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A Malus crabapple chalcone synthase gene, McCHS, regulates red petal color and flavonoid biosynthesis.

Tai D, Tian J, Zhang J, Song T, Yao Y - PLoS ONE (2014)

Bottom Line: The results showed McCHS have their special expression patterns in each tested cultivars, and is responsible for the red coloration and color variation in crabapple petals, especially for color fade process in 'Radiant'.Furthermore, tobacco plants constitutively expressing McCHS displayed a higher anthocyanins accumulation and a deeper red petal color compared with control untransformed lines.We conclude that the endogenous McCHS gene is a critical factor in the regulation of anthocyanin biosynthesis during petal coloration in Malus crabapple.

View Article: PubMed Central - PubMed

Affiliation: Department of Plant Science and Technology, Beijing University of Agriculture, Beijing, China; College of Horticulture, Shanxi Agricultural University, Taigu, Shanxi, China.

ABSTRACT
Chalcone synthase is a key and often rate-limiting enzyme in the biosynthesis of anthocyanin pigments that accumulate in plant organs such as flowers and fruits, but the relationship between CHS expression and the petal coloration level in different cultivars is still unclear. In this study, three typical crabapple cultivars were chosen based on different petal colors and coloration patterns. The two extreme color cultivars, 'Royalty' and 'Flame', have dark red and white petals respectively, while the intermediate cultivar 'Radiant' has pink petals. We detected the flavoniods accumulation and the expression levels of McCHS during petals expansion process in different cultivars. The results showed McCHS have their special expression patterns in each tested cultivars, and is responsible for the red coloration and color variation in crabapple petals, especially for color fade process in 'Radiant'. Furthermore, tobacco plants constitutively expressing McCHS displayed a higher anthocyanins accumulation and a deeper red petal color compared with control untransformed lines. Moreover, the expression levels of several anthocyanin biosynthetic genes were higher in the transgenic McCHS overexpressing tobacco lines than in the control plants. A close relationship was observed between the expression of McCHS and the transcription factors McMYB4 and McMYB5 during petals development in different crabapple cultivars, suggesting that the expression of McCHS was regulated by these transcription factors. We conclude that the endogenous McCHS gene is a critical factor in the regulation of anthocyanin biosynthesis during petal coloration in Malus crabapple.

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Relative expression profiles of endogenous tobacco anthocyanin regulation factors during flower development.Real-time PCR was used to analyze the transcripts in transgenic tobacco flowers, and all real time-PCR reactions were normalized using the Ct value corresponding to the NtActin gene (GQ339768). Error bars correspond to the standard error of the mean ± SE of three replicate reactions. Analysis of relative expression levels of control and McCHS-ox lines and different letters above the bars indicate significantly different values (P<0.05) calculated using one-way analysis of variance (ANOVA) followed by a Duncan's multiple range test.
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pone-0110570-g008: Relative expression profiles of endogenous tobacco anthocyanin regulation factors during flower development.Real-time PCR was used to analyze the transcripts in transgenic tobacco flowers, and all real time-PCR reactions were normalized using the Ct value corresponding to the NtActin gene (GQ339768). Error bars correspond to the standard error of the mean ± SE of three replicate reactions. Analysis of relative expression levels of control and McCHS-ox lines and different letters above the bars indicate significantly different values (P<0.05) calculated using one-way analysis of variance (ANOVA) followed by a Duncan's multiple range test.

Mentions: The qRT-PCR results confirmed a massive elevation of McCHS transcript levels in the transgenic lines, and the absence of expression in the control plants, as expected. The accumulation of anthocyanins was consistent with the relative expression profile of McCHS in McCHS-overexpressing tobacco petals. Moreover, the over-expression of McCHS in tobacco significantly promoted the expression of the downstream endogenous genes NtF3H, NtF3′H, NtDFR, NtANS, and NtUFGT (Figure 7E). To further explore the molecular regulation mechnism in transgenic tobacco, we tested the expression of several endogenous tobacco anthocyanin regulation factors. Interestingly, the results showed the massive accumulated McCHS can alter the expressions of these transcription factors. The expression levels of NtAn1a, NtTTG1, MYB305, NtMYC2a and NtMYC2b were increased, and NtAn1b and NtTTG2 were decreased by overexpressed McCHS (Figure 8).


