谷子糖转运蛋白基因SiSTPs的鉴定及其参与谷子抗逆胁迫响应的研究

doi:10.3724/SP.J.1006.2022.14080

作物学报 ›› 2022, Vol. 48 ›› Issue (4): 825-839.doi: 10.3724/SP.J.1006.2022.14080

• 作物遗传育种·种质资源·分子遗传学 • 上一篇下一篇

谷子糖转运蛋白基因SiSTPs的鉴定及其参与谷子抗逆胁迫响应的研究

晋敏姗¹(), 曲瑞芳¹, 李红英¹^,³, 韩彦卿²^,³, 马芳芳¹^,³, 韩渊怀¹^,³, 邢国芳¹^,³^,^*()

¹山西农业大学农学院, 山西太谷 030801
²山西农业大学植物保护学院, 山西太谷 030801
³杂粮种质创新与分子育种山西省重点实验室, 山西太原 030000

收稿日期:2021-04-28 接受日期:2021-07-12 出版日期:2022-04-12 网络出版日期:2021-08-11
通讯作者: 邢国芳
作者简介:E-mail: jms1995mashy@163.com
基金资助:
山西省重点研发计划项目(201803D221008-4);国家自然科学基金项目(32070366);山西省应用基础研究项目资助(201801D221298)

Identification of sugar transporter gene family SiSTPs in foxtail millet and its participation in stress response

JIN Min-Shan¹(), QU Rui-Fang¹, LI Hong-Ying¹^,³, HAN Yan-Qing²^,³, MA Fang-Fang¹^,³, HAN Yuan-Huai¹^,³, XING Guo-Fang¹^,³^,^*()

¹College of Agronomy, Shanxi Agricultural University, Taigu 030801, Shanxi, China
²College of Plant Protection, Shanxi Agricultural University, Taigu 030801, Shanxi, China
³Shanxi Key Laboratory of Germplasm Innovation and Molecular Breeding of Minor Crop, Taiyuan 030000, Shanxi, China

Received:2021-04-28 Accepted:2021-07-12 Published:2022-04-12 Published online:2021-08-11
Contact: XING Guo-Fang
Supported by:
Key Research and Development Program of Shanxi Province(201803D221008-4);National Natural Science Foundation of China(32070366);Applied Basic Research Project of Shanxi Province(201801D221298)

摘要/Abstract

摘要：

糖转运蛋白(sugar transporter proteins, STPs)是一类主要转运己糖的单糖转运蛋白, 在作物的生长发育和抗逆过程中发挥着重要作用。谷子是绿色旱作农业的主栽作物, 也是C₄光合作用机理和禾本科抗逆基因挖掘的模式植物, 目前有关谷子STP基因功能的研究还鲜有报道。因此, 本研究利用生物信息学方法, 对包括谷子在内的6种禾本科作物的STPs基因进行全基因组鉴定, 重点对谷子SiSTPs基因的理化性质、染色体定位、系统进化、基因结构、保守结构域、组织表达模式, 并对其参与谷子干旱和低磷胁迫以及白发病抗性响应进行了深入分析。在谷子、狗尾草、高粱、玉米、小麦和水稻中分别鉴定出STP基因家族成员24、26、23、22、33和27个, 系统进化分析聚为4类。谷子的24个SiSTP基因不均匀的分布在7条染色体上, 其编码氨基酸的大小为499~581 aa, 这些SiSTP蛋白都具有Sugar_tr (PF00083)保守结构域, 并且在启动子区存在大量的光响应以及逆境响应元件, 分析发现谷子SiSTPs基因在进化过程中受到了强烈的纯化选择压力, 存在明显的组织表达特异性, 且受光诱导, 不同成员在低磷胁迫、干旱胁迫和白发病侵染过程中表达存在差异。本研究为阐明谷子STP蛋白的功能以及响应逆境胁迫机理提供理论基础。

关键词: 谷子, 糖转运蛋白基因, 系统进化, 表达分析, 胁迫应答

Abstract:

