• Title/Summary/Keyword: Soybean callus

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The Factors on Somatic Embryogenesis of Soybean [Glycine max. (L.) Merrill]

  • Kim, Kyong-Ho;Kim, Hag-Sin;Oh, Young-Jin;Suh, Sug-Kee;Kim, Tae-Soo;Park, Ho-Kee;Park, Moon-Soo;Kim, Seok-Dong;Yeo, Up-Dong
    • Journal of Plant Biotechnology
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    • v.2 no.3
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    • pp.123-128
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    • 2000
  • To enhance in vitro plantlet regeneration efficiency of soybean through embryogenesis, the culture conditions such as material part and size of immature seed, 2,4-D, pH and solidifying agents for somatic embryogenesis were investigated. Somatic embryogenesis was induced from the immature embryo, immature cotyledon and embryonic axis explants of the immature seed on MS medium supplemented with 2.0 mg/L 2,4-D. The highest rate (up to 22.9%) of somatic embryogenesis was obtained from the immature cotyledon, following embryonic axis and the immature embryo. The rate varied with the developmental stages of seed. The maximum rate (25.4%) of embryogenesis was obtained from 3-4 mm length of the seed (after 25 days of flowering). The optimum concentration of 2,4-D for embryogenesis was 10 mg/L. The optimum pH was at 5.8 and solidifying agent for medium was better with 0.4% gelrite than with agar. For rapid multiplication of shoot tips from the germinating somatic embryos, they were cultured on MS medium containing 2 mg/L indole-3-butyyic acid (IBA) and 1 mg/L 6-benzyladenine (BA). After then somatic embryos with one and three cotyledons were transferred to the growth regulator free medium. The medium exhibited the higher rate (ca. 50%) of development than the multiplication medium.

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Effect of No-Tillage on Soybean Yield and Weed Emergence in Drained Paddy Field Condition in Jeonnam Province (전남지역 논 조건에서 무경운 재배가 콩의 생육, 수량 및 잡초 발생에 미치는 영향)

  • Kim Dong-Kwan;Chon Sang-Uk;Heo Buk-Gu
    • The Korean Journal of Community Living Science
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    • v.17 no.3
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    • pp.89-97
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    • 2006
  • This study was conducted to compare soybean growth and yield and the degree of weed emergence according to no-tillage and conventional tillage system in two different drained paddy fields, loam of Chilgok series and silty clay loam of Deokpyeong series. In both soil conditions, the maturing time of the soybeans by the no-tillage system was two days earlier than that by the conventional tillage system. In the loam of Chilgok series, the stem length of the soybeans in the no-tillage system was 5.7 cm longer than that in the conventional tillage system. The miss-planted rate and diseased plants of black root rot (Calonectria iliacola) in the no-tillage system were 9.2% and 2.8% lower, respectively than those in the conventional tillage system. Also, the nodulation and seed yield in the no-tillage system were 32% and 13% more, respectively, than those in the conventional tillage system. In the silty clay loam of Deokpyeong series, the stem length of the soybeans in the no-tillage system was 4.6cm shorter than in the conventional tillage system. The diseased plants of black root rot (Calonectria iliacola) in the no-tillage system were 4.2% lower than those in the conventional tillage system. Also, no significant difference in the seed yield between the no-tillage and conventional tillage systems was observed. On the other hand, there was a lower occurrence of weed in the no-tillage system than in the conventional tillage system, and the income increased by 19% owing to yield increase and reduced management costs.

