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    A cytogenetic study of three Japanese cultivars of momordica charantia L.
    (2016-01-01)
    Kido, Michiko
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    Morikawa, Atsumi
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    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<sup>+</sup>DAPI<sup>-</sup>) satellites among cultivars. Four CMA<sup>+</sup>DAPI<sup>-</sup> 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<sup>+</sup>DAPI<sup>-</sup> 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
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    Arunyanart, Sumay
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    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
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    Phansiri, Salak
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    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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    Changes in the activity of enzymes associated with enzymatic browning and chemical composition during Musa sapientum Linn. 'Kluai Khai' banana fruit ripening
    (2023-07-01)
    Makboriboon, N.
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    The results showed that the initial firmness of banana fruit harvested at the raw stage was 7 N and decreased to less than 2 N at overripe and very ripe stages. The total soluble solids increased throughout the ripening period, whereas the titratable acidity maintained a high level during the ripening process. The banana peel stayed green during the raw stage, and the yellow color formed between the unripe stage to the overripe stage. The lightness of the banana peel increased from the raw stage to the ripe stage, decreased at the overripe stage, and then dropped sharply at the very ripe stage. This color change was linked to the browning incidence of the fruit peel and an increasing browning area and soluble browning pigment content. The PPO and POD showed a low activity at the raw stage and then increased with advancing ripening and maintained a significantly higher level than the raw fruit. The findings of this study exhibited that the ripening stages of the 'Kluai Khai' banana influence the firmness, peel color, and browning incidence of the fruit peel and the polyphenol oxidase and peroxidase activities.
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    Induced mutation of curcuma hybrid cv. sweetmemory through tissue culture by ethyl methanesulphonate (EMS)
    (2025-01-01)
    Yoosumran, V.
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    Ruamrungsri, S.
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    Akarapisarn, A.
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    The ethyl methanesulphonate (EMS) is induced mutation in Curcuma hybrid cv. sweetmemory. The shoot size 1 cm were cultured on Murashige and Skoog medium (MS) supplemented with 2 mg/l BA for plantlet induction. After 8 weeks of culture, it was observed that the shoots soaked in 0% EMS at 60 and 120 minutes survived 100%. In additions it was found the highest percentage of shoot emergence, number of shoots, and plant height. While plants treated with EMS solution had decreased survival and growth rates. Thereafter, existing plantlets were potted, and growth was observed. It turned out that the plants none receiving EMS solution (control) had the survival rate and growth rate were better than those of the plants receiving EMS solution. Results showed that control plants receiving 0% EMS at 60 minutes gave highest content of chlorophyll a as 4.06 µg/cm<sup>2</sup>, chlorophyll b of 2.08 µg/cm<sup>2</sup> and carotenoid of 3.09 µg/cm<sup>2</sup> but there was none statistical difference with the plants that received EMS solution. The control plants had larger stomata sizes than those that received the EMS solution. However, there was no difference in the number of chloroplasts among treatments. The living plantlets from all treatments were tested Random amplified polymorphic DNA (RAPD) marker and genomic DNA was extracted from fresh leaves. A dendrogram was constructed polymorphic bands using the NTSYSpc program (version 2.10p) showing 2 clusters which separated with similarity coefficients ranging from 0.74-1.00.
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    Gene actions of resistance to anthracnose (Colletotrichum acutatum) of six F1 hybrid chili cultivars at the mature green fruit stage
    (2023-01-01)
    Sangarun, P.
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    Tsai, W. S.
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    Techawongstien, S.
    Anthracnose is an economically important fungal disease affecting the yield and fruit quality of chili in the tropics. This study aimed to assess the anthracnose disease reaction in six hybrid chili pepper cultivars compared with their parental lines. Two male resistant lines were crossed with three commercial female parents by North Carolina mating design II. All chili genotypes were grown in the greenhouse in a randomized complete block design with three replications, 10 plants each. Five mature green fruit plant<sup>-1</sup> were harvested and inoculated with C. acutatum by using the microinjection method. ANT4 had smaller lesions than ANT10. Additionally, the F1-hybrids derived from ANT4 showed smaller disease lesions (7.47-9.82 mm) than on those derived from ANT10 (9.40-11.04 mm). However, the percentage heterosis of anthracnose disease traits in ‘Jindanil 80’ (JD) and ANT4 gave the lowest value to other parents. Hybrid JD × ANT4 showed the lowest anthracnose lesion size. However, the heterosis of cross the NM × ANT10 was the most negative at -43.68. The mode of gene action was found to be additive rather than involving dominant effects, with a narrow-sense heritability of 50.9% and broad sense heritability of 70.1%. Therefore, the resistant parent ANT4 cultivar is crucial for use as a resistant elite line for producing good F1 hybrid tolerance to anthracnose disease.
