs基因参与了胁迫应答过程并能够积极响应干旱。

关键词: 大麻, HD-Zip基因, 干旱胁迫, 表达分析

Abstract:

HD-Zip transcription factors play an important regulatory role in stress resistance in plants,in order to reveal the characteristics and potential functions of the HD-Zip gene family in the hemp genome,and to provide important clues for further study on the regulation of CsHDZ gene under drought stress.Here this study based on the reference sequence in the hemp whole genome database,perform a genome-wide analysis of HD-Zip gene family by bioinformatics methods in hemp,and their physical and chemical characteristics,evolutionary relationship,gene structure,the cis-acting elements and transcript patterns in drought stress were analyzed.The results showed that a total of 20 hemp HD-Zips (CsHDZs),they were non-evenly distributed on seven chromosomes,and classified into four distinctive subfamilies.Their amino acid sequence size and isoelectric point were 204—837 aa and 4.54—9.04,they were all unstable proteins,and subcellular localization analysis indicated that HD-Zip protein was targeted to the nucleus.Analysis of protein conserved motifs showed that there was a total of 10 conserved motifs in CsHDZs,of which motif 1 and motif 9 were shared by all numbers.In addition,the regulatory element analysis showed that a variety of elements related to abiotic stresses were identified in the promoter region of CdHDZs.The results of Real-time Fluorescence Quantitative analysis showed that the transcription level of CsHDZ1/8/10 gene had no significant difference in the early stage of drought,but was significantly upregulated in the late stage.The results indicated that CsHDZs gene was involved in the stress response process and could actively respond to drought.

Key words: Hemp, HD-Zip gene, Drought stress, Expression analysis

葛雯冬, 王腾辉, 高美玲, 范震宇, 王玉书. 工业大麻HD-Zip基因家族鉴定与干旱胁迫下的表达分析[J]. 华北农学报, 2024, 39(1): 55-62. doi: 10.7668/hbnxb.20194303.

GE Wendong, WANG Tenghui, GAO Meiling, FAN Zhenyu, WANG Yushu. Genome-Wide Identification and Expression Analysis of the HD-Zip Gene Family under Drought Stress in Hemp[J]. Acta Agriculturae Boreali-Sinica, 2024, 39(1): 55-62. doi: 10.7668/hbnxb.20194303.

http://www.hbnxb.net/CN/Y2024/V39/I1/55

图/表 9

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基因名称 Gene name	正向引物 (5'—3') Forward primer (5'—3')	反向引物 (5'—3') Reverse primer (5'—3')
EF1a	ACCAAGATTGACAGGCGTTC	CCTTCTTCTCCACAGCCTTG
CsHDZ1	TCATCATCACCCTCCAACCTCTC	CATCTTCTCCTCCATTGCCTTCTTC
CsHDZ8	CACCAACCGAACCAACCACAG	GGTCTTCAAACATCGTCCTCTCTC
CsHDZ10	AGTGGAATGGATGAGAATGCTGTTG	TAGGGTGCGATTTGGACTGGAG
CsHDZ17	GCGTCGTTTCAGCCTCAGTTTG	GGAGCTTGGTCGTCGGAGAAG
CsHDZ18	CGAGCCAAAGAAGAAGATTCAGTTG	TGTTTCGATTTCCATCTTGCTCTTC

基因名称 Gene name	正向引物 (5'—3') Forward primer (5'—3')	反向引物 (5'—3') Reverse primer (5'—3')
EF1a	ACCAAGATTGACAGGCGTTC	CCTTCTTCTCCACAGCCTTG
CsHDZ1	TCATCATCACCCTCCAACCTCTC	CATCTTCTCCTCCATTGCCTTCTTC
CsHDZ8	CACCAACCGAACCAACCACAG	GGTCTTCAAACATCGTCCTCTCTC
CsHDZ10	AGTGGAATGGATGAGAATGCTGTTG	TAGGGTGCGATTTGGACTGGAG
CsHDZ17	GCGTCGTTTCAGCCTCAGTTTG	GGAGCTTGGTCGTCGGAGAAG
CsHDZ18	CGAGCCAAAGAAGAAGATTCAGTTG	TGTTTCGATTTCCATCTTGCTCTTC

