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Insights into the Mechanisms Underlying Ultraviolet-C Induced Resveratrol Metabolism in Grapevine (V. amurensis Rupr.) cv. "Tonghua-3".

Yin X, Singer SD, Qiao H, Liu Y, Jiao C, Wang H, Li Z, Fei Z, Wang Y, Fan C, Wang X - Front Plant Sci (2016)

Bottom Line: Tonghua-3 following UV-C treatment, to accelerate research into resveratrol metabolism.Comparative RNA-Seq profiling of UV-C treated and untreated grape berries resulted in the identification of a large number of differentially expressed genes.This transcriptome data set provides new insight into the response of grape berries to UV-C treatment, and suggests candidate genes, or promoter activity of related genes, that could be used in future functional and molecular biological studies of resveratrol metabolism.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Crop Stress Biology in Arid Areas, College of Horticulture, Northwest A&F UniversityYangling, China; Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China, Ministry of Agriculture, Northwest A&F UniversityYangling, China.

ABSTRACT
Stilbene compounds belong to a family of secondary metabolites that are derived from the phenylpropanoid pathway. Production of the stilbene phytoalexin, resveratrol, in grape (Vitis spp.) berries is known to be induced by ultraviolet-C radiation (UV-C), which has numerous regulatory effects on plant physiology. While previous studies have described changes in gene expression caused by UV-C light in several plant species, such information has yet to be reported for grapevine. We investigated both the resveratrol content and gene expression responses of berries from V. amurensis cv. Tonghua-3 following UV-C treatment, to accelerate research into resveratrol metabolism. Comparative RNA-Seq profiling of UV-C treated and untreated grape berries resulted in the identification of a large number of differentially expressed genes. Gene ontology (GO) term classification and biochemical pathway analyses suggested that UV-C treatment caused changes in various cellular processes, as well as in both hormone and secondary metabolism. The data further indicate that UV-C induced increases in resveratrol may be related to the transcriptional regulation of genes involved in the production of secondary metabolites and signaling, as well as several transcription factors. We also observed that following UV-C treatment, 22 stilbene synthase (STS) genes exhibited increases in their expression levels and a VaSTS promoter drove the expression of the GUS reporter gene when expressed in tobacco. We therefore propose that UV-C induction of VaSTS expression is an important factor in promoting resveratrol accumulation. This transcriptome data set provides new insight into the response of grape berries to UV-C treatment, and suggests candidate genes, or promoter activity of related genes, that could be used in future functional and molecular biological studies of resveratrol metabolism.

No MeSH data available.


Related in: MedlinePlus

(A) Histochemical analysis of GUS (β-galactosidase) activity after UV-C treatment of transiently transformed tobacco leaves carrying the GUS coding region fused to the VaSTS promoter. Numbers above the bars indicate the GUS activity fold of UV-C treated GUS activity relative to that of mock-inoculated sample, and its significant difference was assessed by a one-sided paired t-test (P < 0.05 [*]). (B) Activity of the VaSTS promoter in tobacco at different time points following UV-C irradiation was determined by quantitative real-time RT-PCR. GUS transcript accumulation was monitored in mock-treated leaves (dashed lines) and UV-C treated leaves (solid lines). Transcript levels were expressed as relative values normalized to the transcript level of the ubiquitin gene. Results are means of triplicate experiments; error bars indicate SD.
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Figure 8: (A) Histochemical analysis of GUS (β-galactosidase) activity after UV-C treatment of transiently transformed tobacco leaves carrying the GUS coding region fused to the VaSTS promoter. Numbers above the bars indicate the GUS activity fold of UV-C treated GUS activity relative to that of mock-inoculated sample, and its significant difference was assessed by a one-sided paired t-test (P < 0.05 [*]). (B) Activity of the VaSTS promoter in tobacco at different time points following UV-C irradiation was determined by quantitative real-time RT-PCR. GUS transcript accumulation was monitored in mock-treated leaves (dashed lines) and UV-C treated leaves (solid lines). Transcript levels were expressed as relative values normalized to the transcript level of the ubiquitin gene. Results are means of triplicate experiments; error bars indicate SD.

Mentions: To test the light-inducible activity of the VaSTS promoter, a 1570 bp fragment upstream from the transcriptional start site was fused to the GUS reporter gene. The construct was introduced into tobacco leaves and these were tested for GUS activity after UV-C irradiation treatment (Figure 8). A CaMV35S::GUS (pC35SGUS) construct was used as a positive control, and a promoter-less construct (pC0380GUS) served as the negative control. The response of the VaSTS promoter was initially analyzed by histochemical GUS staining 24 h after UV-C treatment. As shown in Figure 8A, compared with levels in untreated leaves, GUS activity was significantly greater in UV-C treated leaves. The CaMV35S promoter showed no significant induction after UV-C treatment, while the negative control and wild-type only exhibited low background levels of GUS activity regardless of the treatment (Figure S8).


