• Title/Summary/Keyword: Sterile medium

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Interspecific Hybridization between Populus caspica L. × P. deltoids L 62/154 Using in vitro Embryo Development and Germination

  • Ali, Jafari mofidabadi;Mansooreh, Kamandloo;Hamid, Selamti
    • Journal of Forest and Environmental Science
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    • v.33 no.3
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    • pp.197-201
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    • 2017
  • Populus. caspica L. is an Iranian indigenous poplar species which naturally distributed in the northern part of country. Unfortunately, overuse has removed many of the stems of better form, so that natural stands now usually appear small and crook. Therefore genetic variation for selection of new superior clone of this species is needed. Conventional hybridization system is currently used to induce genetic variation in poplar species but incompatibility barriers have been observed between them. In vitro ovule embryo culture was used to overcome incompatibility obstacle for interspecific hybridization between Populus caspica L. with Populus deltoids L.62/75. Female flowers of Populus caspica L. have artificially been pollinated with pollen grain of P. deltoides 62/75 in one direction using twig and pot crossing system. Ovaries at different ages (7, 14 and 21 days after pollination) were disinfected through 70% ethanol for 1 minute, 5% of sodium-hypochlorite solution for fifteen min followed by three time rising with sterile distil-water. Isolated ovaries were then transferred to MS hormone free medium containing 30 and 60 g/L sucrose for embryo development and germination. Collected data have been analyzed by two factorial experimental designs. The results indicated that there were significant differences between age of embryos for development and germination at ${\alpha}=0.01%$. Highest embryo germination (45%) was observed from 21 days old ovaries. No significant differences were observed between MS culture media containing 30 and 60 g/L for percentages of ovary-embryo germination and number of germinated embryo per ovary at ${\alpha}=0.05%$. Fourteen percentage of embryo germination obtained in MS medium supplemented with 60 g/L sucrose, while only 35% of isolated ovaries were able to germinate in MS containing 30 g/L sucrose. Induced plantlets in 4 cm height were transferred into pots containing soilless (1:1:1 peat, per lit and vermiculite) medium for acclimatization. After successful acclimatization, plants were delivered to nursery.

Optimal Strategy for Ethanol Production in Repeated Fed-batch Operation Using Flocculent Sacchromyces cerevisiae (응집성 Sacchromyces cerevisiae 를 이용한 반복 유가식 ethanol 생산에서의 최적 운전전략)

  • Lee, Sang-Eun;Yeon, Ji-Hyeon;Seo, Yong-Chang;Kang, Do-Hyung;Lee, Hyeon-Yong;Jung, Kyung-Hwan
    • KSBB Journal
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    • v.25 no.2
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    • pp.179-186
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    • 2010
  • We investigated the optimal strategy for ethanol production using flocculent Sacchromyces cerevisiae ATCC 96581. Considering the characteristic of flocculent yeast, a repeated fed-batch ethanol fermentation was designed, in which non-sterile glucose powder was fed every 12 hours and, after cell flocculation, new feeding medium was exchanged every 24 or 36 hours. We particularly compared this fermentation process with those when cell flocculation was not carried out. Finally, the maximal total ethanol production was 825 g-ethanol during 120 hours, in which the time interval of withdrawal-fill of feeding medium was 24 hours and cell flocculation was carried out.

Residual Activity and Effect of Soil Applied Herbicides on Succeeding Crops in Vegetable Field 3. Growth Inhibition of Test Plant According to Concentration Level of Herbicides in Sterile Agar Medium (주요채소용제초제(主要菜蔬用除草劑)의 토양중(土壤中)에서의 잔효(殘效)과 후작물(後作物)에 미치는 영향(影響) 제(第)3보(報) 무균한천배지(無菌寒天培地)에 있어서 제초제(除草劑)의 농도수준(濃度水準)에 따른 검정식물(檢定植物)의 생육저해(生育沮害))

