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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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The transcript levels of seven anthocyanin biosynthetic genes and three TFs in the ray florets of three chrysanthemum cultivars used in this study.The CmACT gene was used to normalize expression of the genes. Non-template reactions were set as the negative control for each gene. The vertical bars represent S.E. of three biological replicates.
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pone.0143892.g003: The transcript levels of seven anthocyanin biosynthetic genes and three TFs in the ray florets of three chrysanthemum cultivars used in this study.The CmACT gene was used to normalize expression of the genes. Non-template reactions were set as the negative control for each gene. The vertical bars represent S.E. of three biological replicates.

Mentions: Accumulation of mRNAs for three TFs, CmMYB6, CmbHLH1 and CmbHLH2, and seven anthocyanin biosynthetic genes in the ray florets of three cultivars were analyzed using QPCR. All biosynthetic genes were expressed at the highest level in ‘Z1’ and were positively correlated with anthocyanin accumulation in these three cultivars, although the transcriptional level of CmCHI (Chalcone isomerase) was similar in ‘Z2’ and ‘Z3’ (Fig 3). The transcript levels of CmDFR and CmUFGT (UDP flavonoid glucosyl transferase) were very low in ‘Z3’ compared with those in ‘Z1’, and CmANS (Anthocyanidin synthase) was undetectable in ‘Z3’ (Fig 3). Accumulation of mRNA for each of the three TFs was highest in ‘Z1’ and lowest in ‘Z3’ (Fig 3) as found for the biosynthetic genes.


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)

The transcript levels of seven anthocyanin biosynthetic genes and three TFs in the ray florets of three chrysanthemum cultivars used in this study.The CmACT gene was used to normalize expression of the genes. Non-template reactions were set as the negative control for each gene. The vertical bars represent S.E. of three biological replicates.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4664390&req=5

pone.0143892.g003: The transcript levels of seven anthocyanin biosynthetic genes and three TFs in the ray florets of three chrysanthemum cultivars used in this study.The CmACT gene was used to normalize expression of the genes. Non-template reactions were set as the negative control for each gene. The vertical bars represent S.E. of three biological replicates.
Mentions: Accumulation of mRNAs for three TFs, CmMYB6, CmbHLH1 and CmbHLH2, and seven anthocyanin biosynthetic genes in the ray florets of three cultivars were analyzed using QPCR. All biosynthetic genes were expressed at the highest level in ‘Z1’ and were positively correlated with anthocyanin accumulation in these three cultivars, although the transcriptional level of CmCHI (Chalcone isomerase) was similar in ‘Z2’ and ‘Z3’ (Fig 3). The transcript levels of CmDFR and CmUFGT (UDP flavonoid glucosyl transferase) were very low in ‘Z3’ compared with those in ‘Z1’, and CmANS (Anthocyanidin synthase) was undetectable in ‘Z3’ (Fig 3). Accumulation of mRNA for each of the three TFs was highest in ‘Z1’ and lowest in ‘Z3’ (Fig 3) as found for the biosynthetic genes.

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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