Sugar transporter proteins (STPs), a class of monosaccharide transporters that mainly transport hexose, play an important role in the growth, development, and stress resistance of crops. Foxtail millet is the main cultivated crop in green dryland agriculture, and also is the model plant for C₄ photosynthesis mechanism and stress resistance gene mining of Gramineae crops. However, no systematical study of SiSTPs gene has been performed in foxtail millet. In this study, we identified the whole genome of six Gramineae crops including foxtail millet by bioinformatics method, and focused on the physicochemical properties, the chromosomal localization, the systematic evolution, gene structure, and the conserved domain. Moreover, the relative expression level of SiSTPs gene and its resistance to the infection of Sclerospora graminicola under drought stress and low phosphate stress in foxtail millet were investigated. The results showed that a total of 24, 26, 23, 22, 33, and 27 STP gene family members were individually identified in Setaria italica, Setaria viridis, Sorghum bicolor, Zea mays, Triticum aestivum, and Oryza sativa, which were divided into four clades by phylogenetic analysis. The 24 SiSTP genes were distributed unevenly on 7 chromosomes, and the size of the encoded amino acids ranged from 499 aa to 581 aa. The SiSTP all had the Sugar_tr (PF00083) conserved domain. The cis-acting elements included a large number of light-responsive elements and stress-responsive elements. These SiSTPs was subjected to strong purification and selection pressure during the evolutionary process of foxtail millet, and had obvious tissue expression specificity and photoinduced phenomena. Different SiSTPs showed different temporal and spatial expression and response to the drought stress, the low phosphate stress, and Sclerospora graminicola infection. Our results provide a theoretical basis for elucidating the function and response mechanism to stress of SiSTP.

Key words: foxtail millet, sugar transporter proteins, systematic evolution, expression analysis, stress response

晋敏姗, 曲瑞芳, 李红英, 韩彦卿, 马芳芳, 韩渊怀, 邢国芳. 谷子糖转运蛋白基因SiSTPs的鉴定及其参与谷子抗逆胁迫响应的研究[J]. 作物学报, 2022, 48(4): 825-839.

JIN Min-Shan, QU Rui-Fang, LI Hong-Ying, HAN Yan-Qing, MA Fang-Fang, HAN Yuan-Huai, XING Guo-Fang. Identification of sugar transporter gene family SiSTPs in foxtail millet and its participation in stress response[J]. Acta Agronomica Sinica, 2022, 48(4): 825-839.

图/表 14

表1

图1

表2

表3

谷子STPs蛋白二级结构预测和亚细胞定位"

序号 Number	名称 Gene name	α-螺旋 α-helix (%)	延伸链 Extended strand (%)	β-折叠 β-turn (%)	无规则卷曲Random coil (%)	亚细胞定位 Subcellular localization	保守结构域 Domain
1	SiSTP1	52.12	15.96	4.62	27.31	细胞质膜Plasma membrane	MFS_STP
2	SiSTP2	48.57	16.38	5.71	29.33	细胞质膜Plasma membrane	MFS_STP
3	SiSTP3	51.16	15.25	4.83	28.76	细胞质膜Plasma membrane	MFS_STP
4	SiSTP4	50.39	16.09	6.59	26.94	细胞质膜Plasma membrane	MFS_STP
5	SiSTP5	50.59	16.40	4.55	28.46	细胞质膜Plasma membrane	MFS_STP
6	SiSTP6	49.62	15.71	5.36	29.31	细胞质膜Plasma membrane	MFS_STP
7	SiSTP7	50.40	16.47	5.36	27.78	细胞质膜Plasma membrane	MFS_STP
8	SiSTP8	47.31	16.54	5.58	30.58	细胞质膜Plasma membrane	MFS_STP
9	SiSTP9	50.09	16.51	5.31	28.08	细胞质膜Plasma membrane	MFS_STP
10	SiSTP10	48.96	17.39	4.73	28.92	细胞质膜Plasma membrane	MFS_STP
11	SiSTP11	47.15	17.49	4.72	30.65	细胞质膜Plasma membrane	MFS_STP
12	SiSTP12	51.70	16.23	5.41	26.65	细胞质膜Plasma membrane	MFS_STP
13	SiSTP13	50.29	17.35	4.68	27.68	细胞质膜Plasma membrane	MFS_STP
14	SiSTP14	49.41	16.99	4.88	28.71	细胞质膜Plasma membrane	MFS_STP
15	SiSTP15	51.46	15.40	5.46	27.68	细胞质膜Plasma membrane	MFS_STP
16	SiSTP16	51.17	16.02	4.88	27.93	细胞质膜Plasma membrane	MFS_STP
17	SiSTP17	51.17	16.73	5.64	26.46	细胞质膜Plasma membrane	MFS_STP
18	SiSTP18	47.00	13.86	4.68	34.46	细胞质膜Plasma membrane	MFS_STP
19	SiSTP19	49.32	16.44	5.68	28.75	细胞质膜Plasma membrane	MFS_STP
20	SiSTP20	47.12	16.92	5.38	30.58	细胞质膜Plasma membrane	MFS_STP
21	SiSTP21	51.05	15.62	5.71	27.62	细胞质膜Plasma membrane	MFS_STP
22	SiSTP22	53.10	15.31	4.46	27.13	细胞质膜Plasma membrane	MFS_STP
23	SiSTP23	45.96	15.83	5.68	32.53	细胞质膜Plasma membrane	MFS_STP
24	SiSTP24	50.00	16.28	6.20	27.52	细胞质膜Plasma membrane	MFS_STP