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Isolation and Culture of Protoplasts from Hypocotyl-derived Callus of Soybean (Glycine max) (대두 (Glycine max) 부배유 유래 칼루스의 원형질체 분리 및 배양)

  • 이광웅
    • Journal of Plant Biology
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    • v.28 no.3
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    • pp.233-241
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    • 1985
  • The isolation and culture of protoplasts from hypocotyl-derived calluses of Glycine max (L.) Merr. cv. Jangyeop were obtained by digestion for 6 hrs in an enzyme solution containing 3.5% cellulase, 1.5% macerozyme, 10% sorbitol and 0.1% CaCl2.2H2O at pH 5.8. Newly formed cell wall of protoplasts cultured in MS agar medium containing 10 $\mu$M $\alpha$-naphthaleneacetic acid (NAA) and 32 $\mu$M N6-benzylaminopurine (BAP) could be observed after 24 hrs culture. The first cell division of the protoplasts was observed after 3 days of culture; cell clusters after 2 weeks of culture. When transferred to solid media, the protoplasts formed cell clusters gave rise to proliferating calluses.

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Phytotoxic Effects of Xanthium occidentale Extracts and Residues on Seedling Growth of Several Plant Species

  • Chon Sang-Uk
    • Plant Resources
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    • v.8 no.2
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    • pp.116-121
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    • 2005
  • Compositae plants are known to contain biologically active substances that are allelopathic to agricultural crops as well as weed species. Aqueous or methanol extracts and plant residues from leaves of Xanthium occidentale were assayed against alfalfa (Medicago sativa) to determine its allelopathic effects, and the results showed that the extracts applied onto filter paper significantly inhibited seed germination as well as root growth of alfalfa. Aqueous leaf extracts of $40g\;L^{-1}$ were completely inhibitory on root growth of alfalfa, while root growths of barley (Hordeum vulgare L.) and soybean (Glycine max L.) were less sensitive. Leaf residue incorporation at $100g\;kg^{-1}$ into soil on seedling growth of barnyard grass (Echinochloa crus-galli Beauv. var. oryzicola Ohwi) inhibited both shoot and root fresh weights of barnyard grass by 94 and $96\%$, respectively. Methanol extracts from BuOn and EtOAc fractions of X. occidentale reduced alfalfa root growth more than did those from hexane and water fractions. The results based on bioassay of extracts and residues show that X. occidentale had potent an allelopathic activity against other plant species.

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Amino Acid Biosynthesis and Gene Regulation in Seed (종자내 아미노산 합성 조절 유전자에 관한 연구)

