• Journal of Plant Biotechnology
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• v.50
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• pp.89-95
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• 2023

In order to investigate the effect of carbon sources on somatic embryogenesis and cotyledon number variations in carrot, embryogenic callus were cultured in the medium supplemented with various concentrations of sucroseor glucose, and with combination of 2% sucrose and various concentrations of mannitol or sorbitol. The maximum number of somatic embryos formed per flask (1,555.70) was obtained in the medium supplemented with 2% sucrose rather than glucose alone or a combination of mannitol or sorbitol and 2% sucrose, and the number of somatic embryos was decreased with the increasing of mannitol or sorbitol concentration. The frequencies of somatic embryos with two cotyledons were 35.14% for sucrose, 19.88% for glucose, 32.55% for mannitol + 2% sucrose, and 38.59% for sorbitol + 2% sucrose, respectively, and the frequencies of abnormal somatic embryos having 3 or more cotyledons were 64.86% for sucrose, 80.12% for glucose, 67.44% for mannitol + 2% sucrose, and 61.41% for sorbitol + 2% sucrose, respectively. Particularly, the frequency of somatic embryos with two cotyledons (59.16%) was the highest in the 2% sucrose medium compared to the frequency of abnormal somatic embryogenesis with three or more cotyledons, and the frequency gradually decreased with increasing concentration of glucose, mannitol or sorbitol. According to these results, it was found that the ratio of abnormal somatic embryo was higher than the normal somatic embryo in carrot, and was shown that somatic embryogenesis and the cotyledon number was affected by the concentrations of sucrose, glucose as carbon source, and mannitol and sorbitol as osmotic agents in culture medium.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.55 no.3
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• pp.187-194
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• 2010

The soybean stay green mutant genotype (SSG) derived from the nuclear gene, d1d2, and cytoplasmic gene, cytG, inhibit the breakdown of chloroplast in the leaves, pod walls, seed coats, and embryos during maturity. Soybean seed with black seed coat and green cotyledon (SBG) are preferred than black seed coat with yellow cotyledon (SBY) especially for cooking with rice and as source of traditional food in Korea. The researchers evaluated the seed's chlorophyll content of SSG and introduced SSG to the SBG variety breeding program. The seed chlorophyll content of SSG with d1d2 was $39.93{\sim}60.80\;{\mu}g/g$ and SSG with cytG $38.08{\sim}39.89\;{\mu}g/g$. The Korean SBG variety which was derived from SSG with cytG, contains $16.35{\sim}37.73\;{\mu}g/g$. The composition of seed chlorophyll differs according to the genetic background of SSG genotype. Inheritance study showed that cotyledon color was segregated 15:1 (yellow:green) at $F_2$ seed indicating two recessive genes control green cotyledon as revealed by previous study. Only less than 3% soybean lines showed black seed coat with green cotyledon among crosses SBY and SSG (d1d2). Results showed that SSG with d1d2 can be used as a good source for SBG with high chlorophyll content in the seed cotyledon, but due to the complex genetic behavior, breeding resource of SBG with d1d2 should be prepared to improve the breeding efficiency for development SBG variety.

• Korean Journal Plant Pathology
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• v.11 no.4
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• pp.292-297
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• 1995

We developed an in vitro assay method for evaluating plant growth promotion and providing an evidence that the growth promotion is rendered by growth enhancing factors. The amendment of culture filtrates of Trichoderma harzianum T95 and Gliocladium virens G872 and G872B in Murashige and Skoog (MS) agar medium enhanced the cotyledon growth of cucumber in terms of fresh weight and primary leaf area of cucumber cotyledon cuttings, of which the treatment of G. virens G872B was the most effective. The mycelial culture filtrate of G872B was more effective in the growth promotion than its conidial germling filtrate. The addition of 1% sucrose in MS mineral medium with 0.1% culture filtrates of the antagonists (T95 and G872B) was optimum for enhancing the effect of the filtrates on the growth of cotyledon cuttings in vitro. When cucumber seeds treated with G872B, Pseudomonas putida Pf3 or the G872B-Pf3 mixture were planted in a greenhouse, the rate of seed germination, biomass of shoot and root, and yield of cucumber fruits were increased, especially by G872B or the G872B-Pf3 mixture. Correspondingly, cucumber fruit yields in early to middle-cycles of harvest were significantly greater in the plots of G872B than the control and Pf3-treated plots, and the final yield was highest in the plots of the G872B-Pf3 mixture applications.

