Identification of the MYB Gene Family and Functional Analysis of Key Genes Related to Blue Flower Coloration in Agapanthus praecox

doi:10.16420/j.issn.0513-353x.2022-0226

Acta Horticulturae Sinica ›› 2023, Vol. 50 ›› Issue (6): 1255-1268.doi: 10.16420/j.issn.0513-353x.2022-0226

• Genetic & Breeding·Germplasm Resources·Molecular Biology • Previous Articles Next Articles

Identification of the MYB Gene Family and Functional Analysis of Key Genes Related to Blue Flower Coloration in Agapanthus praecox

HE Weizhi¹^,², LEI Weiqi¹, GUO Xiangxin¹, LI Ruilian¹^,³, CHEN Guanqun¹^,^*()

¹Department of Landscape Architecture，School of Design，Shanghai Jiao Tong University，Shanghai 200240，China
²School of Agriculture and Biology，Shanghai Jiao Tong University，Shanghai 200240，China
³Shanghai Administration Affairs Center of Parks，Shanghai 200023，China

Received:2022-12-29 Revised:2023-05-05 Online:2023-06-25 Published:2023-06-27
Contact: * （E-mail：chengq@sjtu.edu.cn）

Abstract

Abstract:

MYB gene family members of Agapanthus praecox were identified by bioinformatics methods and the function of key genes that regulate blue flower coloration was verified. In total，123 ApMYBs were identified based on full-length transcriptome sequencing data，including 60 R2R3-MYBs，four 3R-MYBs，two 4R-MYBs and 57 1R-MYBs. Then，the R2R3-MYBs that regulate flavonoid biosynthesis were further analyzed. R2R3-MYBs were divided into 22 subgroups（A1-A22），together with A18（S4），A17（S5）and A16（S7）subgroups were all related to flower coloration，except for S6 subgroup. ApMYB4，ApMYB6，ApMYB7，ApMYB12，ApMYB111 and ApMYB123 were all localized in the nuclei. Whereas，ApMYB123 was especially concentrated in the nucleolus. During blue flower coloration，ApMYB12 was significantly up-regulated，while ApMYB111 was opposite. White flowers showed increased expression of ApMYB4，ApMYB6 and ApMYB7 compared to blue flowers，indicating them as the negative regulators of coloration. A specific expression peak of ApMYB123 was arisen in bloom stage of white flowers，suggesting biosynthesis of proanthocyanidins. Further ectopic expression of ApMYB12 in tobacco found that the petal color turned lighter than wild type plants. In addition，the anthocyanin content reduced by 54.55%-62.50% while the flavonols content increased by 125%-170%. These results demonstrated that ApMYB12 regulated the biosynthesis of auxiliary pigment flavonols，thereby regulating the coloration of blue flower.

Key words: Agapanthus praecox, transcription factor, MYB gene family, blue flower coloration, flavonol

CLC Number:

S682.32

HE Weizhi, LEI Weiqi, GUO Xiangxin, LI Ruilian, CHEN Guanqun. Identification of the MYB Gene Family and Functional Analysis of Key Genes Related to Blue Flower Coloration in Agapanthus praecox[J]. Acta Horticulturae Sinica, 2023, 50(6): 1255-1268.