  • ;;;;;Fumio Takaiwa
    • Proceedings of the Botanical Society of Korea Conference
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    • 1996.07a
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    • pp.61-74
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    • 1996
  • Human and monogastric animals can not synthesize 10 out of the 20 amino asids and therefor need to obtain these from their diet. The plant seed is a major source of dietary protein. It is particular important in their study to increase nutritional quality of the seed storage proteins. The low contents of lysine, asparagine and threonenein various cereal seeds and of cystein and methionine. In legume seeds is due to the low proportions of these amino acids in the major storage proteins, we have tried to apply the three strategies; (1) mutagenesis and selection of specific amino acid analogue resistance, (2) cloning and expression study of lysine biosynthesis related gene, (3) transfomation of lysine rich soybean glycinin gene. The 5-methyltryptophan (5MT) resistant cell lines, SAR1, SAR2 and SAR3 were selected from anther derived callus of rice (Oryza sativa L. "Sasanishiki"). Among these selected cell lines, two (SAR1 and SAR3) were able to grow stably at 200 mg/L of 5MT. Analysis of the freed amino acids in callus shows that 5MT resistant cells (SAR3) accumulated free tryptophan at least up to 50 times higher than those that of the higher than of SAS. These results indicated that the 5MT resistant cell lines are useful in studies of amino acid biosynthesis. Tr75, a rice (Oryza sativa L., var. Sasanishiki) mutant resistant to 5MT was segregated from the progenies of its initial mutant line, TR1. The 5MT resistant of TR75 was inherited in the M8 generations as a single dominant nuclear gene. The content of free amino acids in the TR75 homozygous seeds increased approximately 1.5 to 2.0 fold compared to wild-type seeds. Especially, the contents of tryptophan, phenylalanine and aspartic acid were 5.0, 5.3 and 2.7 times higher than those of wild-type seeds, respectively. The content of lysine is significantly low in rice. The lysine is synthesized by a complex pathway that is predominantly regulated by feedback inhibition of several enzymes including asparginase, aspatate kinase, dihydrodipicolinat synthase, etc. For understanding the regulation mechanism of lysine synthesis in rice, we try to clone the lysine biosynthetic metabolism related gene, DHPS and asparaginase, from rice. We have isolated a rice DHPS genomic clone which contains an ORF of 1044 nucleotides (347 amino acids, Mr. 38, 381 daltons), an intron of 587 nucleotides and 5'and 3'-flanking regions by screening of rice genomic DNA library. Deduced amino acid sequence of mature peptide domain of GDHPS clone is highly conserved in monocot and dicot plants whereas that of transit peptide domain is extremely different depending on plant specie. Southern blot analysis indicated that GDHPS is located two copy gene in rice genome. The transcripts of a rice GDHPS were expressed in leaves and roots but not detected in callus tissues. The transcription level of GDHPS is much higher in leaves indicating enormous chloroplast development than roots. Genomic DNA clones for asparaginase genes were screened from the rice genomic library by using plaque hybridization technique. Twelve different genomic clones were isolated from first and second screening, and 8 of 12 clones were analyzed by restriction patterns and identified by Southern Blotting, Restriction enzyme digestion patterns and Southern blot analysis of 8 clones show the different pattern for asparaginase gene. Genomic Southern blot analysis from rice were done. It is estimated that rice has at least 2-3 copy of asparaginase gene. One of 8 positive clones was subcloned into the pBluescript SK(+) vector, and was constructed the physical map. For transformation of lysine rich storage protein into tobacco, soybean glycinin genes are transformed into tobacco. To examine whether glycinin could be stably accumulated in endosperm tissue, the glycinin cDNA was transcriptionally fused to an endosperm-specific promotor of the rice storage protein glutelin gene and then introduced into tobacco genomic via Agrobacterium-mediated transformation. Consequently the glycinin gene was expressed in a seed-and developmentally-specific manner in transgenic tobacco seeds. Glycinin were targeted to vacuole-derived protein bodies in the endosperm tissue and highly accumulated in the matrix region of many transgenic plant (1-4% of total seed proteins). Synthesized glycinin was processed into mature form, and assembled into a hexamer in a similar manner as the glycinin in soybean seed. Modified glycinin, in which 4 contiguous methionine residues were inserted at the variable regions corresponding to the C - teminal regions of the acidic and basic polypeptides, were also found to be accumulated similarly as in the normal glycinin. There was no apparent difference in the expression level, processing and targeting to protein bodies, or accumulation level between normal and modified glycinin. glycinin.

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The Establishment of Nitrogen Fixation by Cultured Cell-Rhizobium Association Through Tissue Culture Technique in Soybean (대두조직배양세포(大豆組織培養細胞) - Rhizobium에 의(依)한 질소고정력(窒素固定力))

  • Kang, Sang Jae;Park, Woo Churl
    • Current Research on Agriculture and Life Sciences
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    • v.4
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    • pp.27-35
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    • 1986
  • This experiment was carried out to elucidate the factor of nitrogenase formation and to establish the nitrogen fixation system in mixed culture of cultured cells and rhizobia through tissue culture technique using three soybean varieties, Hwangkeum, Namcheon and D 68-0099 as host plants. The results obtained were as follows; The callus was induced in embryo and radicle, but not in hypocotyl. The most favorable callus induction was caused by the individual application of 2,4-D and NAA at the concentration of 2mg/1 and 4mg/1, respectively, but in case of treating both 2,4-D and kinetin, that was done at the concentration of 0.2mg(2,4-D)/0.05mg(kinetin)per liter. The growth of cultured cell was good at the concentration of 2.0mg(2,4-D)/1 and 0.2mg(2,4-D)/0.05mg(kinetin)per liter. When cultured cells were inoculated with R. japonicum 019 and 011, their growthes were considerably inhibited. The addition of single amino acid inhibited the growth of cultured cells. Hwangkeum was inhibited considerably by methionine and leucine. The inhibition of growth by single amino acid can be abolished by the addition of certain amino acids. The differentiation of adventitious root was good at the concentration of 2.0mg 2,4-D and 0.2mg 2,4-D/0.05mg kinetin per liter. Of three host plants tested with 25 R. japonicum strains, Hwangkeum had affinity for 10 strains, Namcheon for 7 strains and D68-0099 for none. The nitrogen fixing abilities of Hwangkeum and Namcheon caused by cultured cell-Rhizobium association were high in strain 019, 007, and in 007 mixed with 119, respectively.

