• Title/Summary/Keyword: resveratrol synthase gene

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Expression of resveratrol synthase gene and accumulation of resveratrol in transgenic potatoes (Solanum tuberosum L.)

  • Yi, Jung Yoon;Seo, Hyo Won;Yun, Song Joong;Ok, HyunChoong;Park, YoungEun;Cho, Ji Hong;Cho, HyunMook
    • Korean Journal of Breeding Science
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    • v.41 no.4
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    • pp.385-390
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    • 2009
  • A resveratrol synthase (RS) gene was isolated from peanut (Arachis hypogaea, L. cv. Jinpoong) plants. This gene was placed under the control of the cauliflower mosaic virus 35S promoter (CaMV35S) and introduced into two Korean varieties of potato (Solanum tuberosum L. cvs. Jasim and Jowon) plants by Agrobacterium-mediated gene transfer. Putative transformants were screened by PCR with primers designed from CaMV 35S promoter, NOS terminator and RS gene. Most of selected transgenic potato plants showed the amplification of expected fragments by PCR of genomic DNA with gene-specific primers, while they were absent in untransformed control plants. Expression of the resveratrol synthase gene was also examined by northern blot analysis. The transformants showed a band which was lacking in the control plant, confirming that the introduced gene is transcribed into mRNA in the transformants. The strength of the band, which reflected the level of mRNA expression, differed among the individual transformants. Among the transformants obtained, the highest trans-resveratrol content in the transgenic young leaves of purple-fleshed "Jashim" was $2.11{\mu}gg^{-1}$ fresh weight and that in the microtubers in vitro of purple fleshed "Jashim" was $8.31{\mu}gg^{-1}$ fresh weight. This amount of resveratrol may have a positive biological effect on human health.

Inhibitory Effects of Resveratrol and Piceid against Pathogens of Rice Plant, and Disease Resistance Assay of Transgenic Rice Plant Transformed with Stilbene Synthase Gene

  • Yu, Sang-Mi;Lee, Ha Kyung;Jeong, Ui-Seon;Baek, So Hyeon;Noh, Tae-Hwan;Kwon, Soon Jong;Lee, Yong Hoon
    • Research in Plant Disease
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    • v.19 no.3
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    • pp.177-182
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    • 2013
  • Resvestrol has been known to inhibit bacterial and fungal growth in vitro, and can be accumulated in plant to concentrations necessary to inhibit microbial pathogens. Hence, stilbene synthase gene has been used to transform to synthesize resveratrol in heterologous plant species to enhance resistance against pathogens. In the present study, we investigated the antimicrobial activities of resveratrol and piceid to bacterial and fungal pathogens, which causing severe damages to rice plants. In addition, disease resistance was compared between transgenic rice varieties, Iksan 515 and Iksan 526 transformed with stlibene synthase gene and non-transgenic rice varieties, Dongjin and Nampyeong. Minimum inhibitory concentration of resveratrol for Burkolderia glumae was 437.5 ${\mu}M$, and the mycelial growth of Biplaris oryzae was slightly inhibited at concentration of 10 ${\mu}M$. However, other bacterial and fungal pathogens are not inhibited by resveratrol and piceid. The expression of the stilbene synthase gene in Iksan 515 and Iksan 526 did not significantly enhanced resistance against bacterial grain rot, bacterial leaf blight, sheath blight, and leaf blight. This study is the first report on the effect of resveratrol and piceid against pathogens of rice plant, and changes of disease resistance of transgenic rice plants transformed with stilbene synthase gene.

