• Title/Summary/Keyword: Soil DNA

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Breeding of the native vegetables using the biotechnology

  • Iwamoto, Yuzuri
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2005.11a
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    • pp.106-111
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    • 2005
  • For breeding of a new rootstock for eggplant production, somatic hybrids between two species, Solanum integrifolium and S. sanitwongsei were obtained through protoplast fusion. The former species has been commonly used for rootstock for eggplant production in Japan. Eggplants on these rootstocks are more productive than ungrafted plants, but are susceptible to bacterial wilt caused Ralstonia solanacearum. While the latter species is resistant, the growth of eggplants on this rootstock is rather slow and low yield. Protoplast of both species were isolated from cotyledons, and inactivated with iodoacetamide or UV-irradiation, then fused electrically. The fused products were then cultured. Regenerated plantlets were then transplanted on soil then maintained in a green house. The plants were classified into four groups. Those in the first group showed morphological characters intermediate of the parentalspecies. The plants bore fruit with viable seeds. The plants showed a chromosome number of 2n=48, the sum of those of the parental species, and are suggested to be symmetric fusion products. While plants in the other groupswas less vigorous and showed chromosome number 2n= 68 to 72 suggesting asymmetric fusion products by genomic in situ hybridization(GISH). Isozyme pattern of shikimate dehydrogenase (SKDH; EC 1.1.1.25), isocitrate dehydrogenase (IDH; EC 1.1.1.41) and phosphoglucomutase (PGM; EC 2.7.5.1) showed that 24 regenerated plants in three groups were somatic hybrids. Analysis of random amplified polymorphic DNA (RAPD) showed that 43 S. integrifolium-specific and 57 S. sanitwongsei-specific bands were all found in 24 plants. Both somatic hybrids and its S1 plants were found to be resistant to bacterial wilt, and eggplant grafted these plants using for rootstocks were more productive than grafted mother plants. Now, S1 progenies are used for commercial eggplant production in Osaka Prefecture.

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Identification of Fluorescent Pseudomonads Producing Siderophore and Construction of Siderophore Biosynthesis Defective Mutant (Siderophore를 생성하는 Fluorescent Pseudomonads의 분리, 동정 및 돌연번이 유기)

  • Park, Yeal;Kim, Hyun Hee;Myeong-gu Yeo;Young-woo Seo;Han-cheol Koh;Young-gi Yang;Hyeon-Sook Cheong;Sung-jun Kim
    • Korean Journal of Microbiology
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    • v.30 no.4
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    • pp.286-290
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    • 1992
  • The present study was performed to isolate the fluorescent pseudomonads from Kwang-Ju soil and to construct a mutant strain defective in siderophore biosynthesis. The siderophore-secreting pseudomonads were screened on Blue agar (Chrome Azuol S agar) plates and one strain of them was designated to Pseudominas fluorescens (P. fluorescens) PY002. To construct a mutant defective in siderophore biosynthesis, P. fluorescens PY002 was randomly mutagenized with a transposon Tn5. The location of Tn5 integrated into chromosomal of the mutants strain was determined by Southern blot analysis. The mutagenized strain showed non-fluorescent on a King's B agar plate and were defective in iron (III) acquisition ability.

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Isolation and Characterization of Strain of Bacillus thuringiensis subsp. kenyae Containing Two Novel cry1-Type Toxin Genes

  • Choi, Jae-Young;Li, Ming Shun;Shim, Hee-Jin;Roh, Jong-Yul;Woo, Soo-Song;Jin, Byung-Rae;Boo, Kyung-Saeng;Je, Yeon-Ho
    • Journal of Microbiology and Biotechnology
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    • v.17 no.9
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    • pp.1498-1503
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    • 2007
  • To identify novel crystal proteins, Bacillus thuringiensis 2385-1 was isolated from Korean soil samples and characterized. The H-serotype of 2385-1 was identical to that of subsp. kenyae (H4a4c), and its crystal toxin was bipyramidal-shaped. However, 2385-1 showed a much higher toxicity towards Plutella xylostella and Spodoptera exigua larvae than subsp. kenyae. In addition, the crystal protein profile and plasmid DNA pattern of 2385-1 differed from those of subsp. kenyae. To verify the crystal protein gene types of 2385-1, a PCR-RFLP analysis was performed, and the results revealed that 2385-1 contained two novel cry1-type crystal protein genes, cryl-5 and cry1-12, in addition to the crylJal gene. The deduced amino acid sequences of cryl-5 and cry1-12 showed a 97.9% and 75.7% sequence similarity with the CrylAb and CrylJa crystal proteins, respectively. Among the novel crystal proteins, Cry1-5 showed a high toxicity towards P. xylostella and S. exigua larvae. In conclusion, B. thuringiensis 2385-1 is a new isolate in terms of its gene types, and should be a promising source for an insecticide to control lepidopteran larvae.