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Optimization of Genetic Transformation Conditions for Korean Soybean Cultivars (국내 콩(Glycine max) 품종 형질전환 초기조건 확립)

  • Lee Ki-Jung;Seo Jen-Kyung;Lee Hye-Young;Jeon Eun-Hee;Shin Sang-Hyun;Lee Jai-Heon;Kim Doh-Hoon;Ko Jong-Min;Hahn Won Young;Baek In-Youl;Oh Boung-Jun;Chung Young-Soo
    • Journal of Life Science
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    • v.16 no.2 s.75
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    • pp.289-296
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    • 2006
  • In order to establish highly efficient gene transfer condition at early stage of soybean transformation, various experiments were performed and compared their efficiencies by transient GUS analysis; those conditions are genotype determination of Korean soybean cultivars for amenability to Agro-infection, appropriate agar and selective agent concentration, orientation of explant placement, hormone pre-culture, and liquid selection condition. In the genotype screen of Korean soybean varieties, 14 amenable genotypes were selected. For efficient Agrobacterium washing, cefotaxime was chosen and hygromycin at the concentration of 10 and 15 ppm was used as selection agent in the media. Agar concentration was slightly better in 0.6% and 0.8% for both shoot and callus formation, and explant placement with adaxial side down showed high frequency of GUS expression. For wounding treatment, oriental needle was efficient than scalpel for shoot formation and gene transfer. To increase the frequency of gene transfer, hormone pre-treatment was applied. BA at the concentration of 5 and 10 ppm resulted in better survival at the late stage of selection in shoot elongation media. Selection in liquid media after hormone pre-treatment seemed to be effective to remove the escaped non-transformants at early stage of procedure. Considering the results obtained, Eunhakong could be the first choice as a material for soybean transformation among Korean soybean genotypes.

Characteristics and Possible Early Harvesting Time of Early Maturing Soybean Cultivars in Southern Korea (남부지방에서 조생종 콩 품종의 특성과 조기수확 한계기)

  • Kim, Dong-Kwan;Son, Dong-Mo;Chon, Sang-Uk;Lee, Kyung-Dong;Kim, Kyong-Ho
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.53 no.2
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    • pp.125-130
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    • 2008
  • This study analyzed the growth, seed quality, and yield of major early-maturing soybean cultivars by comparing them in order to utilize the research results in the selection of early-maturing soybean cultivars in multi-cropping farms in the Southern area. This field trial was conducted at Naju region (latitude $35^{\circ}04'N$, longitude $126^{\circ}54'E$), Jeonnam, with planting on June 15. The maturing date for Keunol-kong and Hwaseong-put-kong was found to be around September 12, which was earlier than other cultivars. Thus, there were advantages to introducing a cropping system as well as having good seed quality and high yield. On the other hand, the maturing date for Saeol-kong and Sinrok-kong was found to be around September 20, which was a little bit late; however, the seed quality of the cultivars was good and they had a high yield. Therefore, if we want to sow the following crops of soybeans around mid-September, Keunol-kong and Hwaseong-put-kong are advantageous, while for the seeding around late September, Saeol-kong and Sinrok-kong would be good. This study was also performed to identify the limitation time for early harvesting by reviewing seed quality and yield of major early-maturing soybean cultivars according to early harvesting. When harvesting Keunol-kong on September 6, which was six days earlier than the optimal harvesting time (September 12), there was no difference in seed weight, yield, or seed quality than those of the harvested at the optimum maturing time. As for Saeol-kong, when harvesting on September 18, which was six days earlier than the optimal harvesting time (September 24), there was no difference in seed weight, yield, or seed quality than those of the harvested at the optimum maturing time. Therefore, the stable limitation time for early harvesting of Keunol-kong and Saeol-kong was concluded to be six days earlier than the optimal harvesting time.

