• Title/Summary/Keyword: In vitro regeneration

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MET1-Dependent DNA Methylation Represses Light Signaling and Influences Plant Regeneration in Arabidopsis

  • Shim, Sangrea;Lee, Hong Gil;Seo, Pil Joon
    • Molecules and Cells
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    • v.44 no.10
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    • pp.746-757
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    • 2021
  • Plant somatic cells can be reprogrammed into a pluripotent cell mass, called callus, which can be subsequently used for de novo shoot regeneration through a two-step in vitro tissue culture method. MET1-dependent CG methylation has been implicated in plant regeneration in Arabidopsis, because the met1-3 mutant exhibits increased shoot regeneration compared with the wild-type. To understand the role of MET1 in de novo shoot regeneration, we compared the genome-wide DNA methylomes and transcriptomes of wildtype and met1-3 callus and leaf. The CG methylation patterns were largely unchanged during leaf-to-callus transition, suggesting that the altered regeneration phenotype of met1-3 was caused by the constitutively hypomethylated genes, independent of the tissue type. In particular, MET1-dependent CG methylation was observed at the blue light receptor genes, CRYPTOCHROME 1 (CRY1) and CRY2, which reduced their expression. Coexpression network analysis revealed that the CRY1 gene was closely linked to cytokinin signaling genes. Consistently, functional enrichment analysis of differentially expressed genes in met1-3 showed that gene ontology terms related to light and hormone signaling were overrepresented. Overall, our findings indicate that MET1-dependent repression of light and cytokinin signaling influences plant regeneration capacity and shoot identity establishment.

Suppressive effect of Curcuma xanthorrhiza oil on plaque and gingivitis (천연추출물 Curcuma xanthorriza oil 함유치약의 치태 및 치은염 억제효과)

  • Hong, Ji-Youn;Kim, Sang-Nyun;Ha, Won- Ho;Chang, Sug-Youn;Jang, In-Kwon;Park, Ji-Eun;Jung, Sung-Won;Um, Yoo-Jung;Choe, Seong-Ho;Kim, Chong-Kwan
    • Journal of Periodontal and Implant Science
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    • v.35 no.4
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    • pp.1053-1071
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    • 2005
  • To find out the suppressive effect of natural extract Curcuma xanthorrhiza on $IL-1{\beta}$ and MMP-2 derived from periodontal ligament cells through in vitro study and to confirm its effect on plaque and gingivitis through clinical study, Curcuma xanthorrhiza containing toothpaste was used and following results were produced. 1. In vitro study, type IV collagenase MMP-2 production was inhibited dose-dependently in the group treated with Curcuma xanthorrhiza compared to the control group. 2. In vitro study, the production of $IL-l{\beta}$ which is one of the inflammatory mediators associated with periodontitis was inhibited dose-dependently in the group treated with Curcuma xanthorrhiza. 3. On the third week, the plaque index of the groups treated with or without Curcuma xanthorrhiza containing toothpastes were both increased significantly compared to the baseline(p<0.05). 4. On the third week, the gingival index of the group treated with Curcuma xanthorrhiza containing toothpaste was not significantly different from baseline. However, the group treated without Curcuma xanthorrhiza containing toothpaste showed a significant increase of gingival index at shielded area(p<0.05). 5. The gingival index of the group without Curcuma xanthorrhiza containing toothpaste showed a significant increase in the sites without tooth brushing when compared to sites with tooth brushing(p<0.05). However. there was no significant difference for the group with Curcuma xanthorrhiza containing toothpaste in sites either with or without tooth brushing. 6. The Bleeding on probing for the group without Curcuma xanthorrhiza containing toothpaste showed no significant difference even when tooth brushing was done. However, for the group with Curcuma xanthorrhiza containing toothpaste, bleeding on probing was significantly reduced compared to baseline when tooth brushing was done(p<0.05).

