Preparation and Some Properties of Active Protoplasts of Bacillus megaterium
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SUMMARY: A method for the preparation of protoplasts of Bacillus megaterium is described. The protoplasts obtained were very active metabolically and lysates of them were rich in polyribosomes. Methods for the detection of polyribosomes and of nascent peptides are detailed.Keywords:
Bacillus megaterium
Protoplast
Polysome
Bacillus (shape)
Introduction What are Plant Protoplasts? Isolation of Plant Protoplasts Mechanical and Enzymatic Isolation Procedures Source Material for Protoplast Isolation Culture of Isolated Plant Protoplasts Nutritional Requirements of Protoplasts and Culture Media Experimental Systems for Protoplast Culture Plating Density and the Use of Nurse Cells Innovative Approaches to Protoplast Culture Chemical Supplements for Protoplast Culture Media: Surfactants and Antibiotics Manipulation of Respiratory Gases Physical Procedures to Stimulate Protoplast Growth in Culture Protoplast-to-Plant Systems Exploitation of Protoplast-to-Plant Systems: Somatic and Gametosomatic Hybridisation Transformation by DNA Uptake into Isolated Protoplasts Transformation of Protoplasts by Isolated DNA Induction of DNA Uptake into Protoplasts Factors Influencing Protoplast Transformation Application of DNA Uptake into Protoplasts: Stable and Transient Gene Expression Studies Somaclonal Variation: A Simple Form of Genetic Engineering? Miscellaneous Studies with Plant Protoplasts Concluding Remarks Bibliography
Protoplast
Somaclonal variation
Plant cell
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Bacillus megaterium is a gram - positive rod-shaped endospore-forming bacteria. They are used as an effective soil modifier, having the ability to disperse phosphorus. In turn, phosphorus plays an important role in several physiological and biochemical processes of plants, such as photosynthesis, the conversion of sugar to starch, and the transfer of genetic traits. Based on the current state of research in the development of biofertilizers, it can be argued that the biocomposition of Bacillus megaterium bacteria will lead to an increase in the availability of a number of macronutrients for plants – the main elements of nutrition. This article presents the results of screening studies of previously created collection of bacterial strains of Bacillus megaterium from the Museum of the Department of microbiology, virology, epizootology and VSE of Ulyanovsk SAU. The developed scheme of isolation and identification of Bacillus megaterium bacteria is given. The species identity of 6 strains of B. megaterium was confirmed. The results of the research are compared with the characteristic phenotypic properties of the studied species described in "Bergey's Manual of Systematic Bacteriology" (2015). 210 soil samples were taken to isolate new strains of microorganisms. Initial identification allowed us to isolate 58 strains of bacteria of the genus Bacillus, 16 were assigned to Bacillus megaterium. The percentage of contamination of soil samples of this type of bacteria was 7.6 %. As the result of research, the collection of bacterial strains of studied microorganisms was expanded and their taxonomic affiliation was determined. The resulting collection will be the object of research at the following stages of work on the development of biocomposition based on bacteria to increase the coefficient of absorption of mineral components of fertilizers.
Bacillus megaterium
Bacillus (shape)
Isolation
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The production of extracellular beta-amylase by some Bacillus cereus, Bacillus megaterium and Bacillus polymyxa [corrected] strains was investigated, and the maximal yields of the enzyme were 3.6; 9.3 and 20.4 U/mL of the culture fluid, respectively (U, 1 mumol of maltose equivalent per min at 30 degrees C). Several cultivation media were used for beta-amylase production. Bacillus cereus and some strains of Bacillus megaterium gave good yields of beta-amylase only in medium with the addition of nutrient broth. However, beta-amylase produced during growth in protein rich medium (nutrient broth) was highly unstable, probably due to inactivation by proteolytic enzymes co-existing in the culture fluid. Bacillus polymyxa [corrected] strains can produce good yields of beta-amylase on a semi-synthetic medium consisting of inorganic salts, potato starch and inexpensive soybean extract instead of costly peptone and meat extract. The most potential beta-amylase producer was the strain Bacillus polymyxa [corrected] NCIB 8524. The tested Bacillus megaterium and Bacillus polymyxa [corrected] strains were apparently differentiated by temperature cultivation (30 and 37 degrees C) suitable for beta-amylase amylase yield.
Bacillus megaterium
Bacillus (shape)
Alpha-amylase
Paenibacillus polymyxa
Bacillaceae
Bacillus amyloliquefaciens
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The paper introduces the progress in protoplast research of Compositae plants with emphasis on protoplast isolation and culture,factors affecting protoplast regeneration,variances of protoplast-regenerated plants,protoplast utilization,and points out currently-existing problems and future research priorties.