Figures/Tables 9

Table 1 Primer list for PCR

基因名称 Gene name	正向引物序列（5′-3′） Forward primer sequence	反向引物序列（5′-3′） Reverse primer sequence
ApActin	AACACCACCAGACTTATGT	GCCGTTGAACTTGATGAG
ApMYB4	CTACTGGAACACGCACATCAA	CTCTCGTCGGTAGAATCCTCAT
ApMYB6	AGTTGTCGGCTTCGTTGGA	TGTCTGTTCTTCCTGGTAGTCTT
ApMYB7	GACCTATCCATTAGCCTTCCTTATC	CTGTTGTTCTTGTCGCTGTCA
ApMYB12	GACCTCGACCAAGACTTGTTAC	ATCTAGCACCTTCTCCTCAACTT
ApMYB111	GGCACGGTATCAGCGGTAT	TTCTCATGCTCGTCTCTTGTTG
ApMYB123	CCTTATCATCCGACTCCACTCT	TTGCTGAGGTGACTGTTCCA
pHB-ApMYB4	CTCTCTCTCAAGCTTGGATCCATGGGTAGGTCTCCATGTTG	CATACTAGTGAGCTCCTGCAGAGCATTATACCCATCCTGTA
pHB-ApMYB6	CTCTCTCTCAAGCTTGGATCCATGGGTAGATCTCCTTGTTG	CATACTAGTGAGCTCCTGCAGCTTCATCTGAAAACTTCTAT
pHB-ApMYB7	CTCTCTCTCAAGCTTGGATCCATGGGAAGGTCCCCATGTTG	CATACTAGTGAGCTCCTGCAGTATCCATCCTTCCTCCAACG
pHB-ApMYB12	CTCTCTCTCAAGCTTGGATCCATGGGAAGGGCTCCATGTTG	CATACTAGTGAGCTCCTGCAGCAACACATCTGAAAGAAGCC
pHB-ApMYB111	CTCTCTCTCAAGCTTGGATCCATGGGGAGAGTTCCATGTTG	CATACTAGTGAGCTCCTGCAGTAATTTGCACCCACTCTCCC
pHB-ApMYB123	CTCTCTCTCAAGCTTGGATCCATGGGGAGAGCTCCATGCTG	CATACTAGTGAGCTCCTGCAGAATCAACAGAGGCTCTGCAG
NtPAL	ATTGAGGTCATCCGTTCTGC	ACCGTGTAACGCCTTGTTTC
Nt4CL	TCATTGACGAGGATGACGAG	TGGGATGGTTGAGAAGAAGG
NtCHS	TTGTTCGAGCTTGTCTCTGC	AGCCCAGGAACATCTTTGAG
NtCHI	GTCAGGCCATTGAAAAGCTC	CTAATCGTCAATGCCCCAAC
NtFLS	TTTGGCACTTGGTGTTGTGG	ACTTGACATCATACCAATGG
NtDFR	AACCAACAGTCAGGGGAATG	TTGGACATCGACAGTTCCAG
NtUFGT	ATGTTGAAGGGCTAAAAGAAAGAGC	CAAGTCCCAGCTGATACATATTCCC
NtGAPDH	GGTGTCCACAGACTTCGTGG	GACTCCTCACAGCAGCACCA

Fig. 1 Phylogenetic tree of R2R3-MYB proteins between Agapanthus praecox（A）and Arabidopsis thaliana（S）

Fig. 2 Consensus sequence of R2R3-MYB domains in Agapanthus praecox The bit score of Y-axis indicated the conservation rate for each X-axis amino acid position in the sequence. Asterisks indicate the conserved residues Trp（W）in the MYB domain.

Fig. 3 The anthocyanin contents during flower development in blue-flowered‘Big Blue’and white-flowered‘Bridal Bouquet’ S1：Bud emergence period；S2：Bud elongation period；S3：Pre-opening period. The different lowercase letters respectively indicate the significant difference in the anthocyanin content（P < 0.05）.

Fig. 4 Heat map of expression（A）and protein conserved motifs（B）of MYB genes related to flavonoid biosynthesis during flower coloration in blue-flowered‘Big Blue’and white-flowered‘Bridal Bouquet’ S1：Bud emergence period；S2：Bud elongation period；S3：Pre-opening period.

Fig. 5 The subcellular localization of MYB proteins related to flavonoid biosynthesis（Bar = 50 μm）

Fig. 6 Flower color phenotypes, flavonol and anthocyanin content in petal and leaf of ApMYB12-transgenic tobacco lines（7#，8#）and wild type ** represents the significant difference between wild-type and transgenic lines（P < 0.01）.

Fig. 7 The flavonoid biosynthesis pathway of ApMYB12-transgenic tobacco lines and wild type

Fig. 8 The expression of key genes in the flavonoid biosynthesis pathway of of ApMYB12-transgenic tobacco lines（7#，8#）and wild type Different lowercase letters indicate significant differences between wild-type and transgenic lines（P < 0.05）.

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Identification of the MYB Gene Family and Functional Analysis of Key Genes Related to Blue Flower Coloration in Agapanthus praecox

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