• Title/Summary/Keyword: MS medium types

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Transformation of Chinese Cabbage Glutathione Reductase (GR) gene into Lettuce (Lactuca sativa L.) with Particle Bombardment (유전자총을 이용한 상추 내로의 배추 Glutathione Reductase (GR)유전자의 도입)

  • 정재동;이부자;이효신;김창길
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.6
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    • pp.475-478
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    • 2000
  • The cytosolic glutathione reductase(GR) gene of chinese cabbage was introduced into Lettuce (Lactuca sativa L.) with particle bombardment method. When cotyledon explants were treated with osmoticum-conditioning medium (0.6 M sorbitol/mannitol) 4 hours prior to and 16 hours after bombardment, it was identified by GUS assay that this condition was the most efficient one for introduction of foreign genes into cotyledon tissue of lettuce with particle bombardment. The stable integration of a GR gene was confirmed by the PCR analysis. 0.3, 0.6, 1.5 kbp PCR fragments of transgenes were obtained by three types of primers designed on the basis of GR sequence. To know whether the expression of the GR gene of pBKs-GR 1 can be stably maintained in the next generation, T$_2$selfing seeds obtained from the transformed mother plants were sowed on MS medium supplemented with 200 mg/L kanamycin sulfate. 70% of seedlings showed resistance to kanamycin.

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Plant Regeneration from Explant Types and Cultivars of Boxthorn (Lycium chinense Mill.) (구기자나무의 절편체 부위와 품종 간 재분화 특성)

  • Kim, Dong-Chan;Chung, Hae-Joon;Min, Byung-Hoon;Yang, Deok-Chun
    • Journal of Plant Biotechnology
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    • v.29 no.1
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    • pp.15-18
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    • 2002
  • Callus and shoot formation of leaf explants from in vitro propagated shoots and field grown plants depending on the position of leaf, and four boxthron cultivars were investigated. Callus formation of explants from both in vitro shoot and field grown plants as easily achieved at the cut surfaces of explants but the callus formed from leaf of in vitro shoots was hardened as the duration of culture was proceed. Calli were effectively induced from leaves detached from the middle position of both in vitro and in vitro plants on MS medium containing 0.5 mg/L NAA with 0.2 mg/L BA, and the growth of calli were better in field grown leaves than in vitro grown leaves. Shoot formation were effectively induced from leaves detached from the upper position in vitro plants, and the middle parts of in vitro plants on MS medium containing 0.01 mg/L (NAA with 0.2 mg/ BA. There was difference on the frequency of shoot formation among four different cultivars; 'Jindojaerae' was the best for shoot formation followed by 'Cheonyang', 'Younghagukija' and 'Cheongyangjaerae'.

Analysis of ground reaction force contributing to horizontal velocity factors in short distance 100M race (육상 단거리 100m 수평속도 요인에 기여하는 지면반력분석)

  • Choi, Su-Nam
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.15 no.4
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    • pp.2134-2141
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    • 2014
  • This study was to analyze ground reaction force according to Crouching Start type at the starting point of 100M race. The subjects of this study were 8 women sprinters and we analyzed their ground reaction force by classifying the distance between start blocks as three types. The followings are the results of the study. According to maximum horizontal ground reaction force analysis result, in the left foot placed in front, BS among excellent group and MS in non-excellent group showed the biggest reaction force value. In the right foot placed at the back, MS in both groups showed the biggest reaction force value. MS in the right foot of the excellent group was the biggest (0.83 BW). According to maximum vertical ground reaction force analysis result, in the left foot placed in front, ES among excellent group and BS in non-excellent group showed the biggest reaction force value. In the right foot placed at the back, BS among excellent group and MS in non-excellent group showed the biggest reaction force value.

