藜麦SOD家族基因的鉴定及其对混合盐碱胁迫的响应

doi:10.13304/j.nykjdb.2022.0558

中国农业科技导报 ›› 2024, Vol. 26 ›› Issue (1): 28-39.DOI: 10.13304/j.nykjdb.2022.0558

藜麦SOD家族基因的鉴定及其对混合盐碱胁迫的响应

邓玉荣¹(), 韩联¹, 王金龙¹, 韦兴翰¹, 王旭东¹, 赵颖¹^,², 魏小红¹^,²(), 李朝周¹()

^1.甘肃农业大学生命科学技术学院，兰州 730070
^2.甘肃农业大学，甘肃省作物遗传改良与种质创新重点实验室，甘肃省干旱生境作物学重点实验室，兰州 730070

收稿日期:2022-07-05 接受日期:2022-10-08 出版日期:2024-01-15 发布日期:2024-01-08
通讯作者: 魏小红,李朝周
作者简介:邓玉荣 E-mail：2940834096@qq.com
基金资助:
甘肃农业大学省级大学生创新创业训练计划项目(S202010733077);甘肃农业大学干旱生境作物学重点实验室开放基金课题项目(GSCS-2021-10)

Identification of SOD Family Genes in Chenopodium quinoa and Their Response to Mixed Saline-alkali Stress

Yurong DENG¹(), Lian HAN¹, Jinlong WANG¹, Xinghan WEI¹, Xudong WANG¹, Ying ZHAO¹^,², Xiaohong WEI¹^,²(), Chaozhou LI¹()

^1.College of Life Science and Technology，Gansu Agricultural University，Lanzhou 730070，China
^2.Gansu Key Lab of Crop Genetic & Germplasm Enhancement，Gansu Provincial Key Lab of Aridland Crop Science，Gansu Agricultural University，Lanzhou 730070，China

Received:2022-07-05 Accepted:2022-10-08 Online:2024-01-15 Published:2024-01-08
Contact: Xiaohong WEI,Chaozhou LI

摘要/Abstract

摘要：

超氧化物歧化酶（superoxide dismutase，SOD）是植物抗氧化系统的关键酶，在保护植物免受生物和非生物胁迫方面发挥重要作用。以拟南芥SOD为基础，通过序列比对在藜麦基因组中鉴定出12个SOD基因，分别定位于细胞核、微体及线粒体，在11条染色体上不均匀分布，其编码蛋白质三级结构显示Cu/Zn-SODs与Fe-SODs为同源二聚体，Mn-SODs为同源四聚体。CqSOD基因内含子/外显子分布模式不尽相同，内含子数介于4～7个，保守基序差异明显。系统发育关系显示，SOD蛋白可分为Cu/Zn-SODs、Fe-SODs及Mn-SODs 3个亚族。此外，所有的CqFe-SODs及CqMn-SODs启动子区都含有脱落酸激素反应顺式元件，CqSOD12与11个CqSOD蛋白及4个CqCAT蛋白存在相互作用。表达谱分析表明，12个CqSOD基因对混合盐碱及硝普钠均有较强响应。研究结果为SOD基因在植物发育和胁迫响应中的作用及分子机制研究奠定基础。

关键词: 藜麦, SOD家族基因, 盐碱胁迫, 生物信息学分析, 表达分析

Abstract:

Superoxide dismutase （SOD） is a key enzyme in the antioxidant system of plants and plays a critical role in protecting plants from various biological and abiotic stresses. In this paper， using bioinformatics methods， 12 SOD genes were identified in quinoa by sequence alignment based on Arabidopsis SOD， which localized in the nucleus， microsomes and mitochondria， and distributed heterogeneously on 11 different chromosomes， and the tertiary structure of their encoded proteins showed that Cu/Zn-SODs were homodimers with Fe-SODs and Mn-SODs were homotetramers.The intron/exon distribution patterns of the CqSOD genes were different， with the number of introns ranging from 4 to 7， and the conserved motifs differing significantly. The phylogenetic relationships showed that the SOD proteins could be divided into 3 subfamilies： Cu/Zn-SODs， Fe-SODs and Mn-SODs. In addition， all CqFe-SODs and CqMn-SODs promoter regions contained abscisic acid（ABA）-responsive cis-elements， and CqSOD12 interacted with 11 CqSOD proteins and 4 CqCAT proteins. Expression profiling showed that the 12 CqSOD genes were strongly responsive to both mixed saline-alkali and nitroprusside. Above results laid the foundation for research on the role and molecular mechanism of SOD genes in plant development and stress response.

Key words: Chenopodium quinoa, SOD family genes, salt-alkaline stress, bioinformatics analysis, expression analysis

中图分类号:

S519

邓玉荣, 韩联, 王金龙, 韦兴翰, 王旭东, 赵颖, 魏小红, 李朝周. 藜麦SOD家族基因的鉴定及其对混合盐碱胁迫的响应[J]. 中国农业科技导报, 2024, 26(1): 28-39.

