玉米ACO基因家族生物信息学及表达模式分析

doi:10.3864/j.issn.0578-1752.2024.07.008

Abstract

Abstract:

【Objective】The objective of this study is to perform the genome-wide identification of the maize ACO (1-aminocyclopropane- 1-carboxylate oxidase) gene family, analyze its expression patterns in different organs and developmental stages of maize, as well as in response to exogenous hormones and pathogen infection, and to lay the foundation for clarifying the function of the maize ACO gene family.【Method】Using bioinformatics methods, the ACO was identified in the genome of maize B73 inbred line, and its gene structure, protein physicochemical properties, phylogenetic relationships among family members, and conserved motifs were analyzed. The expression patterns of the ZmACO gene family were analyzed using real-time fluorescence quantitative PCR (qRT-PCR) technology.【Result】Except for ZmACO11, all members of the ZmACO family have Fe²⁺ binding sites and substrate ascorbic acid binding sites. The phylogenetic tree showed that ZmACO2 and ScACO are in the same branch and have a close genetic relationship, with a Bootstrap value of 98. The gene expression analysis indicated that ZmACO2, 5, 9, 15, 20 and 35 were actively expressed at various developmental stages and exhibited dominant expression in leaves, so the six genes mentioned above were selected for the next step of testing. Spraying ethephon resulted in fluctuations in the expression of all six genes mentioned above, the expression level of ZmACO2 was significantly affected, with a variation multiple of about 8 times. The expression levels of these six genes fluctuated within 0-24 h of ethephon treatment. But after 24 h of treatment, all gene expression levels were close to 0. After salicylic acid treatment, the expression level of ZmACO5 was significantly affected, with a variation multiple of about 2 times. The expression levels of other genes were close to 0 at 24 h after treatment. The expression levels of ZmACO9, 35 fluctuated between 3 to 12 h, and the expression levels of ZmACO2, 15, 20 showed a downward trend. In response to biological stress, the expression levels of ZmACO5, 9 showed the greatest changes after inoculation with the Setosphaeria turcica, and on the 10th day after inoculation, the expression levels of these two genes increased by 50 and 60 times, respectively, compared to the control group. After inoculation with the Cochlibolus heterostrophus, the expression level of ZmACO5 changed significantly, with a variation multiple of 40-90 times. After inoculation with Rhizoctonia solani, the expression levels of ZmACO5, 35 showed the greatest changes, reaching 200 times on the 3rd day of inoculation.【Conclusion】The expression changes of ZmACO2, 5, 20 and 35 are most active during the growth and development of maize; The application of exogenous ethephon and salicylic acid can significantly affect the expression level of ZmACO genes. The expression level of ZmACO genes significantly changes after bacterial infection in maize, which is closely related to the response to biological stress.

Key words: maize ACO gene family, qRT-PCR, gene expression, ethephon, salicylic acid, Setosphaeria turcica, Cochlibolus heterostrophus, Rhizoctonia solani

WANG ChengZe, ZHANG Yan, FU Wei, JIA JingZhe, DONG JinGao, SHEN Shen, HAO ZhiMin. Bioinformatics and Expression Pattern Analysis of Maize ACO Gene Family[J].Scientia Agricultura Sinica, 2024, 57(7): 1308-1318.

