• Proceedings of the Plant Resources Society of Korea Conference
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• 2023.04a
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• pp.34-34
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• 2023

화본과 식물인 홍띠(Imperata cylindrica 'Rubra') 식물체의 기관분화 단계별 시료를 작성하여 이들 몇가지 기관분화에 관련된 유전자의 각 단계별 기관분화체에서 유전자존재와 발현여부를 조사하였다. 또한 식물의 기관분화에 관련된 유전자 정보를 얻기 위하여 일부 유전자의 특성을 정리하였다. 조사된 기관분화 관련 유전자중 탈분화 관련 유전자로는 FIF, RAP2-4 (WIND1) 유전자 등이, shoot 분화 관련 유전자로는 WUS, 부정근 분화관련 유전자로는 OsSCR, WOX11 등, 체세포배발생 관련 유전자로는 BBM1, SERK1, LEC1B, MEA 유전자 등이다. 이들 유전자중 RAP2-4(WID1), FIE, BBM1, SERK1, OsSCR, WOX11, WUS, LEC1B 유전자 등 8종의 기관분화 관련 유전자를 대상으로 화본과 식물의 기관분화의 각 단계별 기관분화체를 작성하여 PCR을 통하여 유전자(gDNA)의 존재여부를 확인한 결과 공시 유전자 모두 홍띠의 각단계 기관분화체에서 존재하였다. 또한 상기 유전자를 사용하여 화본과 식물의 각단계별 기관분화체에 대하여 유전자 발현을 확인한 결과 각단계 기관분화체에서 모두 발현하였다. 5종류 총 15개체의 기관분화 단계별 분화체에서 캘러스 발생 유전자인 FIE는 모식물체 1번을 제외한 14개의 식물체에서 모두 관찰되었으며, 뿌리 발생 유전자인 WOX11은 15개의 모든 식물체에서 탐색되었으며, 체세포 발생 유전자인 LEC1B는 15개 식물체에서 모두 발현하였으나 비교적 약하게 발현하였다. 이상과 같이 본 연구에서는 식물 기내발생시 기관분화관련 유전자의 동향을 파악하여 식물발생학의 기초자료를 구축하고자 하였다.

• Korean Journal of Plant Tissue Culture
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• v.28 no.1
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• pp.7-13
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• 2001

Calli were induced from diploid and haploid tobacco after 4 weeks and maintained on MS medium with combination of 2.0 mg/L 2,4-D,0.1 mg/L BAP and 2.0 mg/L kinetin. Suspension cells were screened through 65 $\mu$m-nylon mesh and 100 $\mu$m-mesh, then they were smeared on selection medium combined with cadmium and PFP by using the low melting agarose of 0.8%. After 30days smeared cultures of the medium the cell was treated with 500 $\mu$M and 1000 $\mu$M to select the resistant cell line were selected. Plant regeneration was induced from the selected cell lines on medium with 0.5, 1.5, 2.0 mg/L BAP and on media with combination of auxin and BAP under 500 $\mu$M and 1000 $\mu$M cadmium. At this time, plant regeneration was achived on cadmium free medium. In case of haploid, occurred from the cell line which is selected in medium with cadmium and PFP. In case of diploid regeneration occurred is in the medium with cadmium alone. The plantlet regenerated from cadmium resistant calli grew well in cadmium 500 $\mu$M. Protein pattern of leaf, root, stem of regenerated plants was analyzed. The quantum was 6.5188 ug/mg.fr.wt in the leaf of plant, 5.3611 ug/mg.fr.wt in the stem, 3.0213 ug/mg.fr.wt in the root. On the other hand, 5.9652 ug/mg.fr.wt. in the leaf of control, 3.5974 ug/mg.fr.wt in the stem of the control, 4.3766 ug/mg.fr.wt. in the root of the control. The one dimension bends regenerated from cadmium resistant calli resistant to cadmium in leaf were 49 involving 198.7KD etc. Disappeared were 4 involving 160.5KD etc, The protein bends were combinized were 3 involving 83.4KD etc. The bends resistant to cadmium stress in stem were 41 involving 4.3KD etc. Disappeared were 5 involving 114.8KD etc. The protein bends combinized were 6 involving 128.7KD etc. The bends which had the resistance to cadmium stress in root is 27 in volving 166,9KD etc. The bends which disappeared were 198.7KD etc. There were 5 involving 83.4KD etc.

