Accumulation of Anthocyanins in Red-flowered Strawberry‘Meihong’Petals and Expression Analysis of MYB Gene

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

Acta Horticulturae Sinica ›› 2021, Vol. 48 ›› Issue (8): 1470-1484.doi: 10.16420/j.issn.0513-353x.2020-0477

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Accumulation of Anthocyanins in Red-flowered Strawberry‘Meihong’Petals and Expression Analysis of MYB Gene

HONG Yanhong, YE Qinghua, LI Zekun, WANG Wei, XIE Qian, CHEN Qingxi(), CHEN Jianqing()

College of Horticulture,Fujian Agriculture and Forestry University,Fuzhou 350002,China

Received:2021-01-22 Revised:2021-02-22 Online:2021-08-25 Published:2021-09-06
Contact: CHEN Qingxi,CHEN Jianqing E-mail:cqx0246@fafu.edu.cn;jianqingchen@fafu.edu.cn

Abstract

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

CLC Number:

S668.1

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.

Figures/Tables 11

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

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

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

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

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

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

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.

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.

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

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

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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Accumulation of Anthocyanins in Red-flowered Strawberry‘Meihong’Petals and Expression Analysis of MYB Gene

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