Somatic Embryogenesis in Immature Zygotic Embryo Cultures of Korean Soybean (Glycine max L.) Culitivars and Effect of 2,4-Dichlorophenoxyacetic Acid on Somatic Embryo Morphology (한국 품종 대두(Glycine max L) 미숙배로부터 체세포배발생과 배지의 2,4-Dichlorophenoxyacetic Acid 농도가 체세포배의 형태에 미치는 영향)

  • Pil S Choi;Yoong Y. SOH;Duck Y.Choi;ang R. LIU
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.1
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    • pp.7-14
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    • 1994
  • Immature zygotic embryos of five Korean soybean cultivars cultured on Murashige and Skoog's (MS) medium supplemented with various concentrations of 2,4-Dichlorophenoxyacetic acid (2,4-D) produced somatic embryos without forming an intervening callus. The highest frequency (up to 90%) of somatic embryo formation was obtained when zygotic embryos were cultlued on medium containing 1 to 2 mga 2, 0-D in four cultivars. However the frequency was highly variable to the cultivars. Transversely sliced primary somatic embryo halves were also capable of forming secondary embryos at frequencies of up to 70% when cultured on medium containing 0.1 to 1 mg/L 2,4-D. Somatic embryos formed on zygotic embryos cultured on medium containing 0.1 to 0.2 mg/L 2,4-D had two cotyledons more frequently than one horn-type cotyledon and those on medium containing 0.5 to 4mg/L 2,4-LD had a horm-type cotyledon at a prominently higher freequency. However somatic embryos on medium containing 10mg/L or higher concentrations of 2,4-D were usually shunted at the globular stage even after transfer to medium containing lower concentrations of 2,4-D or other growth regulators. non somatic embryos with one or two cotyledons or a hem-type cotyledon were transferred to medium containing $GA_3$, those with two cotyledons converted to plantlets at a higher frequency (25%) than the others.

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Herbicidal and Insecticidal Potentials of 5-Aminolevulinic acid, a Biodegradable Substance (생분해성 생리활성물질 5-aminolevulinic acid의 제초 및 살충활성)

  • Chon, Sang-Uk
    • The Korean Journal of Pesticide Science
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    • v.11 no.1
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    • pp.52-58
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    • 2007
  • ALA (5-aminolevulinic acid) has been proposed as a tetrapyrrole-dependent photodynamic herbicide and insecticide by the action of the protoporphyrinogen IX oxidase (Protox IX). The present study was conducted to determine growth responses of plant and insects to ALA, biodegradable biopesticidal substance. In the paddy condition experiment, plant height and shoot fresh weight of barnyardgrass (Echinochloa crus-galli) was more reduced by ALA than rice plants, even though both plant species show great phytotoxicity. Hairy crabgrass (Digitaria sanguinalis), a monocot weed, was more sensitive to ALA at 5mM under upland condition when ALA applied on the foliage, compared with soybean (Glycine max) as a dicot crop. ALA solutions were tested for their insecticidal and larvicidal activities against Spodaptera exigua (Hubner) and Tetranychus urticae Koch. by foliar application and leaf-dipping method. The result showed higher insecticidal activity of ALA at 10mM and its mixture with insecticide luferon against S. exigua. Strongest insecticidal activity against T. urticae was observed from the ALA solution at 10mM 72 days after application. This results show that ALA solution had potent herbicidal and insecticidal activities against agricultural pests even though their activities were lower than those of synthetic pesticides.