Saetiew, Kanjana

Preferred name

Saetiew, Kanjana

Alternative Name

Saetiew, K.

Main Affiliation

School of Agricultural Technology

kanjana.sa@kmitl.ac.th

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A cytogenetic study of three Japanese cultivars of momordica charantia L.
(2016-01-01)
Kido, Michiko
;
Morikawa, Atsumi
;
Saetiew, Kanjana
;
Hoshi, Yoshikazu
Using a base-specific staining method, three Japanese cultivars of Momordica charantia were investigated. The present method using enzymatic digestion produced a high level of well-spread metaphase chromosome complements. The cultivars investigated showed the same somatic chromosome number 2n=22. In condensation behavior from prophase to metaphase, most chromosomes of all cultivars in this study had early condensing segments at the proximal regions. Moreover, at prophase or earlier stage of metaphase, most chromosomes observed had decondenced segments at the distal regions in both arms. Judging by the determination of centromere positions, the chromosome complements at mitotic metaphases in three cultivars consisted of 22 metacentric chromosomes. Moreover, similar sizes and shapes of the chromosomes indicated that the karyotypes of M. charantia cultivars were symmetrical. In contrast, fluorescent staining showed different sizes and numbers of chromycin A<inf>3</inf> (CMA) positive and 4′,6-diamidino-2-phenylindole (DAPI) negative (CMA+DAPI-) satellites among cultivars. Four CMA+DAPI- satellites were observed in M. charantia Abashi-goya and M. charantia Naga-goya, while two satellites were obseved in M. charantia Shiro-goya. All satellites were located at one end of sat-chromosomes. In Shiro-goya. a large size difference of satellites was found between two satchromosomes, which might be homologous chromosomes with polymorphism. Moreover, faintly CMA+DAPI- sites were shown at the proximal regions or primary constrictions of most chromosomes in all cultivars.
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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 of antisense dihydroflavonal 4-reductase (DFR) into sacred lotus 'Buntharik' using Agrobacterium-mediated gene transfer
(2014-03-13)
Saetiew, K.
;
Leethaweesup, W.
;
Parinthawong, N.
;
Arunyanart, S.
Transformation of antisense dihydroflavonal 4-reductase (DFR) into 'Buntharik' sacred lotus (Nelumbo nucifera Gaertn.) using Agrobacterium-mediated gene transfer was experimented. The A. tumefaciens strain EHA105 harbored two binary vectors. The pCAMBIA2301anti-DFR plasmid contained neomycin phosphotransferase (NPTII) as a selectable marker gene, the antisense DFR as an inserted gene and the β-glucuronidas gene (GUS) as a reportor gene. The pBI121anti-DFR plasmid contained NPTII gene and the antisense DFR. Twomonths-old calli regenerated from apical buds were used as plant materials. The plant materials were soaked in the Agrobacterium suspension for 10 or 30 min. The calli were co-cultivated for two days in the darkness. Treated calli were selected on solid MS medium containing 50 mg L-1 kanamycin and 250 mg L-1 cefotaxime. The calli were transferred to the same medium every two weeks for eight weeks. The survived calli regenerated shoots on MS medium containing a combination of 0.54 μM NAA and 4.44 μM BA. Both plasmids gave the best callus growth, maximum callus size and the survival percentage when soaked for 10 min. GUS bioassay was used to verify the presence of GUS gene in petioles and leaves of transgenic plants. It was found that petioles and leaves of transformants which were transformed with pCAMBIA2301anti-DFR by soaking for 30 min showed the highest percentage of blue spots on explants. Of all 14 PCR positive clones, 5 clones showed GUS gene, 11 clones contained NPTII gene and 6 clones had DFR gene.

Saetiew, Kanjana

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