In-vitro Production of Glutathione Using Yeast ATP Regeneration System and Recombinant Synthetic Enzymes from Escherichia coli. (효모의 ATP 재생산계와 대장균 유래의 재조합 생산효소를 이용한 in vitro 글루타치온 생산)

  • 고성영;구윤모
    • Microbiology and Biotechnology Letters
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    • v.26 no.3
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    • pp.213-220
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    • 1998
  • An ATP regeneration system was used for the production of glutathione which was synthesized by a sequential action of ${\gamma}$-glutamyl-cysteine synthetase and glutathione synthetase. The synthetases above were produced in the recombinant E. coli (TG1/pDG7) with the highest specific production yield of 31 mg glutathione/g wet cell. Bakers yeast was considered to have economically a better ATP regeneration system although the glutathione production yield was lower than that of acetate kinase. It was also observed that the ATP regeneration system of bakers yeast was superior to that of Saccharomyces cerevisiae ATCC24858. The yield of glutathione production with bakers yeast was 36% with the ATP concentration of 5 mM. To avoid the cysteine limitation during the early phase of glutatione production, an extra cysteine was added at 2 hours after reaction and the production yield increased 1.91 times. The effectiveness of bakers yeast as an ATP regeneration system was proved by several sets of extra feeding experiments. The product inhibition by glutathione above 14 mM was also observed.

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Effect of Basal Medium and Plant Growth Regulator on in vitro Plant Regeneration from Axillary Buds of Walnut New Cultiver "Sinlyeong"

  • Kwon, Young Hee;Lee, Joung Kwan;Kim, Hee Kyu;Kim, Kyung Ok;Park, Jae Seong;Huh, Yoon Sun
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.10a
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    • pp.15-15
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    • 2019
  • The walnut (Juglans regia L.), a member of the Juglandaceae, is native to the mountain ranges of central Asia. This species of walnut is valued commercially for its nuts and in some areas for its timber. The seeds of walnut are recalcitrant and it has strong integument dormancy and their germination is irregular, making its natural propagation difficult. Low percentage of seed germination and long propagation cycle are the main problems of propagation. This study was conducted medium composition on in vitro plantlet regeneration from axillary buds of walnut. It has proved to be the most generally applicable and reliable method of in vitro propagation. Micropropagation culture that axillary buds are excised aseptically enables faster multiplication of plants. The axillary buds of walnut new cultivar "Sinlyeong" were cultured on two basal media which contained the different plant growth regulators depending on the respective shooting and rooting stage. After 12 weeks, the shoot generation rate was 85.3%, the shoot number and its length were 1.9/explant and 2.7 cm in the most favorable medium composition. The percentage of rooting was 25.4%. From these results, it was found the optimum basal medium and plant growth regulator for in vitro plant regeneration from axillary buds of walnut new cultivar "Sinlyeong". However, we have continued to search the other medium additives to enhance the rate of walnut root.

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In vitro regeneration from cotyledon explants in figleaf gourd (Cucurbita ficifolia Bouch$\'{e}$), a rootstock for Cucurbitaceae

  • Kim, Kyung-Min;Kim, Chang-Kil;Han, Jeung-Sul
    • Plant Biotechnology Reports
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    • v.4 no.2
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    • pp.101-107
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    • 2010
  • An efficient plant regeneration system has been developed for figleaf gourd (Cucurbita ficifolia Bouch$\'{e}$), which is exclusively used as a rootstock for cucumber. The protocol is based on results obtained from a series of culture experiments involving different parts of the cotyledons and various media. The culture of cotyledon explants was critical for the enhancement of shoot regeneration frequency. The lower parts of the cotyledon excised at the plumule base were found to display a markedly enhanced production of adventitious shoots compared to other cotyledon regions. Culture in silver nitrate-supplemented Murashige and Skoog (MS) medium was not beneficial for shoot regeneration and suppressed root regeneration. Efficient shoot regeneration was obtained on MS medium containing 1.0 $mg\;l^{-1}$ zeatin and 0.1 $mg\;l^{-1}$ indole-3-acetic acid. Regenerated shoots successfully elongated and rooted in medium containing 0.1 $mg\;l^{-1}$ 1-naphthalene-acetic acid after 10-15 days of subculturing. The plantlets were satisfactorily acclimatized in a greenhouse and grew into normal plants without any morphological alterations.

