玉米OPR基因家族的泛基因组鉴定与表达模式分析

doi:10.7668/hbnxb.20194096

摘要/Abstract

摘要：

12-氧-植物二烯酸还原酶(OPR)为黄素单核苷酸(FMN)依赖的氧化还原酶,是合成茉莉酸的关键酶,其对植物生长发育及防御调控具有重要作用。为研究OPR基因在玉米中的抗病作用,利用生物信息学方法,在31个玉米不同自交系中对OPR家族成员进行鉴定,并分析受玉米大斑病菌侵染后的表达规律。结果表明,在玉米B73品系中鉴定出了8个OPR基因,这些基因不均匀地分布于7条染色体上,且其表达蛋白富含酸性氨基酸。进一步分析发现,玉米OPR家族只有1种结构域Oxidored_FMN,并且所有成员均包含已鉴定的10个蛋白质保守基序。利用MEGA软件对玉米、小麦、水稻和拟南芥的OPR家族成员进行系统发育关系分析,发现这些植物OPR基因家族进化关系具有明显差异,其中玉米和水稻的进化关系最为接近。运用OrthoFinder软件对其同源组分析发现,玉米OPR基因家族具有高度保守性,所有OPR基因均为核心基因,但其功能略有差别。进一步利用河北省植物生理与分子病理学重点实验室前期的RNA-seq数据,对玉米B73 品系OPR基因响应大斑病菌侵染的表达模式进行分析,并通过qRT-PCR对基因表达模式进行验证,发现这些基因在病菌侵染玉米过程中呈现3种不同的表达模式。本研究系统的鉴定了玉米泛基因组OPR家族基因,以及确定了其在应对玉米大斑病菌侵染后的表达模式。

关键词: 玉米, OPR家族, 系统进化, 泛基因组, 表达模式

Abstract:

12-oxo-phytodienoic(OPR) acid reductase is a flavin mononucleotide-dependent oxidoreductase,a key enzyme for the synthesis of jasmonic acid,which is important for plant growth, development and defence regulation.In order to study the disease resistance effect of OPR gene in maize, bioinformatics methods were used to identify OPR family members in 31 different maize inbred lines and the expression pattern after infestation by S. turcica was analyzed.The results showed that eight OPR genes were identified in maize line B73 and they were unevenly distributed on seven chromosomes.In addition,these expressed proteins of OPR genes were rich in acidic amino acids.Further analysis revealed that the maize OPR family contained only one structural domain,Oxidored_FMN,and all OPR members contained 10 protein conserved motifs identified.Phylogenetic relationship analysis of OPR family members from maize,wheat,rice and Arabidopsis using MEGA software revealed significant differences in the evolutionary relationships of OPR gene families in these plants.The OPR proteins of maize and those of rice were the closest in evolution.Homology analysis using OrthoFinder software revealed that the maize OPR gene family was highly conserved,with all OPR genes being core genes,but with slightly different functions.According to the previous RNA-seq data of Hebei key Laboratory of Plant Physiology and Molecular Pathology,we analyzed the expression pattern of OPR genes of maize B73 in response to S.turcica infection and verified the gene expression pattern of OPR genes through qRT-PCR.It was found that there were three different expression patterns in the process of S.turcica infection.This study systematically identified the maize pan-genomic OPR family genes and their expression patterns in response to S.turcica infection.

Key words: Maize, OPR family, Phylogenetic, Pan-genome, Expression pattern

李雯霏, 李红霞, 刘玉卫, 巩校东, 魏淑珍, 谷守芹. 玉米OPR基因家族的泛基因组鉴定与表达模式分析[J]. 华北农学报, 2023, 38(5): 51-59. doi: 10.7668/hbnxb.20194096.

