• Korean Journal of Plant Resources
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• v.24 no.2
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• pp.139-149
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• 2011

RAPD (random amplified polymorphic DNA) analysis was examined to detect variation of in vitro cultured 30 rhizomes of Cymbidium goeringii Lindley and Cymbidium kanran Makino, with long-term (8 years) subculture, respectively. Out of 151 DNA bands detected, the 40 were polymorphic with a polymorphic rate 26.4% in the C. goeringii. Out of 155 DNA bands detected, the 56 were polymorphic with a polymorphic rate 36.1% in the C. kanran. Genetic similarity matrix (GSM) shows from 0.825 to 1.00 with an average of 0.944 in the rhizomes of C. goeringii and 0.812 to 1.00 with an average of 0.913 in the C. kanran. According to the clustering analysis, C. goeringii was divided into 1 group and 2 independent individuals and its structure of clustering was simple than that of C. kanran. The higher polymorphism and the decreased GSM were showed in the long-term in vitro cultured C. goeringii and C. kanran supplemented with growth regulators. The results provide as fundamental data to develop a new materials for plant breeding and resources plant.

• Korean Journal of Agricultural Science
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• v.12 no.1
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• pp.22-30
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• 1985

Protoplasts of Pisum sativum L. were isolated and cultured from leaf mesophyll tissue. The successful yield of protoplast was obtained in an enzyme solution of 2% 'Onozuka R-10' and 2 % 'Macerozyme R-10' contained 6mM $CaCl_2{\cdot}2H_2O$ within 4 hours. They were divided in B5 culture medium supplemented with 2mg/l kinetin, 1mg/l 2, 4-D and 0.2% Difcobacto agar. Divisions of the protoplasts were continued and led to colony formation for 1 months. The colony from protoplasts of pea mesophyll tissue was formed to callus after subculture in a medium contained macronutrients and amino acids of BII medium and micronutrients and vitamins of B5 medium, and also supplemented with 2mg/l kinetin 2mg/l NAA.

• Journal of Life Science
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• v.25 no.2
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• pp.189-196
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• 2015

Bacterial diversity was studied in the rhizosphere of Suaeda japonica Makino, which is native to Suncheon Bay in South Korea. Soil samples from several sites were diluted serially, and pure isolation was performed by subculture using marine agar and tryptic soy agar media. Genomic DNA was extracted from 29 pure, isolated bacterial strains, after which their 16S rDNA sequences were amplified and analyzed. Phylogenetic analysis was performed to confirm their genetic relationship. The 29 bacterial strains were classified into five groups: phylum Firmicutes (44.8%), Gamma proteobacteria group (27.6%), Alpha proteobacteria group (10.3%), phylum Bacteriodetes (10.3%), and phylum Actinobacteria (6.8%). The most widely distributed genera were Bacillus (phylum Firmicutes), and Marinobacterium, Halomonas, and Vibrio (Gamma proteobacteria group). To confirm the bacterial diversity in rhizospheres of S. japonica, the diversity index was used at the genus level. The results show that bacterial diversity differed at each of the sampling sites. These 29 bacterial strains are thought to play a major role in material cycling at Suncheon Bay, in overcoming the sea/mud flat-specific environmental stress. Furthermore, some strains are assumed to be involved in a positive interaction with the halophyte S. japonica, as rhizospheric flora, with induction of growth promotion and plant defense mechanism.

• Korean Journal of Plant Tissue Culture
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• v.26 no.2
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• pp.103-107
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• 1999

The effects of MS salt strength, sucrose, and cultural conditions on bulblet formation and growth were investigated to optimize the conditions for micropropagating bulblets from in vitro bulb scales of Lilium Oriental Hybrid 'Casa Blanca'. There was no difference on bulblet formation in the range of 1/2~2 $\times$ strength of MS salt, but it was inhibited remarkably in 3 $\times$ strength of MS salt. The growth of regenerated bulblets was most stimulated on MS basal medium. Favorable bulblet formation and its growth from bulb scales were achieved when grown on the media with 1 : 2 or 1 : 3 in the ratio of NH$_4$^+ : NO$_3$^- , as well as on MS basal medium (NH$_4$^+ : NO$_3$^- = about l : 2). Therefore, MS basal medium was very suitable for bulblet formation and growth from bulb scales. Bulblet formation was inhibited but its growth was stimulated with increase sucrose concentration in the medium. The growth of regenerated bulblets was very effective on the media with 9~12% sucrose. Addition of activated charcoal (AC) to the medium inhibited bulblet formation from bulb scales, but enhanced the growth of regenerated bulblets. Especially, the medium containing 1 g/L AC was most effective on the growth of bulblets. No difference was found on bulblet formation and growth from bulb scales under light and dark conditions. In vitro micropropagation of L. Oriental Hybrid 'Casa Blanca' was supposed very reasonable to enhance the growth of the bulblets after forming of bulblets from in vitro bulb scales, and then, subculture the bulb scales from the grown bulblets on MS medium with 9% sucrose and 1 g/L AC.