Complete In Vitro Conversion of n-Xylose to Xylitol by Coupling Xylose Reductase and Formate Dehydrogenase

  • Jang, Sung-Hwan;Kang, Heui-Yun;Kim, Geun-Joong;Seo, Jin-Ho;Ryu, Yeon-Woo
    • Journal of Microbiology and Biotechnology
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    • v.13 no.4
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    • pp.501-508
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    • 2003
  • Artificial coupling of one enzyme with another can provide an efficient means for the production of industrially important chemicals. Xylose reductase has been recently discovered to be useful in the reductive production of xylitol. However, a limitation of its in vitro or in vivo use is the regeneration of the cofactor NAD(P)H in the enzyme activity. In the present study, an efficient process for the production of xylitol from D-xylose was established by coupling two enzymes. A NADH-dependent xylose reductase (XR) from Pichia stipitis catalyzed the reduction of xylose with a stoichiometric consumption of NADH, and the resulting cofactor $NAD^+$ was continuously re-reduced by formate dehydrogenase (FDH) for regeneration. Using simple kinetic analyses as tools for process optimization, suitable conditions for the performance and yield of the coupled reaction were established. The optimal reaction temperature and pH were determined to be about $30^{\circ}C$ and 7.0, respectively. Formate, as a substrate of FDH, affected the yield and cofactor regeneration, and was, therefore, adjusted to a concentration of 20 mM. When the total activity of FDH was about 1.8-fold higher than that of XR, the performance was better than that by any other activity ratios. As expected, there were no distinct differences in the conversion yields of reactions, when supplied with the oxidized form $NAD^+$ instead of the reduced form NADH, as a starting cofactor for regeneration. Under these conditions, a complete conversion (>99%) could be readily obtained from a small-scale batch reaction.

Advances in in vitro culture of the Brassicaceae crop plants

  • Park, Jong-In;Ahmed, Nasar Uddin;Kim, Hye-Ran;Nou, Ill-Sup
    • Journal of Plant Biotechnology
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    • v.39 no.1
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    • pp.13-22
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    • 2012
  • Plant regeneration has been optimized increasingly by organogenesis and somatic embryogenesis using a range of explants with tissue culture improvements focusing on factors, such as the age of the explant, genotype, media supplements and $Agrobacterium$ co-cultivation. The production of haploids and doubled haploids using microspores has accelerated the production of homozygous lines in Brassicaceae crop plants. Somatic cell fusion has facilitated the development of interspecific and intergeneric hybrids in sexually incompatible species of $Brassica$. Crop improvement using somaclonal variation has also been achieved. Transformation technologies are being exploited routinely to elucidate the gene function and contribute to the development of novel enhanced crops. The $Agrobacterium$-mediated transformation is the most widely used approach for the introduction of transgenes into Brassicaceae, and $in$ $vitro$ regeneration is a key factor in developing an efficient transformation method in plants. Although many other Brassicaceae are used as model species for improving plant regeneration and transformation systems, this paper focuses on the recent technologies used to regenerate the most important Brassicaceae crop plants.

Efficient Bulblet Regeneration and Growth from Bulb Scale of Hyacinthus orientalis L. cv. Pink Pearl Cultured in vitro (히아신스(cv. Pink Pearl)의 인편 기내 배양시 효과적인 자구의 재생과 생장)

  • Lee, Kyung-Soon;Chung, Chung-Han;Chung, Yong-Mo;Kim, Doh-Hoon;Jeong, Soon-Jae;Nam, Jae-Sung;Kim, Gyung-Tae;Yi, Young-Byung
    • Journal of Life Science
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    • v.17 no.10
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    • pp.1336-1340
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    • 2007
  • The regeneration and growth of bulblets from the bulb scale segments of Hyacinthus orientalis L. cv. Pink Pearl were more efficient in IBA than IAA at the same concentrations (1.0 and 3.0 mg/l). The normal (base-down) orientation of explants was more effective for bulblet regeneration and root growth than the inverted (base-up) orientation. The growth of bulblets and roots was increased higher in the perlite than the agar medium. These results suggested that the alternate culture system, first cultured in the agar medium for bulblet regeneration, and then in the perlite medium for bulblet growth, may be more useful for efficient in vitro culture of hyacinth (H. orientalils) cv. Pink Pearl.