板栗R2R3-MYB基因家族的鉴定及表达分析

doi:10.13304/j.nykjdb.2024.0477

摘要/Abstract

摘要：

R2R3-MYB转录因子是植物中最大的MYB亚家族，广泛参与调节植物生长发育和胁迫响应。对板栗R2R3-MYB基因家族进行全基因组鉴定，分析其在染色体上分布、亚细胞定位、系统进化关系、蛋白基序、基因结构、保守结构域，最后通过转录组和RT-qPCR分析CmR2R3-MYB基因在干旱胁迫下的表达。结果表明，共鉴定出141个CmR2R3-MYB基因，不均等地分布于12条染色体与1条contig片段。系统进化树将其分为6个亚家族。有7个CmR2R3-MYB基因在干旱胁迫下差异表达，其中CmMYB89表达量上调，CmMYB32、CmMYB95、CmMYB91、CmMYB119、CmMYB131及CmMYB104表达量下调，表明这些基因可能与板栗抗旱性密切相关。以上研究结果为进一步探究R2R3-MYB基因家族在板栗响应干旱胁迫方面的作用奠定了理论基础。

关键词: 板栗, 干旱, R2R3-MYB基因家族

Abstract:

R2R3-MYB transcription factors are the largest MYB subfamily in plants and are widely involved in regulating plant growth and development and stress response. R2R3-MYB gene family of Chinese chestnut was genome-widely identified， and the chromosome distribution of these genes， subcellular localization prediction， phylogenetic relationships， protein motif， gene structure and sequence alignment of conserved domains were analyzed. Finally， the expression of the CmR2R3-MYB gene family under drought stress was analyzed by transcriptome and RT-qPCR. The results showed that total 141 CmR2R3-MYB genes were unequally distributed on 12 chromosomes and 1 contig fragment. And the phylogenetic tree divided them into 6 groups. There were 7 CmR2R3-MYB genes differentially expressed under drought stress. Among them， CmMYB89 was upregulated， while CmMYB32， CmMYB95， CmMYB91， CmMYB119， CmMYB131 and CmMYB104 were downregulated， which indicated that these 7 genes could be involved in drought resistance of Chinese chestnut. Above results provided theoretical foundations for further exploring the role of the R2R3-MYB gene family in response to drought stress of Chinese chestnut.

Key words: Chinese chestnut, drought, R2R3-MYB gene family

中图分类号:

S664.2

刘亚明, 杨涌, 康肖肖, 王萌, 王东升, 张海娥. 板栗R2R3-MYB基因家族的鉴定及表达分析[J]. 中国农业科技导报, 2025, 27(8): 47-59.

Yaming LIU, Yong YANG, Xiaoxiao KANG, Meng WANG, Dongsheng WANG, Haie ZHANG. Identification and Expression Analysis of R2R3-MYB Gene Family in Chinese Chestnut[J]. Journal of Agricultural Science and Technology, 2025, 27(8): 47-59.

图/表 13

表1 试验所用引物序列

Table 1 Primer sequences used in the study

基因Gene	上游引物Forward primer （5’-3’）	下游引物Reverse primer （5’-3’）
CmMYB119	TGGTCAGCAATCGCAGCT	TGTTTCTGGGTTGGCTTGGT
CmMYB91	GCAGACTCCGCTGGACAA	GCTGCAATTGCTGACCACC
CmMYB89	AGCTGCAGGGCTTCTTCG	CCCTGGCAATCTCCCAGC
CmMYB131	TGGTGACCCTCAATGCCC	CCGGATCACATGCGTGGA
CmMYB104	TGCGGAGAGGACCATGGA	ATTCCAACGGCCCTCACC
CmMYB32	GGCTTTGACACTTGCGCC	AACTCTCCGACCTCCGCT
CmMYB95	GAGCAGCCCATCCGGATC	TCTCGCCACGGGTCTGTA
Actin	ATTCACGAGACCACCTACA	TGCCACAACCTTAATCTTCAT

图1 基因在染色体上的分布情况注：1~12为染色体；13为contig片段。

Fig. 1 Distribution of the genes on the chromosomeNote：1~12 are chromosomes； 13 is contig fragment.

图2 CmR2R3-MYB蛋白的系统进化树

Fig. 2 Phylogenetic tree of CmR2R3-MYB proteins

图3 CmMYB蛋白的保守基序

Fig. 3 Conserved motifs of CmMYBproteins

表2 不同类群的Motif

Table 2 Motifs of different groups

类群Group	基序Motif
Group 1	motif 6，motif 10
Group 2	motif 6， motif 7， motifm 10
Group 3	motif 6， motif 7， motif 10
Group 4	motif 6， motif 10
Group 5	motif 6， motif 8~10
Group 6	motif 5， motif 8

图4 R2、R3的保守结构域序列比对注：*表示保守氨基酸。

Fig. 4 Sequence alignment of conserved R2 and R3 domainsNote： * indicates conserved amino acid.

图5 基因家族成员顺式作用元件数量及其在启动子上的分布

Fig. 5 Cis-acting elements of gene family members and distribution on promoters

图6 CmR2R3-MYB基因家族成员的共线性及复制类型A：基因组内共线性；B：基因家族复制类型；C：替换率

Fig. 6 Collinearity analysis among CmR2R3-MYB gene family members and duplication typesA； Collinearity in genome； B： Type of gene family duplications； C： Replacement rate

图7 板栗与其他种间R2R3-MYB基因家族的共线性分析A：板栗与拟南芥、水稻；B：板栗与美洲栗、日本栗

Fig. 7 Collinearity analysis of R2R3-MYB gene family between chestnut and other speciesA： Castanea mollissima with Arabidopsis thaliana， Oryza sativa； B： Castanea mollissima with Castanea dentata and Castanea crenata

图8 CmR2R3-MYB基因密码子的RSCU值

Fig. 8 RSCU values of codon of CmR2R3-MYB genes

图9 CmR2R3-MYB基因家族密码子偏好性分析A：中性绘图分析；B：ENC-plot分析；C：PR2-plot分析

Fig. 9 Codon bias analysis of CmR2R3-MYB gene familyA： Neutrality plot analysis； B： ENC-plot analysis； C： PR2-plot analysis

图10 干旱胁迫下CmR2R3-MYB转录组分析A：干旱胁迫后转录组分析的火山图；B：CmR2R3-MYB基因家族中差异表达基因的表达热图

Fig. 10 Transcriptome analysis of CmR2R3-MYB under drought stressA： Volcano map of transcriptome analysis after drought stress； B： Heat map of differentially expressed CmR2R3-MYB genes

图11 CmR2R3-MYB差异表达基因在不同处理的相对表达量注：**和***分别表示与对照相比在P<0.01和P<0.001水平差异显著。

Fig. 11 Relative expressions of differentially CmR2R3-MYB expressed genes under different treatmentsNote：** and *** indicate significant differences compared with CK at P<0.01 and P<0.001 levels， respectively.

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