表3

图2

表4

图3

图4

图5

表5

图6

图7

图8

图9

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基因 Gene name	上游引物 Forward sequence (5′-3′)	下游引物 Reverse sequence (5'-3')
SiSTP4	ACCATGCTCTGCCACTTCAA	TCTTCCAGAACCAGTGCGAC
SiSTP9	TGACATCGGCATCTCTGGTG	GATGTTGAGCATCCCACGGA
SiSTP19	TGAAGTTCTTCCCGTCGGTG	AGCTGGAACCCGATGTTGAG
SiSTP23	GACTTGCGGTCATCTCCCTC	TGAGGCGACCTCTCTGATGA
β-Actin	CAACAAGATGGATGCCACCAC	GAGATTGGGACGAAGGCAATC

名称 Gene name	基因名称 Gene ID	氨基酸数目 Number of amino acids	等电点Isoelectric point	相对分子质量Molecular weight	不稳定指数 Instability index	脂肪系数Aliphatic index	平均疏水指数 Grand average of hydropathicity
SiSTP1	Seita.1G079800.1	520	9.10	57,150.45	36.01	101.27	0.596
SiSTP2	Seita.1G204900.1	525	8.77	56,604.51	45.82	105.52	0.619
SiSTP3	Seita.1G205000.1	518	9.34	55,004.35	38.13	105.83	0.619
SiSTP4	Seita.2G000400.1	516	8.36	56,378.41	34.38	102.46	0.523
SiSTP5	Seita.2G079100.1	506	9.34	55,238.58	37.97	96.03	0.414
SiSTP6	Seita.2G167300.1	522	9.13	55,270.80	37.54	100.00	0.552
SiSTP7	Seita.2G176800.1	504	9.06	54,941.45	33.51	107.56	0.528
SiSTP8	Seita.2G205100.1	520	9.41	56,007.80	39.32	105.04	0.512
SiSTP9	Seita.2G352200.1	527	9.20	57,372.78	31.35	106.41	0.579
SiSTP10	Seita.4G029600.1	529	9.75	56,006.24	34.25	102.76	0.497
SiSTP11	Seita.5G204000.1	509	9.44	55,305.53	37.53	109.39	0.698
SiSTP12	Seita.6G043500.1	499	6.52	53,829.04	36.97	106.23	0.650
SiSTP13	Seita.7G007500.1	513	9.47	55,508.58	34.04	110.84	0.707
SiSTP14	Seita.7G119800.1	512	9.81	55,140.95	31.73	100.27	0.574
SiSTP15	Seita.7G120000.1	513	9.73	55,566.19	38.80	100.94	0.545
SiSTP16	Seita.7G120100.1	512	8.97	55,632.07	36.42	103.98	0.569
SiSTP17	Seita.7G120200.1	514	9.32	55,147.58	36.22	104.40	0.572
SiSTP18	Seita.7G120300.1	534	9.91	57,645.50	38.48	99.23	0.432
SiSTP19	Seita.7G120800.1	511	9.67	53,895.89	31.18	102.97	0.611
SiSTP20	Seita.9G122300.1	520	9.41	55,987.81	39.48	105.98	0.518
SiSTP21	Seita.9G187300.1	525	9.19	56,067.71	33.69	104.82	0.581
SiSTP22	Seita.9G322100.1	516	9.51	57,307.68	45.13	108.66	0.466
SiSTP23	Seita.9G488700.1	581	9.36	64,227.29	34.11	107.02	0.399
SiSTP24	Seita.9G579300.1	516	9.19	56,549.60	38.76	109.75	0.567