Yurong DENG, Lian HAN, Jinlong WANG, Xinghan WEI, Xudong WANG, Ying ZHAO, Xiaohong WEI, Chaozhou LI. Identification of SOD Family Genes in Chenopodium quinoa and Their Response to Mixed Saline-alkali Stress[J]. Journal of Agricultural Science and Technology, 2024, 26(1): 28-39.

图/表 10

参考文献 34

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处理 Treatment	盐分组成Salt composition
处理 Treatment	氯化钠NaCl/ （200 mmol·L^-1）	硫酸钠Na₂SO₄/ （200 mmol·L^-1）	碳酸氢钠NaHCO₃/ （200 mmol·L^-1）	碳酸钠Na₂CO₃/（200 mmol·L^-1）	硝普纳SNP/ （150 μmol·L^-1）
CK	0	0	0	0	-
A	1	1	0	0	-
B	1	2	1	0	-
C	1	9	9	1	-
D	1	1	1	1	-
E	9	1	1	9	-
CK+	0	0	0	0	+
A+	1	1	0	0	+
B+	1	2	1	0	+
C+	1	9	9	1	+
D+	1	1	1	1	+
E+	9	1	1	9	+

处理 Treatment	盐分组成Salt composition
处理 Treatment	氯化钠NaCl/ （200 mmol·L^-1）	硫酸钠Na₂SO₄/ （200 mmol·L^-1）	碳酸氢钠NaHCO₃/ （200 mmol·L^-1）	碳酸钠Na₂CO₃/（200 mmol·L^-1）	硝普纳SNP/ （150 μmol·L^-1）
CK	0	0	0	0	-
A	1	1	0	0	-
B	1	2	1	0	-
C	1	9	9	1	-
D	1	1	1	1	-
E	9	1	1	9	-
CK+	0	0	0	0	+
A+	1	1	0	0	+
B+	1	2	1	0	+
C+	1	9	9	1	+
D+	1	1	1	1	+
E+	9	1	1	9	+

基因 Gene	正向引物 Forward primer（5’-3’）	反向引物 Reverse primer（5’-3’）	退火温度 Tm/℃	产物长度 Amplicon size/bp
CqActin	CCCTCACCACTTTCCGATCT	TCCTCACCCTCACCCATTTT	62.6	62
CqSOD01	ACTGGGAATGTCTCGGGTCT	GTAGCGGTACCATCATCCCC	61.6	141
CqSOD02	AGGAGATGGCCCAACAACTG	GGCGAACTTCGTCTTCAGGA	61.8	89
CqSOD03	TGCTGGTGGAAGATTGGCTT	TGTGGTGACTCGGTGAACTG	62.7	107
CqSOD04	CGGAAGATGAAGTCCGGCAT	CACAAGGGCTCTACCGACAA	61.9	138
CqSOD05	TTCAGAGAGACATGCGGGTG	CAGCATGCACCACAATAGCC	62.3	111
CqSOD06	TCATCTCCGGCGCCAATAAC	GCCATGAAGACCAGGAGTGA	62.0	87
CqSOD07	GGGGCCTAAACACTTTTCGC	TCCAGGTTGCATGGATTCCC	63.1	125
CqSOD08	GGGGCCTAAACACTTTTCGC	CGGCTCCAAGGCATCAAATG	63.5	86
CqSOD09	GGAGTCACATTGGGGAGAGC	TCCAGGTTGCATGGATTCCC	62.2	90
CqSOD10	CGTACGACTATGGCGCTCTT	ACATGACCTCCGCCATTGAA	61.4	118
CqSOD11	GCTGGGCTTGGCTTGTTTAC	TGCTCCCAAACGTCGATAGT	63.5	140
CqSOD12	CGTACGACTATGGCGCTCTT	GATTCACATGACCTCCGCCA	63.2	114

基因 Gene	正向引物 Forward primer（5’-3’）	反向引物 Reverse primer（5’-3’）	退火温度 Tm/℃	产物长度 Amplicon size/bp
CqActin	CCCTCACCACTTTCCGATCT	TCCTCACCCTCACCCATTTT	62.6	62
CqSOD01	ACTGGGAATGTCTCGGGTCT	GTAGCGGTACCATCATCCCC	61.6	141
CqSOD02	AGGAGATGGCCCAACAACTG	GGCGAACTTCGTCTTCAGGA	61.8	89
CqSOD03	TGCTGGTGGAAGATTGGCTT	TGTGGTGACTCGGTGAACTG	62.7	107
CqSOD04	CGGAAGATGAAGTCCGGCAT	CACAAGGGCTCTACCGACAA	61.9	138
CqSOD05	TTCAGAGAGACATGCGGGTG	CAGCATGCACCACAATAGCC	62.3	111
CqSOD06	TCATCTCCGGCGCCAATAAC	GCCATGAAGACCAGGAGTGA	62.0	87
CqSOD07	GGGGCCTAAACACTTTTCGC	TCCAGGTTGCATGGATTCCC	63.1	125
CqSOD08	GGGGCCTAAACACTTTTCGC	CGGCTCCAAGGCATCAAATG	63.5	86
CqSOD09	GGAGTCACATTGGGGAGAGC	TCCAGGTTGCATGGATTCCC	62.2	90
CqSOD10	CGTACGACTATGGCGCTCTT	ACATGACCTCCGCCATTGAA	61.4	118
CqSOD11	GCTGGGCTTGGCTTGTTTAC	TGCTCCCAAACGTCGATAGT	63.5	140
CqSOD12	CGTACGACTATGGCGCTCTT	GATTCACATGACCTCCGCCA	63.2	114

