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A Novel bHLH Transcription Factor Involved in Regulating Anthocyanin Biosynthesis in Chrysanthemums (Chrysanthemum morifolium Ramat.).

Xiang LL, Liu XF, Li X, Yin XR, Grierson D, Li F, Chen KS - PLoS ONE (2015)

Bottom Line: CmbHLH2 significantly upregulated the CmDFR promoter and triggered anthocyanin accumulation when co-expressed with CmMYB6.Yeast one-hybrid analyses indicated that CmbHLH2 was able to bind directly to the CmDFR promoter.These results suggest that CmbHLH2 is the essential partner for CmMYB6 in regulating anthocyanin biosynthesis in chrysanthemum.

View Article: PubMed Central - PubMed

Affiliation: College of Agriculture & Biotechnology, Zhejiang University, Zijingang Campus, Hangzhou, 310058, PR China.

ABSTRACT
Chrysanthemums (Chrysanthemum morifolium Ramat.) exhibit a variety of flower colors due to their differing abilities to accumulate anthocyanins. One MYB member, CmMYB6, has been verified as a transcription regulator of chrysanthemum genes involved in anthocyanin biosynthesis; however, the co-regulators for CmMYB6 remain unclear in chrysanthemum. Here, the expression pattern of CmbHLH2, which is clustered in the IIIf bHLH subgroup, was shown to be positively correlated with the anthocyanin content of cultivars with red, pink and yellow flower colors, respectively. CmbHLH2 significantly upregulated the CmDFR promoter and triggered anthocyanin accumulation when co-expressed with CmMYB6. Yeast one-hybrid analyses indicated that CmbHLH2 was able to bind directly to the CmDFR promoter. Moreover, yeast two-hybrid assays indicated protein-protein interaction between CmbHLH2 and CmMYB6. These results suggest that CmbHLH2 is the essential partner for CmMYB6 in regulating anthocyanin biosynthesis in chrysanthemum.

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Phylogenetic analysis of 178 bHLH members including 162 from Arabidopsis, 14 from other plants, and 2 from chrysanthemum.The two bHLHs from chrysanthemum are marked with solid circles, while those related to anthocyanin biosynthesis regulation in Arabidopsis and other species are marked with open circles. Genes IDs are as follows: NtAn1b (Nicotiana tabacum, GenBank accession number AEE99258), NtAN1-like (N. tomentosiformis, AEE99260), NtAn1a (N. tabacum, AEE99257), AN1 (Petunia hybrid, AF260918), MrbHLH1 (Myrica rubra, JX629461), LjTT8 (Lotus japonicus, BAH28881), MdbHLH3 (Malus domestica, HM122458), DvIVS (Dahlia variabilis, AB601005), AmDELILA (Antirrhinum majus, AAA32663), R-S (Zea mays, X15806), L-c (Z. mays, NM001111869), MrbHLH2 (M. rubra, JX629462), MdbHLH33 (M. domestica, DQ266451), VvMYC (Vitis vinifera, EU447172).
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pone.0143892.g001: Phylogenetic analysis of 178 bHLH members including 162 from Arabidopsis, 14 from other plants, and 2 from chrysanthemum.The two bHLHs from chrysanthemum are marked with solid circles, while those related to anthocyanin biosynthesis regulation in Arabidopsis and other species are marked with open circles. Genes IDs are as follows: NtAn1b (Nicotiana tabacum, GenBank accession number AEE99258), NtAN1-like (N. tomentosiformis, AEE99260), NtAn1a (N. tabacum, AEE99257), AN1 (Petunia hybrid, AF260918), MrbHLH1 (Myrica rubra, JX629461), LjTT8 (Lotus japonicus, BAH28881), MdbHLH3 (Malus domestica, HM122458), DvIVS (Dahlia variabilis, AB601005), AmDELILA (Antirrhinum majus, AAA32663), R-S (Zea mays, X15806), L-c (Z. mays, NM001111869), MrbHLH2 (M. rubra, JX629462), MdbHLH33 (M. domestica, DQ266451), VvMYC (Vitis vinifera, EU447172).

Mentions: Based on the similarities with bHLHs verified as being related to anthocyanin biosynthesis in other species, one partial sequence bHLH member was mined from the chrysanthemum EST database. The full ORF, which was obtained by RACE and named as CmbHLH2 (GenBank KT724056), contained 1842 nucleotides and encoded 614 amino acids. A second bHLH member isolated from chrysanthemum, CmbHLH1, which was reported to be involved in the response to iron deficiency [30] was also selected. Phylogenetic analysis of these two members and 174 bHLHs including 158 from Arabidopsis and 14 from other plants, showed that CmbHLH2 was clustered in the subgroup containing sequences related to the regulation of anthocyanin biosynthesis, while CmbHLH1 was in a different subgroup (Fig 1).