A Malus crabapple chalcone synthase gene, McCHS, regulates red petal color and flavonoid biosynthesis.

Tai D, Tian J, Zhang J, Song T, Yao Y - PLoS ONE (2014)

Relative expression profiles of endogenous tobacco anthocyanin regulation factors during flower development.Real-time PCR was used to analyze the transcripts in transgenic tobacco flowers, and all real time-PCR reactions were normalized using the Ct value corresponding to the NtActin gene (GQ339768). Error bars correspond to the standard error of the mean ± SE of three replicate reactions. Analysis of relative expression levels of control and McCHS-ox lines and different letters above the bars indicate significantly different values (P<0.05) calculated using one-way analysis of variance (ANOVA) followed by a Duncan's multiple range test.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4214706&req=5

pone-0110570-g008: Relative expression profiles of endogenous tobacco anthocyanin regulation factors during flower development.Real-time PCR was used to analyze the transcripts in transgenic tobacco flowers, and all real time-PCR reactions were normalized using the Ct value corresponding to the NtActin gene (GQ339768). Error bars correspond to the standard error of the mean ± SE of three replicate reactions. Analysis of relative expression levels of control and McCHS-ox lines and different letters above the bars indicate significantly different values (P<0.05) calculated using one-way analysis of variance (ANOVA) followed by a Duncan's multiple range test.
Mentions: The qRT-PCR results confirmed a massive elevation of McCHS transcript levels in the transgenic lines, and the absence of expression in the control plants, as expected. The accumulation of anthocyanins was consistent with the relative expression profile of McCHS in McCHS-overexpressing tobacco petals. Moreover, the over-expression of McCHS in tobacco significantly promoted the expression of the downstream endogenous genes NtF3H, NtF3′H, NtDFR, NtANS, and NtUFGT (Figure 7E). To further explore the molecular regulation mechnism in transgenic tobacco, we tested the expression of several endogenous tobacco anthocyanin regulation factors. Interestingly, the results showed the massive accumulated McCHS can alter the expressions of these transcription factors. The expression levels of NtAn1a, NtTTG1, MYB305, NtMYC2a and NtMYC2b were increased, and NtAn1b and NtTTG2 were decreased by overexpressed McCHS (Figure 8).

Bottom Line: The results showed McCHS have their special expression patterns in each tested cultivars, and is responsible for the red coloration and color variation in crabapple petals, especially for color fade process in 'Radiant'.Furthermore, tobacco plants constitutively expressing McCHS displayed a higher anthocyanins accumulation and a deeper red petal color compared with control untransformed lines.We conclude that the endogenous McCHS gene is a critical factor in the regulation of anthocyanin biosynthesis during petal coloration in Malus crabapple.

View Article: PubMed Central - PubMed

Affiliation: Department of Plant Science and Technology, Beijing University of Agriculture, Beijing, China; College of Horticulture, Shanxi Agricultural University, Taigu, Shanxi, China.

ABSTRACT
Chalcone synthase is a key and often rate-limiting enzyme in the biosynthesis of anthocyanin pigments that accumulate in plant organs such as flowers and fruits, but the relationship between CHS expression and the petal coloration level in different cultivars is still unclear. In this study, three typical crabapple cultivars were chosen based on different petal colors and coloration patterns. The two extreme color cultivars, 'Royalty' and 'Flame', have dark red and white petals respectively, while the intermediate cultivar 'Radiant' has pink petals. We detected the flavoniods accumulation and the expression levels of McCHS during petals expansion process in different cultivars. The results showed McCHS have their special expression patterns in each tested cultivars, and is responsible for the red coloration and color variation in crabapple petals, especially for color fade process in 'Radiant'. Furthermore, tobacco plants constitutively expressing McCHS displayed a higher anthocyanins accumulation and a deeper red petal color compared with control untransformed lines. Moreover, the expression levels of several anthocyanin biosynthetic genes were higher in the transgenic McCHS overexpressing tobacco lines than in the control plants. A close relationship was observed between the expression of McCHS and the transcription factors McMYB4 and McMYB5 during petals development in different crabapple cultivars, suggesting that the expression of McCHS was regulated by these transcription factors. We conclude that the endogenous McCHS gene is a critical factor in the regulation of anthocyanin biosynthesis during petal coloration in Malus crabapple.

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