LI Wenfei, LI Hongxia, LIU Yuwei, GONG Xiaodong, WEI Shuzhen, GU Shouqin. Pan-genomic Identification and Expression Pattern Analysis of the OPR Gene Family in Maize[J]. Acta Agriculturae Boreali-Sinica, 2023, 38(5): 51-59. doi: 10.7668/hbnxb.20194096.

http://www.hbnxb.net/CN/Y2023/V38/I5/51

图/表 11

表1 玉米B73 OPR家族的理化性质

Tab.1 Physical and chemical properties of maize B73 OPR family

基因序号 Gene ID	氨基酸数量/个 Number of amino acids	分子质量/ku Molecular weight	理论等电点 Theoretical isoelectric point(pI)
Zm00001eb373030	373	41.592 98	6.47
Zm00001eb373050	376	41.901 20	5.85
Zm00001eb279210	377	41.616 81	5.33
Zm00001eb131870	372	41.393 61	5.37
Zm00001eb040310	399	44.103 77	8.53
Zm00001eb082050	391	43.489 38	6.76
Zm00001eb177330	399	44.220 88	7.15
Zm00001eb356160	383	42.216 62	5.54

图2 玉米B73 OPR基因家族系统发育树及基因结构

Fig.2 Phylogenetic tree and gene structure of maize B73 OPR gene family

图1 玉米B73 OPR家族染色体定位

Fig.1 Chromosome mapping of maize B73 OPR family

图3 玉米B73 OPR家族蛋白保守基序

Fig.3 Conserved motifs of maize B73 OPR family proteins

图4 物种间OPR家族进化树

Fig.4 Phylogenetic tree of OPR family among the tested species

表2 玉米31个品系中OPR家族的数量

Tab.2 Number of OPR families in 31 maize inbred lines

玉米品系 Maize strains	OPR蛋白个数 Number of OPR proteins	玉米品系 Maize strains	OPR蛋白个数 Number of OPR proteins
B73	11	Ky21	12
B97	16	M37W	11
CML52	14	M162W	12
CML69	12	Mo17	11
CML103	10	Mo18W	18
CML228	10	Ms71	11
CML247	13	NC350	16
CML277	9	NC358	13
CML322	15	Oh7B	11
CML333	15	Oh43	13
EP1	8	P93	16
F7	8	PH207	8
HP301	14	Tx303	12
Il14H	12	Tzi8	14
Ki3	9	W22	10
Ki11	11

表3 玉米OPR家族同源组

Tab.3 Homologous group of maize OPR family

Orthogroup	OG0000000	OG0000001	总计 Total
B73	9	2	11
B97	10	6	16
CML103	10	2	12
CML228	8	2	10
CML247	11	2	13
CML277	7	2	9
CML322	11	4	15
CML333	10	5	15
CML52	12	2	14
CML69	6	2	8
EP1	6	2	8
F7	6	2	8
HP301	10	4	14
Il14H	10	2	12
Ki11	9	2	11
Ki3	7	2	9
Ky21	9	3	12
M162W	8	4	12
M37W	9	2	11
Mo17	9	2	11
Mo18W	16	2	18
Ms71	9	2	11
NC350	12	4	16
NC358	11	2	13
Oh43	9	4	13
Oh7B	9	2	11
P39	12	4	16
PH207	6	2	8
Tx303	10	2	12
Tzi8	11	3	14
W22	8	2	10
总计 Total	290	83	373

图5 玉米泛基因组OPR家族进化树

Fig.5 Evolutionary tree of maize pan-genome OPR family

图6 玉米B73 和B73Ht的OPR基因响应大斑病菌表达模式热点图

Fig.6 Heatmap of expression pattern of maize B73 and B73Ht OPR gene in response to S.turcica

图7 玉米B73叶片受玉米大斑病菌侵染后提取总RNA的完整性检测 1-2.侵染时间为0 h;3.侵染时间为24 h;4.侵染时间为72 h。

Fig.7 Integrity detection of total RNA extracted from leaves of maize B73 infected by S.turcica 1-2.Infection time of 0 h;3.Infection time of 24 h;4.Infection time of 72 h.

图8 玉米B73的8个OPR蛋白受玉米大斑病菌侵染的表达水平分析不同小写字母表示差异显著,P<0.05。

Fig.8 Expression level analysis of eight OPR proteins of maize B73 infected by S.turcica Different lowercase letters indicate significant difference,P<0.05.

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