• Title/Summary/Keyword: PGRs

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Codon Optimization, Soluble Expression and Purification of PE_PGRS45 Gene from Mycobacterium tuberculosis and Preparation of Its Polyclonal Antibody Protein

  • Xu, Tao;Li, Minying;Wang, Chutong;Yuan, Meili;Chang, Xianyou;Qian, Zhongqing;Li, Baiqing;Sun, Meiqun;Wang, Hongtao
    • Journal of Microbiology and Biotechnology
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    • v.31 no.11
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    • pp.1583-1590
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    • 2021
  • Studies have demonstrated that PE_PGRS45 is constitutively expressed under various environmental conditions (such as nutrient depletion, hypoxia, and low pH) of the in vitro growth conditions examined, indicating that PE_PGRS45 protein is critical to the basic functions of Mycobacterium tuberculosis. However, there are few reports about the biochemical function and pathogenic mechanism of PE_PGRS45 protein. The fact that this M. tuberculosis gene is not easily expressed in E. coli may be mainly due to the high content of G+C and the use of unique codons. Fusion tags are indispensable tools used to improve the soluble expression of recombinant proteins and accelerate the characterization of protein structure and function. In the present study, His6, Trx, and His6-MBP were used as fusion tags, but only MBP-PE_PGRS45 was expressed solubly. The purification using His6-MBP tag-specific binding to the Ni column was easy to separate after the tag cleavage. We used the purified PE_PGRS45 to immunize New Zealand rabbits and obtained anti-PE_PGRS45 serum. We found that the titer of polyclonal antibodies against PE_PGR45 was higher than 1:256000. The result shows that purified PE_PGRS45 can induce New Zealand rabbits to produce high-titer antibodies. In conclusion, the recombinant protein PE_PGRS45 was successfully expressed in E. coli and specific antiserum was prepared, which will be followed by further evaluation of these specific antigens to develop highly sensitive and specific diagnostic tests for tuberculosis.

Optimization of Embryogenic Callus Induction and Plant Regeneration in Orchid Coelogyne cristata

  • Naing, Aung Htay;Lim, Ki-Byung
    • Horticultural Science & Technology
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    • v.29 no.3
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    • pp.260-266
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    • 2011
  • An efficient protocol was established for high frequency somatic embryogenesis through a callus culture of Coelogyne cristata. The best frequency of callusing was obtained from a PLB segment (3-5 mm) cultured on MS medium supplemented with coconut water (CW) and a combination of both 3 $mg{\cdot}L^{-1}$ of 2,4-D and BA. When the calli were sub-cultured on the MS medium without any PGRs, the average number of somatic embryos were higher than those with PGRs treatment. NAA is the most critical factor among PGRs, which dramatically hindered for the formation of a somatic embryo. The efficacy of the addition of coconut powder (CP) for somatic embryogenesis was almost the same in all treatments. However, the number of somatic embryos formed distinctly depended on age of the callus. The somatic embryos converted into healthy plants with well-developed shoots on the same medium. Plantlets showed the best responses of root and shoot growth when transferred to $\frac{1}{2}$ MS medium containing 1.5 $g{\cdot}L^{-1}$ of activated charcoal. All plants with above 3.0-cm-high were successfully acclimatized in the greenhouse.

In vitro Multiplication of Hosta Tratt. Species Native to Korea by Shoot-tip Culture (경정배양에 의한 한국 자생 비비추속 식물의 기내증식)

  • Choi, Han;Yang, Jong Cheol;Ryu, Sun Hee;Yoon, Sae Mi;Kim, Sang Yong;Lee, Seung Youn
    • Korean Journal of Plant Resources
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    • v.32 no.1
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    • pp.53-62
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    • 2019
  • The purpose of this study was to establish the in vitro propagation system by shoot tip culture of six Hosta species native to Korea (Hosta capitata (Koidz.) Nakai, H. clausa Nakai, H. jonesii M.G.Chung, H. minor (Baker) Nakai, H. venusta F.Maek., and H. yingeri S.B.Jones) for mass proliferation and a new cultivar development. The shoot tips of each Hosta species were cultured on MS medium containing eight combinations of 0.5, 1.0, 2.0, 4.0 mg/L BA with 0.1 mg/L NAA, 0.1, 0.5, 1.0, 2.0 mg/L TDZ with 0.1 mg/L NAA, and without any PGRs (control). They were investigated on callus, somatic embryo, crown bud, differentiation and growth of shoot and root, total fresh weight after 8 weeks of culture. In all six Hosta species, callus and somatic embryo induction rate and multiple shooting rate of the PGRs treatment group were higher than that of the control group. The highest number of differentiated shoots were obtained on medium supplemented with 2.0 ㎎/L TDZ in H. capitata (5.4), 1.0 mg/L TDZ in H. clausa and H. jonesii (3.3 and 5.8, respectively), 0.5 mg/L BA in H. minor (11.1), 1.0 mg/L BA and 0.1 mg/L TDZ in H. venusta (8.1), and 0.5 mg/L TDZ in H. yingeri (9.8). In somatic embryo formation, the PGRs treatment group of H. jonesii and H. yingeri were more effective than the control group, and the effects were relatively less in H. capitata, H. clausa Nakai, H. minor, H. venusta. Crown bud formation of four Hosta species (H.capitata, H. clausa, H. jonesiig, and H. yingeri) were also higher in the PGRs treatment group than in the control group. Crown bud formation of four Hosta species (H.capitata, H. clausa, H. jonesiig, and H. yingeri) were also higher in the PGRs treatment group than in the control group. H. clausa showed no significant effect on callus and shoot differentiation regardless of the type and concentration of cytokinin, but slightly increased in formation of crown bud in TDZ.

