• Proceedings of the Plant Resources Society of Korea Conference
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• 2002.11a
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• pp.29-30
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• 2002

The goal of this research was to develop the effectiveness of in vitro culture method for A. formosanus and study the environment in vitro conditions affecting on growth. The first series of experiments were examined to investigate the response of three different basal media, MS (Murashige and Skoog, 1962), Knudson (KC; Knudson, 1946) and modified hyponex on growth and multiplication during in vitro culture. Multiple shoot proliferation was induced in shoot tip explants on Hyponex (H3) media supplemented with BA (1 mg1$\^$-1/) or TDZ (1-2 mg1$\^$-1/). Addition of activated charcoal (1%) to the TDZ containing medium promoted rapid shoot tip proliferation (11.1 shoots per explant) but the same medium had an opposite effect resulting in poor proliferation in the nodal explants. However, the regenerated shoots had slow growth rate and failed to elongate. This problem was overcome by transferring the shoot clumps to a hormone free H3 media supplemented with 2% sucrose and 0.5% activated charcoal. Using bioreactor culture for scaling up was also shown the best way for multiple shoot induction and growth of this plant. The second series of experiments was studied to investigate the effect of physical environment factors on growth of in vitro plantlets. The Anoectochilus formosanus plantlets were cultured under different air exchange rate (0.1, 0.9, 1.2h$\^$-1/), without sucrose or supplement 20g.1$\^$-1/ (photoautotrophic or photomixotrophic, respectively), and different photosynthesis photon flux (40, 80, 120 ,${\mu}$mol.m$^2$.s$\^$-1/- PPF). Under non-enrichment CO$_2$ treatment, slow growth was observed in photoautotrophical condition as compared with photomixotrophical condition on shoot height, fresh weigh and dry weight parameters; High air exchange (1.2.h-l) was found to be inadequate for plant growth in photomixotrophical condition. On the contrary, under CO$_2$, enrichment treatment, the plant growth parameters were sharply (visibly) improved on photoautotrophic treatments, especially on the treatment with air exchange rate of 0.9.h-1. The growth of plant in photoautotrophic condition was not inferior compared with photomixotrophic, and the best growth of plantlet was observed in treatment with low air exchange rate (0.9.h-1). Raising the PPF level from 80 to 120${\mu}$mol.m$\^$-2/.s$\^$-1/ decreased the plant height, particularly at 120${\mu}$mol.m$\^$-2/.s$\^$-1/ in photoautotrophic condition, fresh weight and dry weight declined noticeably. At the PPF of 120${\mu}$mol.m$\^$-2/,s$\^$-1/, chlorophyll contents lowed compared to those grown under low PPF but time courses of net photosynthesis rate was decreased noticeably. Light quality mainly affected morphological variables, changes of light quality also positively affected biomass production via changes in leaf area, stem elongation, chlorophyll content. Plant biomass was reduced when A. formosanus were grown under red LEDs in the absence of blue wavelengths compare to plants grown under supplemental blue light or under fluorescent light. Stem elongation was observed under red and blue light in the present experiment. Smaller leaf area has found under blue light than with other lighting treatments. Chlorophyll degradation was more pronounced in red and blue light compared with white light or red plus blue light which consequent affected the photosynthetic capacity of the plant. The third series of experiment were studied to investigate the effect of physical environment factors on growth of ex vitro plants including photosynthesis photon flux (PPF), light quality, growing substrates, electrical conductivity (EC) and humidity conditions. In the present experiments, response of plant on PPF and light quality was similar in vitro plants under photosynthesis photon flux 40${\mu}$mol.m,$\^$-2/.s$\^$-1/ and white light or blue plus red lights were the best growth. Substrates testing results were indicated cocopeat or peat moss were good substrates for A. formosanus growth under the greenhouse conditions. In case of A. formosanus plants, EC is generally maintained in the range 0.7 to 1.5 dS.m-1 was shown best results in growth of this plant. Keeping high humidity over 70% under low radiation enhanced growth rate and mass production.

