• Title/Summary/Keyword: Regeneration efficiency

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Optimization of Regeneration Condition Under Agrobacterium-mediated Transformation in in vitro Cultured Korean Soybean

  • Kantayos, Vipada;Bae, Chang-Hyu
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.36-36
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    • 2018
  • Soybean is the important crop in Asian countries as protein source, oil production and animal feed. Improving soybean using genetic transformation is the principal tool in nowadays. Developing herbicide resistant transgenic soybean plants through Agrobacterium-mediated transformation has been worked in many previous studied. However, the transformation efficiency is still low. Many attempts try to find the optimum media condition for plant regeneration after infection. After transformation, the plant regeneration is very important condition to promote growth of transgenic plant. In this study, we optimized a regeneration condition for two Korean soybean cultivar, Dawonkong and Pungsannamulkong using cotyledon, cotyledonary nodes and hypocotyl as explant. The results showed that shoot regeneration of cotyledonary nodes on B5 medium containing 2 mg/L 6-benzylaminopurine showed the highest percentage of regeneration in Dawonkong (75.8%) while Pungsannamulkong presented high number of shoots 2.12 shoots per explant. For transformation condition, co-cultivation in 7 days showed a high number of GUS positive expression. Most of explants can survived under media including 5 mg/L of glufocinate which refers phosphinotricin for 2-week selection. Washing with 400 mg/L of cefotaxime in several times and selection in plant regeneration media with 400 mg/L of cefotaxime can prevent bacteria growth, effectively.

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Effects of Hormone and Several Factors on the Regeneration and Transformation rate of Potato Cultivars Bred in Korea (국내 육성 감자의 재분화와 형질전환 효율에 미치는 생장조절제의 조성 및 몇 가지 요소)

  • Yi, Jung-Yoon;Seo, Hyo-Won;Cho, Ji-Hong;Lee, Shin-Woo;Yun, Han-Dae
    • Journal of Plant Biotechnology
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    • v.30 no.1
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    • pp.27-33
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    • 2003
  • The optimal condition of in vitro regeneration and transformation were investigated for newly bred potato varieties in Korea. Leaf and internodal stem tissues of 'Chubak', 'Namsuh', 'Jasim', 'Jopung' and 'Jowon' were used to investigate the influence of growth regulator on regeneration efficiency The effect of phenolic compound acetosyringone on gene transformation efficiency was also investigated. Leaf tissue of 'Jowon' and internodal stem tissues of 'Jopung' were showed high regeneration efficiency on M5 medium supplemented with 0.1 mg/L GA₃, 2.0 mg/L Zeatin and 0.01 mg/L NAA. The other three potato cultivars were showed low regeneration efficiency less than 25%. The effect of acetosyringone on Agrobacterium-mediated gene transformation with leaf and internodal stem tissues were noticeable. By adding the 75 μM of acetosyringone during the Agrobacterium innoculation, the transformation efficiency was increased up to 1.5∼4.0 fold compare to non-treated control. In case of 'Jowon' the transformation efficiency was 87.9% in leaf tissue and 'Jopung' was 68.4% in internodal stem tissues.

De novo Regeneration of Fertile Common Bean (Phaseolus vulgaris L.) Plants

  • Albino Margareth M.C.;Vianna Giovanni R.;Falcao Rosana;Aragao Francisco J.L.
    • Journal of Plant Biotechnology
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    • v.7 no.4
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    • pp.267-272
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    • 2005
  • Common bean (Phaseolus vulgaris L.) plants were regenerated via organogenesis from mature embryonic axes, cultured on MS medium supplemented with ildole-3-ecetic acid (IAA) and thidiazuron (TDZ) for one week in the dark. Embryonic axillary regions were excised, longitudinally cut to split the both sides, and cultured for two weeks on MS medium supplemented with IAA and TDZ. The combination 0.5 mg $l^{-1}$ TDZ/0.5 mg $l^{-1}$ IAA presented the higher efficiency in shoot regeneration and the combination 0.5 mg $l^{-1}$ TDZ/0.25 mg $l^{-1}$ IAA presented the higher efficiency in conversion of shoots to plants. Regenerating explants were transferred to MS medium containing 1 mg $l^{-1}$ BAP for shoot development. All elongated shoots were rooted in vitro, presented normal phenotype and produced viable seeds. Histological analysis confirmed the mode of regeneration as de novo shoot organogenesis.

