• Title/Summary/Keyword: cotyledon

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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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Growth Responses of Petunia as Affected by Several Pollutants in Vitro (Petunia 조직배양시 몇가지 공해물질이 생장반응에 미치는 영향)

  • 백기엽;최성렬;이재욱;최주견
    • Journal of Plant Biology
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    • v.27 no.3
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    • pp.139-147
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    • 1984
  • This in vitro study was employed to clarify the effects of several pollutants i.e. $SO_2$, fluoride, cadmium(Cd), aluminum(Al) and NaCl, on the organogenesis and growth responses of shoot-tip, stem and multiple-buds segments derived from hypocotyl or cotyledon culture of petunia seedlings. ${Na_2}{SO_3}$levels of more than 200$\mu{g}$/ml had significantly reduced organogenesis, growth, and chlorophyll content. The injuries caused by ${Na_2}{SO_3}$, concentration of more than 400$\mu{g}$/ml were alleviated by increasing hydrogenion concentration of medium, indicating some relationship between two factors. Organogenesis was not affected by the fluoride concentration up to 100ppm in the media, but the growth and chlorophyll content were greatly reduced by the fluoride. The effect of Cd depended on the explant sources used for the culture; 1.0ppm was effective for fresh weight increase in shoot tip culture, and 3.0ppm in stem segments culture. Organogenesis and growth were greatly reduced by more than 10.0 Cd treatment. Growth and formation of shoots were better with Na conc. of 0.3% compared to control, but those of roots were inhibited. Na concentration goes over 1.0%, organogenesis and subsequent growth were inhibited, and chlorophyll synthesis was drastically reduced. Chlorophyll content was increased on the medium supplemented with Al 50$\mu{g}$/ml compared to control. However the formation and growth of shoots were greatly inhibited with more than 400$\mu{g}$/ml and roots were not produced at all.

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Antioxidant Potentials and Quantification of Flavonoids in Mung Bean (Vigna radiata L.) Seeds

  • Kim Dong-Kwan;Kim Jung-Bong;Chon Sang-Uk;Lee Ya-Seong
    • Plant Resources
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    • v.8 no.2
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    • pp.122-129
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    • 2005
  • Mung bean (Vigna radiata L.) is an increasingly important human food source, as well as a new functional agent, mainly due to its potent antioxidant activity. This study was conducted to determine antioxidant activity of fractions from mung bean seeds by measuring DPPH radical scavenging activity and to quantify the flavonoids by means of HPLC analysis. Vitexin and isovitexin were present in both ethanol and water extracts in highest amount. Flavonoids, vitexin and isovitexin were quantified from 195 germplasms of mung beans and their concentrations varied by 4.7 fold. Especially, the breeding line KM99004-4B-2 (Suwon28/KM94004), which has grown in Jeollanamdo Agricultural Research and Extension Services, showed the highest amount (15.88 mg/g) of total flavonoids. The vitexin portion was averaged $70.73{\pm}1.38\%$. High positive correlation $(r=0.96^{***})$ between vitexin and isovitexin contents showed. However, the flavonoid content showed very low correlation with the 24 growth and ecological characteristics. Seed coats of mung beans had the highest flavonoid amount, showing $50\~70$ times more than cotyledons. Flavonoid contents in the seed, the cotyledon, and the seed coat were decreased as the seed imbibition time increased.

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Studies on the Free Amino Acids Metabolism in Germinating Mung Bean by Paper Chromatography. (Part $1{\sim}2$) (Parkt 1). Variation of free Amino acids and Amides contents in germinating Mung Bean (페파크로마토그라피에 依한 發芽綠豆의 遊離아미노酸代謝의 硏究 (第 1 報${\sim}$第 2 報) (第 1 報) 綠豆發芽에 따르는 遊離아미노酸 及 아미드의 變動에 關하여)

  • Kim, Tae-Rin;Song, Chang-Won
    • Journal of the Korean Chemical Society
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    • v.5 no.1
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    • pp.9-14
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    • 1961
  • Ethanol extracts of Mung Bean seeds and seedings were analysed by 2-dimensional and circular paper partion chromatography for Nitrogen compounds as a part of the study on the Amino acids metabolism. In the seeds, 18 ninhydrin positive substances were present, before germination, but the number increased to 21 after germination. There were 3 unknown substances and one of it formed newly after germination. After 2-days germination, the amount of amides, such as Asparagine and Glutamine. where increased very large which were very small amount before it. Those were accumulated more in dark place than in light and the amount of Asparigine were more than that of Glutamine. Through the germination, there were large amount of Glutarmic acid, Aspartic acid and Alanine which seems to be concerned in transamination reaction in seedings. Valine, Leucine, and Phenylalanine increased to considerable amount after germination. This is very remarkable fact as those Amino acids were reported to be concerned in transamination reaction recently. ${\gamma}$-amino butyric acid was detected in both Cotyledon and Embroy through the germination. It seemed that there is no any Nitrogen Metabolism in the unbroken seed even if it is preserved very long period.