Callus Induction and Plant Regeneration in Hordeum spontaneum (Hordeum spontaneum에서의 callus 유기와 식물체 양생)

  • Jong-Yol Choi
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.32 no.4
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    • pp.466-470
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    • 1987
  • Immature embryos of Hordeum spontaneum were cultured on B5 and CI medium (Cheng's modified MS) to induce callus formation. CI media containing lmg/$\ell$ and 2mg/$\ell$ 2,4-D were more effective than B5 medium with lmg/$\ell$ 2,4-D for the initiation of callus. Total 883 calli were induced from 1,060 immature embryos plated. Callus induction frequency was 83%. Calli were transferred to differentiation media after one subculture to regenerate plants. Forty six plants were regenerated from 608 calli. Seventeen plants were chlorophyll deficient. There was no significant difference for plant regeneration among genotypes and media effects in calli which had been induced from immature embryos of seven types of trisomies. The overall regeneration frequency was 7.6%.

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Brief Introduction of Research Progresses in Control and Biocontrol of Clubroot Disease in China

  • He, Yueqiu;Wu, Yixin;He, Pengfei;Li, Xinyu
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.45-46
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    • 2015
  • Clubroot disease of crucifers has occurred since 1957. It has spread to the whole China, especially in the southwest and nourtheast where it causes 30-80% loss in some fields. The disease has being expanded in the recent years as seeds are imported and the floating seedling system practices. For its effective control, the Ministry of Agriculture of China set up a program in 2010 and a research team led by Dr. Yueqiu HE, Yunnan Agricultural University. The team includes 20 main reseachers of 11 universities and 5 institutions. After 5 years, the team has made a lot of progresses in disease occurrence regulation, resources collection, resistance identification and breeding, biological agent exploration, formulation, chemicals evaluation, and control strategy. About 1200 collections of local and commercial crucifers were identified in the field and by artificiall inoculation in the laboratories, 10 resistant cultivars were breeded including 7 Chinese cabbages and 3 cabbages. More than 800 antagostic strains were isolated including bacteria, stretomyces and fungi. Around 100 chemicals were evaluated in the field and greenhouse based on its control effect, among them, 6 showed high control effect, especially fluazinam and cyazofamid could control about 80% the disease. However, fluzinam has negative effect on soil microbes. Clubroot disease could not be controlled by bioagents and chemicals once when the pathogen Plasmodiophora brassicae infected its hosts and set up the parasitic relationship. We found the earlier the pathogent infected its host, the severer the disease was. Therefore, early control was the most effective. For Chinese cabbage, all controlling measures should be taken in the early 30 days because the new infection could not cause severe symptom after 30 days of seeding. For example, a biocontrol agent, Bacillus subtilis Strain XF-1 could control the disease 70%-85% averagely when it mixed with seedling substrate and was drenching 3 times after transplanting, i.e. immediately, 7 days, 14 days. XF-1 has been deeply researched in control mechanisms, its genome, and development and application of biocontrol formulate. It could produce antagonistic protein, enzyme, antibiotics and IAA, which promoted rhizogenesis and growth. Its The genome was sequenced by Illumina/Solexa Genome Analyzer to assembled into 20 scaffolds then the gaps between scaffolds were filled by long fragment PCR amplification to obtain complet genmone with 4,061,186 bp in size. The whole genome was found to have 43.8% GC, 108 tandem repeats with an average of 2.65 copies and 84 transposons. The CDSs