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Functional Analysis of a Pomegranate (Punica granatum L.) MYB Transcription Factor Involved in the Regulation of Anthocyanin Biosynthesis.

ABSTRACT: BACKGROUND:Pomegranate fruit (Punica granatum L.) is a rich source of anthocyanin pigments resulting in vibrant colours and anti-oxidant contents. Although the intensity and pattern of anthocyanin biosynthesis in fruit are strongly influenced by R2R3-MYB transcription factors, little is known about the regulation and role of MYB in anthocyanin pathway of pomegranate. OBJECTIVES:The present study was conducted to elucidate the relationship between the expression of MYB transcription factor and the anthocyanin accumulation during the colour development phase of pomegranate fruits. MATERIALS AND METHODS:In this work, R2R3-MYB transcription factor (PgMYB) was isolated and characterized from pomegranate skin through RACE-PCR. The expression of PgMYB gene was monitored in three distinct pomegranate accessions with distinctive skin colour and pattern by semi-quantitative RT-PCR. RESULTS:The results indicated a strong association between skin colour in mature pomegranate fruits with the PgMYB transcripts. The highest expression level of PgMYB gene was observed in Poost Siyah Yazd (dark purple skin) throughout the ripening process. Furthermore, comparison of PgMYB amino acid sequences with those of R2R3-MYB family in grapevine, eucalyptus, peach, cacao, populus and Arabidopsis demonstrated that this protein shares high similarity (75-85% amino acid identity) with their conserved MYB domain. Computational structure prediction of PgMYB showed that the three conserved amino acids (Asn, Lys and Lys) are present in the same position of the MYB domain. CONCLUSIONS:It is speculated that PgMYB gene influences the fruit colour and could be used to improve the accumula-tion of anthocyanin pigments in the pomegranate fruit.

SUBMITTER: Khaksar G

PROVIDER: S-EPMC5434983 | biostudies-literature | 2015 Mar

REPOSITORIES: biostudies-literature

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Functional Analysis of a Pomegranate (<i>Punica granatum</i> L.) MYB Transcription Factor Involved in the Regulation of Anthocyanin Biosynthesis.

Khaksar Ghazale G Sayed Tabatabaei Badraldin Ebrahim BE Arzani Ahmad A Ghobadi Cyrus C Ebrahimie Esmaeil E

Iranian journal of biotechnology 20150301 1

<h4>Background</h4>Pomegranate fruit (<i>Punica granatum</i> L.) is a rich source of anthocyanin pigments resulting in vibrant colours and anti-oxidant contents. Although the intensity and pattern of anthocyanin biosynthesis in fruit are strongly influenced by R2R3-MYB transcription factors, little is known about the regulation and role of MYB in anthocyanin pathway of pomegranate.<h4>Objectives</h4>The present study was conducted to elucidate the relationship between the expression of MYB trans ...[more]

PMID: 28959277

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Project description:BACKGROUND: Red coloration of fruit is an important trait in apple, and it is mainly attributed to the accumulation of anthocyanins, a class of plant flavonoid metabolites. Anthocyanin biosynthesis is genetically determined by structural and regulatory genes. Plant tissue pigmentation patterns are mainly controlled by expression profiles of regulatory genes. Among these regulatory genes are MYB transcription factors (TFs), wherein the class of two-repeats (R2R3) is deemed the largest, and these are associated with the anthocyanin biosynthesis pathway. Although three MdMYB genes, almost identical in nucleotide sequences, have been identified in apple, it is likely that there are other R2R3 MYB TFs that are present in the apple genome that are also involved in the regulation of coloration of red color pigmentation of the skin of apple fruits. RESULTS: In this study, a novel R2R3 MYB gene has been isolated and characterized in apple. This MYB gene is closely related to the Arabidopsis thaliana AtMYB3, and has been designated as MdMYB3. This TF belongs to the subgroup 4 R2R3 family of plant MYB transcription factors. This apple MdMYB3 gene is mapped onto linkage group 15 of the integrated apple genetic map. Transcripts of MdMYB3 are detected in all analyzed tissues including leaves, flowers, and fruits. However, transcripts of MdMYB3 are higher in excocarp of red-skinned apple cultivars than that in yellowish-green skinned apple cultivars. When this gene is ectopically expressed in Nicotiana tabacum cv. Petite Havana SR1, flowers of transgenic tobacco lines carrying MdMYB3 have exhibited increased pigmentation and accumulate higher levels of anthocyanins and flavonols than wild-type flowers. Overexpression of MdMYB3 has resulted in transcriptional activation of several flavonoid pathway genes, including CHS, CHI, UFGT, and FLS. Moreover, peduncles of flowers and styles of pistils of transgenic plants overexpressing MdMYB3 are longer than those of wild-type plants, thus suggesting that this TF is involved in regulation of flower development. CONCLUSIONS: This study has identified a novel MYB transcription factor in the apple genome. This TF, designated as MdMYB3, is involved in transcriptional activation of several flavonoid pathway genes. Moreover, this TF not only regulates the accumulation of anthocyanin in the skin of apple fruits, but it is also involved in the regulation of flower development, particularly that of pistil development.