Basic Static Characteristics of a Closed and a Regeneration Cycles for the OTEC System (해양온도차발전 Closed and Regeneration Cycle의 기본 정특성)

  • Cha, Sang-Won;Kim, You-Taek;Mo, Jang-Oh;Lim, Tae-Woo;Lee, Young-Ho
    • Journal of Advanced Marine Engineering and Technology
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    • v.36 no.8
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    • pp.1151-1157
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    • 2012
  • Ocean Thermal Energy Conversion(OTEC) technology is one of the new and renewable energy that utilizes the natural temperature gradient that exists in the tropical ocean between warm surface water and the deep cold water, to generate electricity. The selection of working fluid and the OTEC cycle greatly influence the effect on the system operation, and it's energy efficiency and impacts on the environment. Working fluids of the OTEC are ammonia, R22, R407C, and R410A. In this paper, we compared boiling pressure to optimize OTEC system at $25^{\circ}C$. Also, this paper showed net-power and efficiency according to working fluids for closed cycle and regeneration cycle.

The Effect of agrobacterium Density on Transformation Efficiency in Apple (Agrobacterium 농도가 사과 형질전화 효율에 미치는 영향)

  • Seong, Eun-Soo;Cha, Ji-Eun;Kim, Jeong-Hee;Park, Seong-Whan;Yu, Chang-Yeon;Song, Kwan-Jeong
    • Journal of Plant Biotechnology
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    • v.30 no.3
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    • pp.215-219
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    • 2003
  • This study was conducted to find optimum bacterial density for improving the efficiency of transformation mediated by Agrobacterium tumefaciens in apples. Regeneration(15%) and transformation frequency(10%) were increased in resuspension-culture density $A_{600}$ 1.3 from preculture density $A_{600}$ 0.7 of Agrobacterium tumefaciens in ′Fuji′. In ′Gala′, 20% regeneration and 16% transformation frequency were observed at optimum bacterial density $A_{600}$ 0.7 form preculture density $A_{600}$ 1.3. ′Mclntosh as well as "Gala" were 25%regeneration and 10% transformation frequency. Hence a frequency optimum condition of bacterial density for the efficient transformation of apple could be depend on apple genotypes.

Isolation, Regeneration and PEG-Induced Fusion of Protoplasts of Pleurotus pul-monarius and Pleurotus florida

  • Eyini, M.;Rajkumar, K.;Balaji, P.
    • Mycobiology
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    • v.34 no.2
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    • pp.73-78
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    • 2006
  • Inter-specific hybridization between Pleurotus pulmonarius and P. florida was attempted through PEG-induced protoplast fusion to select a fusant. The protocol for protoplast release, regeneration and fusion in these two Pleurotus species was standardized using the variables controlling the process. The mixture of mycolytic enzymes, i.e. commercial cellulase, crude chitinase and pectinase, KCl (0.6 M) as osmotic stabilizer, pH 6 of the phosphate buffer and an incubation time of 3 hours resulted in the maximum release of protoplasts from 3-day-old mycelia of P. florida ($5.3{\sim}5.75{\times}10^{7}$ protoplasts/g) and P. pulmonarius ($5.6{\sim}6{\times}10^{7}$ protoplasts/g). The isolated protoplasts of P. florida regenerated mycelium with 3.3% regeneration efficiency while P. pulmonarius showed 4.1% efficiency of regeneration. Polyethyleneglycol (PEG)-induced fusion of protoplasts of these two species resulted in 0.28% fusion frequency. The fusant produced fruiting bodies on paddy straw but required a lower temperature of crop running ($24{\pm}2^{\circ}C$) than its parents which could fruit at $28{\pm}2^{\circ}C$. The stable fusant strain was selected by testing for the selected biochemical markers i.e. Carbendazim tolerance and utilization of the lignin degradation product, vanillin.

Comparison of the LNT Regeneration Methods in 2.2L Common Rail Direct Injection Diesel Engine (2.2L 직분사 디젤 엔진에서 LNT 촉매 재생을 위한 환원제 분사 방법 비교)

  • Nam, Chungwoo;Han, Manbae
    • Transactions of the Korean Society of Automotive Engineers
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    • v.23 no.2
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    • pp.169-177
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    • 2015
  • In this study we investigated the regeneration methods for the lean $NO_x$ trap (LNT) catalyst in a 2.2L direct injection diesel engine. The regeneration methods were 1) in-cylinder post fuel injection and 2) external fuel injection strategy. The in-cylinder post fuel injection method uses in-cylinder injectors with the addition of the post fuel injection to supply enough reductants such as CO, $H_2$, THC. The external fuel injection method was enabled by installing a fuel injector with a wide spray angle before the LNT catalyst. Through the engine experiment, the $NO_x$ conversion efficiency, the amount of reductant exhaust gases, fuel consumption, and temperature behavior in the LNT catalyst were evaluated and compared for the two regeneration methods.