were predicted as 3,853 in which 112 CDSs were predicted to secondary metabolite biosynthesis, transport and catabolism. Among those, five NRPS/PKS giant gene clusters being responsible for the biosynthesis of polyketide (pksABCDEFHJLMNRS in size 72.9 kb), surfactin(srfABCD, 26.148 kb, bacilysin(bacABCDE 5.903 kb), bacillibactin(dhbABCEF, 11.774 kb) and fengycin(ppsABCDE, 37.799 kb) have high homolgous to fuction confirmed biosynthesis gene in other strain. Moreover, there are many of key regulatory genes for secondary metabolites from XF-1, such as comABPQKX Z, degQ, sfp, yczE, degU, ycxABCD and ywfG. were also predicted. Therefore, XF-1 has potential of biosynthesis for secondary metabolites surfactin, fengycin, bacillibactin, bacilysin and Bacillaene. Thirty two compounds were detected from cell extracts of XF-1 by MALDI-TOF-MS, including one Macrolactin (m/z 441.06), two fusaricidin (m/z 850.493 and 968.515), one circulocin (m/z 852.509), nine surfactin (m/z 1044.656~1102.652), five iturin (m/z 1096.631~1150.57) and forty fengycin (m/z 1449.79~1543.805). The top three compositions types (contening 56.67% of total extract) are surfactin, iturin and fengycin, in which the most abundant is the surfactin type composition 30.37% of total extract and in second place is the fengycin with 23.28% content with rich diversity of chemical structure, and the smallest one is the iturin with 3.02% content. Moreover, the same main compositions were detected in Bacillus sp.355 which is also a good effects biocontol bacterial for controlling the clubroot of crucifer. Wherefore those compounds surfactin, iturin and fengycin maybe the main active compositions of XF-1 against P. brassicae. Twenty one fengycin type compounds were evaluate by LC-ESI-MS/MS with antifungal activities, including fengycin A $C_{16{\sim}C19}$, fengycin B $C_{14{\sim}C17}$, fengycin C $C_{15{\sim}C18}$, fengycin D $C_{15{\sim}C18}$ and fengycin S $C_{15{\sim}C18}$. Furthermore, one novel compound was identified as Dehydroxyfengycin $C_{17}$ according its MS, 1D and 2D NMR spectral data, which molecular weight is 1488.8480 Da and formula $C_{75}H_{116}N_{12}O_{19}$. The fengycin type compounds (FTCPs $250{\mu}g/mL$) were used to treat the resting spores of P. brassicae ($10^7/mL$) by detecting leakage of the cytoplasm components and cell destruction. After 12 h treatment, the absorbencies at 260 nm (A260) and at 280 nm (A280) increased gradually to approaching the maximum of absorbance, accompanying the collapse of P. brassicae resting spores, and nearly no complete cells were observed at 24 h treatment. The results suggested that the cells could be lyzed by the FTCPs of XF-1, and the diversity of FTCPs was mainly attributed to a mechanism of clubroot disease biocontrol. In the five selected medium MOLP, PSA, LB, Landy and LD, the most suitable for growth of strain medium is MOLP, and the least for strains longevity is the Landy sucrose medium. However, the lipopeptide highest yield is in Landy sucrose medium. The lipopeptides in five medium were analyzed with HPLC, and the results showed that lipopeptides component were same, while their contents from B. subtilis XF-1 fermented in five medium were different. We found that it is the lipopeptides content but ingredients of XF-1 could be impacted by medium and lacking of nutrition seems promoting lipopeptides secretion from XF-1. The volatile components with inhibition fungal Cylindrocarpon spp. activity which were collect in sealed vesel were detected with metheds of HS-SPME-GC-MS in eight biocontrol Bacillus species and four positive mutant strains of XF-1 mutagenized with chemical mutagens, respectively. They have same main volatile components including pyrazine, aldehydes, oxazolidinone and sulfide which are composed of 91.62% in XF-1, in which, the most abundant is the pyrazine type composition with 47.03%, and in second place is the aldehydes with 23.84%, and the third place is oxazolidinone with 15.68%, and the smallest ones is the sulfide with 5.07%.