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    The effect of BA on inducing shoots of Philodendron erubescent 'Pink Princes' in vitro
    (2023-11-01)
    Chiewchan, N.
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    The study investigated the influence of plant growth regulator on the induction of direct shoots from node explants of Philodendron erubescens 'Pink Princes' in vitro condition. The result showed that the most of width, length, and height was observed in MS + BA 0.5 mg/l, measuring 3.40 cm × 3.22 cm × 1.96 cm and 1.74 g fresh weight. The highest number of shoots was consistently observed 60.44 shoots per nodal MDA content (1.91 nmol/g FW) and phenolic content (9.51 mg GAE/g FW) are lowest. The explant was cultured on MS medium performed highest chlorophyll content (chl A 213.96 μg/g FW, chl B 162.54 μg/ FW, total chlorophyll 304.64 μg/FW) and carotenoid content (2.61 μg/g FW) Moreover, they were the largest plant with green leaves when compared to another treatment.
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    Effect of ethyl methanesulfonate on growth and morphology of Doritis pulcherrima Lindl.
    (2026-01-01)
    Supreeworakij, S.
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    Results showed that 0.5 mg/l BA and 1 mg/l NAA promoted the greatest seedling growth, with increased leaf number, leaf length, plant height, and root development. In the third experiment, protocorm-like bodies were treated with EMS at 0, 0.5, and 1.0% for 60 or 90 minutes. Survival declined with increasing concentration and time, with LD₅₀ values of 0.82% at 60 minutes and 0.54% at 90 minutes. Chlorophyll a, Chlorophyll b, and carotenoids were not significantly affected, but stomatal size changed. Stomatal width and length increased under 1.0% EMS, while density decreased from 220.17 to 149.47 stomata/mm². EMS treatment for 90 minutes showed an increased density relative to 60 min. RAPD analysis using 10 primers revealed high polymorphism indicating EMS induced genetic variation. Cluster analysis divided 22 samples into 9 groups at a similarity coefficient of 0.89, confirming genetic diversity despite no visible morphological changes.
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    Lotus flower extract as a natural anti-browning agent for fresh romaine lettuce (Lactuca sativa L. var. longifolia)
    (2023-11-01)
    Pradabkun, N.
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    Yeamsuriyotai, K.
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    The effect of lotus (Nelumbo nucifera Gaertn.), butterfly pea (Clitoria ternatea) and Siam tulip (Curcuma sessilis) aqueous extract on reduction of polyphenol oxidase (PPO) and peroxidase (POD) in romaine lettuce (Lactuca sativa L. var. longifolia) were evaluated. Results showed that in vitro lotus flower aqueous extract exerted the highest reduction of PPO and POD activities on the romaine lettuce. The effectiveness of in vivo application of lotus flower aqueous extract to control browning cut stem ends of fresh romaine was examined. Romaine lettuce harvested as a whole plant and cut off at the stem. Cut stem ends of romaine lettuce were dipped in various concentrations of lotus flower extract for 5 min and packaged in polypropylene plastic bags. Weight loss, color, browning index as well as PPO and POD activities were evaluated during 12 days of storage at temperature of 10±2°C and relative humidity 50±5%. Cut stem ends dipped in 0.5% aqueous extract of lotus flower resulted in inhibition of browning of cut stem ends and decrease in PPO and POD activities. Thus, exogenous 0.5% aqueous lotus flower extract treatment could be a useful application to alleviate browning in cut stem ends of fresh romaine lettuce.
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    Transformation of antisense dihydroflavonal 4-reductase (DFR) into sacred lotus 'Buntharik' using Agrobacterium-mediated gene transfer
    (2014-03-13) ;
    Leethaweesup, W.
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    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.