Column regeneration for Partisil/Partisphere ion-exchange columns (Partisil/Partisphere 이온 교환 컬럼 재생 가이드)

  • Mark Fever;Gemma Howse
    • FOCUS: LIFE SCIENCE
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    • no.1
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    • pp.5.1-5.3
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    • 2024
  • The document discusses the regeneration of Partisil/Partisphere ion-exchange columns in chromatography. It mentions that column efficiency can diminish with use due to the accumulation of sample and/or mobile phase impurities at the head of the column. This can lead to a change in back pressure, lower column efficiency, and sometimes a change in selectivity. The document outlines a procedure that may restore column performance. The document also provides everyday practices to enhance the lifetime of a column. These include using only high-purity HPLC solvents and buffers, using freshly prepared mobile phases and buffers, filtering mobile phases to remove particulates, using appropriate sample clean-up procedures, using a guard column or pre-column filter, and working within the pressure and flow rate limitations of the column. For the regeneration of Partisil/Partisphere SAX, SCX, WAX, and WCX columns, the document suggests passing 20 column volumes of various mobile phases through the column. These include a buffer wash, distilled water, an acid wash, a chelating wash, a methanol wash, and a buffer for separation. The document emphasizes that not all of these wash steps are required for every column clean-up and that some chromatographers require only a combination of certain steps.

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Nitrogen Removal using Zeolite at On-site Wastewater Treatment System (소규모 오수처리 시스템에서의 제올라이트에 의한 질소 제거)

  • 방천희;권순국
    • Magazine of the Korean Society of Agricultural Engineers
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    • v.44 no.2
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    • pp.161-173
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    • 2002
  • Recently, absorbent biofilters, which are inexpensive and easy to manacle, have been supplied to the rural areas, but have limitations in removing the nutrients effectively. Accordingly, as an alternative plan. natural zeolites were arranged in front or at the rear of the absorbent biofilters, and their removal efficiency for nitrogen and, ultimately, their applicability to the on-site wastewater treatment system were studied. Furthermore, the same experiments were carried out on artificial zeolites, made from coal ashes at National Honam Agricultural Experiment Station, to compare natural zeolites with artificial ones. Treated wastewater through the Absorbent Biofilter showed 22.6% nitrogen removal efficiency, while 64.6% was attained when natural Zeolites were placed in front of the absorbent biofilters (Zeolite-Aerobic process). As an addition, phosphorus was also efficiently removed. On the other hand, Aerobic-Zeolite process, which arranged natural zeolites at the rear of the biofilters, did not have significantly higher nitrogen removal as compared to the treatment using only the absorbent biofilters. Furthermore, upon regeneration of the natural zeolite, the ion exchange rate was fecund to increase over 10% as compared to before regeneration. Our results show that natural zeolites, applied to the on-site wastewater treatment system through the Zeolite-Aerobic process, not only increase the removal efficiency of nutrients, but, by choosing the appropriate regeneration time, can also be cast-effective. Artificial zeolites, on the other hand, though more efficient in removing nutrients, cannot be regenerated and, therefore, are not cost-effective.

Factors for high frequency plant regeneration in tissue cultures of Indian mustard (Brassica juncea L.)

  • Bhuiyan, Mohammed Shafi Ullah;Min, Sung-Ran;Choi, Kwan-Sam;Lim, Yong-Pyo;Liu, Jang-Ryol
    • Journal of Plant Biotechnology
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    • v.36 no.2
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    • pp.137-143
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    • 2009
  • An efficient system for high frequency plant regeneration was established through investigating various factors such as plant growth regulator combinations, explant types and ages, and addition of $AgNO_3$ influenced on shoot regeneration in Brassica juncea L. cv. BARI sarisha-10. Murashige and Skoog (MS) medium supplemented with 0.1 mg/L NAA (1-naphthaleneacetic acid) and 1 mg/L BA (6-benzyladenine) showed the maximum shoot regeneration frequency (56.67%) among the different combinations of NAA and BA. Explant type, explant age, and addition of $AgNO_3$ also significantly affected shoot regeneration. Of the four type of explants (cotyledon, hypocotyl, root, and leaf explants)- cotyledon explants produced the highest shoot regeneration frequency and hypocotyls explants produced the highest number of shoots per explant, whereas root explants did not produce any shoot. The cotyledonary explants from Four-day-old seedlings showed the maximum shoot regeneration frequency and number of shoots per explant. Shoot regeneration frequency increased significantly by adding $AgNO_3$ to the medium. Two mg/L $AgNO_3$ appeared to be the best for shoot regeneration with the highest shoot regeneration frequency (86.67%) and number of shoots per explant (7.5 shoots). Considerable variation in shoot regeneration from cotyledonay explants was observed within the B. juncea L. genotypes. The shoot regeneration frequency ranged from 47.78% for cv. Shambol to 91.11% for cv. Rai-5. In terms of the number of shoots produced per explant, B. juncea L. cv. Daulot showed the maximum efficiency. MS medium supplemented with 0.1 mg/L NAA showed the highest frequency of rooting. The regenerated plantlets were transferred to pot soil and grown to maturity in the greenhouse. All plants were fertile and morphologically identical with the source plants.