• Journal of the Korea Organic Resources Recycling Association
• /
• v.24 no.4
• /
• pp.77-83
• /
• 2016

The objectives of this study was to investigate the difference between organic-farming and conventional-farming soils relatives to soil chemical properties and microbial flora. Fifteen soil sampling sites were chosen from the certified organic upland farm, considered with its location, crop and application of organic compost types. Soil chemical properties were analyzed by standard methods established by National Institute of Agricultural Sciences, Rural Development Administration. For the soil chemical properties, the values of pH were ranged from 4.5 to 7.3. The values of electrical conductivity (EC) in the sampling sites were below 2 dS/m of convention cultivation soil. For analyzing the microbial flora, the bacillus(16S rDNA) and cladothricosis(18S rDNA) were analyzed by using PCR-DGGE (Denaturing Gradient Gel Electrophoresis) in the soil of 15 sampling sites. Cluster analysis of biodiversity index was performed by using pattern of DGGE. DGGE patterns and clustering analysis of bacterial DNA from soil extracts revealed that the bacterial community was differentiated between less than 5 years and more than 5 years depending on the cultivation history. But there was no consistent tendency between cultivation history and regional trend in the case of molds. Therefore, it would be very effective to analyze bacterial clusters of organically cultivated soils in long - term cultivated soil for more than 5 years.

• Journal of Korean Society of Forest Science
• /
• v.58 no.1
• /
• pp.1-7
• /
• 1982

Megagametophyte and embryo tissue of Pinus densiflora were subjected to study the inheritance of glutamate-oxalate transaminase(GOT) and leucine aminopeptidase(LAP), and linkage relationship among isozyme loci coding both enzymes by starch gel zone-electrophoresis. Four zones of activity were observed for GOT. No variation was found in the fastest migrating zone (GOT-A). Electrophoretic phenotypes of the other two zones (GOT-B and GOT-C) showed 1:1 segregation ration, suggesting that each zone is controlled by a single locus. Foru and three alleles were identified at both loci respectively. The isozyme pattern of the fourth zone(GOT-D), migrated cathodally, coincided precisely with that of GOT-C. Whether the two zones are controlled by the same locus or by two tightly linked loci remained unknown. In all three variant GOT zones, heterozygoes embryos produced triple band patterns, indicating that GOT isozyme in Pinus densiflora is a dimer. Two zones of activity stained for LAP were found. The segregation of the two zones (LAP-A and LAP-B) suggested that tow loci control each of both isozymes. Two and three alleles were identified at both loci. GOT-B and LAP-B were found to be tightly linked, showing an average recombination frequency of 12.5 percent. Slight deviation from independent assortment was observed between GOT-B and GOT-C, with recombination frequency of 41 percent.

• Korean Journal of Microbiology
• /
• v.40 no.4
• /
• pp.342-347
• /
• 2004

A bacterial strain YC4963 with antifungal activity against Colletotrichum orbiculare, a causal organism of cucumber anthracnose was isolated from the rhizosphere soil of Siegesbeckia pubescens Makino in Korea. Based on physiological and biochemical characteristics and 16S ribosomal DNA sequence analysis, the bac­terial strain was identified as Pseudomonas aurantiaca. The bacteria also inhibited mycelial growth of several plant fungal pathogens such as Botrytis cinerea, Fusarium oxysporum and Rhizoctonia solani on PDA and 0.1 TSA media. The antifungal activity was found from the culture filtrate of this isolate and the active compound was quantitatively bound to XAD adsorption resin. The antibiotic compound was purified and identified as phenazine-l-carboxylic acid on the basis of combined spectral and chemical analyses data. This is the first report on the production of phenazine-l-carboxylic acid by Pseudomonas aurantiaca.