Optimization of efficient protocorm-like body (PLB) formation of Phalaenopsis and Dendrobium hybrids

  • Soe, Khaing Wah;Myint, Khin Thida;Naing, Aung Htay;Kim, Chang Kil
    • Current Research on Agriculture and Life Sciences
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    • v.32 no.4
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    • pp.179-183
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    • 2014
  • Optimization of the protocorm-like body (PLB) formation of Phalaenopsis and Dendrobium hybrids was performed by determining the effects of plant growth regulators (PGRs) and different parts and division sizes of the PLB. For both genera, the base part was the best for the proliferation of PLBs, yielding the highest number of PLBs on a PGR-free medium for Phalaenopsis and medium containing $0.1{\mu}M$ NAA and $10.0{\mu}M$ BAP for Dendrobium. As regards the division size, four-division sections resulted in a higher PLB formation efficiency for Phalaenopsis, while two-division sections produced a higher PLB formation efficiency for Dendrobium. It is expected that these findings will be applicable to efficient PLB formation of other Phalaenopsis and Dendrobium orchids.

Chemical Mowing of A Fairway Zoysiagrass Turf (Zoysia japonica Steud.) Using New Generation of PGRs (식물생장조절제를 이용한 들잔디의 화학적 예초관리에 관한 연구)

  • 김경남;남상용;김용선
    • Journal of the Korean Institute of Landscape Architecture
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    • v.28 no.3
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    • pp.1-12
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    • 2000
  • 식물생장조절제(Plant Growth Regulators, PGRs)를 이용한 예초관리 방법이 들잔디 엽색 및 품 질에 미치는 영향을 규명함으로 잔디 적응력 관점에서 적합한 PGR 종류 및 살포수준을 파 악하고자 본 연구를 시작하였다. 실험은 들잔디 생육이 왕성한 시기인 6월(실험 I)과 7월(실험 II) 2회에 걸쳐 각각 다른 장소에서 실시하였다. 공시약제 처리구는 Type I PGR인 amidochlor 3 수준 및 mefluidide 3 수준 과 Type II PGR인 trinexapacethyl 4 수준, 그리고 무처리구를 포함하여 전체 11 처리구였 고, 실험구 배 치는 난괴법 4 반복으로 하였다. 공시한 3종류 PGR 모두 잔디엽색 및 품질에 통계적으로 유의한 차이 가 있었지만, PGR 종류.살포수준 및 생육 상태에 따라 처리효과는 다르게 나타났다. 잔디 엽색 및 품질 은 PGR 살포후 경과기간에 따라 점진적으로 억제되는 경향이었다. 처리 1주후 잔디품질은 대조구에 비 해 큰 차이가 없었지만, 2주에서 4주까지는 PGR 종류 및 살포수준에 따라 유의하게 감소하 여 0.08 ml/ $m^2$ 이상의 trinexapac-ethyl 처리구 및 1.20 ml/$m^2$ 이상의 mefluidide 처리구에서는 가장 크 게 나타났다. 처리 4주부터 대부분 처리구에서 잔디품질이 적정수준 이상으로 회복되기 시작하였다. 본 실험을 통해 잔디관리 수준에서 저관리 잔디지역은 0.08 ml/$m^2$ 수준의 trinexapac-ethyl 처리구가 장기간 생장억제 관점에서 실용성과 경제성 측면에서 들잔디 관리에 효율적인 것으로 판단되었고, 고관리 잔 디지역은 0.04~0.08ml/$m^2$ 수준의 trinexapac-ethyl 처리구 및 0.60~1.20 ml/$m^2$ 수준의 amidochlor 처 리구가 적합 한 것으로 판단되었지만, 계절에 따라 PGR 효과정도가 다르게 나타났다. 따라서, 잔디 관리 시 잔디의 생육상태, 예초 절감 기간, 잔디품질의 기대수준 및 적용할 수 있는 관리수준에 따라 적합한 PGR 종류 선정과 함께 살포시기 및 살포량을 효율적으로 결정하는 것이 필요한 것으로 사료되었다.