• Korean Journal of Plant Tissue Culture
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• v.26 no.2
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• pp.93-97
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• 1999

Shoot tips with 2 leaf primordia were cultured to induce shoot primordia in MS liquid medium supplemented with several concentrations of BA and hIAA under the conditions of 10,000 lux illuminations for 24 h and of vertical shaking of 2 rpm in carrot. Two F$_1$ hybrids and two male sterility lines were used. Shoot primordia were only induced in the medium supplemented with 2.0 mg/L of BA and 0.2 mg/L of NAA. Genotypic specificity and seasonal effect of donor parents on shoot primordia induction were not observed and average 15-20% of the planted dornes developed to shoot primordia. The induced shoot primordia were successfully propagated by subculture in the same medium. However, they were grown into three different types during multiplication, that is, the type with multiple small shoots on the surface, the type of without any shoot, and the type of callus. Shoot primordia clusters with small shoots on the surface differentiated multiple shoots successfully in 1/2 MS solid medium supplemented with 0.2 to 1.0 mg/L of IAA and 0.2 to 1.0 mg/L of kinetin. New shoot primordia with small shoots were well formed when pieces bigger than 2 mm in diameter of the out layer of the shoot primordia cluster with small shoots were subcultured. No differences of multiplication and shooting ability and chromosomal variation of shoot primordia were observed until the 13th sub-culture.

• Journal of Embryo Transfer
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• v.28 no.4
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• pp.373-379
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• 2013

Spermatogonial stem cells (SSCs) developed into sperms through spermatogenesis have been utilized as a useful tool in the field of regenerative medicine and infertility. However, a small number of highly qualified SSCs are resided in the seminiferous tubule of testis, resulted in developing effective in-vitro culture system of SSCs for solving simultaneously quantitative and qualitative problems. Presently, SSCs can be enriched on testicular stromal cells (TSCs), but there are no systematic researches about TSC culture. Therefore, we tried to optimize culture condition of TSCs derived from mouse with different strains. For these, proliferation and viability were measured and compared by culturing ICR outbred or DBA/2 inbred mouse-derived TSCs at 35 or $37^{\circ}C$. In case of ICR strain, primary TSCs cultured at $37^{\circ}C$ showed significantly higher proliferation and viability than those at $35^{\circ}C$ and significant increase of proliferation and viability in sub-passaged TSCs was detected in the $35^{\circ}C$ culture condition. Moreover, sub-passage of primary TSCs at $35^{\circ}C$ induced no significant effects on proliferation and viability. In contrast, in case of DBA/2 strain, significantly improved proliferation were detected in the primary TSCs cultured at $35^{\circ}C$, which showed no significant difference in the viability, compared to those at $37^{\circ}C$. Furthermore, sub-passaged TSCs cultured at $37^{\circ}C$ showed no significant differences in proliferation and viability, compared to those at $35^{\circ}C$. However, with significant decrease of proliferation induced by sub-passage of primary TSCs at $35^{\circ}C$, no significant effects on proliferation and viability were resulted from sub-passage of primary TSCs at $37^{\circ}C$. From these results, culture temperature of primary TSCs derived from outbred and inbred strain of mouse could be separately optimized in primary culture and subculture.

• Korean Journal of Weed Science
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• v.15 no.2
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• pp.148-153
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• 1995

Glyphosate (N-[phosphonomethyl]glycine) applied to the assimilate-exporting leaves or sprayed to the whole plants of tomato(Lycopersicon esculentum Mil var. Moneymaker) induced the rapid inhibition of 5-enolpyruvyl skimic acid 3-phosphate synthase(EPSP-synthase). It shows that EPSP-synthase activity precedes chlorophyll loss. There is no difference in EPSP-synthase activity between in vivo tomato meristem and cell suspension culture if glyphosate is not applied. The EPSP-synthase activity is in a range of 4 to 6 nkat per mg protein. The inhibition of EPSP-synthase action is induced within 36 h after glyphosate application while the Chl contents were reduced 48 h after the application. In cell suspension culture of tomato and Corydalis (Corydalis sempervirens), a sublethal concentration of glyphosate retards the fresh weight increase and prolonged lag phase. The fresh weight is reached maximal about 14 days after the subculture in the presence of glyphosate. The inhibitory effect of glyphosate on EPSP-synthase is remarkably induced in lag phase.

