红花草莓‘莓红’花瓣花色苷积累及其MYB基因的表达分析

doi:10.16420/j.issn.0513-353x.2020-0477

摘要/Abstract

摘要：

以红花草莓品种‘莓红’为研究材料,观察花瓣发育过程中色泽及花色苷变化规律,同时对4个发育时期花瓣进行转录组测序,基于相关性筛选花色苷合成途径的关键功能基因及转录调控因子,结果表明：（1）花瓣发育过程中,花色苷总含量呈先升后降趋势,且在半开期达到最高;（2）花色苷合成途径关键功能基因FaPAL、Fa4CL-1、Fa4CL-2、FaDFR、Fa3GT表达水平与花色苷含量正相关;（3）R2R3-MYB基因FaMYB6和FaMYB90与花色苷含量呈正相关;（4）Pearson相关性分析表明FaMYB6与FaPAL、FaDFR显著正相关,FaMYB90与FaPAL、Fa4CL-1、Fa4CL-2、FaDFR、Fa3GT极显著正相关,FaMYB82则与上述5个结构基因之间均极显著负相关。综上,‘莓红’草莓花瓣发育过程中,花色苷积累引起花瓣颜色变红,FaMYB82、FaMYB6、FaMYB90可能通过调节FaPAL、Fa4CL、FaDFR和Fa3GT的表达来调控花瓣色泽形成。

关键词: 草莓, 红花, 花瓣, 花色苷, 生物合成, R2R3-MYB 转录因子

Abstract:

‘Meihong’which is a red-flowered strawberry variety was dynamically observed the anthocyanin contents during petals development. Moreover,we performed transcriptome sequencing on four development periods of petals for screening key functional genes and transcription factors in the anthocyanin synthesis pathway. The results showed that：(1) During the lifecycle of petal,the content of anthocyanin is sharply increased and reached the highest at the petal half-open period,and then decreased;(2) The key functional genes of the anthocyanin synthesis pathway,FaPAL,Fa4CL-1,Fa4CL-2,FaDFR,and Fa3GT,were positively correlated with the accumulation of anthocyanins in their expression patterns;(3) FaMYB6 and FaMYB90 were performed positively correlation with anthocyanin content;(4) Pearson correlation analysis showed that FaMYB6 was significantly positively correlated with FaPAL and FaDFR;FaMYB90 had a significant positive correlation with FaPAL,Fa4CL-1,Fa4CL-2,FaDFR and Fa3GT;FaMYB82 was significantly negatively correlated with FaPAL,Fa4CL-1,Fa4CL-2,FaDFR and Fa3GT. In summary,FaMYB82,FaMYB6,FaMYB90 regulate the transcriptional of anthocyanin synthesis genes facilitated the accumulation of anthocyanin caused the color formation of the petals in‘Meihong’strawberry.

Key words: strawberry, red-flower, petal, anthocyanin, biosynthesis, R2R3-MYB transcription factor

中图分类号:

S668.1

洪燕红, 叶清华, 李泽坤, 王威, 谢倩, 陈清西, 陈建清. 红花草莓‘莓红’花瓣花色苷积累及其MYB基因的表达分析[J]. 园艺学报, 2021, 48(8): 1470-1484.

HONG Yanhong, YE Qinghua, LI Zekun, WANG Wei, XIE Qian, CHEN Qingxi, CHEN Jianqing. Accumulation of Anthocyanins in Red-flowered Strawberry‘Meihong’Petals and Expression Analysis of MYB Gene[J]. Acta Horticulturae Sinica, 2021, 48(8): 1470-1484.

图/表 11

图1 ‘莓红’和‘蒙特瑞’草莓花朵发育的7个时期

Fig. 1 The seven developmental stages of flowers on‘Meihong’and‘Monterey’strawberries