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Studies on the Processing of Crude Drugs(IX) -Preparing Standardization and Regulation of Stir-Frying Glycyrrhzia root(1)- (한약수치에 관한 연구(제 9보) -초감초(炒甘草) 제법의 표준화 및 규격화(1)-)

  • Choi, Hyuck-Jai;Lee, Woo-Jung;Park, Sung-Hwan;Song, Bo-Whan;Kim, Dong-Hyun;Kim, Nam-Jae
    • Korean Journal of Pharmacognosy
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    • v.36 no.3 s.142
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    • pp.209-219
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    • 2005
  • In this study, we carried out the preparing standardization and regulation of processed Glycyrrhizae Radix (PGR) which have been widely used in oriental medicines. Glycyrrhizae Radix(GR) have been generally prepared by the stir-frying, or mix-frying with honey for the purpose of decreasing sweetness and augmenting vitality. Firstly, we tried to standardize PGR prepared by the stir-frying. We purchased 14 kinds of PGR and non-processed GR(NPGR) at oriental physician's offices and oriental pharmacies on a nation scale, respectively. The amounts of dry on loss, water extract, diluted ethanol extract, ether extract, total ash, acid insoluble ash, glycyrrhizin(GL), glycyrrhetic acid(GA) and liquiritin(LQ) of them were examined. The amounts of dry on loss, GL and LQ in commercial PGRs showed remarkable decrease, while GA showed increased as compared with NPGR. In order to standardize preparing method of PGR, the effect of heating time on physico-chemical parameters and biological activities were examined. Physico-chemical parameters such as dry on loss, extract amount, GL and LQ contents in PGRs showed decrease, however, GA was increased with heating time as compared with NPGR. Also, GA, obtained from heat-treated GR, was found as an artifact in PGRs. PGR was more effective than NPGA in vitro test of DPPH scavenging effect and TBA-Rs reducing effect. PGR and NPGR showed potent hepatoprotective effect on $CCl_4-intoxicated$ rats. Especially, PGR prepared by 80 min of heating was the most effective. Considering these results, the optimal condition for PGR preparation was $150^{\circ}C$ for 80 min.

Effects of Medium Compositions and Plant Growth Regulators on in vitro Organogenesis in Cultured Explants of Platycodon grandiflorum Species

  • Kwon, Soo Jeong;Roy, Swapan Kumar;Kim, Hye-Rim;Moon, Young-Ja;Yoon, Ki-Hong;Woo, Sun Hee;Boo, Hee Ock;Koo, Jin-Woog;Kim, Hag Hyun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.62 no.3
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    • pp.259-274
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    • 2017
  • Platycodon grandiflorum (Bell flower) is an important plant that has traditionally been used as herbal medicine for the treatment of cough, phlegm, sore throats, lung abscesses, chest pains, dysuria, and dysentery. The present study was initiated to investigate the feasibility of inducing shoot and root organogenesis in cultured explants of P. grandiflorum in a range of culture media and through use of various plant growth regulators (PGRs). The plantlets (Stem containing one node) were isolated and cultured on different concentrations of Murashige and Skoog (MS) medium supplemented with PGRs. We found that proliferation and elongation of shoots and roots could be achieved on 1/4 MS for P. grandiflorum with wild and green petals and on 1/8 MS for P. grandiflorum with double petals. The highest levels of development and elongation of adventitious shoots and roots were observed when petal explants were cultured on 1/4 MS (pH 3.8) supplemented with 5% sucrose. Increasing the agar concentration reduced shoot growth and rooting potential; nevertheless, the highest number of shoots and roots was observed on 0.6% agar. In the case of growth regulators, 1/4 MS supplemented with $1mg\;L^{-1}$ 6-benzylaminopurine (BA) was found to be best for shooting, although higher concentrations of BA tended to reduce shoot and root elongation. The highest number of shoots was achieved on $0.5mg{\cdot}L^{-1}$ thidiazuron (TDZ) from double petal explants grown on 1/8 MS. However, root and shoot elongation were found to decrease when TDZ concentrations were increased. Low concentrations of kinetin, naphthalene acetic acid, indole acetic acid, and 3-indole butyric acid induced shoot and root proliferation and elongation. Taken together, our study showed that low concentrations of PGRs induced the greatest root formation and elongation, showing that the optimal concentration of PGRs for shoot proliferation was species-dependent.