• Journal of Life Science
• /
• v.25 no.7
• /
• pp.826-832
• /
• 2015

This study describes the effects of activated charcoal on the removal of salts, detergents, and pigments from protein extracts of ginseng leaves and roots. Incubation of protein extracts with 5% (w/v) activated charcoal (100-400 mesh) for 30 min at 4℃ almost removed the salts and detergents including NP-40 as can be observed on SDS-PAGE. In addition, analysis of chlorophyll content showed significant depletion of chlorophyll (~33%) after activated charcoal treatment, suggesting potential effect of activated charcoal on removal of pigments too along with the salts and detergents. 2-DE analysis of activated charcoal treated protein samples showed better resolution of proteins, further indicating the efficacy of activated charcoal in clearing of protein samples. In case of root proteins, although not major differences were observed on SDS-PAGE, 2-DE gels showed better resolution of spots after charcoal treatment. In addition, both Hierarchical clustering (HCL) and Principle component analysis (PCA) clearly separated acetone sample from rest of the samples. Phenol and AC-phenol samples almost overlapped each other suggesting no major differences between these samples. Overall, these results showed that activated charcoal can be used in a simple manner to remove the salts, detergents and pigments from the protein extracts of various plant tissues.

• Korean Journal of Veterinary Research
• /
• v.36 no.4
• /
• pp.795-807
• /
• 1996

This study was conducted to identify the effects of caffeine on the lipid and protein components or blood chemistry levels of the serum as well as the total homogenate, mitochondrial and microsomal fraction of the rat(Sprague-Dawley, female) liver. Acute test were conducted to determine those effects. The acute test was conducted by dividing rats into 7 groups according to the time lapsed after a single oral administration of 100mg/kg caffeine(that is control, 2hrs, 4hrs, 8hrs, 24hrs, 48hrs and 72hrs lapsed group). The concentrations of glucose, urea nitrogen, uric acid, creatinine, total protein, albumin, A/G ratio, triglyceride, total cholesterol, HDL-cholesterol, free fatty acid, phospholipid as well as the activities of alanine aminotransferase(ALT), aspartate aminotransferase(AST) and alkaline phosphatase(ALP) were measured in the serum of each experimental groups. The concentrations of the carbonyl group, malondialdehyde(MDA) and the patterns of the SDS-PAGE(Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis) were analyzed to determine the oxidative damages and metabolic changes on the lipid and protein components in the serum, and total homogenate, mitochondrial and microsomal fractions of the rat liver. The results obtained from this study were summarized as follows; 1. The concentrations of serum glucose were significantly higher(p<0.01) between 4(143.0mg/dl) and 8hrs(138.0mg/dl) in comparison to that of the control(101.1mg/dl) after a single oral administration of caffeine(100mg/kg). While on the other, there were no significant differences in the concentrations of urea nitrogen, uric acid, creatinine, total protein, albumin and albumin/globulin(A/G) ratio in comparison to those of the control. 2. The concentrations of total cholesterol and HDL-cholesterol in serum were significantly higher(p<0.01) between 4(77.4mg/dl, total cholesterol) and 8hrs(64.7mg/dl, HDL-cholesterol) in comparison to those of the control(62.8, 46.7mg/dl) after a single oral administration of caffeine(100mg/kg). On the other hand, the concentrations of triglyceride in serum were significantly lower(p<0.01) after 8hrs(38.8mg/dl) in comparison to that of the control(66.5mg/dl). 3. The activities of AST in serum was significantly higher(p<0.05) from 2hrs(149U/L) to 8hrs(178U/L) in comparison to the control(112U/L) after a single oral administration of caffeine(100mg/kg). The activities of ALT in serum were significantly higher(p<0.01) at 4(45.5U/L), 24(49.3U/L), 48(46.8U/L) and 72 hrs(42.3U/L) in comparison to that of the control(39.7U/L) after a single oral administration of caffeine(100mg/kg). On the other hand, there were no significant differences in the activities of ALP in comparison to that of the control. 4. The concentrations of free fatty acid in serum were significantly higher(p<0.01) at 8hrs(65.0mg/dl) in comparison to that of the control(37.6mg/dl) after a single oral administration of caffeine(100mg/kg). However, there were no significant differences in the concentrations of carbonyl group and malondialdehyde within serum, and liver homogenate, mitochondrial and microsomal fractions in comparison to that of the control. 5. The patterns of SDS-PAGE in serum, mitochondrial and microsomal fraction of the liver showed no significant differences.

