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TRINH NGOC AI,AUNG HTAY NAING,김창길 한국식물생명공학회 2016 JOURNAL OF PLANT BIOTECHNOLOGY Vol.43 No.1
Anthocyanin accumulation and plant growth were examined in petunia (NT and T2 transgenic plants) by determining the effects of different sources of light and varying light/dark cycles. Red light significantly enhanced anthocyanin content of B-peru+mPAP1; however, it had a negative effect on anthocyanin production in RsMYB1 plants. In general, white light was found to be reasonable for anthocyanin accumulation in all plants. In case of light/dark cycles, application of seven days of light:14 days of dark significantly enhanced anthocyanin content. We found that anthocyanin content detected in transgenic plants expressing anthocyanin regulatory transcription factor genes (B-peru+mPAP1 or RsMYB1) was higher than that in NT plants in all treatments. Plant growth was also influenced by the different light sources and dark/light cycles. Taken together, our results suggest that light source and light/dark cycle play an important role in anthocyanin production and plant growth. The choice of the optimal conditions is also important for anthocyanin production and plant growth depending on NT or transgenic plants carrying anthocyanin regulatory transcription factors.
Ai, Trinh Ngoc,Naing, Aung Htay,Kim, Chang Kil The Korean Society of Plant Biotechnology 2016 식물생명공학회지 Vol.43 No.1
Anthocyanin accumulation and plant growth were examined in petunia (NT and $T_2$ transgenic plants) by determining the effects of different sources of light and varying light/dark cycles. Red light significantly enhanced anthocyanin content of B-peru+mPAP1; however, it had a negative effect on anthocyanin production in RsMYB1 plants. In general, white light was found to be reasonable for anthocyanin accumulation in all plants. In case of light/dark cycles, application of seven days of light:14 days of dark significantly enhanced anthocyanin content. We found that anthocyanin content detected in transgenic plants expressing anthocyanin regulatory transcription factor genes (B-peru+mPAP1 or RsMYB1) was higher than that in NT plants in all treatments. Plant growth was also influenced by the different light sources and dark/light cycles. Taken together, our results suggest that light source and light/dark cycle play an important role in anthocyanin production and plant growth. The choice of the optimal conditions is also important for anthocyanin production and plant growth depending on NT or transgenic plants carrying anthocyanin regulatory transcription factors.
Ai, Trinh Ngoc,Naing, Aung Htay,Arun, Muthukrishnan,Jeon, Su Min,Kim, Chang Kil International Society for Horticultural Science 2017 Scientia horticulturae Vol. No.
<P><B>Abstract</B></P> <P>Anthocyanin production enhanced by heterologous expression of R2R3 MYB transcription factors has been studied. However, little is known about validated information on the ability of <I>RsMYB</I> (an MYB gene from radish) to enhance anthocyanin production in a heterologous system. In the present study, heterologous expression of <I>RsMYB1</I> in <I>Petunia hybrida</I> ‘Mirage Rose’ enhanced anthocyanin production in vegetative and floral tissues such as leaves, stems, roots, and petals by transcriptional activation of anthocyanin biosynthetic genes and endogenous antocyanin regulatory genes. Line PM6 expressed higher transcript levels of <I>RsMYB1</I> than line PM2 and regulated transcript levels of the investigated genes largely than line PM2, whereas those regulated in wild type (WT) plants were the lowest. In addition, transcript levels of the genes detected using qualitative real-time polymerase chain reaction were found to be higher in petals, followed by leaves, stems and roots. Taken together, our results suggest that <I>RsMYB1</I> enhances anthocyanin production in vegetative and floral tissues of this cultivar, thus, we expect that heterologous expression of <I>RsMYB1</I> would help to modify flower color of other ornamental plants as well.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Expression of <I>RsMYB1</I> in <I>Petunia</I> enhanced anthocyanin production in vegetative and floral tissues. </LI> <LI> RsMYB1 enhances transcript levels of several anthocyanin biosynthetic genes and endogenous genes. </LI> <LI> Transcript levels of <I>RsMYB1</I> and investigated genes were found to be higher in leaves, flowers, followed by stems and roots. </LI> </UL> </P>
Ai, Trinh Ngoc,Arun, Muthukrishnan,Naing, Aung Htay,Lim, Sun-Hyung,Kim, Chang Kil Elsevier 2016 Scientia horticulturae Vol.200 No.-
