陆地棉BGLU基因家族成员的全基因组鉴定与表达分析

doi:10.13304/j.nykjdb.2021.0699

摘要/Abstract

摘要：

植物BGLU基因在生物和非生物胁迫防御中起着重要作用，利用生物信息学手段对陆地棉（Gossypium hirsutum） BGLU家族成员进行全基因组鉴定，对序列特征、保守域结构、染色体定位、启动子元件等家族特征进行了分析，并进一步结合转录组数据和qRT-PCR分析了BGLU基因家族的病菌胁迫响应模式。结果表明，共鉴定出53个陆地棉BGLU基因，根据系统发育进化关系分为6个亚族，部分基因在病菌胁迫下特异性表达。荧光定量和转录组测序结果表明，GhBGLU5、GhBGLU14、GhBGLU18、GhBGLU26、GhBGLU34在抗病棉花中的表达水平显著高于感病品种，推测这些基因正向调控棉花黄萎病抗性。上述结果为进一步分析棉花BGLU家族基因的功能以及分子机制提供一定的理论基础。

关键词: 陆地棉, BGLU基因家族, 系统发育, 表达分析

Abstract:

BGLU genes play crucial role in the defense of biotic and abiotic stresses， this study identified the members of the BGLU family in cotton via bioinformatics methods. The sequence features， conserved domain structure， chromosome location， promoter elements and other family features were analyzed， and the transcriptome data and qRT-PCR were used to analyze the pathogenic stress response pattern of BGLU family genes. The results showed a total of 53 BGLU genes were identified and divided into 6 sub-families according to the phylogenetic relationship in upland cotton， and some genes were specifically expressed under pathogen stress. qPCR analysis and RNAseq data showed that GhBGLU5， GhBGLU14， GhBGLU18， GhBGLU26 and GhBGLU34 maintained higher expression level in resistant and susceptible cottons， implying that these genes positively regulated cotton Verticillium wilt resistance. This study provided theoretical basis for further analyzing the functions and molecular mechanisms of the BGLU genes in cotton.

Key words: Gossypium hirsutum, BGLU gene family, phylogenetic analysis, expression profiling

中图分类号:

S562

张曼, 王志城, 刘正文, 王国宁, 王省芬, 张艳. 陆地棉BGLU基因家族成员的全基因组鉴定与表达分析[J]. 中国农业科技导报, 2023, 25(2): 48-59.

Man ZHANG, Zhicheng WANG, Zhengwen LIU, Guoning WANG, Xingfen WANG, Yan ZHANG. Genome-Wide Identification and Analysis of BGLU Genes Family in Gossypium hirsutum[J]. Journal of Agricultural Science and Technology, 2023, 25(2): 48-59.

图/表 10

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引物名称 Primer name	引物序列 Primer sequence（3’-5’）
GhUBQ14-F	CAACGCTCCATCTTGTCCTT
GhUBQ14-R	TGATCGTCTTTCCCGTAAGC
GhBGLU9-RT-F	GGCGGTGATGGACACTGGA
GhBGLU9-RT-R	GCCACCCTTATTAGCCATCC
GhBGLU19-RT-F	CGTAGAACATGATGATTCCAACACT
GhBGLU19-RT-R	CCACCTTCGTTCACACCTCCTT
GhBGLU05-RT-F	GTTTTATTTTCGGTGCCGGAT
GhBGLU05-RT-R	GATGTTGTCTAACGAGAATGTCCC
GhBGLU23-RT-F	CGAGGGAGCAGTGAAGGAGGAT
GhBGLU23-RT-R	TATCTTCAACATAGCGGTGGTACT
GhBGLU26-RT-F	AAGATGGAAAAGGCCTAAGCAAC
GhBGLU26-RT-R	GCATCATCTCCATTCTCATTGTTTT
GhBGLU34-RT-F	TGTGTTCTATCTGTTGTTCCTTTGG
GhBGLU34-RT-R	CTGTATCTCCATTGCTTCCATCCAT
GhBGLU52-RT-F	CTGAAACAAAGCCTGGGTAATGGTA
GhBGLU52-RT-R	CTTCATACTGGTAAGCAGAGGAGGC
GhBGLU35-RT-F	GTGAATGAAGGAAACAAGGGAGATA
GhBGLU35-RT-R	AAGCATCCATTCCCAGATCTTTCAT
GhBGLU14-RT-F	GTTCTTCTTCTTACTCAACTTCATTTC
GhBGLU14-RT-R	TTCATACTGGTAAGCGGAAGAGGC