• Korean Journal of Pharmacognosy
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• v.5 no.2
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• pp.111-124
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• 1974

The radioactive compound sodium $acetate-U-C^{14}\;(C^{14}-acetate)$ was administered to two- and four-year-old July and September American ginseng (Araliaceae, Panax quinquefolium L.) plants and cuttings. The $C^{14}-acetate$ uptake was approximately 99%. The autoradiochromatograms suggest that the saponins isolated by preparative thin-layer chromatography contained impurities, especially those isolated from the leaf and stem extracts. The root and fruit methanol extracts yielded relatively pure saponins. The large amounts of panaquilin B and its proximity to panaquilin C on preparative thin-layer plates resulted in some admixing. The average concentration (% plant dry weight) of semi-purified saponins were high in the leaves (13.8%), as compared to fruits (9.8%), stems (7.9%) and roots (6.3%). The average percentage of $C^{14}-acetate$ incorporation into panaquilins was 4.8%. The average percentage of $C^{14}-acetate$ incorporation into panaquilins B and C was higher (1.40% and 1.13%, respectively) than that into panaquilins C, (d), G-1 and G-2 (0.75%, 0.65%, 0.13% and 0.53%, respectively). Panaquilin synthesis may be depending upon the part, collection period and age of the plant. The average percentage of $C^{14}-acetate$ incorporation into panaquilin B is high in roots (0.58%) and stems (0.48%); that into panaquilins C and (d) high in leaves (0.40% and 0.45%, respectively); and that into panaquilin E high in roots and leaves (0.55% and 0.50%, respectively). Panaquilin G-2 was synthesized in all parts of plants. The panaquilins appear to be biosynthesized more actively in July than September (exception-panaquilin G-1). Panaquilins B, C and G-1 may be biosynthesized more actively in four-year-old plants and panaquilins (d) and E more actively in two-year-old plants. The results from expectance with cuttings suggest that the panaquilins are synthesized de novo in the above-ground parts of ginseng plants, and that panaquilin G-1 may be synthesized de novo in the leaf. It is known from the tissue culture studies that panaquilins are produced by leaf, stem and root callus tissues and cailus-root cultures of American and Korean ginseng plants. Panaquilins may actively be synthesized de novo in most any cell or organ of the ginseng plants. It was verified that $C^{14}-acetate$ was incorporated into the panaxadiol portions of the panaquilins of two-year-old plants (sp. act. 0.56 mmcCi/mg) and four-year-old plants $(sp.\;act.\;0.54\;m{\mu}Ci/mg)$.

• Horticultural Science & Technology
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• v.35 no.1
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• pp.121-130
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• 2017

This study was carried out to investigate a method for producing cultured virus - free ovules for breeding high - quality Citrus cultivars. Ovules from the immature fruits of three citrus cultivars native to Jeju (Dongjeongkyool, Cheongkyool, and Jikak) and two cultivars of Citrus unshiu Marc. (Miyagawa wase and Haryejosaeng) that were thought to be infected with Citrus tristeza virus (CTV) were cultured on MS2 medium (Murashige - Skoog [MS] basal medium containing $500mg{\cdot}L^{-1}$ malt extract, $50g{\cdot}L^{-1}$ sucrose, $1.0 mg{\cdot}L^{-1}$ kinetin, and $8g{\cdot}L^{-1}$ agar). After four weeks of culture, 10, 21, 13, 5, and 7 somatic embryos and 2, 4, 2, 4, and 5 white callus cells (surrounding green somatic embryos) were obtained from Dongjeongkyool, Cheongkyool, Jikak, Miyagawa wase, and Haryejosaeng, respectively. After six weeks of culture, somatic embryos were obtained from cultured cells grown on MT basal medium supplemented with malt extract ($500mg{\cdot}L^{-1}$), lactose ($70g{\cdot}L^{-1}$), and agar ($16g{\cdot}L^{-1}$). Over 60% of the somatic embryos from citrus cultivars native to Jeju developed into normal plants on MS basal medium supplemented with malt extract ($500mg{\cdot}L^{-1}$), sucrose ($50g{\cdot}L^{-1}$), and agar ($8g{\cdot}L^{-1}$) after 10 weeks of culture. Normal plants were regenerated from two Citrus unshiu Marc. cultivars on MT basal medium supplemented with sorbitol (1.0 M), galactose (1.0 M), $GA_3$ ($1.0mg{\cdot}L^{-1}$), and Gelrite ($3g{\cdot}L^{-1}$). The absence of virus in plants generated from cultured ovules was confirmed by RT - PCR and antigen - antibody reactions. Therefore, virus - free Citrus cells can be obtained for breeding high - quality citrus cultivars using the biotechnological technique evaluated in this study.