基序 Motif	序列 Sequence	长度 Width (aa)	所在基因个数 Number of genes
Motif 1	LCAFKYGIFLFFAAWVVVMTVFVAAFLPETKGVPJEEM	38	24
Motif 2	NGAAVNVAMLIVGRILLGVGVGFTNQAVPLYLSEMAPARWRGALNIGFQL	50	24
Motif 3	QYCKYDSQLLTAFTSSLYLAGLVASLVASRVTRRFGRKASM	41	24
Motif 4	CVYVAGFGWSWGPLGWLVPSEIFPLEIRS	29	24
Motif 5	DYGGKVTFFVVLTCLVAASGGLIFGYDIGISGGVTSMDPFLKKFFPEVYR	50	23
Motif 6	IPVFQQLTGINVIMFYAPVLFRTVGFGSDA	30	24
Motif 7	WGWRLSLGLAAVPAAVITLGALFLPDTPNSLILR	34	23
Motif 8	VSIVTVDRLGRRKLFLQGGAQMJICQVAVGWILGAKFGASG	41	24
Motif 9	RLVWRRHWYWKRFVRDEPDEA	21	24
Motif 10	FRNJLRRRYRPQLVM	15	24
Motif 11	FITJGILAANLINYGTNKIPG	21	23
Motif 12	DVDAEFKDJVAASEEARAVEH	21	24
Motif 13	NLLFTFVIAQAFLSM	15	24
Motif 14	GRPDEARAVLQRIRG	15	23
Motif 15	SLMSAVITGLVNLFA	15	23

比对基因_1 Gene_1	比对基因_2 Gene_2	非同义替换 Ka	同义替换 Ks	非同义替换/同义替换 Ka/Ks
SiSTP1	OsSTP3	0.11781299	0.32157912	0.36635770
SiSTP3	OsSTP12	0.33694743	0.60239717	0.55934430
SiSTP4	OsSTP17	0.09187094	0.37007634	0.24824862
SiSTP5	OsSTP20	0.13761001	0.47474108	0.28986328
SiSTP6	OsSTP24	0.13021302	0.34325476	0.37934802
SiSTP8	OsSTP25	0.04544091	0.23400541	0.19418743
SiSTP9	OsSTP21	0.04068573	0.31030313	0.13111608
SiSTP10	OsSTP16	0.10871793	0.30090572	0.36130230
SiSTP11	OsSTP1	0.10108616	0.28074655	0.36006199
SiSTP12	OsSTP22	0.07095251	0.33044715	0.21471666
SiSTP15	OsSTP13	0.21042393	0.38939049	0.54039311
SiSTP17	OsSTP6	0.37053540	0.62711506	0.59085712
SiSTP23	OsSTP8	0.03796308	0.30304824	0.12527075
SiSTP24	OsSTP7	0.11469825	1.12797658	0.10168496
SiSTP9	AtSTP1	0.26072995	3.45057903	0.07556122
SiSTP9	AtSTP12	0.32117875	2.34003570	0.13725378

谷子糖转运蛋白基因SiSTPs的鉴定及其参与谷子抗逆胁迫响应的研究

Identification of sugar transporter gene family SiSTPs in foxtail millet and its participation in stress response

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