• Horticultural Science & Technology
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• v.30 no.6
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• pp.650-656
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• 2012

The aims of the experiment, conducted over three years in the Central Europe field conditions, were (1) to investigate the effect of growing period (plantings in the middle and at the end of August: $1^{st}$ and $2^{nd}$ term, respectively) on yield and chemical composition of the non-heading Chinese cabbage (Brassica rapa var. chinensis) cultivars 'Taisai', 'Pak Choy White', and 'Green Fortune', and (2) to develop regression models to evaluate the changes in crop yields as a function of weather conditions. A highest marketable yield was obtained from 'Taisai' (65.71 and 77.20 $t{\cdot}ha^{-1}$), especially in the $2^{nd}$ term of production. Low yield, observed for 'Pak Choy White' was due to its premature bolting. Almost 39% ($1^{st}$ term) and 70% ($2^{nd}$ term) of plants of this cultivar formed inflorescence shoots before harvest. The highest dry matter level was observed in the leaf petioles of 'Taisai', while 'Green Fortune' was the most abundant of carotenoids and L-ascorbic acid. The content of soluble sugars was the lowest for 'Pak Choy White'. In a phase of harvest maturity, more of the analyzed constituents were gathered by plants from earlier plantings, and differences were as follows: 4.7% (dry matter), 26.3% (carotenoids) and 22.1% (L-ascorbic acid), in comparison to $2^{nd}$ term of production. Significant increase of soluble sugars level was observed for plants from later harvest. The regression model for marketable yield of Chinese cabbage cultivar 'Taisai' as a function of maximum air temperature can predict the yield with accuracy 68%. The models for yield or bolting of 'Pak Choy White', based on extreme air temperatures and sunshine duration, were more precise (98%). It should be pointed out that Taisai could be recommended for later growing period in Central Europe conditions with regard to maximum yield potential. 'Green Fortune' was notable for its uniform yielding. To obtained plants of higher nutritional value, earlier time of cultivation should be suggested. Described models can be successfully applied for an approximate simulation of Chinese cabbage yielding.

• Korean Journal of Plant Resources
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• v.16 no.2
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• pp.160-167
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• 2003

In order to investigate the effects of developmental stage of embryos and plant growth regulators on mass production of Zantedeschia spp. Southern Light, immature zygotic embryos of Zantedeschia spp. Southern Light were cultured on Murashige and Skoog(1962) basal media or containing 2,4-D, NAA and BA. Globular embryos did not grow on any of the 2,4-D, NAA and BA combinations. The most suitable stage of immature zygotic embryo culture on the induction callus and multiple shoot was at early cotyledonary embryo stage, and at this stage of embryos were germinated up to 87.5%. The whitish watery callus and yellowish compact nodular callus produced on all 2,4-D, NAA and BA media. The best combination for inducing embryogenic callus was 0.5 mgL NAA and 1.0 mg/L BA. Whitish watery calli have been subcultured for more than 8 months and have retained their producing ability, Plant regeneration was only obtained by direct shoot development and yellowish compact nodular calli. Abundant plantlets were regenerated from cotyledonary stage of embryo culture on MS medium supplemented with 0.5 mg/L NAA and 1.0 mg/L BA. Supplementation of the media with 10％ coconut water showed as the best concentration for plant differentiation from direct developed of shoots. The number of regenerated plants from one embryo could be seperated 25-35s plantlets. All yellowish compact callus-derived plantlets were transferred to pots containing a mixture of vermiculite, perlite and sand(1:1;1 v/v) and 100% of divided plantlets were phenotypically normal.

• Korean Journal of Medicinal Crop Science
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• v.3 no.3
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• pp.207-216
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• 1995

This study was conducted to establish technologies for plug seedling production using somatic embryos-derived regenerants and their seeds in Bupleurum falcatum L. Among distilled water, GA (0.1mg/l) and putrescine (0.lmg/l) treated to regenerants for acclimatization, GA was most effective to develop shoots and roots, 1/2X MS medium and NAA 0.1mg/l + BA 0. 5mg/l enhanced the growth rates of the regenerants and increased dry weight. Activated charcoal effected to grow markedly leaves and roots of the regenerants at the level of 0.4 %. Regenerants increased their plant height, root length and dry weight at $30^{\circ}C$. Plug seedlings originated from seeds of the tissue culture regenerants showed the maxium growth on the mixture of peatmoss soil (2) and mountain sand (1) .Root length, leaf area and dry weight of plug seedlings increased significantly when No.1, 2 and 3 of Wondergrow solution were mixed in the ratio of 1.3 - 0.9 - 0.1. Light supplement (4%) and high tem­perature $(30^{\circ}C)$ promoted the growth of plug seedlings as well as dry weight. Ninety days seedlings were more vigorous and adaptable for transplanting than other seedlings.