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Identification and Changes of Physiologically Active Substances During Chilling Storage of Dehisced Ginseng Seeds (저온저장중 개갑인삼종자내의 생리활성물질 동정 및 변화)

  • Kwon, Woo-Saeng;Baek, Nam-In;Lee, Jung-Myung
    • Journal of Ginseng Research
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    • v.21 no.1
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    • pp.13-18
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    • 1997
  • Identification and changes of physiologically active substances during chilling storage of dehisced ginseng (Panax ginseng C. A. Meyer) seeds were analyzed using various preparatory separation methods and purification columns; Dowex 50W and silica gel columns. Seven components with Rf values of 0.20, 0.40, 0.58, 0.66, and 0.70 In solvent system, $CHCl_3$:MeOH=3:1 (v/v), Rf values of 0. 63 and 0.74 in solvent system, $CHCl_3$:MeOH:$H_2O$:=7:3:1 (v/v) were obtained through Dowex 50W and silica gel column chromatographies. Two components with Rf values of 0.20 and 0.63 in the all chilling treatments were detected in the extract obtained through both chromatographies, and only the former component was gradually increased till 4 weeks of chilling storage and then rapidly decreased from 8 to 16 weeks. UV spectra of Rf values of 0.66 and 0.56 were similar to that of cytokinin, but their physiological activities were not found. Rf values of 0.20 showed activity by radish cotyledon expansion bioassay. The component with Rf value of 0.20 was revealed to have a naphthalene in the proposed chemical structure by various NMR techniques.

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Somatic Embryogenesis and Plant Regeneration from Immature Zygotic Embryo Culture of Wasabia japonica Matsum. (고추냉이의 미숙배배양으로부터 체세포배 발생과 식물체 재분화)

  • 은종선;고정애;김영선;김명준
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.4
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    • pp.207-211
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    • 1995
  • Immature zygotic embryos from immature seeds of Wasabia japonica (cv Dalma) were isolated and cultured on modified MS medium supplemented with 2,4-D, IAA, and BA. Immature zygotic embryos were classified into torpedo shape and cotyledon stage. The highest rates of callus formation were obtained of 1.0mg/L IAA(torpedo stage, 90.0%)and 1.0mg/L 2,4D plus 0.1mg/L BA(cotyledany stage,84.3%). Somatic embryos after 60 days of culture. These numerous somatic embryo could be seperated and subcultured on the same media for further propagation. After 90 days of culture, most somatic embryos were developed well organized embryos which were able to produce into whole plants.

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Influence of formulated organic Plant tissue culture medium in the shoot regeneration study of Brassica juncea (l.) - Indian mustard

  • Kashyap, Suman;Tharannum, Seema;R, Taarini
    • Journal of Plant Biotechnology
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    • v.46 no.2
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    • pp.114-118
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    • 2019
  • Efficient protocol for plant shoot regeneration of Brassica juncea L. CZERN was established by using organic media components and growth stimulating factors of the vermicompost and coelomic fluids. Formulated organic plant tissue culture media (Vermicompost (30%) extracts supplemented with 20 mL/L coelomic fluid) have shown maximum shoot regeneration when compared with the Murashige and Skoog (MS) medium, which were supplemented with 1 mg/L 6-benzyladenine (BA) and 0.1 mg/L of Naphthaleneacetic acid (NAA). Cotyledon explants produced the highest shoot regeneration frequency from fourday-old germinated seedlings in comparison with non-germinated seedlings. The vermicompost extracts have proved to be the best organic plant growth media to induce shoots from cotyledons compared to the MS media. Statistically significant difference (P = 0.008) for the root length, shoot length (P=0.000350) and the leaves (P=0.375) of the mustard plantlets were analyzed successfully. The survival rate was 98% in the mustard cotyledons on the Vermicompost extract media and 63% on MS media respectively. The coelomic fluid also is much suitable to induce shoots from cotyledons at lower concentrations. It was also shown that the vermicompost extract, which comprised of humic acids along with coelomic fluid, affected shoot regeneration from the cotyledons. An efficient and organic shoot regeneration study was standardized and it can be applicable in the improvement of the economically important crops.

Inheritance of Shoot-Lacking Phenomenon in Interspecific Hybrids of Squash (호박 종간잡종에서 생장점 결여현상의 유전)

  • Hong, K.H.;Huh, Y.C.;Woo, Y.H.;Lee, K.H.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.12 no.1
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    • pp.47-52
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    • 2010
  • Squash cultivars of Cucurbita moschata have good fresh quality, but their viny growth habit hinders wide cultivation under structure. On the other hand, 'Zucchini' varieties of C. pepo have bush growth habit and better growth ability under the low temperature condition than C. moschata, but have inferior flesh quality. To combine desirable characteristics of the two species, many attempts have been exerted to obtain interspecific hybrids. A shootless phenomenon was found among segregating populations; the cotyledon expanded normally but further shoot growth. The inheritance of the character was investigated in F2 and BC1F2 populations. The segregation ratios indicated that the shoot-lacking characteristics is expressed by double recessive gene pairs.