引物名称 Primer name	引物序列 Primer sequence（3’-5’）
GhUBQ14-F	CAACGCTCCATCTTGTCCTT
GhUBQ14-R	TGATCGTCTTTCCCGTAAGC
GhBGLU9-RT-F	GGCGGTGATGGACACTGGA
GhBGLU9-RT-R	GCCACCCTTATTAGCCATCC
GhBGLU19-RT-F	CGTAGAACATGATGATTCCAACACT
GhBGLU19-RT-R	CCACCTTCGTTCACACCTCCTT
GhBGLU05-RT-F	GTTTTATTTTCGGTGCCGGAT
GhBGLU05-RT-R	GATGTTGTCTAACGAGAATGTCCC
GhBGLU23-RT-F	CGAGGGAGCAGTGAAGGAGGAT
GhBGLU23-RT-R	TATCTTCAACATAGCGGTGGTACT
GhBGLU26-RT-F	AAGATGGAAAAGGCCTAAGCAAC
GhBGLU26-RT-R	GCATCATCTCCATTCTCATTGTTTT
GhBGLU34-RT-F	TGTGTTCTATCTGTTGTTCCTTTGG
GhBGLU34-RT-R	CTGTATCTCCATTGCTTCCATCCAT
GhBGLU52-RT-F	CTGAAACAAAGCCTGGGTAATGGTA
GhBGLU52-RT-R	CTTCATACTGGTAAGCAGAGGAGGC
GhBGLU35-RT-F	GTGAATGAAGGAAACAAGGGAGATA
GhBGLU35-RT-R	AAGCATCCATTCCCAGATCTTTCAT
GhBGLU14-RT-F	GTTCTTCTTCTTACTCAACTTCATTTC
GhBGLU14-RT-R	TTCATACTGGTAAGCGGAAGAGGC

基因名称 Gene name	基因ID号 Gene ID	氨基酸数目 Number of amino acids	分子量 Molecular weight/kD	等电点 pI	信号肽 Signal peptide	亚细胞定位 Subcellular localization
GhBGLU01	Ghir_A01G020950	359	42.213	8.740	35~36	细胞质膜Plasma membrane
GhBGLU02	Ghir_A01G021100	495	57.410	6.123	23~24	液泡Vacuole
GhBGLU03	Ghir_A01G021370	502	58.129	7.219	23~24	液泡Vacuole
GhBGLU04	Ghir_A02G012630	390	44.336	5.309	38~39	细胞质Cytoplasmic
GhBGLU05	Ghir_A05G007460	476	54.519	5.022	21~22	液泡Vacuole
GhBGLU06	Ghir_A06G012310	491	55.875	9.314	23~24	溶酶体Lysosomal
GhBGLU07	Ghir_A06G015860	527	59.958	6.555	26~27	细胞质膜Plasma membrane
GhBGLU08	Ghir_A06G015870	494	56.087	6.519	21~22	液泡Vacuole
GhBGLU09	Ghir_A07G008330	331	37.350	9.361	19~20	胞外Extracellular
GhBGLU10	Ghir_A07G008340	497	56.916	8.338	19~20	液泡Vacuole
GhBGLU11	Ghir_A08G024630	520	58.768	7.434	28~29	细胞质Cytoplasmic
GhBGLU12	Ghir_A08G025080	371	41.575	6.883	26~27	细胞质膜Plasma membrane

基因名称 Gene name	基因ID号 Gene ID	氨基酸数目 Number of amino acids	分子量 Molecular weight/kD	等电点 pI	信号肽 Signal peptide	亚细胞定位 Subcellular localization
GhBGLU01	Ghir_A01G020950	359	42.213	8.740	35~36	细胞质膜Plasma membrane
GhBGLU02	Ghir_A01G021100	495	57.410	6.123	23~24	液泡Vacuole
GhBGLU03	Ghir_A01G021370	502	58.129	7.219	23~24	液泡Vacuole
GhBGLU04	Ghir_A02G012630	390	44.336	5.309	38~39	细胞质Cytoplasmic
GhBGLU05	Ghir_A05G007460	476	54.519	5.022	21~22	液泡Vacuole
GhBGLU06	Ghir_A06G012310	491	55.875	9.314	23~24	溶酶体Lysosomal
GhBGLU07	Ghir_A06G015860	527	59.958	6.555	26~27	细胞质膜Plasma membrane
GhBGLU08	Ghir_A06G015870	494	56.087	6.519	21~22	液泡Vacuole
GhBGLU09	Ghir_A07G008330	331	37.350	9.361	19~20	胞外Extracellular
GhBGLU10	Ghir_A07G008340	497	56.916	8.338	19~20	液泡Vacuole
GhBGLU11	Ghir_A08G024630	520	58.768	7.434	28~29	细胞质Cytoplasmic
GhBGLU12	Ghir_A08G025080	371	41.575	6.883	26~27	细胞质膜Plasma membrane