Insights into the Mechanisms Underlying Ultraviolet-C Induced Resveratrol Metabolism in Grapevine (V. amurensis Rupr.) cv. "Tonghua-3".

Yin X, Singer SD, Qiao H, Liu Y, Jiao C, Wang H, Li Z, Fei Z, Wang Y, Fan C, Wang X - Front Plant Sci (2016)

(A) Histochemical analysis of GUS (β-galactosidase) activity after UV-C treatment of transiently transformed tobacco leaves carrying the GUS coding region fused to the VaSTS promoter. Numbers above the bars indicate the GUS activity fold of UV-C treated GUS activity relative to that of mock-inoculated sample, and its significant difference was assessed by a one-sided paired t-test (P < 0.05 [*]). (B) Activity of the VaSTS promoter in tobacco at different time points following UV-C irradiation was determined by quantitative real-time RT-PCR. GUS transcript accumulation was monitored in mock-treated leaves (dashed lines) and UV-C treated leaves (solid lines). Transcript levels were expressed as relative values normalized to the transcript level of the ubiquitin gene. Results are means of triplicate experiments; error bars indicate SD.
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Related In: Results  -  Collection

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Figure 8: (A) Histochemical analysis of GUS (β-galactosidase) activity after UV-C treatment of transiently transformed tobacco leaves carrying the GUS coding region fused to the VaSTS promoter. Numbers above the bars indicate the GUS activity fold of UV-C treated GUS activity relative to that of mock-inoculated sample, and its significant difference was assessed by a one-sided paired t-test (P < 0.05 [*]). (B) Activity of the VaSTS promoter in tobacco at different time points following UV-C irradiation was determined by quantitative real-time RT-PCR. GUS transcript accumulation was monitored in mock-treated leaves (dashed lines) and UV-C treated leaves (solid lines). Transcript levels were expressed as relative values normalized to the transcript level of the ubiquitin gene. Results are means of triplicate experiments; error bars indicate SD.
Mentions: To test the light-inducible activity of the VaSTS promoter, a 1570 bp fragment upstream from the transcriptional start site was fused to the GUS reporter gene. The construct was introduced into tobacco leaves and these were tested for GUS activity after UV-C irradiation treatment (Figure 8). A CaMV35S::GUS (pC35SGUS) construct was used as a positive control, and a promoter-less construct (pC0380GUS) served as the negative control. The response of the VaSTS promoter was initially analyzed by histochemical GUS staining 24 h after UV-C treatment. As shown in Figure 8A, compared with levels in untreated leaves, GUS activity was significantly greater in UV-C treated leaves. The CaMV35S promoter showed no significant induction after UV-C treatment, while the negative control and wild-type only exhibited low background levels of GUS activity regardless of the treatment (Figure S8).

Bottom Line: Tonghua-3 following UV-C treatment, to accelerate research into resveratrol metabolism.Comparative RNA-Seq profiling of UV-C treated and untreated grape berries resulted in the identification of a large number of differentially expressed genes.This transcriptome data set provides new insight into the response of grape berries to UV-C treatment, and suggests candidate genes, or promoter activity of related genes, that could be used in future functional and molecular biological studies of resveratrol metabolism.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Crop Stress Biology in Arid Areas, College of Horticulture, Northwest A&F UniversityYangling, China; Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China, Ministry of Agriculture, Northwest A&F UniversityYangling, China.

ABSTRACT
Stilbene compounds belong to a family of secondary metabolites that are derived from the phenylpropanoid pathway. Production of the stilbene phytoalexin, resveratrol, in grape (Vitis spp.) berries is known to be induced by ultraviolet-C radiation (UV-C), which has numerous regulatory effects on plant physiology. While previous studies have described changes in gene expression caused by UV-C light in several plant species, such information has yet to be reported for grapevine. We investigated both the resveratrol content and gene expression responses of berries from V. amurensis cv. Tonghua-3 following UV-C treatment, to accelerate research into resveratrol metabolism. Comparative RNA-Seq profiling of UV-C treated and untreated grape berries resulted in the identification of a large number of differentially expressed genes. Gene ontology (GO) term classification and biochemical pathway analyses suggested that UV-C treatment caused changes in various cellular processes, as well as in both hormone and secondary metabolism. The data further indicate that UV-C induced increases in resveratrol may be related to the transcriptional regulation of genes involved in the production of secondary metabolites and signaling, as well as several transcription factors. We also observed that following UV-C treatment, 22 stilbene synthase (STS) genes exhibited increases in their expression levels and a VaSTS promoter drove the expression of the GUS reporter gene when expressed in tobacco. We therefore propose that UV-C induction of VaSTS expression is an important factor in promoting resveratrol accumulation. This transcriptome data set provides new insight into the response of grape berries to UV-C treatment, and suggests candidate genes, or promoter activity of related genes, that could be used in future functional and molecular biological studies of resveratrol metabolism.

No MeSH data available.


Related in: MedlinePlus