• Title/Summary/Keyword: cotyledon

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Plant Regeneration and Expression of Mouse Adenosine Deaminase Gene in Transgenic Hot Pepper (Capsicum annuum L.) Plants (형질전환된 고추( Capsicum annum L.) 식물체의 Mouse Adenosine Deaminas 유전자 발현)

  • 양덕춘;이계연;유영숙;최경화;임학태
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.1
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    • pp.37-41
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    • 1997
  • The in vitro regeneration and genetic transformation systems in hot pepper(Capsicum annuum L.) have not been routinely available, which has been a major limiting factor in the application of new genetic manipulations. An efficient procedure to regenerate whole pepper plants and to generate transgenic plants expressing a foreign gene was established. A relatively high frequency of plant regeneration was observed when hypocotyl and cotyledon explants were cultured on MS medium supplemented with NAA 0.1 mg/L plus zeatin 2.0 mg/L or IBA 10.0 mg/L plus BAP 1.0 mg/L. Addition of AgNO$_3$5 $\mu$M to these media improved the regeneration frequency up to 8%. For plant transformation, hypocotyl and cotyledon explants of hot pepper were precultured on shoot induction media without kanamycin added for 2 days, and then cocultured with Agrobacterium tumefaciens pDY183 for 2 days. Putative transformants were obtained from selection media containing 100 mg/L kanamycin sulfate and 500 mg/L carbenicillin. Putatively selected transformants were confirmed by amplification of selectable marker genes (ADA and NPT II) by polymerase chain reacion. Successful transcripts of ADA gene were detected by Northern blot analysis. Enzyme activity of ADA was also examined by spectrophotometric analysis, and expression of ADA gene in hot pepper suggests the potential application of ADA gene as a selectable marker in plants.

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Spectrophotometric microtitre assay for rapid screening of membrane-disrupting herbicides (분광분석을 이용한 막과산화작용 제초제의 신속한 검정법)

  • Kwon, Ok-Kyung;Cho, Kwang-Yun;Kim, Jin-Seog
    • The Korean Journal of Pesticide Science
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    • v.4 no.1
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    • pp.11-18
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    • 2000
  • This study was conducted to establish an efficient screening method of membrane-disrupting herbicides, based on the result that membrane leakage leading to evolution of carbonyl compounds and aldehydes could be colorimetrically assessed by formation of stable chromophores with MBTH and ferric chloride. Assay conditions including plant material, light intensity, irradiation time, concentrations of reagents for colorization, and reaction time were optimized, and activities of different types of known herbicides were investigated with cucumber cotyledon discs. That result indicated that this assay was sensitive to membrane-disrupting herbicides, such as protoporphyrinogen oxidase inhibitors (acifluorfen, oxyfluorfen, oxadiazon) and an electron interceptor from photosystem I(paraquat). With seven compounds newly synthesized, highly positive correlation was observed between herbicidal activities measured by greenhouse bioassay and MBTH method. Because only a single disc(4 mm diameter) from cucumber cotyledon in 60 ${\mu}L$ of incubation solution was required to assay, a large number of herbicides could be tested simultaneously using 96-well microtitre plates. But the assay efficiency is not likely to significantly increase because of the boiling step during the reaction.

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Induction of Boiling Stable Proteins by Cold and ABA Treatment in Radish Cotyledon and Hypocotyl (무 자엽과 하배축에서 저온과 ABA처리로 유도된 중탕에 강한 단백질 분석)

