Effect of Exogenous NO on Expression of Tomato Antioxidant Enzyme Gene Under Tomato Yellowing Leaf Curl Virus Stress

doi:10.13304/j.nykjdb.2023.0647

Abstract

Abstract:

In order to investigate the effect of exogenous nitric oxide （NO） on the expression of tomato antioxidant gene under the stress of tomato yellow leaf curl virus （TYLCV）， the susceptible tomato Jinpeng 1 was used as the experimental material. Transcriptome sequencing， fluorescent qRT-PCR and bioinformatics analysis were performed under control （CK）， TYLCV （TY） and NO+TYLCV （NO+TY） treatments. The results showed that a total of 55 antioxidase-coding genes were selected in the tomato genome， among which the antioxidase-containing genes with the highest number of exons in different subcellular regions were significantly responsive to TYLCV stress. The number of NO-mediated antioxidant enzyme coding genes in different subcellular compartms showed chloroplast>cell membrane>cytoplasm>peroxisome>vacuole， in which the expression levels of Chl Cu-Zn SOD， Chl MR2， Chl GR， Per MR， Pla CAT1 and Pla CAT7 were significantly up-regulated， and the expression levels of Chl Fe SOD1， Chl Fe SOD2， Cyt GPX， Cyt APX1， Cyt APX2 L-5， Pla CAT3， Pla CAT8 and Vac CAT were significantly down-regulated. Fluorescence qRT-PCR verified that Chl GR， Min Mn SOD and Per CAT2 responded to TYLCV， and Chl Cu-Zn SOD， Pla CAT7， Pla CAT8 and Cyt APX2 L-5 responded to TYLCV and NO. Above results provided theoretical basis for studying the mechanism of NO in improving tomato disease resistance.

Key words: tomato, tomato yellow leaf curl virus, nitric oxide, antioxidant enzyme gene

摘要：

为探究番茄黄化曲叶病毒（tomato yellow leaf curl virus， TYLCV）胁迫下外源一氧化碳（nitric oxide， NO）对番茄抗氧化物酶基因表达的影响，以易感病番茄品种金鹏1号为试验材料，在对照（CK）、TYLCV（TY）和NO+TYLCV（NO+TY）3种处理下，通过转录组测序、荧光qRT-PCR和生物信息学分析进行研究。结果表明，在番茄基因组中共筛选出55个抗氧化酶相关编码基因，其中存在于不同亚细胞区室中含有外显子数目最多的抗氧化酶基因均显著响应TYLCV胁迫。NO介导的抗氧化酶编码基因数量在不同亚细胞区室的分布表现为叶绿体>细胞膜>细胞质>过氧化物酶体>液泡，其中Chl Cu-Zn SOD、Chl MR2、Chl GR、Per MR、Pla CAT1和Pla CAT7的表达量显著上调；Chl Fe SOD1、Chl Fe SOD2、Cyt GPX、Cyt APX1、Cyt APX2 L-5、Pla CAT3、Pla CAT8和Vac CAT的表达量显著下调。实时荧光qRT-PCR验证发现，Chl GR、Min Mn SOD和Per CAT2响应TYLCV表达，Chl Cu-Zn SOD、Pla CAT7、Pla CAT8和Cyt APX2 L-5响应TYLCV和NO表达。以上结果为研究NO在提高番茄抗病性机制中的作用提供理论依据。

关键词: 番茄, 番茄黄花曲叶病, 一氧化氮, 抗氧化酶基因

CLC Number:

S641.2

Jincheng LUO, Xiaolin ZHU, Xiaohong WEI, Xian WANG, Baoqiang WANG, Xuefen DU. Effect of Exogenous NO on Expression of Tomato Antioxidant Enzyme Gene Under Tomato Yellowing Leaf Curl Virus Stress[J]. Journal of Agricultural Science and Technology, 2025, 27(2): 125-135.

罗金城, 朱晓林, 魏小红, 王贤, 王宝强, 杜雪芬. 番茄黄化曲叶病毒胁迫下外源NO对番茄抗氧化物酶基因表达的影响[J]. 中国农业科技导报, 2025, 27(2): 125-135.

Figures/Tables 9

References 35

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基因Gene	引物序列Primer sequence（5’-3’）
Chl Cu-Zn SOD	F： TATGCTGTCACTACCCCTAA，R： TATTGGAGTCAAACCAACCA
Chl GR	F： CGGCATTACGATTTCGATTT，R： ACAATGGCAATTTTGTCAGG
Pla CAT7	F： GTTCCGTTATACTCGTGACA，R： ATTGCCATAACAGGGTGAAT
Pla CAT8	F： ACGTTCATCAGATACCAGTG，R： CCCAGTTATAGCAACCACAT
Cyt APX2 L-5	F： CTGATTTCCTTCGTTTAGCG，R： GAAATTCCGGATCATCAAGC
Min Mn SOD	F： CCAAATTGCACAGTGCCCTC，R： CCCAGCCAAGAGAACCCTTT
Per CAT2	F： TACGGTTGGTGCAAGAGGTC，R： CAGCACAGGTAAGGTGAGCA
UBI	F： TCGTAAGGAGTGCCCTAATGCTGA，R： CAATCGCCTCCAGCCTTGTTGTAA