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Development of Near Isogenic Lines with Various Seed Sizes and Study on Seed Size-related Characteristics in Watermelon (다양한 종자크기를 가진 Near Isogenic 수박 계통 육성 및 종자관련 특성 분석)

  • Kim, Yong-Jae;Yang, Tae-Jin;Park, Young-Hoon;Lee, Yong-Jik;Kang, Sun-Cheol;Kim, Yong-Kwon;Cho, Jeoung-Lai
    • Korean Journal of Breeding Science
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    • v.41 no.4
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    • pp.403-411
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    • 2009
  • We inspected seed sizes of 353 genetic accessions of watermelon to diversify functional utility related to seed size and classified them into six representative groups based on their seed sizes. Each group was named as giant seed (GS), big seed (BS), medium size (NS), small size (SS), micro seed (MS) and tomato seed (TS) from the biggest. As the seed size was getting smaller, decreased seed length and seed width, increased seed number per fruit, and decreased seed weight per fruit were observed, but seed shape did not change significantly. In order to study the effect of seed size on fruit weight and seed germination, we developed three near isogenic lines (NILs) with three different seed sizes, SS, MS and TS, from crossing between two accessions 'NT' and 'TDR', and one NIL with seed size of TS from crossing between two accessions 'S55' and 'TDR'. In the study on the fruit weight of NILs with various seed sizes, NS, SS, MS, and TS NILs produced an average of 6.4, 6.3, 5.9, and 4.2 kg fruits, respectively. The bigger seed types showed the better germination rate. NS type showed the highest germination percentage, while TS showed very low germination percentage. Fermentation treatment for 48 hrs increased the germination percentage on TS type seed, but still remained at a low level. In NS, SS, and MS material, the ratio of embryo/whole seed weight was over 50%, meanwhile that of TS was only 44.4% of which low embryo percentage would be one of the reason of low germination percentage. From this study, we concluded that watermelon has very wide genetic diversity on seed size which is somehow related to fruit sizes and germination rate.

Plant Regeneration and Transformation of Kentucky Bluegrass(Poa pratensis L.) via the Plant Tissue Culture (조직배양을 이용한 Kentucky bluegrass(Poa pratensis L.)의 식물체 재분화 및 형질전화 조건의 검토)

  • Miki Kusano;Koichi Tohyama;Bae, Chang-Hyu;Riu, Key-Zyung;Lee, Hyo-Yeon
    • Journal of Plant Biotechnology
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    • v.30 no.2
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    • pp.115-121
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    • 2003
  • In this study, plant regeneration and Agrobacterium tumefaciens-mediated transformation Kentucky bluegrass(Poa pratensis L.) were evaluated. Three different types of calli were produced depending on the combinations of growth regulators. They were non-friable brown or gray-colored callus (type I), compact, friable and yellow or white-colored callus (typeII), and soft, watery translucent callus with differentiated structure (typeIII). The highest regenerable organogenic callus (typeII) was obtained on the medium containing 1mg/L, 2,4-D and 0.1mg/L BA. Additionally, the production of typeII calli increased significantly when AgNO$_3$ was added to the callus induction and growth medium. The highest frequency of multiple shout formation from typeII callus was obtained on MS medium containing 1mg/L BA and 1mg/L Thidiazuron(TDZ). The organogenic calli(typeII) were inoculated with Agrobacterium tumefaciens strain EHA101 harboring the binary vector pIG121Hm with $\beta$-glucuronidase gene, and various factors were found to influence the transfer-DNA delivery efficiency. The highest transient GUS activity was observed on typeIIcallus. In the present work, we reported the first transient GUS activity of Kentucky bluegrass mediated by Agrobacterium tumefaciens. Our system may contribute to genetic improvement for breed-recalcirtrant grass species, Kentucky bluegrass.