A Novel bHLH Transcription Factor Involved in Regulating Anthocyanin Biosynthesis in Chrysanthemums (Chrysanthemum morifolium Ramat.).

Xiang LL, Liu XF, Li X, Yin XR, Grierson D, Li F, Chen KS - PLoS ONE (2015)

Phylogenetic analysis of 178 bHLH members including 162 from Arabidopsis, 14 from other plants, and 2 from chrysanthemum.The two bHLHs from chrysanthemum are marked with solid circles, while those related to anthocyanin biosynthesis regulation in Arabidopsis and other species are marked with open circles. Genes IDs are as follows: NtAn1b (Nicotiana tabacum, GenBank accession number AEE99258), NtAN1-like (N. tomentosiformis, AEE99260), NtAn1a (N. tabacum, AEE99257), AN1 (Petunia hybrid, AF260918), MrbHLH1 (Myrica rubra, JX629461), LjTT8 (Lotus japonicus, BAH28881), MdbHLH3 (Malus domestica, HM122458), DvIVS (Dahlia variabilis, AB601005), AmDELILA (Antirrhinum majus, AAA32663), R-S (Zea mays, X15806), L-c (Z. mays, NM001111869), MrbHLH2 (M. rubra, JX629462), MdbHLH33 (M. domestica, DQ266451), VvMYC (Vitis vinifera, EU447172).
© Copyright Policy
Related In: Results  -  Collection

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pone.0143892.g001: Phylogenetic analysis of 178 bHLH members including 162 from Arabidopsis, 14 from other plants, and 2 from chrysanthemum.The two bHLHs from chrysanthemum are marked with solid circles, while those related to anthocyanin biosynthesis regulation in Arabidopsis and other species are marked with open circles. Genes IDs are as follows: NtAn1b (Nicotiana tabacum, GenBank accession number AEE99258), NtAN1-like (N. tomentosiformis, AEE99260), NtAn1a (N. tabacum, AEE99257), AN1 (Petunia hybrid, AF260918), MrbHLH1 (Myrica rubra, JX629461), LjTT8 (Lotus japonicus, BAH28881), MdbHLH3 (Malus domestica, HM122458), DvIVS (Dahlia variabilis, AB601005), AmDELILA (Antirrhinum majus, AAA32663), R-S (Zea mays, X15806), L-c (Z. mays, NM001111869), MrbHLH2 (M. rubra, JX629462), MdbHLH33 (M. domestica, DQ266451), VvMYC (Vitis vinifera, EU447172).
Mentions: Based on the similarities with bHLHs verified as being related to anthocyanin biosynthesis in other species, one partial sequence bHLH member was mined from the chrysanthemum EST database. The full ORF, which was obtained by RACE and named as CmbHLH2 (GenBank KT724056), contained 1842 nucleotides and encoded 614 amino acids. A second bHLH member isolated from chrysanthemum, CmbHLH1, which was reported to be involved in the response to iron deficiency [30] was also selected. Phylogenetic analysis of these two members and 174 bHLHs including 158 from Arabidopsis and 14 from other plants, showed that CmbHLH2 was clustered in the subgroup containing sequences related to the regulation of anthocyanin biosynthesis, while CmbHLH1 was in a different subgroup (Fig 1).

Bottom Line: CmbHLH2 significantly upregulated the CmDFR promoter and triggered anthocyanin accumulation when co-expressed with CmMYB6.Yeast one-hybrid analyses indicated that CmbHLH2 was able to bind directly to the CmDFR promoter.These results suggest that CmbHLH2 is the essential partner for CmMYB6 in regulating anthocyanin biosynthesis in chrysanthemum.

View Article: PubMed Central - PubMed

Affiliation: College of Agriculture & Biotechnology, Zhejiang University, Zijingang Campus, Hangzhou, 310058, PR China.

ABSTRACT
Chrysanthemums (Chrysanthemum morifolium Ramat.) exhibit a variety of flower colors due to their differing abilities to accumulate anthocyanins. One MYB member, CmMYB6, has been verified as a transcription regulator of chrysanthemum genes involved in anthocyanin biosynthesis; however, the co-regulators for CmMYB6 remain unclear in chrysanthemum. Here, the expression pattern of CmbHLH2, which is clustered in the IIIf bHLH subgroup, was shown to be positively correlated with the anthocyanin content of cultivars with red, pink and yellow flower colors, respectively. CmbHLH2 significantly upregulated the CmDFR promoter and triggered anthocyanin accumulation when co-expressed with CmMYB6. Yeast one-hybrid analyses indicated that CmbHLH2 was able to bind directly to the CmDFR promoter. Moreover, yeast two-hybrid assays indicated protein-protein interaction between CmbHLH2 and CmMYB6. These results suggest that CmbHLH2 is the essential partner for CmMYB6 in regulating anthocyanin biosynthesis in chrysanthemum.

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