基因名称 Gene name	基因ID号 Gene ID	氨基酸数目 Number of amino acids	分子量 Molecular weight/kD	等电点 pI	信号肽 Signal peptide	亚细胞定位 Subcellular localization
GhBGLU13	Ghir_A09G005920	506	58.313	9.323	20~21	溶酶体Lysosomal
GhBGLU14	Ghir_A10G001000	536	61.583	6.510	29~30	细胞质膜Plasma membrane
GhBGLU15	Ghir_A10G001010	500	57.229	7.590	27~28	液泡Vacuole
GhBGLU16	Ghir_A10G001020	536	61.339	6.976	24~25	溶酶体Lysosomal
GhBGLU17	Ghir_A11G032270	514	58.455	6.337	26~27	细胞质Cytoplasmic
GhBGLU18	Ghir_A11G033190	199	23.264	4.694	40~41	细胞质Cytoplasmic
GhBGLU19	Ghir_A11G033200	195	22.294	6.799	37~38	细胞质Cytoplasmic
GhBGLU20	Ghir_A12G017270	132	15.366	7.963	19~20	胞外Extracellular
GhBGLU21	Ghir_A12G026530	482	55.028	9.470	23~24	细胞质膜Plasma membrane
GhBGLU22	Ghir_A13G008250	498	56.644	5.970	23~24	液泡Vacuole
GhBGLU23	Ghir_A13G008260	136	15.630	9.499	26~27	细胞质膜Plasma membrane
GhBGLU24	Ghir_A13G008270	504	56.842	7.635	24~25	液泡Vacuole
GhBGLU25	Ghir_A13G014930	383	43.308	4.834	22~23	细胞质膜Plasma membrane
GhBGLU26	Ghir_A13G015150	546	62.303	6.561	26~27	细胞质膜Plasma membrane
GhBGLU27	Ghir_D01G022500	445	51.828	8.334	18~19	细胞质膜Plasma membrane
GhBGLU28	Ghir_D01G022510	500	57.851	7.275	19~20	液泡Vacuole
GhBGLU29	Ghir_D01G022660	499	58.085	6.203	23~24	液泡Vacuole
GhBGLU30	Ghir_D01G022670	511	59.569	6.400	23~24	液泡Vacuole
GhBGLU31	Ghir_D03G006350	328	37.578	6.139	27~28	细胞质Cytoplasmic
GhBGLU32	Ghir_D05G007520	482	55.193	5.549	21~22	液泡Vacuole
GhBGLU33	Ghir_D06G012190	437	50.036	9.662	21~22	溶酶体Lysosomal
GhBGLU34	Ghir_D06G016620	483	54.692	6.788	16~17	细胞质膜Plasma membrane
GhBGLU35	Ghir_D06G016640	534	60.754	6.782	21~22	细胞质膜Plasma membrane
GhBGLU36	Ghir_D07G008440	139	15.520	8.918	19~20	胞外Extracellular
GhBGLU37	Ghir_D07G008450	479	54.695	8.085	19~20	溶酶体Lysosomal
GhBGLU38	Ghir_D08G025520	514	57.962	6.515	28~29	细胞质膜Plasma membrane
GhBGLU39	Ghir_D08G025990	463	52.616	7.511	26~27	细胞质膜Plasma membrane
GhBGLU40	Ghir_D09G005490	341	39.515	9.728	24~25	胞外Extracellular
GhBGLU41	Ghir_D10G001780	466	53.328	5.882	29~30	细胞质膜Plasma membrane
GhBGLU42	Ghir_D10G001790	509	58.366	7.255	27~28	细胞质膜Plasma membrane
GhBGLU43	Ghir_D10G001800	518	58.984	6.416	24~25	溶酶体Lysosomal
GhBGLU44	Ghir_D11G032750	514	58.322	6.206	28~29	细胞质膜Plasma membrane
GhBGLU45	Ghir_D11G033820	490	56.008	5.179	38~29	细胞质Cytoplasmic
GhBGLU46	Ghir_D12G017490	121	13.931	7.500	47~48	胞外Extracellular
GhBGLU47	Ghir_D12G026540	482	54.786	9.368	23~24	细胞质膜Plasma membrane
GhBGLU48	Ghir_D12G026620	472	53.808	9.573	23~24	细胞质膜Plasma membrane
GhBGLU49	Ghir_D13G008490	504	56.847	6.680	24~25	液泡Vacuole
GhBGLU50	Ghir_D13G008500	205	23.526	8.424	37~38	细胞质Cytoplasmic
GhBGLU51	Ghir_D13G008510	498	56.553	5.218	23~24	液泡Vacuole
GhBGLU52	Ghir_D13G015630	488	55.822	8.565	22~23	细胞质膜Plasma membrane
GhBGLU53	Ghir_D13G015870	546	61.929	6.658	26~27	细胞质膜Plasma membrane