Agrobacterium-mediated Transformation of Rehmannia glutinosa L. with Resveratrol Gene (RS3) of Peanut

  • Lim, Jung-Dae;Yang, Deok-Chun;Yun, Song-Joong;Chung, Ill-Min;Sung, Eun-Soo;Kim, Myong-Jo;Heo, Kweon;Yu, Chang-Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.2
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    • pp.171-178
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    • 2004
  • The objectives of this study were to establish the genetic transformation system of stilbene synthase in Rehmannia glutinosa. Resveratrol, which is both a phytoalexin with antifungal activity and a phytochemical associated with reduced cancer risk and reduced cardiovascular disease, is synthesized in a limited number of plant species including peanut. Resveratrol synthesis is catalyzed by the enzyme stilbene synthase including resveratrol synthase (RS). Stilbene synthase gene (RS3) obtained from peanut, Arachis hypogaea, Fabaceae has been transferred into chinese foxglove, Rehmannia glutinosa by using Agrobacterium mediated transformation. PCR analysis with RS3 primer confirmed that the targeted gene was introduced into the plant genome, 904 bp in size. Further analyses of identification of transformation using developed other molecular techniques and transgenic plants that RS t-DNA introduced to chinese foxglove (R. glutinosa L) and its reaction product, stilbene such as resveratrol will be isolate and characterize using NMR, MS, and HPLC.

A Molecular Switch for the Induction of Resveratrol Biosynthesis in Grapes

  • Lee, Mi-Sook;Pyee, Jae-Ho
    • Natural Product Sciences
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    • v.10 no.5
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    • pp.248-251
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    • 2004
  • Resveratrol has been reported to possess a variety of biological and pharmaceutical activities. Regardless of its beneficial effects on health, the amount of resveratrol in grapes is very low. In order to induce the resveratrol biosynthesis, the promoter region of a genomic fragment encoding the resveratrol synthase was isolated and a molecular switch was identified which provides us with defining biotic or abiotic inducers that transcriptionally up-regulate the gene expression involved in the resveratrol biosynthesis. We could successfully increase the amount of resveratrol in grapes up to 3-fold by using these environmental factors.

Isolation and Biological Activity of $Resveratrol-3-O-{\beta}-D-Glucoside$ in Transgenic Rehmannia glutinosa L. Transformed by Peanut Resveratrol Synthase Gene (RS3)

  • Lim, Jung-Dae;Yang, Deok-Chun;Yun, Song-Joong;Chung, Ill-Min;Sung, Eun-Soo;Kim, Myong-Jo;Heo, Kweon;Yu, Chang-Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.5
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    • pp.406-414
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    • 2004
  • Resveratrol, which is both a phytoalexin with antifungal activity and a phytochemical associated with reduced cancer risk and reduced cardiovascular disease, is synthesized in a limited number of plant species including peanut. Resveratrol synthesis is catalyzed by the enzyme stilbene synthase including resveratrol synthase (RS). Resveratrol synthase gene (RS3) obtained from peanut, Arachis hypogaea, Fabaceae has been transferred into chinese foxglove, Rehmannia glutinosa by using Agrobacterium mediated transformation. RS t-DNA introduced to chinese foxglove (R. glutinosa L) by transformation and its reaction product, $resveratrol-3-O-{\beta}-D-glucoside$ was isolated and characterized using HPLC. Also its biological effects was tested in inhibition of the lipid peroxidation of mouse LDL by glycosylated stilbenes derivatives obtained from transgenic plants. $Resveratrol-3-O-{\beta}-D-glucoside$ isolated from transgenic R. glutinosa L. showed antimicrobial activity of the growth inhibition zone against Escherichia coli and Salmonella typhimurium. Therefore, this compound can be contributed to be useful as a phytoalexin for plant health as well as a phytochemical for human health.

Induction of Resveratrol Biosynthesis in Grape Skins and Leaves by Ultrasonication Treatment