  • Ryang, H.S.;Moon, Y.H.;Choi, E.S.;Jang, M.S.;Lee, J.H.;Chang, Y.N.
    • Korean Journal of Weed Science
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    • v.11 no.2
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    • pp.117-121
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    • 1991
  • This experiment was conducted to identify the relationships of concentration with phytotoxic rate of dinitroamide and acid amide herbicides which have showed the longest residual period in soil. Four herbicides treated showed greater inhibition on roots than shoots, greater inhibition by herbicides was obtained for Italian ryegrass than for Radish. Nitralin, pendimethalin, and ethalfluralin at 0.01ppm gave about 20% inhibition of Italian ryegrass roots, whereas about 47% inhibition was obtained with napropamide. Fifty percent inhibition($I_{50}$) of roots and shoots of Italian ryegrass was 0.036 and 0.132ppm for nitralin. 0.063 and 0.097ppm for pendimethalin. 0.042 and 0.092ppm for ethalfluralin. 0.027 and 0.071ppm for napropamide respectively. On the other hand, $I_{50}$ of roots and shoots of Radish was 1.028 and over 10ppm for nitralin. 1.925 and 4.885ppm for pendimethalin, 2.669 and over 10ppm for ethalfluralin, and 0.515 and over 10ppm for napropamide respectively. There was positive correlation between the concentration of herbicides and growth inhibition of Italian ryegrass and radish.

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Propagation of Bell Pepper (Capsicum annuum var. grossum) through tissue culture

  • Senarath, WTPSK;Shaw, D.S.;Lee, Kui-Jae;Lee, Wang-Hyu
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2003.04a
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    • pp.131-132
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    • 2003
  • Leaf discs and apical meristems were cultured in Murashige and Skoog (MS) medium supplemented with cytokinin and auxin at different concentrations. Callus production was observed in all tested media after six days of incubation. Callus produced in the presence of high concentration of NAA (2.0mg/1) was fragile in texture and yellow in colour. Highest callus formation was observed from leaf discs in the medium supplemented with 1.0mg/1 NAA and 0.5 mg/l BAP in dark at $25{\pm}1{\circ}C$. Percentage of callus formation was 95% and mean callus fresh weight was 654.88 43.53 mg. Shoots were induced from the callus after 4 weeks in 1/2MS medium supplemented with BAP and kinetin both at 0.5mg/1. When elongated shoots were separated and transferred into multiplication medium (MS+0.5mg/1 BAP+0.5mg/1 kinetin) multiplication rate was 6.4 after 6 weeks. Higher concentrations of BAP caused callus production at the base. Direct shoot induction was observed from apical meristems in MS medium in the presence of 0.175 mg/1 IAA + 2.25mg/1 BAP and 0.175 mg/1 IAA + 3.0 mg/1 BAP in 16 hour day at $25{\pm}1{\circ}C$. Explants (apical meristems) elongated to form a single shoot forming a callus at the base. Adventitious buds were sprouted out from the base. Percentage explants which producing shoots was 28.57 and 65.5 respectively. Multiple shoot induction was also observed in the same media. Highest multiple shoot production was observed in the presence of 0.175 mg/l IAA and 3.0mg/l BAP, Mean number of shoots per explant was 5.36 and the mean shoot length was $16.66{\pm}4.15$mm. Shoots (20 30m length) were tested for root induction. Excised shoots were transferred into rooting media, which contains different concentrations of NAA and IAA. Best rooting performance was observed in 1/2MS medium supplemented with 0.1mg/1 NAA after 10 days of incubation in 16 hr photoperiod at $25{\pm}1{\circ}C$. Mean number of roots per shoot was 6 and the mean root length was 252mm. Rooted plantlets were transferred into sterile coir dust:sand (1:4) mixture and maintained in a humid chamber for two weeks, They were gradually exposed to the natural environment. After three weeks they were transferred to pots containing coir dust:sand (1:2) mixture for further development where the 90% survival was observed.

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Light Influences, Morphogenesis and Protein Content on Callus Differentiation of Cucumber (Cucumis sativus L.) (오이 캘러스 분화시 광의 영향, 형태발생 및 단백질함량)

  • 이은모;조만현;송남현;우인식;이영복;곽상수
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.3
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    • pp.213-217
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    • 2000
  • To investigate the cucumber regeneration from embryogenic calli, shoot tips of aseptically-grown cucumber seedlings were used as explants for establishing tissue cultures. Growth and differentiation of callus were studied by using Murashige and Skoog's (MS) medium containing 0.5 to 2 mg/L 2,4-D. Plantlets were induced from shoot tip culture on the plant growth regulators-free MS medium. Non-embryogenic calli and viscous calli were induced on the medium supplemented with 0.5 to 2 mg/L 2,4-D, but embryogenic callus was not induced on the same medium. Segments (ca. 5∼10 mm) of aseptically-grown hypocotyl from five to seven days old seedlings after germination were placed on MS medium supplemented with 1 mg/L 2,4-D for 50 days. Embryogenic calli and embryoids were induced only from the seedlings grown in dark condition, and hypocotyl was placed on the media explanted in light condition. Foully-five point one percent of white fragile calli and 0.6% yellowish compact calli formed roots. Yellowish callus lines were investigated to have a considerably higher concentration of crude proteins than white callus lines. Plantlets derived from embryogenic calli or embryoids have been transferred to pots containing sterile vermiculite and perlite. Normal fruits were harvested from nutrient culture on aggregated hydroponics in the F-clean house.