• Korean Journal of Medicinal Crop Science
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• v.3 no.2
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• pp.100-106
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• 1995

This study was conducted to find the factors affecting somatic embryogenesis in suspension culture of Rehmania glutinosa and investigate the possibility of artificial seed production by encapsulation of somatic embryos. Linsmeier-Skoog medium was appeared as proper for somatic embryogenesis. Sucrose with $3{\sim}5%$ as carbon sources was good for somatic embryogenesis, and both ammonium and nitrate nitrogen were necesary for normal somatic embryo production. BA with NAA or kinetin with NAA were better than the use of cytokinin alone for both somatic embryogenesis and numbers of somatic embryos. $AgNO_3$ as protectant for vitrification of seedlings in vitro culture had no harmful effect on somatic embryos. Sphericity of encapsulated seeds was good at 3% gel of sodium alginate but germination was better at 2.5% sodium alginate level. Artificial seeds were germinated and developed normal shoots and roots under in vitro condition.

• Journal of Microbiology and Biotechnology
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• v.30 no.5
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• pp.668-680
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• 2020

Bacillus velezensis is an important plant growth-promoting rhizobacterium with immense potential in agriculture development. In the present study, Bacillus velezensis Lle-9 was isolated from the bulbs of Lilium leucanthum. The isolated strain showed antifungal activities against plant pathogens like Botryosphaeria dothidea, Fusarium oxysporum, Botrytis cinerea and Fusarium fujikuroi. The highest percentage of growth inhibition i.e., 68.56±2.35% was observed against Fusarium oxysporum followed by 63.12 ± 2.83%, 61.67 ± 3.39% and 55.82 ± 2.76% against Botrytis cinerea, Botryosphaeria dothidea, and Fusarium fujikuroi, respectively. The ethyl acetate fraction revealed a number of bioactive compounds and several were identified as antimicrobial agents such as diketopiperazines, cyclo-peptides, linear peptides, latrunculin A, 5α-hydroxy-6-ketocholesterol, (R)-S-lactoylglutathione, triamterene, rubiadin, moxifloxacin, 9-hydroxy-5Z,7E,11Z,14Z-eicosatetraenoic acid, D-erythro-C18-Sphingosine, citrinin, and 2-arachidonoyllysophosphatidylcholine. The presence of these antimicrobial compounds in the bacterial culture might have contributed to the antifungal activities of the isolated B. velezensis Lle-9. The strain showed plant growth-promoting traits such as production of organic acids, ACC deaminase, indole-3-acetic acid (IAA), siderophores, and nitrogen fixation and phosphate solubilization. IAA production was accelerated with application of exogenous tryptophan concentrations in the medium. Further, the lily plants upon inoculation with Lle-9 exhibited improved vegetative growth, more flowering shoots and longer roots than control plants under greenhouse condition. The isolated B. velezensis strain Lle-9 possessed broad-spectrum antifungal activities and multiple plant growth-promoting traits and thus may play an important role in promoting sustainable agriculture. This strain could be developed and applied in field experiments in order to promote plant growth and control disease pathogens.

• Journal of Plant Biotechnology
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• v.29 no.1
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• pp.63-68
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• 2002

The possibility for mass production of phytol, inhibitory diterpene against ACAT (Acyl-CoA: Cholesterol acyltransferase) was investigated by using callus culture of lettuce. The callus were induced from lettuce cotyledon explants on MS medium containing 0.5 mg.L$^{-1}$ NAA after 4 week's culture. Adventitious roots were formed from the explants on MS medium containing 0.5 mg.L$^{-1}$ IBA or 1.0 mg.L$^{-1}$ NAA. Adventitious shoots and roots were emerged from the callus when the callus was transferred to MS medium containing auxin alone, or with cytokinin. The plant growth regulators and their concentrations for the organogenesis were 1.0 mg.L$^{-1}$ NAA, 0.1 mg.L$^{-1}$ BA, 0.5 mg.L$^{-1}$ NAA with 0.1 mg.L$^{-1}$ kinetin, or 0.5 g.L$^{-1}$ 2.4-D with 1.0 mg.L$^{-1}$ kinetin. Analyses of chlorophyll contents showed that chlorophyll contents were higher in morphogenic calli than in non-morphogenic calli. However, the chemical analyses of gas chromatography indicated that phytol contents were not proportionate to the chlorophyll contents of callus. The content of phytol was higher in callus than in lettuce cotyledon.ledon.