• Korean Journal of Plant Tissue Culture
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• v.28 no.2
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• pp.61-67
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• 2001

Using mature seed-derived callus, optimal conditions for efficient callus growth and plant regeneration, and regeneration efficiency by callus type were investigated in zoysiagrass (Zoysia japonica steud.). Callus induction was highest when the seeds were cultured on MS medium containing 2 mg/L 2,4-D, 0.2 mg/L BAP, 4 mg/L thiamine-HCl and 100 mg/L $\alpha$-ketoglutaric acid. Callus growth was highest when callus were cultured on MS medium containing 0.5 mg/L 2,4-D, 0.05 mg/L BAP, 4 mg/L thiamine-HCl and 100 mg/L $\alpha$-ketoglutaric acid. Plant regeneration was highest when callus was transferred on MS medium containing 3% maltose and 1 mg/L BAP, or 1 mg/L thidiazuron (TDZ). The combinations and concentrations of 2,4-D and BAP were shown to be critical factors for both the frequency and the type of callus. And four morphologically distinct types of callus were induced from the 2,4-D and BAP treatment. Type I,II and III calli produced shoots upon subculture, while the watery callus, type IV produced roots without shoots. Of four types of callus, type I exhibited the maximum frequency (82%) of shoot regeneration and the minimum frequency (4%) of albinism.

• FLOWER RESEARCH JOURNAL
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• v.18 no.1
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• pp.9-14
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• 2010

This study was conducted to examine the effects of IAA and kinetin on induction of gemmae and gametophytes in Hypnum plumaeforme moss during tissue culture. Explants were obtained from sterilized gametophyte tips cultured on solid basal medium containing Knop's major salts and Nitsch and Nitsch's trace elements. After culture, inoculated gametophyte tip produced protonema firstly, changed to new gametophytes after four weeks. Aseptic gametophytes were chopped and inoculated on the same media containing 0.01, 0.1, 1 and $10{\mu}M$ of IAA and kinetin. As a result, secondary protonemata as well as protonemal gemmae were formed from gametophytes. But protonemal gemmae formation was varied according to the concentration of IAA and kinetin. Lower concentration of IAA and kinetin promoted gemma formation and bud induction. Especially, $0.01{\mu}M$ of kinetin showed the highest frequency of bud and gemma production. All the materials, obtained from $0.01{\mu}M$ kinetin medium, were subcultured to media supplemented with 0.01, 0.1, 1 and $10{\mu}M$ of IAA and kinetin again to induce gametophytes. After subculture, protonema and calli were developed from secondary protonema, and induced gametophytes finally. IAA regulated induction and growth of gametophytes, and kinetin influenced gemma formation and gametophyte induction also. All aspects of development of this moss species were governed by the external growth regulators.

• Journal of Plant Biotechnology
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• v.50
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• pp.56-62
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• 2023

Transgenic lilies have been obtained using Agrobacterium tumefaciens (AGL1) with the plant scale explants, followed by DL-phosphinothricin (PPT) selection. In this study, scales of lily plants cv. "red flame" were transformed with the pCAMBIA3301 vector containing the gus gene as a reporter and the blpR gene as a selectable marker, as well as a gene of interest showing herbicide tolerance, both driven by the CaMV 35S promoter. Using a 20-minute infection time and a 5-day cultivation period, factors that optimized and demonstrated a high transformation efficiency were achieved. With these conditions, approximately 22-27% efficiency was observed for Agrobacterium-mediated transformation in lilies. After transformation with Agrobacterium, scales of lilies were transferred to MS medium without selective agents for 2 weeks. They were then placed on selection MS medium containing 5 mg/L PPT for a month of further selection and then cultured for another 4-8 weeks with a 4-week subculture regime on the same selection medium. PPT-resistant scales with shoots were successfully rooted and regenerated into plantlets after transferring into hormone-free MS medium. Also, most survived putatively transformed plantlets indicated the presence of the blpR gene by PCR analysis and showed a blue color indicating expression of the gus gene. In conclusion, when 100 scales of lily cv. "red flame" are transformed with Agrobacterium, approximately 22-27 transgenic plantlets can be produced following an optimized protocol. Therefore, this protocol can contribute to the lily breeding program in the future.