小麦类硫素基因家族鉴定及表达分析

doi:10.3969/j.issn.1004-1524.20230048

浙江农业学报 ›› 2024, Vol. 36 ›› Issue (4): 729-737.DOI: 10.3969/j.issn.1004-1524.20230048

小麦类硫素基因家族鉴定及表达分析

李晶晶¹(), 李闯², 路亚南², 郑文明²^,^*()

1.河南护理职业学院,河南安阳 455000
2.河南农业大学生命科学学院,小麦玉米作物学国家重点实验室,河南郑州 450046

收稿日期:2023-01-05 出版日期:2024-04-25 发布日期:2024-04-29
作者简介:李晶晶(1989—),女,河南安阳人,硕士,研究方向为生物化学与分子生物学。E-mail:371702918@qq.com
通讯作者: *郑文明,E-mail:wmzheng@henau.edu.cn
基金资助:
河南省重大科技专项(221100110100);小麦玉米作物学国家重点实验室课题“小麦抗病与养分高效种质资源发掘利用”

Identification and expression analysis of Thionin-like gene family in wheat

LI Jingjing¹(), LI Chuang², LU Yanan², ZHENG Wenming²^,^*()

1. Henan Vocational College of Nursing, Anyang 455000, Henan, China
2. State Key Laboratory of Wheat and Maize Crop Science, College of Life Sciences, Henan Agricultural University, Zhengzhou 450046, China

Received:2023-01-05 Online:2024-04-25 Published:2024-04-29
Contact: ZHENG Wenming

摘要/Abstract

摘要：

类硫素(thionin-like,Thil)在植物应对病原微生物侵染的防御反应中具有重要生物学功能。本研究通过拟南芥(Arabidopsis thaliana)的Thil基因序列,利用同源搜寻及生物信息学方法初步鉴定到普通小麦(Triticum aestivum L.)的Thil基因家族。结果表明,Thil基因家族在普通小麦中包含26个成员。基因序列结构分析得到Thil家族存在保守的基序结构,家族各成员基因编码的蛋白质序列包含79~196个氨基酸,大部分蛋白质的氨基酸数量少于140,其中20个蛋白质包含信号肽。顺式作用元件预测分析结果显示,小麦Thil基因的启动子调控元件受到脱落酸等逆境胁迫相应激素的影响,同时基因表达谱呈现出受到病原菌胁迫诱导后的多样性表达。通过进一步qRT-PCR验证分析表明, 普通小麦的TaThil-4A、TaThil-4B和TaThil-4D等3个基因在叶锈病菌早期侵染中被诱导表达。本研究结果可为小麦抗病育种及Thil基因家族的功能鉴定与应用提供参考。

关键词: 小麦, 类硫素, 抗病, 叶锈菌

Abstract:

Thionin-like (Thil) has important biological functions in plant defense response to pathogenic microorganisms. In this study, the Thil gene family of Triticum aestivum L. was initially identified using the homologous search and bioinformatics method based on Thil genes of the model plant Arabidopsis thaliana. A total of 26 Thil genes are included in the wheat genome. Structural analysis of gene sequences showed that the conserved motif structure exists in the Thil family, Thil genes encode proteins of 79-196 amino acids, most proteins have less than 140 amino acids, and 20 Thil proteins contain signal peptides. The cis-acting element prediction indicated that promoter elements of the Thil genes are mainly affected by corresponding hormones about adversity stress such as abscisic acid. Gene expression profiles showed that diversifying expression of the Thil family genes exists in response to pathogenic infection. qRT-PCR analysis showed that the expressions of TaThil-4A, TaThil-4B, and TaThil-4D were induced in the early response to wheat leaf rust fungus. The results of this study can provide a reference for wheat disease resistance breeding in wheat and functional study and application of the Thil gene family.

Key words: wheat, Thionin-like, disease-resistance, Puccinia triticina

中图分类号:

S512.1

李晶晶, 李闯, 路亚南, 郑文明. 小麦类硫素基因家族鉴定及表达分析[J]. 浙江农业学报, 2024, 36(4): 729-737.

LI Jingjing, LI Chuang, LU Yanan, ZHENG Wenming. Identification and expression analysis of Thionin-like gene family in wheat[J]. Acta Agriculturae Zhejiangensis, 2024, 36(4): 729-737.