  • Cho, Bong-Heuy
    • Analytical Science and Technology
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    • v.13 no.3
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    • pp.346-350
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    • 2000
  • Boiling stable proteins of 53 kDa and 29 kDa existed natively in the cotyledons of Bak Kyoung, fall radish (Raphanus raphanistrodes L.) Boiling stable proteins of 36 kDa and 16.5 kDa were newly induced by cold stress and the proteins of 53 kDa and 29 kDa increased during the cold stress. The proteins of 53 kDa were denatured within 2 hrs after removing cotyledons from plants. Boiling stable proteins of 53 kDa existed natively in the hypocotyls as much as in the cotyledons whereas 24 kDa and 18 kDa proteins were increased by stress. Boiling stable proteins of 53 kDa were induced and those of the 25 kDa and 23 kDa were increased by cold treatment and ABA treatment in the cotyledons of Jangchundaehyung F1 spring white (Raphanus raphanistrodes L.). These results showed the differences of induced boiling stable proteins between fall radishes and spring radishes. Cycloheximide inhibited the induction of 25 kDa and 23 kDa proteins during stress. 22 kDa native protein disappeared during ABA treatment and reappeared by cycloheximide treatments. It may be explained that cycloheximide was responsible for the destruction process of proteins in the living organisms. The profile of boiling stable proteins in hypocotyls of spring radishes during stress was same as that of fall redishes.

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Changes in the Accumulation of Isoflavonoids in Soybeans by Stress (Stress에 의한 대두의 Isoflavonoid 화합물 축적변화)

  • Kim, Jin Tae;Kim, Jang Eok
    • Current Research on Agriculture and Life Sciences
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    • v.12
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    • pp.9-22
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    • 1994
  • The changes of the time-dependent accumulation of isoflavone aglycones(daidzein, genistein) and their glucosides(daidzin, genistin) by various stress-inducing treatment on cotyledon of soybeans(Dankyungkong and Paldalkong) were investigated. Levels of isoflavone aglycones and their glucosides in soybeans treated with UDP-glucose, $MgSO_4$ and $HgCl_2$ and infected with phytopathogen appeared to be higher than those of treatment with distilled water. When compared with data for control, the levels of isoflavone glucosides in citrate-treated soybeans were increased but those of their aglycones did not appear appreciable differences. In Paldalkong treated with UDP-glucose, $MgSO_4$ and $HgCl_2$ the levels of isoflavone aglycones and their glucosides were higher than in Dankyungkong. In particular, the accumulation of daidzein in Paldalkong was significantly higher than in Dankyungkong. By infection with Botrytis cinerea, the maximum amount of accumulation of daidzein in two cultivar did not to be a large different, but accumulation time appeared more rapidly in Paldalkong than Dankyungkong. The accumulation amount of daidzein and genistein in Dankyungkong treated with elicitors appeared to be similar at initial stage, but the level of daidzein after 48hours appeared to be higher than that of genistein. In Paldalkong, the level of daidzein was higher than that of genistein at all stage. The level of daidzein in soybeans infected with phytopathogen appeared to be higher than that of genestein.

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Changes of Nitrogen Compounds and Nutritional Evaluation of Soybean Sprout - Part II. Changes of amino acid composition - (콩나물제조중(製造中) 질소화합물(窒素化合物)의 변화(變化)와 그 영양학적(營養學的) 연구(硏究) - 제2보(第二報). 총(總)아미노산조성(組成)의 변화(變化) -)

  • Yang, Cha-Bum
    • Applied Biological Chemistry
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    • v.24 no.2
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    • pp.94-100
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    • 1981
  • For the chemical assessment of soybean sprouts as a protein food, the changes of amino acid compositions were investigated in soybean as well as both in cotyledon and axis at various growing stages. The total amino acid content per soybean sprout was not changed until 2 days, and thereafter decreased. In cotyledon it is decreased but increased in axis. The ratio of essential amino acid to total amino acids decreased rapidly after 4 days, especially in axis. Aspartic acid increased considerably while glutamic acid decreased. The assessment of soybean sprout with chemical score, A/E and A/T, methods showed that the values were low and that there was no consistency among two methods for sprout products. The limiting amino acid was methionine. Both chemical scores by FAO reference protein and requirement pattern showed similar decreasing order of 2 day-sprout>soybean>4 day-sprout>8 day-sprout>6 day-sprout, while both essential amino acid index and requirement index showed 2 day-sprout>soybeans>4 day-sprout> 6 day-sprout>8 day-sprout.