基因Gene	引物序列Primer sequence（5’-3’）
Chl Cu-Zn SOD	F： TATGCTGTCACTACCCCTAA，R： TATTGGAGTCAAACCAACCA
Chl GR	F： CGGCATTACGATTTCGATTT，R： ACAATGGCAATTTTGTCAGG
Pla CAT7	F： GTTCCGTTATACTCGTGACA，R： ATTGCCATAACAGGGTGAAT
Pla CAT8	F： ACGTTCATCAGATACCAGTG，R： CCCAGTTATAGCAACCACAT
Cyt APX2 L-5	F： CTGATTTCCTTCGTTTAGCG，R： GAAATTCCGGATCATCAAGC
Min Mn SOD	F： CCAAATTGCACAGTGCCCTC，R： CCCAGCCAAGAGAACCCTTT
Per CAT2	F： TACGGTTGGTGCAAGAGGTC，R： CAGCACAGGTAAGGTGAGCA
UBI	F： TCGTAAGGAGTGCCCTAATGCTGA，R： CAATCGCCTCCAGCCTTGTTGTAA

抗氧化酶 Antioxidant enzyme	基因数量 Number of genes	亚细胞区室 Subcellular compartment
超氧化物歧化酶SOD	9	细胞质、叶绿体、线粒体、核膜 Cytoplasm， chloroplast， mitochondrion， nucleus membrane
过氧化氢酶CAT	18	细胞质、内质网、过氧化物酶体、细胞膜、液泡 Cytoplasm， endoplasmic reticulum， peroxisome， plasma membrane， vacuole
谷胱甘肽过氧化物酶GPX	6	叶绿体、细胞核、细胞质 Chloroplast， nucleus， cytoplasm
抗坏血酸过氧化物酶APX	9	细胞质、线粒体、叶绿体 Cytoplasm， mitochondrion， chloroplast
单脱氢抗坏血酸还原酶MR	4	叶绿体、细胞质、过氧化物酶体 Chloroplast， cytoplasm， peroxisome
谷胱甘肽还原酶GR	2	叶绿体、细胞膜Chloroplast， plasma membrane
脱氢抗坏血酸还原酶DR	7	细胞质、细胞核、叶绿体、线粒体 Cytoplasm， nucleus， chloroplast， mitochondrion

抗氧化酶 Antioxidant enzyme	基因数量 Number of genes	亚细胞区室 Subcellular compartment
超氧化物歧化酶SOD	9	细胞质、叶绿体、线粒体、核膜 Cytoplasm， chloroplast， mitochondrion， nucleus membrane
过氧化氢酶CAT	18	细胞质、内质网、过氧化物酶体、细胞膜、液泡 Cytoplasm， endoplasmic reticulum， peroxisome， plasma membrane， vacuole
谷胱甘肽过氧化物酶GPX	6	叶绿体、细胞核、细胞质 Chloroplast， nucleus， cytoplasm
抗坏血酸过氧化物酶APX	9	细胞质、线粒体、叶绿体 Cytoplasm， mitochondrion， chloroplast
单脱氢抗坏血酸还原酶MR	4	叶绿体、细胞质、过氧化物酶体 Chloroplast， cytoplasm， peroxisome
谷胱甘肽还原酶GR	2	叶绿体、细胞膜Chloroplast， plasma membrane
脱氢抗坏血酸还原酶DR	7	细胞质、细胞核、叶绿体、线粒体 Cytoplasm， nucleus， chloroplast， mitochondrion

登记号 Gene accession No.	名称 Name	氨基酸数量 Number of amino acids	分子质量 Molecular weight/Da	等电点 Isoelectric point	亲水性 GRAVY
Solyc01g067740.3	Cyt Cu-Zn SOD2	152	15 284.95	5.47	-0.199
Solyc02g021140.3	Chl Fe SOD3	252	29 126.27	6.65	-0.348
Solyc03g062890.3	Cyt Cu-Zn SOD1	156	15 870.86	6.53	-0.022
Solyc03g095180.3	Chl Fe SOD1	303	34 575.69	5.38	-0.627
Solyc06g048410.3	Chl Fe SOD2	278	31 144.47	6.60	-0.270
Solyc06g048420.2	Nucp Fe SOD	109	12 193.83	5.38	-0.322
Solyc06g049080.3	Mit Mn SOD	228	25 314.79	7.13	-0.318
Solyc08g079830.3	Chl SOD copper chaperone 1	311	32 987.61	6.45	-0.027
Solyc11g066390.2	Chl Cu-Zn SOD	217	22 281.92	6.01	-0.018
Solyc01g099420.2	Cyt CAT isozyme 2	106	12 121.88	5.26	0.004
Solyc01g100630.2	ER CAT	427	47 770.03	6.20	-0.008
Solyc01g100640.3	Pla CAT isozyme 1	220	25 603.58	8.37	-0.140
Solyc12g096380.1	Pla CAT1a	599	65 926.19	8.64	0.497
Solyc12g094620.2	Per CAT1	497	57 078.50	6.56	-0.538
Solyc12g011370.2	Pla CAT8	586	64 090.11	8.76	0.539
Solyc11g006710.2	Pla CAT7	584	63 360.16	7.02	0.651
Solyc10g081460.2	Vac CAT	650	67 933.20	6.58	0.628
Solyc10g018600.2	Pla CAT9	569	60 434.61	6.34	0.820
Solyc08g077823.1	Pla CAT6a	305	33 185.14	8.88	0.573
Solyc08g077820.3	Pla CAT6b	229	26 277.39	9.03	0.808