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High frequency somatic embryogenesis and plant regeneration of interspecific ginseng hybrid between Panax ginseng and Panax quinquefolius

  • Kim, Jong Youn;Adhikari, Prakash Babu;Ahn, Chang Ho;Kim, Dong Hwi;Kim, Young Chang;Han, Jung Yeon;Kondeti, Subramanyam;Choi, Yong Eui
    • Journal of Ginseng Research
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    • v.43 no.1
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    • pp.38-48
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    • 2019
  • Background: Interspecific ginseng hybrid, Panax ginseng ${\times}$ Panax quenquifolius (Pgq) has vigorous growth and produces larger roots than its parents. However, F1 progenies are complete male sterile. Plant tissue culture technology can circumvent the issue and propagate the hybrid. Methods: Murashige and Skoog (MS) medium with different concentrations (0, 2, 4, and 6 mg/L) of 2,4-dichlorophenoxyacetic acid (2,4-D) was used for callus induction and somatic embryogenesis (SE). The embryos, after culturing on $GA_3$ supplemented medium, were transferred to hormone free 1/2 Schenk and Hildebrandt (SH) medium. The developed taproots with dormant buds were treated with $GA_3$ to break the bud dormancy, and transferred to soil. Hybrid Pgq plants were verified by random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) analyses and by LC-IT-TOF-MS. Results: We conducted a comparative study of somatic embryogenesis (SE) in Pgq and its parents, and attempted to establish the soil transfer of in vitro propagated Pgq tap roots. The Pgq explants showed higher rate of embryogenesis (~56% at 2 mg/L 2,4-D concentration) as well as higher number of embryos per explants (~7 at the same 2,4-D concentration) compared to its either parents. The germinated embryos, after culturing on $GA_3$ supplemented medium, were transferred to hormone free 1/2 SH medium to support the continued growth and kept until nutrient depletion induced senescence (NuDIS) of leaf defoliation occurred (4 months). By that time, thickened tap roots with well-developed lateral roots and dormant buds were obtained. All Pgq tap roots pretreated with 20 mg/L $GA_3$ for at least a week produced new shoots after soil transfer. We selected the discriminatory RAPD and ISSR markers to find the interspecific ginseng hybrid among its parents. The $F_1$ hybrid (Pgq) contained species specific 2 ginsenosides (ginsenoside Rf in P. ginseng and pseudoginsenosides $F_{11}$ in P. quinquefolius), and higher amount of other ginsenosides than its parents. Conclusion: Micropropagation of interspecific hybrid ginseng can give an opportunity for continuous production of plants.

An Efficient in vitro Micropropagation for Production of Disease-free Bulbs in Korean Native Lilium

  • Song, Jae-Young;Yi, Jung-Yoon;Yoon, Mun-Sup;Lee, Jung-Ro;Lee, Young-Yi
    • Korean Journal of Plant Resources
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    • v.32 no.6
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    • pp.730-734
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    • 2019
  • Cryopreservation is one of the ideal and suitable methods for long-term storage of plant germplasm. The plant contaminated with diseases and pathogens are decreased the multiplication rate, survival rate and high quality of plants after cryopreservation. The aim of this work was to improve a micropropagation method for lily in Korea, which is indigenous plant. In the last process of rinsing scales after surface-sterilization, we tried to control the diseases and pathogens lived within the tissue by rinsing in 0.03% sodium hypochlorite (NaClO) instead of sterile distilled water. Bulb scales of Lilium were cultured in vitro on MS medium supplemented with Plant Preservative Mixture (PPM). The results showed that L. tsingtauense accessions were observed ranged from 53.9 to 100% with a mean value of 76.8% and L. hansonii accessions were checked from 84.5 to 85.5% with a mean of 85% survival rate. The newly small bulb formed from bulb-scales was transferred to MS medium. We checked the presence of microorganisms and survival rate after 3 weeks in culture after examination of bacterial incidences. The results indicated that the non-contamination rate were shown ranged from 75.0 to 94.1% with mean value of 83.2% in L. tsingtauense species, and that L. hansonii were observed 85.1 to 91.7% with mean value of 88.4%. This study will provide a valuable basis for establishment of effective axenic cultures for in vitro micropropagation of Korean native lily species.