Effect of Pinching Time and Plant Growth Retardants on Growth Control of Ardisia pusilla in an Ebb and Flow System (Ebb and Flow 시스템에서 적심시기와 생장조절제 처리에 의한 산호수의 생장조절)

  • Won, Eun-Jeong;Park, Ji-Eun;Park, Yoo-Gyeong;Jeong, Byoung-Ryong
    • Journal of Bio-Environment Control
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    • v.20 no.4
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    • pp.333-340
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    • 2011
  • This study was conducted on the effect of plant growth retardants (PGRs) on growth of potted Ardisia pusilla and Ardisia pusilla var. variegata grown for a short term in an ebb and flow system. Plants were planted in 10 cm plastic pots, containing a mixture of peat moss and perlite (1 : 1, v/v) on 30 June 2005 and were grown until 23 Sep. 2005. The PGRs used were 50, 200, 350, 500 $mg{\cdot}L^{-1}$ daminozide (B- 9), 10, 40, 70, 100 $mg{\cdot}L^{-1}$ paclobutrazol (Boundy), and 5, 15, 25, 35 $mg{\cdot}L^{-1}$ ethephon (Florel). On every irrigation, the nutrient solution containing PGRs was supplied at a 2 cm depth and was kept for 15~20 minutes. The surplus nutrient solution was drained back into the tank for next irrigations. In both cultivars, paclobutrazol gave the most pronounced effect in inhibition of stretchiness. Ardisia pusilla, which was pinched just before initiation of paclobutrazol treatment, resulted in the greatest inhibition of shoot elongation. Daminozide treatments above 200 $mg{\cdot}L^{-1}$ also gave reduction of shoot length. However, ethephon treatments gave no dwarfing effect in Ardisia pusilla. Inhibition of stretchiness was observed even in the lowest concentration treatments. Among the PGRs, paclobutrazol was the most effective in suppressing plant stretchiness.

Effects of Plant Growth Regulators on In Vitro Germination and Organ Formation of Wild Angelica gigas N. (야생 참당귀(Angelica gigas N.)의 기내발아 및 기관유도에 미치는 PGRs의 영향)

  • Lee, Su-Gwang;Cho, Won-Woo;Lee, Song-Hee;Park, Kwang-Woo;Choi, Kyung;Kang, Ho-Duck
    • Korean Journal of Plant Resources
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    • v.25 no.4
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    • pp.456-464
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    • 2012
  • This study was conducted to establish the in vitro optimal condition for seed germination and organogenesis of wild Angelica gigas. The experiment was evaluated the effects of $GA_3$ for pre-treatment with different periods of time (0h, 24h, 48h, 72h) and followed the treatment of seeds by control, scarification and methanol-heating method. As a result, the highest rates (15%) of seed germination was shown under the treatment without soaking of $GA_3$ and methanol-heating treatment. The seed germination was highly increased 60% under the condition of treatment on ultrasonic waves (frequency 80 KHz) with methanol-heating treatment including 0.1 mg/L $GA_3$. The highest callus induction rate was obtained from in vitro germinated stem, root and hypocotyl on the MS medium with 1.0 mg/L NAA and 0.5 mg/L BA. The highest percentages of shooting (50%) and rooting (85%) induction were observed in hypocotyl and root cultured on PGRs free medium and 0.1 mg/L NAA, respectively. In addition, somatic embryogenesis was observed from stem (1.0 mg/L 2,4-D) and hypocotyl (0.1 mg/L NAA).

Seasonal Growth Inhibition of Perennial Ryegrass (Lolium perenne L.) after Applying Two Plant Growth Regulators (생장조정제 처리 후 생육 시기별 퍼레니얼 라이그래스의 억제 효과)

  • Seung Jae Lim;Tae Wooung Kim;Young-Sun Kim
    • Journal of Bio-Environment Control
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    • v.32 no.3
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    • pp.242-248
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    • 2023
  • This study was conducted to evaluate the effects of trinexapac-ethyl (TE) and prohexadione-calcium (PC) on the growth and turf quality of perennial ryegrass (Lolium perenne L.). Treatments were designed as follows; control (non-treatment), TE (TE 0.01 a.i. g·m-2·100mL-1), and PC (PC 0.01 a.i. g·m-2·100mL-1). Compared with control, turf color index and chlorophyll content of TE and PC treatments were not significantly different. As applied plant growth regulators (PGRs), shoot length in the TE and PC was decreased by 8.9-12.4% and 6.7-13.6%, respectively, and clipping yield by 44.4-45.8% and 40.6-40.9%. When evaluated with the growth of shoot length after applying PGRs, residual days of TE and PC were 43.6 and 37.9 days after treatment of PGRs (DAT) in the 1st experiment (July 29-August 26), respectively, and 38.3 and 39.5 DATs in the 2nd experiment (September 30-October 28). These results indicated that an application of TE or PC decreased shoot length and clipping yield in the perennial ryegrass, and their residual time was about 40±3 DATs. Perennial ryegrass applied with PGRs like TE and PC was not significant seasonal difference, and might be not phytotoxic.