• Journal of Plant Biotechnology
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• v.8 no.1
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• pp.15-19
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• 2006

An efficient protocol for in vitro multiple shoot bud induction and plant regeneration from mature green cotyledon derived callus tissues of Acacia sinuata has been developed. Callus formation occurs at all the concentrations of thidiazuron (TDZ) in Murashige and Skoog's (MS) medium, but 0.6 ${\mu}M$ proved to be the best with maximum callus formation frequency. Supplementation of TDZ in combination with indole-acetic acid (IAA) in MS media accelerates shoot bud organogenesis in differentiating callus tissues with 60-70% conversion of shoot buds into shoot Most efficient shoot organogenesis was recorded when TDZ induced calli were subcultured at different concentrations of 6-benzyla-denine (BA). Optimum shoot bud induction and plant regeneration from callus was achieved when 0.6 ${\mu}M$ (TDZ) induced calli were subcultured at 3.0 ${\mu}M$ (BA) where $16.6{\pm}0.74$ shoots/unit callus on obtained. Rooting in in vitro differentiated shoots was achieved when transferred to medium containing different concentration of indole-3-butyric acid (IBA) in full & half strength MS medium. The well rooted plantlets were hardened and transferred to net house with 90% survival rate.

• Proceedings of the Botanical Society of Korea Conference
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• 1993.07a
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• pp.1-36
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• 1993

It is possible to regenerate plants from calli, single cells and protoplasts of numerous species via organogenasis or embryogenesis in cell and tissue culture systems. Also such regeneration of plants can directly occur from cells of explants. However certain plant species has not been yet provided cultures suitable for plant regeneration from cells or tissues. For example, we have to confirm the regenerability of plant from cells before preparing transformed cells for application. Even more, it is very important to notice that regenerated plants in cell and tissue cultures often show structural abnormality. The mojority of those plants is functionally disordered and eventually cases degenerated. One of such examples is vitreous plants which are manifested mainly in the leaves and manifesteds to a lesser extent in the stems and roots. Regenerants in suspension cultures show more frequent vitrification than on gelled media so that relative humidity and water potential are the key factors involved in abnormal morphogenesis in vitro. The other is that somatic embryos formed in media containing BAP or high concentration of sucrose show frequently cotyledon aberrancy such as polycotyledon and born type cotyledon. The embryos with aberrant cotyledon of Codonopsis lanceolata could not germinate or regenerate into plants in many cases. In contrast, the polycotyledon embryos of Aralia cordata germinated in higher percentage than two cotyledonary embryos, but horn type cotyledonary embryos rarely germinated. The major cause of poor germination is the abnormal development of plumule apex meristem.

• Journal of Plant Biotechnology
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• v.34 no.1
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• pp.69-73
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• 2007

A reliable and effective tissue culture system was established for Perilla frutescens (perilla) using different cultivar, explant source, and growth regulator composition in medium. MS medium supplemented with 2.0 mg/L BA and 0.1 mg/L NAA was maximum at shoot induction. Cotyledon explants formed more shoots than hypocotyl explants. The frequency of plant regeneration through organogenesis was higher (22.8%) than that (6.1%) of somatic embryogenesis. Five genotypes of perilla were screened for the feasibility of shoot regeneration, cotyledon explant of 'Manbaek' showed the highest shoot induction at a frequency of 27.3% among the tested cultivars.