  • Hasan, Md. Mohidul;Baek, Kwang-Hyun
    • Horticultural Science & Technology
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    • v.31 no.4
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    • pp.496-502
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    • 2013
  • Grapes (Vitis vinifera) are one of the most important fruits worldwide and are eaten raw or after conversion to jelly, jam, juice and wine. Grape skins are a major source of resveratrol (3,5,4'-trihydroxystilbene), which has the ability to reduce blood sugar as well as anticancer, anti-inflammatory, and other beneficial cardiovascular effects. In this study, we investigated the increased accumulation of resveratrol in grape skin and leaves following ultrasonication treatment, which has been shown to induce resveratrol accumulation in several plants. Various ultrasonication treatment times and incubation periods were employed to identify the optimum conditions for the maximum accumulation of resveratrol. Treatment and further incubation led to increased resveratrol in both grape skins and leaves, with the highest increases of 7.7-fold and 1.9-fold occurring in response to 5 min ultrasonication treatment followed by 6 hour incubation and 15 min ultrasonication treatment followed by 3 hour incubation, respectively. The underlying mechanism for the increased amounts of resveratrol were studied by employing a semi-quantitative RT-PCR to monitor the expression levels of the resveratrol synthase (RS) gene in response to ultrasonication treatment. The RS gene increased the expression in response to ultrasonication treatment, suggesting that up-regulation of the RS gene by ultrasonication treatment triggers increased amounts of resveratrol. Taken together, these data indicate that this simple ultrasonication treatment of grapes can be an efficient post-harvest technology for increasing resveratrol in grape skins in addition to cleaning the fruits.

The overexpression of Arachis hypogaea resveratrol synthase 3 (AhRS3) modified the expression pattern of phenylpropanoid pathway genes in developing rice seeds

  • Lee, Choonseok;Jeong, Namhee;Kim, Dool-Yi;Ok, Hyun-Choong;Choi, Man-Soo;Park, Ki-Do;Kim, Jaehyun
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.167-167
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    • 2017
  • Our previous study for developing seeds of Iksan 526 (I.526), an inbred line of resveratrol-producing transgenic rice line, showed that, in 20 days after heading (DAH) seeds, resveratrol was almost saturated and accumulation of piceid was highest though the expression of Arachis hypogaea resveratrol synthase 3 (AhRS3, GenBank DQ124938) was highest in 31 DAH seeds. In this study, it was investigated how the overexpression of AhRS3 affects phenylpropanoid pathway genes. p-Coumaroyl-CoA is derived from phenylpropanoid pathway and used as a substrate of AhRS3 reaction for resveratrol production. In 6, 13, 20, 31 and 41 (45 for Dongjin) DAH seeds of I526 and Dongjin, a wild type of I.526, respectively, the expression pattern of phenylpropanoid pathway genes, including phenylalanine ammonia-lyase (PAL: LOC_Os02g41630.2, LOC_Os04g43760.1), cinnamate 4-hydroxylase (C4H: LOC_Os05g25640.1), 4-coumarate-CoA ligase (4CL: LOC_Os02g08100.1), cinnamoyl-CoA reductase (CCR: LOC_ Os09g25150.1, LOC_Os08g34280.1), hydroxycinnamoyl-CoA shikimate/quinate hydroxycinnamoyl transferase (HCT: LOC_Os04g42250.2, LOC_Os02g39850.1) and cinnamyl alcohol dehydrogenase (CAD: LOC_Os02g09490.1), was examined using real time (RT)-PCR. Compared to developing seeds of Dongjin, RT-PCR results showed that the expression pattern of phenylpropanoid pathway genes was modified in developing seeds of I.526. In most genes, except for CAD, of I.526 developing seeds, the gene expression was highest in 20 DAH corresponding to biosynthesis of resveratrol and piceid, i.e. the expression of phenylpropanoid pathway genes was gradually increased by 20 DAH and decreased as seeds develop. Especially, in Dongjin, the highest expression of PALs and 4CL was in 6 DAH and their expression was gradually decreased as seeds develop. These genes expression data also exhibited that, in developing seeds of I.526, phenylpropanoid pathway genes were slightly or significantly (in some genes) upregulated compared to Dongjin. Therefore, the overexpression of AhRS3 changed the expression pattern of phenylpropanoid pathway genes in I.526 developing seeds and this modification for gene expression is closely related to biosynthesis of resveratrol and piceid.