• Korean Journal of Plant Resources
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• v.20 no.5
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• pp.389-397
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• 2007

Using the active-increment of Tos17 copies in the genome of Oryza sativa L., there were many studies about induction and selection of new mutants. This study mainly focuses on the induction for retrotransposon(Tos17) activity in the callus of Ilpumbyeo(Oryza sativa L.) according to varied culture period and condition. The objectives of this study are obtaining various mutants($M_1$) through plant regeneration, identification of the mutation relation with Tos17, and subsequent phenotyping of the mutants($M_2,\;M_3$). A total of 371 $M_1$ mutants was obtained. The degree of Tos17 activity obtained regeneration plants with each different culture period was evaluated by Southern blot analysis. The result showed that control Ilpumbyeo rice has 5 numbers of copies and the band numbers obtained 7, 8, 9.5, 12, 6, 13.5, 17.5 from culture period of 1, 2, 3, 5, 6, 7, 8 month, respectively. In this study, the result showed that most effectual culture period for activity of Tos17 in Ilpumbyeo rice is 5 month. Hereafter, collections and analysis of various recombination plants will act on an important factor in multiplication and preservation of $M_2$ and $M_3$ generation. And an urgent and important subject is a development of screening method for selection of diverse mutants.

• Korean Journal of Plant Resources
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• v.26 no.2
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• pp.320-327
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• 2013

This study was carried out to address an efficient in vitro regeneration system from seed-derived callus of Phragmites communis, and to evaluate genetic variations of the regenerants using ISSR markers. Shoot regeneration via calli was greatly influenced by N6 medium compared with MS medium, and plant regeneration frequency was 90% in N6 supplemented with BA 0.25 mg/L and BA 0.5 mg/L. According to ISSR analysis of the thirty regenerants, out of 94 loci detected overall, 16 were identified to be polymorphic with a rate (PR) of 17.0%. The mean gene diversity (h) of different in vitro condition was 0.03 and ranged from 0.008 for N6 with BA 5 mg/L, to 0.040 for MS with IAA 0.1 mg/L+kinetin 2 mg/L. The results indicate that the regenerants have a low genetic variation, and ISSR analysis is effective to detect genetic variation of regenerants.

• Horticultural Science & Technology
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• v.19 no.4
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• pp.459-464
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• 2001

Leaf disks from cultivar 'Kennebec' and one selection line (ND 860-2) were cultured on Murashige-Skoog medium with various combinations of indole acetic acid (IAA) and zeatin riboside. Shoots, roots and callus were induced at various combinations of plant growth regulator levels. The medium containing $3.5mg{\cdot}L^{-1}$ IAA and $4.0mg{\cdot}L^{-1}$ zeatin riboside produced the most plantlets. Rooted regenerants were grown in the greenhouse. The growth of regenerated plants obtained from the MS medium supplemented with $7.0mg{\cdot}L^{-1}$ IAA and $3.0mg{\cdot}L^{-1}$ zeatin riboside was significantly greater than those grown from nodal expalnts. In ND 860-2, a leaf chimera with chlorophyll deficient (light yellow) sectors was found in plants regenerated fiom leaf disks (grown on MS medium supplemented with $3.5mg{\cdot}L^{-1}$ IAA and $3.0mg{\cdot}L^{-1}$ zeatin riboside) but not in plants grown from nodal explants. The phenotypic variability was also observed in the tuber number, size and weight.