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Genetic Transformation of Watermelon (Citrullus vulgaris Schard.) by Callus Induction (캘러스 유도에 의한 수박 형질전환)

  • Kwon, Jung-Hee;Park, Sang-Mi;Lim, Mi-Young;Shin, Yoon-Sup;Harn, Chee-Hark
    • Journal of Plant Biotechnology
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    • v.34 no.1
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    • pp.37-45
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    • 2007
  • The genetic transformation of watermelon by Agrobacterium has been known very difficult and a few successful cases have been reported by obtaining the direct shoot formation. However, since this direct shoot formation is not guaranteed the stable transformation, the stable transformation with reproducibility is required by a different approach such as a callus induced manner. The best conditions for inducing the callus from cotyledon and root explants of watermelon were 2 mg/L zeatin + 0.1 mg/L IAA and 2 mg/L BA + 0.1 mg/L 2,4-D, respectively. The GFP expression in the callus was identified and monitored through fluorescent microscopy after transformation with pmGFP5-ER vector. Paromomycin rather than kanamycin was used for selecting the nptll gene expression because it was more effective to select the watermelon explants. Four different callus types were observed and the solid green callus showed stronger GFP expression. The highest frequency of GFP expression in the callus developed from cotyledon was 9.0% (WM8 inbred line), while the highest frequency from root was 8.3% (WM6 inbred line). The WMV-CP was transformed using the method of GFP transformation and the genetic transformation of WMV-CP was confirmed by PCR and Southern blot analysis. Here we present a system for callus induction of watermelon explant and the callus induced method would facilitate the establishment of stable watermelon transformation.

Effect of Electrolyzed Acidic Water on the Growth of Soybean Sprout. (산성 전해수가 콩나물의 생육에 미치는 영향)

  • 윤동준;이정동;강동진;박순기;황영현
    • Journal of Life Science
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    • v.14 no.5
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    • pp.809-814
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    • 2004
  • To investigate the effect of the electrolyzed acidic water for soybean sprouts growth, the responses of characteristics of soybean sprouts were evaluated. Soybean sprouts grown by the electrolyzed acidic water showed shorter length in total body, root, and hypocotyl, etc. but they were evaluated to be increased in hypocotyl diameter and weight per sprout. Total length of soybean sprouts grown for 5 days by electrolyzed acidic water were much shorter than those by tap water. Soybean sprouts grown by tap water showed rapid growth in length even after 5 days but no more growth in length for those grown by electrolyzed acidic water. The growth of hypocotyl showed the same tendency as total length. No difference in root length among the soybean sprouts grown for 4 ~ 11 days by electrolyzed acidic water while those grown by tap water showed continuous rapid growth in length. The diameter of hypocotyl was thicker in those grown by electrolyzed acidic water than those grown by tap water and increased up 5 days. The weight of cotyledon grown by electrolyzed acidic water showed the proportional increase to the growing days but those grown by tap water showed no increase in hypocotyl weight up to 7 days, but a little bit increase after 11 days with the growth of new buds. The fresh weight per sprout was higher in those grown by electrolyzed acidic water until 7 days than tap water but it was the same weight in 11 days cultivation. The electrolyzed acidic water effected on shortening of hypocotyl and root length, thickening of hypocotyl diameter, and enlarging of cotyledon during soybean sprout cultivation.

Sprout Properties and Lipoxygenase Activity of Lipoxygenase-less Soybean Genotypes (Lipoxygenase 결여 콩 계통의 나물 특성 및 Lipoxygenase 활성)

  • Lee Heung Il;Kim Kwang Chul;Park Eui-Ho
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.50 no.spc1
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    • pp.112-117
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    • 2005
  • This study was conducted to see the feasibility of breeding for sprout soybean cultivar with minimum benny flavor using lipoxygenase-less lines. Lipoxygenase-less cultivar Jinpumkong2 was crossed by lipoxygenase containing Gwangankong, Sebaeknamulkong, and Pureunkong as paternal parent and 24 lipoxygenase-less lines derived from those 3 combinations were selected and those lines were evaluated with their parental cultivars. Germination rate showed no difference between lipoxygenase-less lines and their parental cultivars, however, rates of normal sprout of those and Jinpumkong2 were 63 and $56\%$, and were lower than that of paternal parents. Hypocotyl length of those was same as Jinpumkong2, however, shorter than paternal parents. Texture characteristics including hardness, cutting force and mastication of 96 hour-cultured sprout of lipoxygenase-less lines showed higher value than that of their parental cultivars. Lipoxtgenase isozyme was not detected in the sprout cotyledon of lipoxygenase-less lines, however it was observed in the sprout hypocotyl of all the used genotypes. Though lipoxygenase activity in the seed of lipoxygenase-less lines was lower than that of Jinpumkong2(0.477, ${Delta}A$ 234 nm min-1 mg meal-1),2 lines revealed more than 0.5 value. Lipoxygenase-activity of 2 day-cultured sprout(both cotyledon and hypocotyl) was the highest, decreased in 3 days after culture and re-increased thereafter. Several lipoxygenase-less lines with lower lipoxygenase activity of sprout than Jinpumkong2 were selected and this suggested the possibility of breeding lines for soy-sprout with low benny flavor.