图/表 7

参考文献 31

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基因名称	正向引物(5'→3')	反向引物(5'→3')
Gene name	Forward primer(5'→3')	Reverse primer(5'→3')
TaThil-4A	CCGTCCCAGCCATCGTG	TCTTGAGCATCCGCTTGGAG
TaThil-4B	GCAGCGGAGGATTGGAGAAT	AAGGGTCAAACCAAGCCTCA
TaThil-4D	GCGGAGGAGTGGAGAATCTG	ACGCCTACCAAATCAGGACA
Ta26S	GAAGAAGGTCCCAAGGGTTC	TCTCCCTTTAACACCAACGG

基因名称	正向引物(5'→3')	反向引物(5'→3')
Gene name	Forward primer(5'→3')	Reverse primer(5'→3')
TaThil-4A	CCGTCCCAGCCATCGTG	TCTTGAGCATCCGCTTGGAG
TaThil-4B	GCAGCGGAGGATTGGAGAAT	AAGGGTCAAACCAAGCCTCA
TaThil-4D	GCGGAGGAGTGGAGAATCTG	ACGCCTACCAAATCAGGACA
Ta26S	GAAGAAGGTCCCAAGGGTTC	TCTCCCTTTAACACCAACGG

基因名称	基因ID	染色体位置	氨基酸数量	半胱氨酸数量	编码蛋白	等电点	亚细胞定位	信号肽
Gene name	Gene ID	Chromosome position	Number of amino acids	Number of cysteines	分子量 Molecular mass/ku	pI	Subcellular location	Signal peptide
TaThil-1B;1	TraesCS1B03G1120400LC.1	1B:648189213-648189524	103	5	11.95	10.45	细胞核Nucleus	-
TaThil-1B;2	TraesCS1B03G1155100.1	1B:659630413-659631542	130	13	13.63	4.39	细胞外Extracellular	√
TaThil-2B	TraesCS2B03G0504400LC.1	2B:198179489-198179953	154	6	16.54	11.51	细胞核Extracellular	-
TaThil-3A;1	TraesCS3A03G0552900.1	3A:395791412-395792540	195	13	20.09	4.42	细胞外Extracellular	√
TaThil-3A;2	TraesCS3A03G0553000.1	3A:395835980-395837038	158	14	16.96	8.92	细胞外Extracellular	√
TaThil-3A;3	TraesCS3A03G1150100LC.1	3A:718510414-718510833	139	5	15.37	7.68	细胞外Extracellular	√
TaThil-3B;1	TraesCS3B03G0636600.1	3B:396308838-396309889	173	13	17.98	4.93	细胞外Extracellular	-
TaThil-3B;2	TraesCS3B03G0636800.1	3B:396354614-396355551	168	13	18.10	9.23	细胞外Extracellular	√
TaThil-3D;1	TraesCS3D03G0514500.1	3D:293341730-293342870	196	13	20.30	4.36	细胞外Extracellular	√
TaThil-3D;2	TraesCS3D03G0514700.1	3D:293354488-293355046	155	14	16.48	8.64	细胞外Extracellular	√
TaThil-4A	TraesCS4A03G0005400.1	4A:2778918-2779295	125	10	13.84	8.74	细胞外Extracellular	-
TaThil-4B	TraesCS4B03G0791300.1	4B:587455267-587456199	125	10	13.87	8.74	细胞外Extracellular	-
TaThil-4D	TraesCS4D03G0708500.1	4D:468863939-468864781	125	10	13.87	8.74	细胞外Extracellular	-
TaThil-5A	TraesCS5A03G0585800.1	5A:445399009-445399659	139	15	14.94	7.38	细胞外Extracellular	√
TaThil-5B	TraesCS5B03G0129000.1	5B:56613644-56614396	79	9	8.48	8.09	细胞外Extracellular	√
TaThil-5D;1	TraesCS5D03G0000700.1	5D:504194-504552	110	6	12.00	8.52	细胞外Extracellular	√
TaThil-5D;2	TraesCS5D03G0000900.1	5D:716462-716794	110	6	12.00	8.52	细胞外Extracellular	√
TaThil-6A;1	TraesCS6A03G0166000.1	6A:44857118-44857880	101	13	11.01	7.94	细胞外Extracellular	√
TaThil-6A;2	TraesCS6A03G0803600LC.1	6A:543299654-543299977	107	6	11.55	8.26	细胞外Extracellular	√
TaThil-6B;1	TraesCS6B03G0169000.1	6B:52931251-52932028	124	13	12.75	6.03	细胞外Extracellular	√
TaThil-6B;2	TraesCS6B03G0171400.1	6B:54283983-54284834	109	12	11.76	8.76	细胞外Extracellular	√
TaThil-6D;1	TraesCS6D03G0024500LC.1	6D:4597285-4597644	119	9	13.18	4.97	细胞外Extracellular	√
TaThil-6D;2	TraesCS6D03G0117500.1	6D:33532140-33532823	113	13	11.83	7.44	细胞外Extracellular	√
TaThil-6D;3	TraesCS6D03G0117700.1	6D:33587808-33588272	109	12	11.75	8.63	细胞外Extracellular	√
TaThil-7A	TraesCS7A03G1387900.1	7A:744208691-744208981	96	6	10.18	6.87	细胞外Extracellular	√
TaThil-7D	TraesCS7D03G1304800LC.1	7D:642451602-642451922	106	7	11.38	8.27	细胞外Extracellular	√