• Biotechnology and Bioprocess Engineering:BBE
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• v.6 no.1
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• pp.51-55
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• 2001

This study was conducted to establish a plant cell culture system for the production of medically important secondary metabolites from Xanthium strumarium. The effects of plant growth regulators including NAA, 2,4-D, kinetin, and ABA were examined in terms of callus induction, maintenance of callus and suspension cultures. It was shown that callus was induced upon treatment with NAA while embryo was induced after treatment with 2,4-D. Callus formation was further improved by treatment with ABA and NAA. The level of callusing increased by 17-29% for the seed case, cotyledon, leaf, and hypocotyl and by 96% in the case of the root. Suspension cell lines were established using calli produced from cotyledon, hypocotyl and root and cultured at 25$\^{C}$ under light conditions. The cells grew up to 15g/L with NAA 2ppm, BA 2ppm, and ABA 1ppm treatment. Supernatants of suspension cultures of cell lines derived from coyledon and hypocotyl produced some distinctive secondary metabolites, one of which was identified as 8-epi-tomentosin, which belongs to the xanthanolides. The amounts of 8-epi-tomentosin produced by the cotyledon- and hypocotylderived cell lines were 13.4mg/L and 11.0mg/L, respectively.

• Journal of Plant Biotechnology
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• v.33 no.4
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• pp.263-270
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• 2006

Pongamia pinnata Pierre is a tree legume, having potential in production of raw material for biodiesel. A protocol for in wk propagation of this plant was standardized using seedling explants. Growth regulators (GR) including gibberellic acid $(GA_3),\;N^6-benzylaminopurine(BA)$, thidiazuron (TDZ), and Adenine sulphate (Ads) were tested for optimum germination of seeds. Removal of seed coat prior to germination, controlled fungal growth partially but enhanced bacterial growth. Antibiotic cefotaxime was ineffective in controlling bacterial contamination. Seedling derived nodal explants and cotyledon nodes with attached cotyledons were excised and cultured for induction of shoots. Optimum sprouting and multiplication of shoot buds were obtained in MS medium supplemented with $8.88{\mu}M$ BA. These buds differentiated and rooted on medium devoid of GR. Optimum growth of Pongamia seedling was obtained in cotton plugged culture vessels. Reculturing of the cotyledon node explants produced more shoots from the same site. This process of removing shoots and reculturing of cotyledon node was followed for eight passages yielding 4 to 8 shoots in each cycle. The shoots (75%) rooted on half strength MS basal medium supplemented with 0.22% charcoal. All plants survived on transfer to soil. This is the first report on in vitro regeneration of Pongamia pinnata. This report demonstrates the possibility of coupling more than one parameter in single experiment to hasten the process of standardization. The process of cycling the nodal explant repeatedly for production of large number of shoots from single meristem may find application in genetic transformation experiments wherein meristems are used for transformation.

• Korean Journal of Plant Tissue Culture
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• v.24 no.6
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• pp.341-344
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• 1997

Calli were induced from the cotyledon and the hypocotyl of alfalfa and the callus lines were tested for the resistance to glyphosate in the liquid medium containing 0.01-3.00 mM glyphosate. Some resistant cell lines were selected from the gradual increase of glyphosate concentration and the lines resistant to 10 mM glyphosate were analyzed with EPSPS activity. Vigorous callus proliferation from the cotyledon and the hypocotyl was observed from the MS medium containing 1 mg/L 2,4-D and 0.5 mg/L kinetin. The hypocotyl was thought to be better explant source for callus induction than the cotyledon. $ID_{50}$ (Inhibition Dosage of 50%) to glyphosate was between 0.1 mM and 0.2 mM level. A49-10G and A58-10G cell lines selected as resistant to 10 mM glyphosate had 8.0 and 9.1 fold increased EPSPS activity to those of the control lines, respectively.

• Journal of Korean Society of Forest Science
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• v.15 no.1
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• pp.43-48
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• 1972

The variation of numbers of resin canal in cotyledon at different growing stages are observed and results are summarized as follows: 1. Resin canals of cotyledon are not found in May, but in September the average number was 3.26. 2. Unlike in later-formed needles, cotyledon resin canals were external and the range of resin canal numbers are 1 to 5. 3. Correlation coefficients between growing stages and resin canal numbers are significant. And correlations between resin canal numbers and leaf thickness, or the number and cross-section area are also highly significant. 4. Low correlation between resin canal and cotyledon number is observed.