• Journal of Plant Biotechnology
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• v.47 no.1
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• pp.66-72
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• 2020

Alstroemeria plants were transformed by using an improved particle-gun-mediated transformation system. Friable embryogenic callus (FEC) induced from the leaves with axil tissues of Alstroemeria plant was used as the target tissue. Also, FEC was transformed with the bar gene was used as a selectable marker. In the case of plasmid pAHC25, 7.5% of the twice-bombarded FEC clumps showed blue foci, whereas the clumps with single bombardment showed only 2.3%. Additionally, a 90° rotation with double bombardment led to a higher frequency (6 times) of luciferase gene expression in PBL9780 than the control treatment. After 8 weeks of bombardment, more than 60 independent transgenic lines were obtained for pAHC25 and nearly 150 independent transgenic lines were obtained for PBL9780, all of which were resistant to PPT and demonstrated either GUS or luciferase activity. Regarding effect of osmotic treatment (0.2 M mannitol) with 7 different periods, the highest transient gene expression was obtained in 8 h before and 16 h after transformation in both pAHC25 and PBL9780. Compared with the control, at least three times more GUS foci and photons were observed in this treatment. With respect to different combinations of mannitol and sorbitol with 8 h before and 16 h after transformation, high numbers of transient and stable transgene expressions were observed in both 0.2 M mannitol and 0.2 M sorbitol used in the osmotic pre-culture. This combination showed the highest transformation efficiency in both pAHC25 (8.5%) and PBL9780 (14.5%). In the control treatment, only 10% of the FEC clumps produced somatic embryos. However, by using 0.2 M mannitol and 0.2 M sorbitol, the frequency of somatic embryos increased to 36.5% (pAHC25) and 22.9% (PBL9780). Of the somatic embryos produced, at least 60% germinated. Approximately 100 somatic embryos from these 210 independent transgenic lines from 2 plasmids developed into shoots, which were then transferred to the greenhouse. PCR analysis confirmed the presence of the bar gene. This is the report on the production of transgenic Alstroemeria plants by using particle bombardment with a high efficiency, thereby providing a new alternative for the transferring of gene of interests in Alstroemeria in the breeding program in the future.

• FLOWER RESEARCH JOURNAL
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• v.17 no.1
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• pp.23-28
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• 2009

This study was conducted to examine the effect of plant growth regulators on the formation of adventitious bud callus(ABC) and plant regeneration in shoot tip culture of Zantedeschia spp. 'Florex Gold'. Treatment of $0.1mgL^{-1}\;N$-phenyl-N'-1,2,3-thiadiazol-5-ylurea(thidiazuron, TDZ) was more promotive for formation of ABC in shoot tip culture than 6-benzylaminopurine(BA) treatment, and short shoots were developed. Comparing to treatment of BA, mixing treatment of BA and 1-naphthaleneacetic acid(NAA) inhibited the formation of ABC and multiple shoot. The proliferation of ABC derived from sections(0.3 cm) of ABC produced by shoot tip culture in medium supplemented with $0.1mgL^{-1}\;TDZ$ was more effective in medium with $0.1mgL^{-1}\;TDZ$ or $2.0mgL^{-1}\;BA$ than the other treatments. The shoot regeneration and growth from sections of ABC was more promotive in treatment of $0.001mgL^{-1}\;TDZ$. Also, the root growth from sections of ABC was better in medium with $0.001mgL^{-1}\;TDZ$ or $0.2mgL^{-1}\;BA$. Consequently, in vitro mass production of Zantedeschia spp. 'Florex Gold' can be obtained via indirect organogenesis through plant regeneration and proliferation of ABC which was derived from shoot tip culture at $0.1mgL^{-1}\;TDZ$.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2021.04a
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• pp.24-24
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• 2021

The Cassava (Manihot esculenta Crantz) is one of the major food crops in the tropical or subtropical regions. Recently, clean planting materials of improved cassava cultivars are in high demand. Problems in the propagation of cassava are virus vulnerable and low rates of seed germination. Thus, the study was undertaken to develop an efficient in vitro mass propagation protocol of Manihot esculenta Crantz. So we tried to optimize protocols for mass production from axillary buds of Cassava. Young and actively growing stem segments were excised from adult plants of cassava. Samples were cut into a 3~4 cm nodal segments with axillary buds, and cultivated in the different medium supplemented with various plant growth regulators for 4 weeks. For shoot multiplication, axillary buds approximately 1 cm in length were taken from in vitro derived shoots and subcultured. After 4~6 weeks, the shoot generation rate showed 55.6%. The shoot number and its length was 1.0/explant and 2.3 cm in the most favorable medium composition. The auxin β-indolebutyric acid(IBA) 0~2.0 mg/L was proved to be effective on root development. Plantlets with fibrous roots easily generated tuberous roots in vitro. The tuberous roots were induced only when both kinetin and IBA were used in combination. after 8 weeks, the root generation rate showed 100%. The root number and its length was 17.2/explant and 2.2 cm in the most promising medium composition. Our experiments confirmed that in vitro growth and multiplication of plantlets could depend on its reaction to the different medium composition, and this micropropagation techniques could be a useful system for healthy and vigorous plant production.