Protoplast
Isolation
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Protoplast technique,method of mutation,protoplast fusion technique and the combining of protoplast technique and other biology techniques were reviewed in this paper,the deficiencies and the tendency of the research on protoplast technique was also generalized as well.
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Introduction What are Plant Protoplasts? Isolation of Plant Protoplasts Mechanical and Enzymatic Isolation Procedures Source Material for Protoplast Isolation Culture of Isolated Plant Protoplasts Nutritional Requirements of Protoplasts and Culture Media Experimental Systems for Protoplast Culture Plating Density and the Use of Nurse Cells Innovative Approaches to Protoplast Culture Chemical Supplements for Protoplast Culture Media: Surfactants and Antibiotics Manipulation of Respiratory Gases Physical Procedures to Stimulate Protoplast Growth in Culture Protoplast-to-Plant Systems Exploitation of Protoplast-to-Plant Systems: Somatic and Gametosomatic Hybridisation Transformation by DNA Uptake into Isolated Protoplasts Transformation of Protoplasts by Isolated DNA Induction of DNA Uptake into Protoplasts Factors Influencing Protoplast Transformation Application of DNA Uptake into Protoplasts: Stable and Transient Gene Expression Studies Somaclonal Variation: A Simple Form of Genetic Engineering? Miscellaneous Studies with Plant Protoplasts Concluding Remarks References
Protoplast
Somaclonal variation
Plant cell
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Bacillus megaterium
Bacillus (shape)
Bacillaceae
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Introduction Why Fuse Isolated Plant Protoplasts? Procedures for Protoplast Fusion Chemical Fusion of Protoplasts Electrical Fusion of Protoplasts Products of Protoplast Fusion and the Selection of Somatic Hybrid Tissues and Plants Somatic Hybrid Tissues and Plants Homokaryons and Heterokaryons Procedures for Selecting Hybrid Cells and Plants Characterization of Somatic Hybrid Plants Generation of Asymmetrical Hybrids and Cybrids by Protoplast Fusion Gametosomatic Hybridization Transfer of Unique Traits by Protoplast Fusion Concluding Remarks Bibliography
Protoplast
Somatic fusion
Heterokaryon
Cell fusion
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B3(Bio Best Bacillus)와 RABC(Rotating Activated Bacillus Contactor) 공법은 Bacillus를 이용하여 유기물뿐만 아니라 질소와 인도 효율적으로 제거하는 공법이다. 그러나 현 단계에서 폐수처리 시설에 이용되고 있는 Bacillus의 유기물 제거에 대한 미생물학적인 특성을 과학적으로 입증하지 못하였다. 이에 본 연구에서는 활성슬러지에서 보고되는 균주 Burkholderia cepacia(KCTC 2966)를 대조균으로 하고 Bacillus megaterium(KCTC 3007), Paenibacillus polymyxa(KCTC 3627) 균주와 B3 공법의 분뇨처리장으로부터 분리한 Bacillus속 4개 균주 Bacillus sp.(A12, C21, F12, L1)를 선정하여 0.2% Nutrient Broth, $30^{\circ}C$ 에서 배양하면서 Burkholderia의 Bacillus의 유기물 제거능력을 비교하였다. 접종량에 따른 개체군 차이로 유기물 제거능력이 영향 받을 수 있지만, Bacillus의 유기물 제거능력이 Burkholderia보다 $2\sim5$ 배 뛰어남을 알 수 있었다. 또한, Bacillus megaterium과 Bacillus sp.(C21, F12, N12)의 영양세포와 내생포자의 유기물 제거능력의 차이를 알아본 결과 내생포자의 제거능력이 영양세포보다 더욱 뛰어남을 알 수 있었다. 【The Bio Best Bacillus(B3) and Rotating Activated Bacillus Contactor(RABC) processes, in which Bacillus strains are predominating, are reported to remove nitrogen and phosphorus as well as organic matter effectively. Nevertheless the nutrient removal characteristics of the Bacillus strains have not been studied in detail so far. This study investigated the organic and nutrient removal by Bacillus strains, Bacillus megaterium(KCTC 3007), Paenibacillus polymyxa(KCTC 3627), and Bacillus sp. A12, C21, F12, and L1(isolated from a B3 process), by incubating the strains in 0.2% nutrient broth at $30^{\circ}C$ . Burkholderia cepacia(KCTC 2966), a common activated sludge organism, was used as a reference species for comparison. Although the degradation rate was affected by the population sire, the specific removal rates of organic matter by Bacillus strains were greater by $2\sim5$ times than that of Burkholderia. In particular, the culture bottles inoculated with the endospores of Bacillus megaterium and Bacillus sp. C21, F12, and N12 showed significantly higher degradation rate than those of vegetative cells.】
Bacillus megaterium
Bacillus (shape)
Paenibacillus polymyxa
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Bacillus megaterium
Bacillus (shape)
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