Effect of Cytokinins on Nodal Cultures of Citrus Species (감귤류의 마디배양에서 싸이토키닌류가 기내 대량증식에 미치는 영향)

  • Kyung chul HAN;Youn Hwa HAN
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.3
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    • pp.145-150
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    • 1994
  • This study was conducted to determine the effects of 3 cytokinins (BA,2iP and kinetin) and their concentrations (0, 0.1, 0.5, 1.0, and 2.0 mg/L) on multiple shoot production of Citrus spp. 'Sambokam' and 'Byungkyool' by nodal culture. Nodal explants were obtained from in vitro germinated seedlings of both cultivars. 'Sambokam' produced more multiple shoots than did 'Byungkyool' by nodal culture. Among the 3 cytokinins tested in this study BA supplemented in semi-solid MS basal medium was the most effective stimulator for multiple shoot production, and an optimal concentration was determined to be 1.0 mg/L. Shoot elongation and root formation were inhibited by increasing cytokinin concentration, regardless of cytokinin types. BA at 1.0 mg/L produced the most multiple shoots and the highest number of leaves in 'Sambokam', whereas any cytokinin and concentration studied in this experiment did not affect any scored variables such as shoot and leaf numbers, etc. in 'Byungkyool'.

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In Vitro Shoot Multiplication of Albizzia julibrissin Duraz. (자귀나무(Albizzia julibrissin Duraz.)의 기내 대량증식)

  • Ahn, Ji-Young;Kim, Jung-Hee;Kang, Ho-Duck
    • Korean Journal of Plant Resources
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    • v.19 no.4
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    • pp.524-529
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    • 2006
  • In vitro culture system was established to induce multiple shoots of Albizzia julibrissin Duraz. by investigating the effects of cytokinins. Cotyledon, hypocotyl and root explants were cultured on MS media supplemented with either three different plant growth regulators or their combinations. The most effective cytokinin sources were zeatin 2.0 + TDZ 0.5 mg/L in cotyledon, zeatin 1.0 mg/L in hypocotyl, and BA 0.2 + TDZ 0.01 mg/L in root explant for producing shoots ($5.67\;{\pm}\;1.20$, $19.50\;{\pm}\;3.50$, and $20.50\;{\pm}\;2.47$, respectively). Also, zeatin treatment was tended to induce more shoots rather than the combinations of other cytokinins. In addition, the root induced in 1/2 MS medium without any plant growth regulators was longer and thicker than treatments of IBA, NAA, IAA and 2.4-D as auxins. Overall, the highest average percent of in vitro shoot formation was 73% from three different types of explants with treatment of zeatin (1.0mg/L).

Determination of Siderophore from Bacillus Mojavensis Using Liquid Chromatography quadrupole Time-of-flight Tandem Mass Spectrometry (액체크로마토그래피-사중극 비행시간형 탠덤질량분석기를 이용한 Bacillus mojavensis 균주 속 사이드로포어 규명)

  • Cheon, Hae In;Yeo, Mi Seon;Kim, Kang Min;Kang, Jae Seon;Pyo, Jaesung
    • Journal of Life Science
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    • v.29 no.2
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    • pp.198-201
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    • 2019
  • Recently, it has been reported that Bacillus mojavensis possesses antifungal properties and plant growth-promoting activities, which are similar to the characteristics of siderophore. In this study, the siderophore produced by B. mojavensis was assessed using a solid phase extraction (SPE) cartridge and liquid chromatography quadrupole time-of-flight tandem mass spectrometry (Q-TOF MS/MS). After B. mojavensis was incubated in phenol medium for 16 hr and lyophilized, the sample was dissolved in water and loaded to an SPE cartridge to remove interferences. The cartridge was washed with 5% methanol in water and eluted with 2% formic acid in methanol sequentially. The eluted solution was evaporated under a stream of nitrogen gas and reconstituted in methanol. The reconstituted sample was filtered, and $1{\mu}l$ of the sample was assessed using Q-TOF MS/MS. The mass spectrometer was operated using the positive electrospray ionization mode. Based on the mass spectrum and tandem mass spectrum, the siderophore produced by B. mojavensis was bacillibactin, one of the catechol types of siderophore with a molecular weight of 882.2556. This siderophore analysis could provide a justification for the study of B. mojavensis as a functional food and for pharmaceutical applications.