基因名称	基因ID	染色体位置	氨基酸数量	半胱氨酸数量	编码蛋白	等电点	亚细胞定位	信号肽
Gene name	Gene ID	Chromosome position	Number of amino acids	Number of cysteines	分子量 Molecular mass/ku	pI	Subcellular location	Signal peptide
TaThil-1B;1	TraesCS1B03G1120400LC.1	1B:648189213-648189524	103	5	11.95	10.45	细胞核Nucleus	-
TaThil-1B;2	TraesCS1B03G1155100.1	1B:659630413-659631542	130	13	13.63	4.39	细胞外Extracellular	√
TaThil-2B	TraesCS2B03G0504400LC.1	2B:198179489-198179953	154	6	16.54	11.51	细胞核Extracellular	-
TaThil-3A;1	TraesCS3A03G0552900.1	3A:395791412-395792540	195	13	20.09	4.42	细胞外Extracellular	√
TaThil-3A;2	TraesCS3A03G0553000.1	3A:395835980-395837038	158	14	16.96	8.92	细胞外Extracellular	√
TaThil-3A;3	TraesCS3A03G1150100LC.1	3A:718510414-718510833	139	5	15.37	7.68	细胞外Extracellular	√
TaThil-3B;1	TraesCS3B03G0636600.1	3B:396308838-396309889	173	13	17.98	4.93	细胞外Extracellular	-
TaThil-3B;2	TraesCS3B03G0636800.1	3B:396354614-396355551	168	13	18.10	9.23	细胞外Extracellular	√
TaThil-3D;1	TraesCS3D03G0514500.1	3D:293341730-293342870	196	13	20.30	4.36	细胞外Extracellular	√
TaThil-3D;2	TraesCS3D03G0514700.1	3D:293354488-293355046	155	14	16.48	8.64	细胞外Extracellular	√
TaThil-4A	TraesCS4A03G0005400.1	4A:2778918-2779295	125	10	13.84	8.74	细胞外Extracellular	-
TaThil-4B	TraesCS4B03G0791300.1	4B:587455267-587456199	125	10	13.87	8.74	细胞外Extracellular	-
TaThil-4D	TraesCS4D03G0708500.1	4D:468863939-468864781	125	10	13.87	8.74	细胞外Extracellular	-
TaThil-5A	TraesCS5A03G0585800.1	5A:445399009-445399659	139	15	14.94	7.38	细胞外Extracellular	√
TaThil-5B	TraesCS5B03G0129000.1	5B:56613644-56614396	79	9	8.48	8.09	细胞外Extracellular	√
TaThil-5D;1	TraesCS5D03G0000700.1	5D:504194-504552	110	6	12.00	8.52	细胞外Extracellular	√
TaThil-5D;2	TraesCS5D03G0000900.1	5D:716462-716794	110	6	12.00	8.52	细胞外Extracellular	√
TaThil-6A;1	TraesCS6A03G0166000.1	6A:44857118-44857880	101	13	11.01	7.94	细胞外Extracellular	√
TaThil-6A;2	TraesCS6A03G0803600LC.1	6A:543299654-543299977	107	6	11.55	8.26	细胞外Extracellular	√
TaThil-6B;1	TraesCS6B03G0169000.1	6B:52931251-52932028	124	13	12.75	6.03	细胞外Extracellular	√
TaThil-6B;2	TraesCS6B03G0171400.1	6B:54283983-54284834	109	12	11.76	8.76	细胞外Extracellular	√
TaThil-6D;1	TraesCS6D03G0024500LC.1	6D:4597285-4597644	119	9	13.18	4.97	细胞外Extracellular	√
TaThil-6D;2	TraesCS6D03G0117500.1	6D:33532140-33532823	113	13	11.83	7.44	细胞外Extracellular	√
TaThil-6D;3	TraesCS6D03G0117700.1	6D:33587808-33588272	109	12	11.75	8.63	细胞外Extracellular	√
TaThil-7A	TraesCS7A03G1387900.1	7A:744208691-744208981	96	6	10.18	6.87	细胞外Extracellular	√
TaThil-7D	TraesCS7D03G1304800LC.1	7D:642451602-642451922	106	7	11.38	8.27	细胞外Extracellular	√

小麦类硫素基因家族鉴定及表达分析

Identification and expression analysis of Thionin-like gene family in wheat

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