Project description:BackgroundThe basic leucine zipper (bZIP) transcription factor is one of the most abundant and conserved gene families in eukaryotes. In addition to participating in plant development and growth, bZIP transcription factors play crucial roles in various abiotic stress responses and anthocyanin accumulation. Up to now, analysis of bZIP gene family members in pomegranate (Punica granatum) has not been reported. Three published pomegranate genome sequences provide valuable resources for further gene function analysis.ResultsUsing bioinformatics analysis, 65 PgbZIPs were identified and analyzed from the 'Taishanhong' pomegranate genome. We divided them into 13 groups (A, B, C, D, E, F, G, H, I, J, K, M, and S) according to the phylogenetic relationship with those of Arabidopsis, each containing a different number of genes. The regularity of exon/intron number and distribution was consistent with the classification of groups in the evolutionary tree. Transcriptome analysis of different tissues showed that members of the PgbZIP gene family were differentially expressed in different developmental stages and tissues of pomegranate. Among them, we selected PgbZIP16 and PgbZIP34 as candidate genes which affect anthocyanin accumulation. The full-length CDS region of PgbZIP16 and PgbZIP34 were cloned from pomegranate petals by homologous cloning technique, encoding 170 and 174 amino acids, which were 510 bp and 522 bp, respectively. Subcellular localization assays suggested that both PgbZIP16 and PgbZIP34 were nucleus-localized. Real-time quantitative PCR (qPCR) was used to explore the expression of PgbZIP16 and PgbZIP34 in the petals of three kinds of ornamental pomegranates at the full flowering stage. The results demonstrated that the expression of PgbZIP16 in red petals was 5.83 times of that in white petals, while PgbZIP34 was 3.9 times. The results of transient expression in tobacco showed that consistent trends were observed in anthocyanin concentration and expression levels of related genes, which both increased and then decreased. Both PgbZIP16 and PgbZIP34 could promote anthocyanin accumulation in tobacco leaves. We obtained transgenic strains overexpressing PgbZIP16, and the histochemical staining for GUS activity showed that overexpressed PgbZIP16 seedlings were expressed in the stem. Transgenic experiments indicated that overexpression of PgbZIP16 significantly upregulated UF3GT, ANS and DFR genes in Arabidopsis and enhanced anthocyanin accumulation.ConclusionsThe whole genome identification, gene structure, phylogeny, gene cloning, subcellular location and functional verification of the pomegranate bZIP gene family provide a theoretical foundation for the functional study of the PgbZIP gene family and candidate genes for anthocyanin biosynthesis.

Dataset Information

Functional Analysis of a Pomegranate (Punica granatum L.) MYB Transcription Factor Involved in the Regulation of Anthocyanin Biosynthesis.

Publications

Functional Analysis of a Pomegranate (<i>Punica granatum</i> L.) MYB Transcription Factor Involved in the Regulation of Anthocyanin Biosynthesis.

Similar Datasets

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