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References 39

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引物Primer	序列Sequence (5′-3′)
ZmACO1-F/R	TGAAAACCTTGGGCTGGACA/TCACAAGCTCTGGATGGCTG
ZmACO2-F/R	CAAGATGCGTGCGAGAACTG/CGCTTGTAGTGCTCCTTGGT
ZmACO3-F/R	CATCTCCGGCGGCCTAAAA/AGTTCTTTGTCGAATACTCGGT
ZmACO4-F/R	CTACAACCCGGCTAACGAGG/GAATCTGGGCTCCTTGGCTT
ZmACO5-F/R	TACTTCATCCAGCACCACCC/TTGAACCACGGGTGCTTGCC
ZmACO6-F/R	GAGAAGATGCTGGGCGTGAT/ATAGTGGCTGACCTTGGTGC
ZmACO7-F/R	GATGATGGAGTACCGCAGGG/TGACCTTGGTGCCGTAGAAG
ZmACO8-F/R	AGCTGCTTCGAAGAGGTTATCAA/TGACGCCTACGGAAGGTTTAG
ZmACO9-F/R	TCGGGGACCTCTTGCAGATA/AGATGCCGATGGAAACCCTG
ZmACO10-F/R	AAGGTGGCAAAGTACCCCAG/GCCATCCACCTACCGTCTTT
ZmACO11-F/R	CGTCTTCCGCTCCTACATCG/CAGTGAGGTACAGAGCGAGC
ZmACO12-F/R	AGCTCTGTACCTCACTGGAGA/AAAACGCCTCTTGGTAAGCG
ZmACO15-F/R	TGCCTTCTCCATCCTCAAGTC/CGATGACGGGAACCACCATT
ZmACO20-F/R	GGGAGAGCACCTTCTTCGTC/TTCTCCGACAGCTTCTGCAC
ZmACO31-F/R	AAGAAGGTGTGCTCCGACTG/GATGTCCTCCCAGTCCAGGT
ZmACO35-F/R	CTGGGAGAGCACCTTCTTCG/GATCTCCGATGCGAACTGCT
UBQ9-F/R	TACAGTTCTACAAGGTGGACGAC/GCAGTAGTGGCGGTCGAAGT

反应组分Reaction component	反应体系Reaction system	终浓度Final concentration
2×Fast Super EvaGreen Master Mix	10 μL	1×
正向引物Forward primer	1 μL	0.1-0.5 μmol·L^-1 each
反向引物Reverse primer	1 μL	0.1-0.5 μmol·L^-1 each
cDNA (500 ng·μL^-1) template	1 μL
H₂O	补足至20 μL Make up to 20 μL

名称 Name	基因号 Gene ID	氨基酸长度 Size (aa)	开放阅读框 ORF (bp)	基因定位 Gene mapping	分子量 Molecular weight (Da)	等电点 Isoelectric point
ZmACO1	Zm00001d036955	293	1024	scaffold_6:114257252-114258119(+)	33323.18	6.01
ZmACO2	Zm00001d020686	326	1484	scaffold_7:128315057-128316692(-)	36437.57	5.18
ZmACO3	Zm00001d024852	315	1259	scaffold_10:90656251-90657509(-)	35020.67	5.42
ZmACO4	Zm00001d024851	315	948	scaffold_10:90634003-90634950(-)	35020.67	5.42
ZmACO5	Zm00001d011208	317	1532	scaffold_8:142565914-142567685(+)	34646.05	5.06
ZmACO6	Zm00001d004718	310	1234	scaffold_2:133578506-133580227(+)	34557.35	5.13
ZmACO7	Zm00001d000163	313	1451	scaffold_4:209050198-209052191(-)	35045.80	5.01
ZmACO8	Zm00001d005927	226	681	scaffold_2:193010237-193011059(+)	25466.25	6.22
ZmACO9	Zm00001d039575	376	1526	scaffold_3:8204808-8206696(+)	40606.00	5.77
ZmACO10	Zm00001d015860	234	705	scaffold_5:127015024-127015847(+)	26546.42	6.23
ZmACO11	Zm00001d044978	123	372	scaffold_9:9100281-9102917(-)	13700.64	9.47
ZmACO12	Zm00001d046848	260	793	scaffold_9:108298093-108299643(-)	29077.46	9.52
ZmACO15	Zm00001d024853	356	1599	scaffold_10:90488171-90489769(+)	39417.55	6.03
ZmACO20	Zm00001d052136	323	1330	scaffold_4:180604677-180607029(-)	35551.08	4.97
ZmACO31	Zm00001d024843	314	945	scaffold_10:90456450-90457394(-)	34836.40	5.09
ZmACO35	Zm00001d018211	317	1342	scaffold_5:216633702-216636170(+)	35295.88	4.89

Bioinformatics and Expression Pattern Analysis of Maize ACO Gene Family

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