Saetiew, Kanjana
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Preferred name
Saetiew, Kanjana
Alternative Name
Saetiew, K.
Main Affiliation
Email
kanjana.sa@kmitl.ac.th
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Item type:Publication, Transformation of antisense dihydroflavonal 4-reductase (DFR) into sacred lotus 'Buntharik' using Agrobacterium-mediated gene transfer(2014-03-13); ; 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<sup>-1</sup> kanamycin and 250 mg L<sup>-1</sup> 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. - Some of the metrics are blocked by yourconsent settings
Item type:Publication, Transformation RNA interference-mediated silencing of the Flavanone 3-Hydroxylase gene in Lotus (Nelumbo nucifera Geartn. cv. Buntharik)(2020-07-01); ; 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.