Isolation of an Isocarbophos-Degrading Strain of Arthrobacter sp. scl-2 and Identification of the Degradation Pathway

  • Li, Rong;Guo, Xinqiang;Chen, Kai;Zhu, Jianchun;Li, Shunpeng;Jiang, Jiandong
    • Journal of Microbiology and Biotechnology
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    • v.19 no.11
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    • pp.1439-1446
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    • 2009
  • Isocarbophos is a widely used organophosphorus insecticide that has caused environmental pollution in many areas. However, degradation of isocarbophos by pure cultures has not been extensively studied, and the degradation pathway has not been determined. In this paper, a highly effective isocarbophos-degrading strain, scl-2, was isolated from isocarbophos-polluted soil. The strain scl-2 was preliminarily identified as Arthrobacter sp. based on its morphological, physiological, and biochemical properties, as well as 16S rDNA analysis. The strain scl-2 could utilize isocarbophos as its sole source of carbon and phosphorus for growth. One hundred mg/l isocarbophos could be degraded to a non detectable level in 18 h by scl-2 in cell culture, and isofenphos-methyl, profenofos, and phosmet could also be degraded. During the degradation of isocarbophos, the metabolites isopropyl salicylate, salicylate, and gentisate were detected and identified based on MS/MS analysis and their retention times in HPLC. Transformation of gentisate to pyruvate and fumarate via maleylpyruvate and fumarylpyruvate was detected by assaying for the activities of gentisate 1,2-dioxygenase (GDO) and maleylpyruvate isomerase. Therefore, we have identified the degradation pathway of isocarbophos in Arthrobacter sp. scl-2 for the first time. This study highlights an important potential use of the strain scl-2 for the cleanup of environmental contamination by isocarbophos and presents a mechanism of isocarbophos metabolism.

Studies on the Isolation and Identification of Xylanase and Mannanase Producing Aspergillus niger (Xylanase와 Mannanase를 생산하는 Aspergillus niger의 분리와 동정에 관한 연구)

  • Kim, Byoung-Suk;Cho, Jin-Kook;Song, Jin-Ook;Lee, Hak-Kyo;Hwang, Seong-Gu
    • Journal of Animal Science and Technology
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    • v.51 no.5
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    • pp.427-432
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    • 2009
  • This study was undertaken to screen a high xylanase and mannanase producing microbes. In the first experiment, screening was undertaken against 50 samples of microorganisms having xylanase and mannanase activities from soil and fallen leaves. The screening process has focused on picking out fungi having high xylanase and mannanase activities under the solid-state fermentation. The xylanase and mannanase activities of 6 screened microbes were 0.9~1.6 unit/mL and 0.2~0.4 unit/mL, respectively, under the submerged fermentation condition. However, under the solid-state fermentation, xylanase and mannanase activities were 103.7~220.0 unit/g and 20.1~40.3 unit/g, respectively. Finally one microbe (E-3) was selected and its xylanase and mannanase activities were 197.3 unit/g and 39.9 unit/g, respectively. The morphological and molecular biological classification of E-3 showed 99% homology with the Aspergillus niger.

Physiological properties and transformation of alkaline-tolerant bacteria (알카리내성 세균의 생리적 특성 및 형질전환)

  • 유주현;정용준;정건섭;오두환
    • Microbiology and Biotechnology Letters
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    • v.14 no.3
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    • pp.239-244
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    • 1986
  • To develop the potential use as new host strain for gene cloning, alkaline-tolerant isolates from soil were examined for amylase activity, protease activity, antimicrobial activity and transformability by using plasmid pUB 110. Of these strains, one was selected and identified as Bacillus sp. YA-14. in the enzymatic properties of Bacillus sp. YA-14 the optimal conditions for the reaction of amylase and protease were at pH 0.8 and pH 7.5 respectively. The antimicrobial activity of Bacillus sp. YA-14 was also found. For the transformation, Bacillus sp. YA-14 was cultured to late logarithmic growth phase ai 37$^{\circ}C$ in modified SPI medium (pH 8.0) containing 0.4% MgSO$_4$. The presence of pUB 110 plasmid DNA in transformants was confirmed by electrophoresis and stably maintained in the new host.

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Selection and Antagonistic Mechanism of Pseudomonas fluorescens 4059 Against Phytophthora Blight Disease (고추역병과 시들음병을 방제하는 토착길항세균 Pseudomonas fluorescens 4059의 선발과 길항기작)

  • Jeong, Hui-Gyeong;Kim, Sang-Dal
    • Microbiology and Biotechnology Letters
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    • v.32 no.4
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    • pp.312-316
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    • 2004
  • In oder to select the powerful rhizophere-dorminatable biocontrol agent, we had isolated an indigenous antagonistic bacterium which produced antibiotic and siderophore from a disease suppressive local field soil of Gyungsan, Korea. And we could select the Pseudomosp. 4059 which can strongly antagonize against Fusarium oxysporum and Phytophthora capsici by two kinds of antifungal mechanism that can be caused by the antibiotic of Phenazin, a siderophore and a auxin like subThe selected strain was identified as Pseudomonas fluorescens (biotype A) 4059 by biochemical tests, API $\textregistered$ test, MicroLog TM system and 16S rDNA analysis. The selected antagonistic microorganism, Pseudomosp. 4059 had an antifungal mechanism of antifungal antibiotic and sidrophore. And we were confirmed the antagonistic activity of P fluorescens 4059 with in vitro antifungal test against Phytophthora capsici and in vivo by red-pepper.