<P><B>Abstract</B></P> <P>The present study aimed to determine the role of transcription factors (bHLH and MYB) in enhancing anthocyanin production in <I>Petunia</I> ‘Mirage rose’. Initially, we optimized an <I>Agrobacterium</I>-mediated transformation protocol to over-express transcription factors (<I>B-Peru</I>, <I>mPAP1</I>, and <I>B-Peru</I> + <I>mPAP1</I>) in <I>Petunia.</I> Phosphinothricin (PPT) concentrations of 0.5, 1.0, and 1.5mgl<SUP>−1</SUP> were found to be ideal for the selection and regeneration of transformed shoots at different developmental stages. Clavamox (250mgl<SUP>−1</SUP>) efficiently eliminated <I>Agrobacterium</I> after co-cultivation (2 days) and favored maximum shoot regeneration. In addition, incubation for 30min in <I>Agrobacterium</I> suspension increased the number of transformed cells and resulted in improved regeneration in the selection medium. The transformed plants were successfully developed through a direct organogenesis system. The transformed plants were selected using BASTA<SUP>®</SUP> and the presence of transgenes was assessed using PCR. Visible anthocyanin accumulation was evident only in plantlets (shoots, stem, leaves, and roots) carrying the gene combination <I>B-Peru</I> <I>+</I> <I>mPAP1</I>. The expression of biosynthetic genes involved in the flavonoid pathway was analyzed using quantitative real-time PCR. Expression levels of <I>PAL</I>, <I>CHS</I>, <I>CHI</I>, <I>F3H</I>, <I>DFR</I>, and <I>ANS</I> were higher in the young and mature leaves of plants transformed with <I>B-Peru</I> <I>+</I> <I>mPAP1</I> compared to those transformed with <I>B-Peru</I>, <I>mPAP1,</I> and/or non-transformed plants. Furthermore, the highest level of anthocyanin was recorded in the leaves, stem, and roots of plants transformed with <I>B-Peru</I> <I>+</I> <I>mPAP1</I>. These results validate the combinatorial requirement of <I>B-Peru</I> and <I>mPAP1</I> to enhance anthocyanin content in <I>Petunia hybrida</I> ‘Mirage rose’.</P> <P><B>Highlights</B></P> <P> <UL> <LI> We report an improved <I>Agrobacterium</I>-mediated transformation protocol for <I>Petunia</I>. </LI> <LI> PPT used throughout the selection procedure reduced the total number of escapes. </LI> <LI> Clavamox controlled <I>Agrobacterium</I> after co-cultivation and supported regeneration. </LI> <LI> The protocol was utilized to transfer anthocyanin regulatory genes into <I>Petunia</I>. </LI> <LI> Co-expression of <I>B-peru</I> and <I>mPAP1D</I> significantly improved anthocyanin in <I>Petunia</I>. </LI> </UL> </P>
Trinh, Ngoc Ai,Nguyen, Hien Thi Thanh,Park, Seon Joo The Plant Resources Society of Korea 2012 한국자원식물학회지 Vol.25 No.6
We performed phylogenetic analyses of a total of 21 acessions covering 5 species in the Korean Trigonotis and one outgroup species using nuclear ribosomal ITS and chloroplast rbcL, matK, ndhF sequences. Outgroup were chosen from the closely related genus Lithospermum zollingeri. Both parsimony and Bayesian Inference methods were used to reconstruct the evolutionary history of the group. The evidence collected indicated that phylogenetic relationships among Korean Trigonotis species are unresolved based on nuclear marker (ITS), as the same as based on separated chloroplast sequences. While the phylogenetic relationships of Korean Trigonotis species almost clearly were resolved in combined chloroplast sequences. Thus, the members of Trigonotis coreana can be distinguished to the members of Trigonotis peduncularis in combined cpDNA sequences and Trigonotis nakaii was treated as a synonymed to Trigonotis radicans var. sericea. In addition, the MP and BI analysis showed Trigonotis icumae as sister of the remained Korean Trigonotis species based on combined molecular markers (BI: PP = 1).
Effect of pH on the expression of RsMYB1 that regulates anthocyanin production in Petunia plants
이덕범,TRINH NGOC AI,Aung Htay Naing,김창길 한국식물생명공학회 2018 JOURNAL OF PLANT BIOTECHNOLOGY Vol.45 No.1
We established an in vitro system to investigate transcription levels of the RsMYB1 gene expressed in T2 20-day-old transgenic Petunia plants (three independent lines: PhRs1, PhRs2, and PhRs3), and the association between those transcription levels and anthocyanin production at various pH values (3.0 to 8.0) for a period of 10 days. All the lines treated with pH 5.0-7.0 exhibited increased anthocyanin content and delays in growth compared to the wild-type (WT) seedlings. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis confirmed that the enhancement of anthocyanin production in the transgenic lines was due to the upregulation of RsMYB1 transcription at various pH values. The results suggest that pH value can control expression of RsMYB1 which is associated with anthocyanin production.