基因名称 Gene name	蛋白登录号 Protein ID	氨基酸数量/个 Number of amino acids	分子质量/ku MW	等电点 PI	不稳定指数 Instability Index	疏水系数 GRAVY	亚细胞定位 Subcellular localization
CsHDZ1	XP_030482567.1	309	35.24	6.42	58.49	-0.990	细胞核
CsHDZ2	XP_030483803.1	380	43.56	6.51	54.13	-1.244	细胞核
CsHDZ3	XP_030484314.1	231	26.64	8.37	69.94	-0.823	细胞核
CsHDZ4	XP_030484473.1	328	36.59	4.88	59.46	-0.933	细胞核
CsHDZ5	XP_030484657.1	321	36.24	4.88	47.02	-0.815	细胞核
CsHDZ6	XP_030484728.1	357	39.43	8.57	65.66	-0.795	细胞核
CsHDZ7	XP_030487585.1	247	28.80	9.04	61.62	-1.041	细胞核
CsHDZ8	XP_030489450.1	320	36.80	5.42	56.71	-1.090	细胞核
CsHDZ9	XP_030489869.1	333	36.87	7.30	48.15	-0.826	细胞核
CsHDZ10	XP_030491373.1	837	92.07	5.94	48.23	-0.134	细胞核
CsHDZ11	XP_030491770.1	755	83.58	5.64	48.05	-0.457	细胞核
CsHDZ12	XP_030492567.1	249	27.86	7.71	64.98	-0.847	细胞核
CsHDZ13	XP_030493216.1	347	39.13	4.54	59.28	-0.755	细胞核
CsHDZ14	XP_030496355.1	301	32.98	8.63	55.87	-0.654	细胞核
CsHDZ15	XP_030498752.1	327	38.01	6.07	47.73	-1.128	细胞核
CsHDZ16	XP_030499036.1	214	25.04	6.16	53.76	-0.907	细胞核
CsHDZ17	XP_030499944.1	348	39.34	5.27	59.86	-0.859	细胞核
CsHDZ18	XP_030500137.1	204	24.63	8.87	61.96	-1.624	细胞核
CsHDZ19	XP_030501039.1	330	36.77	8.21	62.81	-0.817	细胞核
CsHDZ20	XP_030510620.1	399	44.59	7.17	59.71	-1.114	细胞核

基因名称 Gene name	蛋白登录号 Protein ID	氨基酸数量/个 Number of amino acids	分子质量/ku MW	等电点 PI	不稳定指数 Instability Index	疏水系数 GRAVY	亚细胞定位 Subcellular localization
CsHDZ1	XP_030482567.1	309	35.24	6.42	58.49	-0.990	细胞核
CsHDZ2	XP_030483803.1	380	43.56	6.51	54.13	-1.244	细胞核
CsHDZ3	XP_030484314.1	231	26.64	8.37	69.94	-0.823	细胞核
CsHDZ4	XP_030484473.1	328	36.59	4.88	59.46	-0.933	细胞核
CsHDZ5	XP_030484657.1	321	36.24	4.88	47.02	-0.815	细胞核
CsHDZ6	XP_030484728.1	357	39.43	8.57	65.66	-0.795	细胞核
CsHDZ7	XP_030487585.1	247	28.80	9.04	61.62	-1.041	细胞核
CsHDZ8	XP_030489450.1	320	36.80	5.42	56.71	-1.090	细胞核
CsHDZ9	XP_030489869.1	333	36.87	7.30	48.15	-0.826	细胞核
CsHDZ10	XP_030491373.1	837	92.07	5.94	48.23	-0.134	细胞核
CsHDZ11	XP_030491770.1	755	83.58	5.64	48.05	-0.457	细胞核
CsHDZ12	XP_030492567.1	249	27.86	7.71	64.98	-0.847	细胞核
CsHDZ13	XP_030493216.1	347	39.13	4.54	59.28	-0.755	细胞核
CsHDZ14	XP_030496355.1	301	32.98	8.63	55.87	-0.654	细胞核
CsHDZ15	XP_030498752.1	327	38.01	6.07	47.73	-1.128	细胞核
CsHDZ16	XP_030499036.1	214	25.04	6.16	53.76	-0.907	细胞核
CsHDZ17	XP_030499944.1	348	39.34	5.27	59.86	-0.859	细胞核
CsHDZ18	XP_030500137.1	204	24.63	8.87	61.96	-1.624	细胞核
CsHDZ19	XP_030501039.1	330	36.77	8.21	62.81	-0.817	细胞核
CsHDZ20	XP_030510620.1	399	44.59	7.17	59.71	-1.114	细胞核

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