Diversities of Arbuscular Mycorrhizal Fungi in Cultivated Field Soils of Korean Ginseng (금산의 인삼 재배 토양에 분포하는 수지상균근균의 다양성)

  • Kil, Yi-Jong;Eo, Ju-Kyeong;Eom, Ahn-Heum
    • The Korean Journal of Mycology
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    • v.40 no.1
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    • pp.1-6
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    • 2012
  • In this study, soil samples were collected from cultivated fields of 1-5 year old Korean ginseng in Geumsan, Korea. Spores of arbuscular mycorrhizal fungi were extracted from soils and identified using morphological characteristics and 18s rDNA sequences of the spores. Total 10 species of AMF were identified: Acaulospora longula, Archaeospora trappei, Glomus caledonium, Glomus etunicatum, Glomus intraradices, Glomus mosseae, Glomus sp., Paraglomus occultum, Paraglomus brasilianum, and Scutellospora heterogama. Relative abundance of spores of A. trappei were increased with increase of cultivation period of the ginseng. However, relative abundance of other species of AMF and Shannon diversity (H') of AMF were significantly decreased with the increase of cultivation periods of the ginseng.

Comparison of Characteristics between Insecticidal and Noninsecticidal Basillus thuringiensis Strains belonging to Serotype H8a8b

  • Roh, Jong-Yul;Je, Yeon-Ho;Park, Hyun-Woo;Chang, Jin-Hee;Jin, Byung-Rae;Lee, Dae-Weon;Ziwen Yang;Kang, Seok-Kwon
    • Journal of Sericultural and Entomological Science
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    • v.40 no.2
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    • pp.126-130
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    • 1998
  • A noninsecticidal strain, Bacillus thuringiensis NTB-88, isolated from Korean soil, had a typical bipyramidal parasporal inclusion and its serotype is identical to B. thuringiensis subspmorrisoni (H8a8b). To elucidate differences between insecticidal and noninsecticidal strains, we compared strain NTB-88 to other toxic B. thuringiensis subsp. morrisoni strains (HD-12 and PG-14). Restriction endonucleases digested plasmid DNA patterns showed that strain NTB-88 was different from lepidopteran-toxic strain, HD-12, but it was similar to dipteran-toxic strain, PG-14. The gene type of strain NTB-88 was different from those of other insecticidal strains, Furthermore, the NH2-terminal amino acid sequence of crystal protein of strain NTB-88 had no relation to those of the previously known $\delta$-endotoxins in other toxic strains as well as HD-12 and PG-14 strains. Therefore, the noninsecticidal crystal protein in strain NTB-88 is novel and its property is different from insecticidal ones.

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Enhanced Biosynthesis of ${\alpha}$-tocopherol in Transgenic Soybean by Introducing ${\gamma}$-TMT gene

  • Kim Young-Jin;Seo Hong-Yul;Park Tae-Il;Baek So-Hyeon;Shin Woon-Chul;Kim Hyun-Soon;Kim Jung-Gon;Choi Yong-Eui;Yun Song-Joong
    • Journal of Plant Biotechnology
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    • v.7 no.3
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    • pp.203-209
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    • 2005
  • This study was conducted to improve tocopherol (vitamin E) composition in soybean (Glycine max) by introducing a gamma-tocopherol methyl transferase (${\gamma}$-TMT) gene via Agrobacterium tumefaciens-mediated transformation. Immature cotyledon explants were cocultivated with Agrobacterium tumefaciens. Putative transgenic embryos were selected from immature cotyledons on MS medium supplemented with 40 mg/L 2,4-D containing 100 mg/L kanamycin, 500 mg/L carbenicillin and 250 mg/L cefotaxime. Plantlets were developed from somatic embryos, and then transferred to soil. Nineteen regenerated plantlets obtained on the selection medium from 1,460 cotyledons. However, only 9 plantlets were confirmed as transformed plants. Integration of the transgene into the soybean genomic DNA was confirmed by PCR and Southern blot analysis. HPLC analysis showed that the content of ${\alpha}$-tocopherol in transgenic soybean seeds (AT-1) was approximately 4-fold higher than that of non-transgenic plants. Conclusively, we obtained the transgenic soybean having increased ${\alpha}$-tocopherol content by the overexpression of ${\gamma}$-TMT transgene.