• Journal of Bio-Environment Control
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• v.22 no.4
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• pp.322-327
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• 2013

This research was conducted to elucidate the appropriate light environment right after grafting to produce vigorous cherry tomato seedlings. Tomato plants were grafted and then treated in 4 different ways: to keep in the natural light (Non), to cover the grafted stem part with aluminum foil to make only that part dark (Part), to put the grafted seedlings in the acclimation room for two (Day-2) or four days (Day-4) to make the whole seedlings in the dark condition. Tube grafting method was used for grafting, in which silicon tube of 1.5mm in diameter was used. The survival rate was the maximum in the treatment Day-2. The SPAD value, seedling quality and yield of $1^{st}$ and $2^{nd}$ cluster were the best in the treatment Part. The treatment Part needs cost more than other treatments but is more economic thanks to higher yield. Therefore it was concluded to be economically feasible to make the grafted stem part dark right after grafting in case of cherry tomato.

• Journal of Plant Biotechnology
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• v.35 no.3
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• pp.235-243
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• 2008

Here we describe a procedure for Chinese cabbage protoplast culture and effects of various treatments. Chinese cabbage protoplasts were isolated from different parts of young seedlings as using an enzyme mixture, of which yield was maximized in seven hours around after digestion. The highest rate of initial cell division followed by micro-callus formation was obtained in the medium with 1.0 mg/L 2,4-D, 0.5 mg/L NAA, and 1.0 mg/L BA when the cotyledon-derived protoplasts were cultured. Initiation of cell division and micro-callus proliferation significantly depended upon Chinese cabbage genotype under a same culture circumstances. The micro-calli developed from cotyledon tissue of Norang-Bom cultivar successfully grew toward callus colonies on the solidified medium with 0.2 mg/L zeatin and 0.1 mM spermidine. The callus colonies generated de novo shoots at the maximum frequency of 4.3% on the medium with 5.0 mg/L BA and 1.0 mg/L NM. Our results might be helpful for further studies to enhance the regeneration efficiency in Chinese cabbage protoplast culture.

• Journal of Life Science
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• v.16 no.3 s.76
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• pp.527-533
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• 2006

The purpose of this study is to improve the culturability of a indica type rice cultivar, IR 36, using DNA marker associated with the ability of plant regeneration in anther and seed culture. The varietal difference of ability of callus formation and plant regeneration was investigated in anther and seed culture of 8 rice cultivars. Three japonica rice cultivars showed to have better culturability than those of tongil and indica type genotypes. But two indica/japonica lines, 'MGRI 079' and 'MGRI 036', which were selected to have good culturability in previous study showed the highest regenerability (20%) in anther culture of 8 rice cultivars. Thirty four $BC_2F_4$ lines were selected by marker screening using RZ400 for 100 $BC_2F_4$ lines derived from a cross $'MGRI\;079/IR\;36^{^*3}'$. The frequency of callus formation of 30 $BC_2F_4$ lines was higher than those of 'IR 36' in anther culture of the selected $BC_2F_4$ lines. The ability of plant regeneration of 15 lines was higher than that of 'IR 36' in the seed culture of 34 $BC_2F_4$ lines. A promising line, $BC_2F_4-28$, was selected to have better culturability in the anther and seed culture of the $BC_2F_4$ lines. The heading date and grain shape of the $BC_2F_4-28$ was similar to 'IR 36'. In seed culture of 50 $BC_4F_3$ lines derived from a rice cross $'MGRI\;079/IR\;36^{^*5}'$, 11 lines including $BC_4F_3-3$ showed to have higher regenerability compared with 'IR 36'. The highest frequency of plant regeneration (11%) was obtained from $BC_4F_3-46$ in seed culture of the $BC_4F_3$ lines.