表1 引物序列

Table 1 Primer sequences

基因名称 Gene Name	基因ID Gene ID	引物序列（5′-3′） Primer sequences
FaMYB82	maker-Fvb6-4-augustus-gene-249.24	F：ATCGTTAGAGGCAACATTACCG
FaMYB82	maker-Fvb6-4-augustus-gene-249.24	R：TCGCAGCTTCCTCTTTACGAA
FaMYB6	maker-Fvb5-1-augustus-gene-245.45	F：AGACCAACGCCTCATCGACT
FaMYB6	maker-Fvb5-1-augustus-gene-245.45	R：CTCCCCGCAATCAAAGACCAC
FaMYB90	maker-Fvb6-3-snap-gene-424.65	F：AACTACCTTCGGCCAAACATC
FaMYB90	maker-Fvb6-3-snap-gene-424.65	R：TCCAGTAGTTCTTCACATCATTACC
FaPAL	maker-Fvb6-2-augustus-gene-174.36	F：CGCCCCAATTCTAAGTCCGTT
FaPAL	maker-Fvb6-2-augustus-gene-174.36	R：CAGAGCCAACAGCAGTACCAT
Fa4CL-1	maker-Fvb5-2-snap-gene-144.44	F：AGTTCAGACCCTTCCTCATGT
Fa4CL-1	maker-Fvb5-2-snap-gene-144.44	R：CTAGCTGGAGGCAAAAAGGAT
Fa4CL-2	maker-Fvb5-3-augustus-gene-134.28	F：GCTGTGTTGATTTCGCATCCT
Fa4CL-2	maker-Fvb5-3-augustus-gene-134.28	R：GCTTCCTAACCACAAATGCCACT
FaF3H-1	maker-Fvb1-3-augustus-gene-64.31	F：CATCCACCCCAAGCTCAT
FaF3H-1	maker-Fvb1-3-augustus-gene-64.31	R：CAATCCTGCACTGCCTCT
FaF3H-2	maker-Fvb1-4-augustus-gene-56.40	F：GCATCGACCCCAAGCTCAT
FaF3H-2	maker-Fvb1-4-augustus-gene-56.40	R：CCAATCCTGCACCGCTTCT
FaDFR	maker-Fvb2-4-snap-gene-258.87	F：CACGCCTATGGATTTCGAGT
FaDFR	maker-Fvb2-4-snap-gene-258.87	R：AGACCTCCTTTCGATGCTCT
FaANS	maker-Fvb5-3-augustus-gene-266.54	F：TCATGCACATAGGCGACACC
FaANS	maker-Fvb5-3-augustus-gene-266.54	R：CAGAAAACCGCCCACGAGAT
Fa3GT	maker-Fvb7-1-augustus-gene-319.41	F：CGCTCTTTTCACGTATGCTT
Fa3GT	maker-Fvb7-1-augustus-gene-319.41	R：CTTGCTGGTGGTTCTAGTAGGT
Fa3,5GT	maker-Fvb7-2-snap-gene-316.69	F：CCACACCTCCGATCTTCACC
Fa3,5GT	maker-Fvb7-2-snap-gene-316.69	R：CGTACCACCCACAAGAACCT
FaUBC10 （内参Reference gene）	maker-Fvb5-2-augustus-gene-25.49	F：TGCCCTGACCATATCGAAGG
FaUBC10 （内参Reference gene）	maker-Fvb5-2-augustus-gene-25.49	R：CTTGCGGTGGTCTCATACTT

图2 红花草莓‘莓红’和白花草莓‘蒙特瑞’不同发育时期花瓣的花色苷总含量

Fig. 2 Total anthocyanin content in petals of red-flowered strawberry‘Meihong’and white-flowered strawberry‘Monterey’at different developmental stages P ≤ 0.01.

图3 ‘莓红’草莓花瓣花色苷合成相关结构基因表达量分析（FPKM > 10,经Z-score标准化）

Fig. 3 Expression analysis of Structural genes related to anthocyanin synthesis in‘Meihong’strawberry petals （FPKM > 10,standardized by Z-score）

图4 ‘莓红’草莓花瓣MYB基因数、类型统计

Fig. 4 Statistics on the number and types of MYB genes in‘Meihong’strawberry petals

图5 ‘莓红’草莓花瓣差异R2R3-MYB基因表达量（A,FPKM > 10,经Z-score标准化）及其与花色苷含量的Pearson相关性（r）分析（B）

Fig. 5 Expression analysis of differential R2R3-MYB genes（A,FPKM > 10,standardized by Z-score）and Pearson correlation analysis between differential R2R3-MYB genes and anthocyanin content（B）in‘Meihong’strawberry petals

图6 R2R3-MYB转录因子的进化树和保守基序分析分支上数值为邻接法1 000次Bootstrap检验的自引导值。

Fig. 6 Phylogenetic tree and conserved motif analysis of R2R3-MYB transcription factor The branch values were self-guided values of Bootstrap test for 1 000 times by neighbor-joining method.

图7 花色苷合成相关9个差异结构基因和3个R2R3-MYB基因的相对表达量以数据最低表达量,即S1时期Fa3GT的表达量,作为参照数据进行数据相对量计算。

Fig. 7 Relative expression levels of nine differential structural genes and three R2R3-MYB genes related to anthocyanin synthesis The lowest expression level of the data（the expression level of Fa3GT in S1 period）was used as the reference data to calculate the relative amount of data.

图8 3个R2R3-MYB基因与花色苷含量的线性关系分析

Fig. 8 Analysis of linear relationship between three R2R3-MYB genes and anthocyanin content

表2 3个R2R3-MYB基因与9个结构基因的Pearson相关性分析

Table 2 Pearson correlation analysis between three R2R3-MYB genes and nine structural genes

基因名称 Gene name	Pearson系数（r）
基因名称 Gene name	FaPAL	Fa4CL-1	Fa4CL-2	FaF3H-1	FaF3H-2	FaDFR	FaANS	Fa3GT	Fa35GT
FaMYB82	-0.90	-0.93	-0.87	-0.28	-0.32	-0.88	-0.70	-0.85	0.81
FaMYB6	0.61	0.54	0.58	-0.27	-0.20	0.62	0.30	0.56	-0.86
FaMYB90	0.92	0.87	0.95	0.31	0.33	0.94	0.70	0.90	-0.79

图9 候选花色苷合成相关基因在白花草莓‘蒙特瑞’花瓣发育过程中的表达模式分析以数据最低表达量,即S1时期Fa3GT的表达量作为参照数据进行数据相对量计算。

Fig. 9 Relative expression levels of nine structural genes and three R2R3-MYB genes related to anthocyanin synthesis in white-flowered strawberry‘Monterey’ The lowest expression level of the data（Fa3GT in S1 period）was used as the reference data to calculate the relative amount of data.

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