基因登录号 Gene accession No.	基因名称 Gene name	氨基酸数 Size/aa	分子量 Molecular weight /kD	等电点 pI	不稳定指数Instability index	脂肪酸指数Aliphatic index	疏水性Hydrophobicity	亚细胞定位Subcellular localization	磷酸化位点数量Phosphorylation site number
AUR62000929	CqSOD01	152	15.26	5.28	15.37	76.91	-0.19	细胞质Cytoplasm	14
AUR62005041	CqSOD02	152	15.27	5.28	15.37	76.91	-0.21	细胞质Cytoplasm	11
AUR62014976	CqSOD03	287	29.96	8.34	41.28	78.61	-0.19	细胞质Cytoplasm	77
AUR62029152	CqSOD04	246	25.26	6.02	28.11	85.28	-0.01	细胞质Cytoplasm	56
AUR62032030	CqSOD05	157	16.01	6.38	22.92	87.52	-0.18	细胞质Cytoplasm	16
AUR62032721	CqSOD06	130	13.41	6.33	16.19	89.23	-0.16	细胞质Cytoplasm	16
AUR62001685	CqSOD07	262	29.89	8.33	38.29	81.15	-0.35	微体Microbody	36
AUR62010480	CqSOD08	280	31.65	6.19	35.50	69.71	-0.46	微体Microbody	25
AUR62020097	CqSOD09	262	30.00	7.73	39.44	82.63	-0.33	微体Microbody	34
AUR62030413	CqSOD10	281	31.66	6.40	36.42	70.85	-0.47	微体Microbody	31
AUR62024917	CqSOD11	295	32.43	8.80	37.17	93.63	-0.12	线粒体Mitochondrial	46
AUR62030627	CqSOD12	233	25.89	6.79	39.36	87.98	-0.35	线粒体Mitochondrial	30

藜麦SOD家族基因的鉴定及其对混合盐碱胁迫的响应

Identification of SOD Family Genes in Chenopodium quinoa and Their Response to Mixed Saline-alkali Stress

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图/表 10

参考文献 34

相关文章 15

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基因 Gene	蛋白质二级结构Secondary structure of protein			染色体定位 Chromosomal localization
基因 Gene	α螺旋α helix	延伸链Extension chain	无规则卷曲Random coil	染色体定位 Chromosomal localization
CqSOD01	4	57	91	Chr12
CqSOD02	4	57	91	Chr05
CqSOD03	86	46	155	Chr15
CqSOD04	47	55	144	Chr00
CqSOD05	0	55	102	Chr11
CqSOD06	4	42	84	Chr07
CqSOD07	71	61	130	Chr07
CqSOD08	93	49	138	Chr13
CqSOD09	93	48	121	Chr18
CqSOD10	94	51	136	Chr16
CqSOD11	96	49	150	Chr01
CqSOD12	89	43	101	Chr04

基因 Gene	激素响应元件 Phytohormone responsiveness element							压力响应元件 Stress responsiveness element			组织特异性表达元件 Tissue-specific expression element
基因 Gene	ABA响应元件 ABRE	CGTCA 基序 CGTCA-motif	P盒 P-box	TATC盒TATC-box	TCA 元件 TCA-element	TGACG基序TGACG-motif	TGA 元件 TGA-element	低温应答元件 LTR	MYB结合位点 MBS	富含TC的重复序列 TC-rich repeats	厌氧诱导作用元件 ARE	CAT盒CAT-box	MYB 结合位点MBSI	O₂位点 O₂-site
CqSOD01		3			1	3	1				1			1
CqSOD02		2		1		2	1	1	1		2			1
CqSOD03	5	2	1			2		2			7
CqSOD04				2	3			1			1
CqSOD05			1		2						3	1		1
CqSOD06	1	3			1	3		2		1	3
CqSOD07	2	2		1	1	2	2		1	1
CqSOD08	1				2					1	1		1	1
CqSOD09	1	1	1		1	1	3				1
CqSOD10	3	4	1		1	4		2			1
CqSOD11	1				2		2	1	1	1	5	1
CqSOD12	3	1				1			1	1	2			1

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