Effect of Plant Growth Regulator Treatments on the Growth and Lateral Root Formation in Soybean Sprouts - I. Effect of Plant Growth Regulator Treatments on the Growth in Soybean Sprouts (생장조절물질(生長調節物質) 처리(處理)가 콩나물의 생육(生育) 및 세근발생(細根發生)에 미치는 영향(影響) - I. 생장조절물질(生長調節物質)의 단용(單用) 및 혼용처리(混用處理)가 콩나물의 생육(生育)에 미치는 효과(效果))

  • Kang, C.K.;Lee, J.M.;Saka, H.
    • Korean Journal of Weed Science
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    • v.9 no.1
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    • pp.56-68
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    • 1989
  • aA series of experiments were conducted to investigate the effect of plant growth regulator treatments on the growth and lateral root formation in soybean sprouts in order to establish the effective method of producing root-less or short-rooted soybean sprouts with larger diameter in the hypocotyl. Major results can be summarized as follows. 1. Soybean sprouts showed fairly uniform elongation rate from 3 to g days after imbibition with daily increase of 3.8cm. The speed of elongation of hypocotyl was reduced whereas that of root accelerated 7 days after imbibition. Lateral roots began to emerge fairly evenly from 5 to 9 days after imbibition with a daily increase of 4.4. 2. Auxins(IAA, IBA, NAA, 2,4-D) inhibited hypocotyl elongation and formation of lateral roots and increased hypocotyl diameter without influencing root length and hook diameter at higher concentrations. The dry weight of cotyledon was increased significantly as compared to that of hypocotyl and root. Among the tested auxins, 2, 4-D was the most effective. 3. BA and 4PU-30 significantly reduced elongation of hypocotyl and root and resulted in the biggest diameter of hypocotyl when treated at higher concentrations. The lowest effective concentration of BA to prevent the formation of larval gal roots was 12.5ppm. The formation of lateral roots could be completely prevented by BA and 4PU-30 treatment but kinetin, zeatin, zeatin riboside resulted in many lateral roots and increased thickness of soybean sprouts with little influence. Cotyledon deformation was found in soybean sprouts treated by 4PU-30. 4. 2, 4-D was the most effective for increasing the hypocotyl diameter while 4PU-30 was the most effective for reducing no. of lateral roots. 5. It can be concluded that among the plant growth regulators tested, BA was effective in reducing root length and increasing hypocotyl diameter. BA 12.5 ppm or 15 ppm may thus be the more practical for production of soybean sprouts. 6. ABA showed no significant effect of growth parameter, however ABA 25 ppm inhibited only no of lateral roots with little influence on the growth of seedling. 7. Ethephon inhibited the elongation of hypocotyl and root and increased hypocotyl diameter at higher concentrations. 8. The combined effect of cytokinins and ethephon was very similar to result of BA treatment alone. As the ethephon concentration increased, hypocotyl diameter and dry weight of cotyledon tended to increase.

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Establishment of an Efficient Agrobacterium Transformation System for Eggplant and Study of a Potential Biotechnologically Useful Promoter

  • Claudiu Magioli;Ana Paula Machado da Rocha;Pinheiro, Marcia-Margis;Martins, Gilberto-Sachetto;Elisabeth Mansur
    • Journal of Plant Biotechnology
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    • v.2 no.1
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    • pp.43-49
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    • 2000
  • An efficient and reliable Agrobacterium transformation procedure based on TDZ (thidiazuron)-induced organogenesis was established and applied to six Brazilian eggp1ant varieties. Optimum transgenic plants recovery was achieved upon the study of the following parameters affecting transformation efficiency, using F-100 variety as a model: i) explant source; ii) pre-culture period; iii) physical state of the pre-culture medium and iv) coculture conditions. The highest frequency of kanamycin-resistant calli derived from leaf explants (5%) was obtained without a pre-culture period and co-cultivation for 24 h in liquid medium followed by five days on solid RM (regeneration medium). For cotyledon explants, best results were achieved upon a pre-culture of 24 h in liquid RM and a co-cultivation period of 24 h in liquid RM followed by three days in solid RM, resulting in a transformation Sequency of 22.7%. Kanamycin-resistant organogenic calli were also obtained from cultivars Emb, Preta Comprida, Round nose Shaded, Campineira and Florida Market. The expression pattern of an epidermis-specific promoter was studied using transformants expressing a chimaeric construct comprised by the promoter Atgrp-5 transcriptionally fused to the coding region of the gus gene. The expression pattern was similar to that previously observed in tobacco and Arabidopsis thaliana, with preferential expression at the epidermis and the stem phloem. These results support the idea that the Atgrp-5 promoter can be used to drive defense genes in these tissues, which are sites of pathogen interaction and spread, in programs for the genetic improvement of eggplant.

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