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The overexpression of Arachis hypogaea resveratrol synthase 3 (AhRS3) modified the expression pattern of phenylpropanoid pathway genes in developing rice seeds

  • Lee, Choonseok;Jeong, Namhee;Kim, Dool-Yi;Ok, Hyun-Choong;Choi, Man-Soo;Park, Ki-Do;Kim, Jaehyun
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.105-105
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    • 2017
  • Our previous study for developing seeds of Iksan 526 (I.526), an inbred line of resveratrol-producing transgenic rice line, showed that, in 20 days after heading (DAH) seeds, resveratrol was almost saturated and accumulation of piceid was highest though the expression of Arachis hypogaea resveratrol synthase 3 (AhRS3, GenBank DQ124938) was highest in 31 DAH seeds. In this study, it was investigated how the overexpression of AhRS3 affects phenylpropanoid pathway genes. p-Coumaroyl-CoA is derived from phenylpropanoid pathway and used as a substrate of AhRS3 reaction for resveratrol production. In 6, 13, 20, 31 and 41 (45 for Dongjin) DAH seeds of I526 and Dongjin, a wild type of I.526, respectively, the expression pattern of phenylpropanoid pathway genes, including phenylalanine ammonia-lyase (PAL: LOC_Os02g41630.2, LOC_Os04g43760.1), cinnamate 4-hydroxylase (C4H: LOC_Os05g25640.1), 4-coumarate-CoA ligase (4CL: LOC_Os02g08100.1), cinnamoyl-CoA reductase (CCR: LOC_Os09g25150.1, LOC_Os08g34280.1), hydroxycinnamoyl-CoA shikimate/quinate hydroxycinnamoyl transferase (HCT: LOC_Os04g42250.2, LOC_Os02g39850.1) and cinnamyl alcohol dehydrogenase (CAD: LOC_Os02g09490.1), was examined using real time (RT)-PCR. Compared to developing seeds of Dongjin, RT-PCR results showed that the expression pattern of phenylpropanoid pathway genes was modified in developing seeds of I.526. In most genes, except for CAD, of I.526 developing seeds, the gene expression was highest in 20 DAH corresponding to biosynthesis of resveratrol and piceid, i.e. the expression of phenylpropanoid pathway genes was gradually increased by 20 DAH and decreased as seeds develop. Especially, in Dongjin, the highest expression of PALs and 4CL was in 6 DAH and their expression was gradually decreased as seeds develop. These genes expression data also exhibited that, in developing seeds of I.526, phenylpropanoid pathway genes were slightly or significantly (in some genes) upregulated compared to Dongjin. Therefore, the overexpression of AhRS3 changed the expression pattern of phenylpropanoid pathway genes in I.526 developing seeds and this modification for gene expression is closely related to biosynthesis of resveratrol and piceid.

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Disease Occurrence in Transgenic Rice Plant Transformed with Silbene Synthase Gene and Evaluation of Possible Horizontal Gene Transfer to Plant Pathogens

  • Yu, Sang-Mi;Jeong, Ui-Seon;Lee, Ha Kyung;Baek, So Hyeon;Kwon, Soon Jong;Lee, Yong Hoon
    • Research in Plant Disease
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    • v.20 no.3
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    • pp.189-195
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    • 2014
  • Genetic engineering is being used to enhance disease resistance and nutritional value of crops including rice plant. Considering the fast-growing agricultural biotechnology and rapidly increasing global area of transgenic crops, the risk evaluation on environment is necessary. In this study, we surveyed the difference of disease occurrence between transgenic rice variety, Iksan526 transformed with peanut stilbene synthase gene and non-transgenic rice varieties, Dongjin and Nampyeong in the field. Moreover, the possibility of gene transfer from transgenic rice to bacterial and fungal pathogens was investigated. The results of this study indicated that there was no significant difference in the occurrence and severity of the diseases between Iksan526 and Dongjin or Nampyeong. In addition, the results suggested that rice pathogen, such as Xanthomonas oryzae pv. oryzae, Rhizoctonia solani and Magnaporthe grisea did not take up stilbene synthase and bar genes under natural conditions. Moreover the transformed DNA was not transferred to the pathogens even in repetitive contacts.