Study on functional improvement of peanut sprouts by LEDs

  • Shin, So-Hee;Choi, So-Ra;Song, Eun-Ju;Song, Young-Eun;Choi, Min-Kyung;Han, Hyun-Ah;Lee, Ki-Kwon;Lee, In-Sok;Chung, Nam-Jin
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.277-277
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    • 2017
  • The research was carried out to investigate a total polyphenol content, antioxidant activity, amino acid and resveratrol content of peanut sprouts (cotyledon, epicotyl, leaf, hypocotyl, root), in different light (white, blue, red, F-red, UV-A, UV-B, UV-C) conditions for 24 hours. Peanut seeds were sown on a $27.5{\times}15.9{\times}13cm$ tray and grown at the $25^{\circ}C$ under the dark condition for 14 days. Total polyphenolic contents of epicotyl and leaf were about 288mg GAE/100g in blue light. The DPPH radical scavenging of cotyledon and hypocotyl were 1.3~1.5 times (63%) and 2 times (40%) compared to control (43%, 19%), respectively. As to ABTS activity, its activity was increased by all LEDs treatment, Especially, the highest ABTS activity of the hypocotyl and leaf was shown as 99.1% in blue light. The essential amino acid content of hypocotyl and leaf was increased 1.9 times in the UV-B, 1.6 times in red, and 1.5 times in F-red, respectively. The non-essential amino acid content was increased by all LEDs treatment in hypocotyl and leaf. The content of resveratrol was increased by 1.3 times in UV-B compared to that of other tissues. Assessing inclusively, this study showed that there was a significantly positive effect between increase of physiological substance activity and LED light treatment, resulting in stably producing peanut sprouts. Therefore, a material treated with LEDs is thought to be useful as a functional food resources.

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Factors Influencing Somatic Embryo Induction and Plant Regeneration in Aralia elata Seem. (두릅(Aralia elata)의 체세포배 유도, 발아 및 식물체 재분화에 미치는 요인)

  • 문흥규;오경은;손성호
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.4
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    • pp.275-280
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    • 1999
  • In order to find optimum conditions for somatic embryogenesis from different individual (2-year-old) in Aralia elata were cultured on MS medium supplemented with 1.0 mg/L 2,4-D, 3% sucrose, and 0.3% gelrite. We also investigated the effect of MS medium salt concentration, BA and ABA on the embryo germination and plant regeneration. While noticeable difference was observed on somatic embryo induction among different individual tree, no apparent difference was seen in both germination and regeneration frequencies. Compared with nonembryogenic calli, embryogenic calli tended to look yellow and/or pale brown in color, slowly growing and soft in their texture. Regardless of BA or ABA treatment, half-strength MS salt medium proved to be better than full strength MS medium in both embryo germination and plant regeneration. Both hypocotyl and cotyledon developments were slightly promoted by adding 0.1 mg/L BA. However, its effect on germination and regeneration seemed inferior to control. ABA treatment on somatic embryos at their torpedo and early cotyledonary stages resulted in poor response in germination and regeneration. Although most regenerated plantlets varied greatly in cotyledon number and shape, they could be developed into normal plants after 4 weeks in culture. More than 95% plantlets were acclimatized in an artificial soil mixture, successfully transplanted to nursery bed and grew normally without any phenotypic abnormalty.

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