Parinthawong, Nonglak

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Parinthawong, Nonglak
Alternative Name
Parinthawong, N.
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Email
nonglak.pa@kmitl.ac.th

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    Isolation and expression analysis of the Flavanone 3-Hydroxylase genes in Lotus (Nelumbo nucifera gaertn.), Waterlily (Nymphaea sp.) and transient silencing in Waterlily
    (2017-04-01)
    Chaipanya, Chaivarakun
    ;
    Saetiew, Kanjana
    ;
    Arunyanart, Sumay
    ;
    Parinthawong, Nonglak
    Flavanone 3-hydroxylase (F3H) converts flavanones from dihydroflavonols, which leads to production of flavonoid compounds via the anthocyanin biosynthesis pathway in plants. In this study, the F3H gene was isolated from lotus (Nelumbo nucifera Gaertn.) cv. Buntharik (white petal lotus), cv. Satabankacha (pink petal lotus), and waterlily (Nymphaea sp.) var. St. Louis Gold by reverse transcription PCR (RT-PCR). The open reading frames (ORF) of three cultivars’ genes were 1,134 bp in length, encoding a predicted protein of 377 amino acids. Their nucleotide sequences were identical, and the amino acid sequence shared high homology to F3H from different plant species. Expression of F3H was specifically regulated in petals and stamens, while less expression was found in leaf tissue of waterlily variety St. Louis Gold. The correlation of F3H expression according to specific colouration was performed in waterlily. The F3H gene was more highly expressed in decreasing order of red, purplish blue, and yellow petals when compared using semi quantitative PCR (sqPCR). Gene regulation according to flowering stage and pigmentation was determined in lotus. The F3H expression was slightly diminished in petals of cultivar Satabankacha at the fully-opening stage, whereas it was detected in the cultivar Buntharik only when white petals were tinted with pink. An RNAi gene-silencing vector, pJA8F3H, encoding a hairpin F3H RNA, was introduced to waterlily petals using the Agrobacterium infiltration method, and the F3H expression was analysed at 1 and 3 days post infiltration (dpi) by sqPCR. The results showed that the F3H expression was down-regulated at 3 dpi in flowers tested of the red petal variety and purplish blue petal variety compared to controls. The results confirmed that pJA8F3H is efficient and could be used as a transformation vector to transiently suppress F3H expression in waterlily or lotus.
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    Tissue culture and transformation of the antisense DFR gene into lotus (Nelumbo nucifera Gaertn.) through particle bombardment
    (2013-09-24)
    Buathong, Raveevatoo
    ;
    Saetiew, Kanjana
    ;
    Phansiri, Salak
    ;
    Parinthawong, Nonglak
    ;
    Arunyanart, Sumay
    The effects of explant and plant growth regulators on callus induction and plant regeneration and particle bombardment-induced transformation were examined in lotus (Nelumbo nucifera Gaertn.) cv. Buntharik. The apical buds from embryos (~3. mm) formed an embryogenic callus when cultured on MS medium supplemented with 40. μM 1-napthaleneactic acid (NAA) and 0.5. μM 1-phenyl-3-(1,2,3-thiadiazol-5-yl)-urea (TDZ) for 8 weeks. All of the embryogenic calluses were transferred to MS medium supplemented with 0, 40, 50 and 60. μM 6-benzyladenine (BA) and the highest number of shoots was achieved in the medium supplemented with 50. μM BA after 8 weeks of culture. The transformation of lotus using a particle bombardment device was examined with the shoot clusters from embryogenic apices. The pCAMBIA2301anti-DFR plasmid contained β. -glucuronidase (GUS) as a reporter gene, neomycin phosphotransferase (NPTII) as a selectable marker, and antisense dihydroflavonol 4-reductase (anti-. DFR) as the gene of interest. The particle bombardment procedure with a helium gas pressure of 1100. psi and a target distance of 6. cm yielded the highest number of blue spots (4.8 spots/shoot cluster). However, stable transformations, as confirmed by polymerase chain reaction and reverse transcription polymerase chain reaction analysis, were only achieved using a helium gas pressure of 1100. psi and a target distance of 9. cm. Our work illustrates that the stable transformation of lotus is possible, and we suggest that this methodology can be used to target and modify specific charactics of this plant. © 2013 Elsevier B.V.
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    Transformation RNA interference-mediated silencing of the Flavanone 3-Hydroxylase gene in Lotus (Nelumbo nucifera Geartn. cv. Buntharik)
    (2020-07-01)
    Saetiew, K.
    ;
    Konee, C.
    ;
    Parinthawong, N.
    ;
    Arunyanart, S.
    RNA interference mediated-silencing of the F3H gene in the lotus (Nelumbo nucifera Geartn.) cv. Buntharik inhibited the anthocyanin pathway after gene transformation. The plasmid pJA8F3H contained the inverted repeats of F3H fragments, bar gene as a selectable marker in plants, which is resistant to basta (glufosinate amonium). The transformation of particle bombardment was obtained with multiple shoots of lotus. F3H gene expression analysis showed that expression of F3H decreased in transgenic lotus when compared with the control. On the first and the second test, 5 transgenic lotus of the relative expression F3H gene was detected with 0.0496 and 0.466 times, respectively. The inhibition of gene expression decreased when they were cultured for a long time. The expression F3H gene was detected by RT-PCR, and real-time PCR resulted in consistent decreasing in the transgenic lotus compared with the non-transgenic lotus, which was able to inhibit the expression F3H gene.