Enhanced Lovastatin Production by Solid State Fermentation of Monascus ruber

  • Xu Bao-Jun;Wang Qi-Jun;Jia Xiao-Qin;Sung Chang-Keun
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.1
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    • pp.78-84
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    • 2005
  • The purpose of this study was to optimize the solid state cultivation of Monascus ruber on sterile rice. A single-level-multiple-factor and a single-factor-multiple-level experimental design were employed to determine the optimal medium constituents and to optimize carbon and nitrogen source concentrations for lovastatin production. Simultaneous quantitative analyses of the ${\beta}$-hydroxyacid form and ${\beta}$-hydroxylactone for of lovastatin were performed by the high performance liquid chromatography (HPLC) method with a UV photodiode-array (PDA) detector. The total lovastatin yield ($4{\sim}6\;mg/g$, average of five repeats) was achieved by adding soybean powder, glycerol, sodium nitrate, and acetic acid at optimized levels after 14 days of fermentation. The maximal yield of lovastatin under the optimal composition of the medium increased by almost 2 times the yield observed prior to optimization. The experimental results also indicated that the ${\beta}$-hydroxylactone form of lovastatin (LFL) and the ${\beta}$-hydroxyacid form of lovastatin (AFL) simultaneously existed in solid state cultures of Monascus ruber. while the latter was the dominant form in the middle-late stage of continued fermentation. These results indicate that optimized culture conditions can be used for industrial production of lovastatin to obtain high yields.

Hybridization and Use Of Grapes as an Oviposition Substrate Improves the Adaptation of Olive Fly Bactrocera oleae (Rossi) (Diptera: Tephritidae) to Artificial Rearing Conditions

  • Sohel, Ahmad;Viwat, Wornoayporn;Polychronis, Rempoulakis;Emily A., Fontenot;Ul Haq, Ihsan;Carlos, Caceres;Hannes F., Paulus;Marc J.B., Vreysen
    • International Journal of Industrial Entomology and Biomaterials
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    • v.29 no.2
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    • pp.198-206
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    • 2014
  • The olive fly Bactrocera oleae (Rossi) is the key pest for olive cultivation worldwide. Substantial effort has been invested in the development of the sterile insect technique (SIT) to control this pest. One of the limitations to develop SIT technology for olive fruit fly is the low ability of wild females to lay eggs in other medium than olive fruits, and their slow adaptation to oviposition in artificial substrates. In the present study, fruit grapes were used as an alternative egg collection medium to harvest eggs and young larvae from freshly colonized wild strains originating from France, Italy, Spain and Croatia. The larvae were allowed to develop into the fruits until the second instar, before they were extracted out and further reared on a standard artificial diet. Furthermore, F1 to F4 female flies were alternatively offered wax bottles to oviposit. Finally, the performance of hybrid strains created from crosses between wild and long colonised flies was assessed. The results showed that females of all 4 wild strains readily oviposited eggs in grapes and from the F2 generation onward, females from all strains were adapted to laying eggs in wax bottles. No difference was observed in eggs and pupae production among all strains tested. The findings are discussed for their implications on SIT application against olive fruit fly.

Development of Preservation Prediction Chart for Long Term Storage of Fermented Cucumber (발효오이의 산패예견표의 개발)

  • Kim, Jae-Ho;Breidt, Fred
    • Journal of Life Science
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    • v.17 no.12
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    • pp.1616-1621
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    • 2007
  • Off-flavors and odors in fermented cucumbers result from the growth of undesirable microorganisms during the secondary fermentation. Under laboratory conditions using a sterile fermented cucumber slurry medium, the spoilage fermentations were reproduced. Using this system the salt and pH conditions that allow the spoilage to occur were determined by varying the NaCl concentration and pH of the slurry medium. At pH 3, no spoilage was observed, regardless of the salt concentration, while at pH 3.5, pH 4, and pH 4.5, spoilage occurred in the 0 and 2% NaCl samples. For pH 5.0 samples, spoilage products were seen for all NaCl treatments. Based on these results the Preservation Prediction Chart was developed. The Chart may be used for selection of proper pH value and salt concentration for long term storage of fermented cucumber.