• Current Research on Agriculture and Life Sciences
• /
• v.4
• /
• pp.55-64
• /
• 1986

Polyphenol oxidase in japanese pear (Pyrus communis var. mansamkil) was isolated, partially purified and its some properties were investigated. Polyacrylamide disc gel electrophoresis indicated two bands with polyphenol oxidase activity in the extract from acetone dry powder of par flesh. These two polyphenol oxidases (PPO A and PPO B) were purified through acetone precipitation and diethylaminoethyl cellulose column chromatography. PPO A and B were purified 7.8 fold and 8.7 fold by the present procedure, respectively. The Rm values of partially purified PPO A and B were estimated to be 0.58 and 0.68, respectively. The optimum temp, and pH of PPO A activity were $33^{\circ}C$ and pH 7.0, while those of PPO B were $30^{\circ}C$ and pH 4.2, respectively. Two PPO were unstable over the temperature of $60^{\circ}C$. The substrate specificity of pear PPO showed high affinity toward o-diphenolic compounds, especially catechol in PPO A and chlorogenic acid in PPO B, but inactive toward m-diphenol, p-diphenol and monophenols. PPO A showed affinity toward the trihydroxyphenolic compound. $Zn^{{+}{+}}$ activated the PPO A activity but $Fe^{{+}{+}}$ inhibited PPO B activity, while $Fe^{{+}{+}}$ and $Zn^{{+}{+}}$ activated the PPO B activity, while $Fe^{{+}{+}}$ and $Zn^{{+}{+}}$ activated the PPO B activity but $K^+$, $Mg^{{+}{+}}$, $Ca^{{+}{+}}$ and $Hg^{{+}{+}}$ inhibited at 10mM concentration. $Cu^{{+}{+}}$ activated the enzyme action at low concentrations but inhibited at high concentration. Inhibition studies indicated that L-ascorbic acid, L-cysteine and thiourea were most potent. The Km values of PPO A and PPO B for catechol were 20mM and 14.3mM, respectively.

• Clinical and Experimental Reproductive Medicine
• /
• v.35 no.2
• /
• pp.99-110
• /
• 2008

Objectives: Preimplantation genetic diagnosis (PGD) has become an assisted reproductive technique for couples carrying genetic conditions that may affect their offspring. Osteogenesis imperfecta (OI) is an autosomal dominant disorder of connective tissue characterized by bone fragility and low bone mass. At least 95% of cases are caused by dominant mutations in the COL1A1 or COL1A2. In this study, we report on our experience clinical outcomes with 5 PGD cycles for OI in two couples. Methods: Before clinical PGD, we assessed the amplification rate and allele drop-out (ADO) rate of alkaline lysis and nested PCR protocol using heterozygous patient's single lymphocytes in the pre-clinical diagnostic tests for OI. We performed 5 cycles of PGD for OI by nested PCR for the causative mutation loci, COL1A1 c.2452G>A and c.3226G>A, in case 1 and case 2, respectively. The PCR products were analyzed by agarose gel electrophoresis, restriction fragment length polymorphism (RFLP) analysis with HaeIII restriction enzyme in the case 1 and direct DNA sequencing. Results: We confirmed the causative mutation loci, COL1A1 c.2452G>A in case 1 and c.3226G>A in case 2. In the pre-clinical tests, the amplification rate was 94.2% and ADO rate was 22.5% in case 1, while 98.1% and 1.9% in case 2, respectively. In case 1, a total of 34 embryos were analyzed and 31 embryos (91.2%) were successfully diagnosed in 3 PGD cycles. Eight out of 19 embryos diagnosed as unaffected embryos were transferred in all 3 cycles, and in the third cycle, pregnancy was achieved and a healthy baby was delivered without any complications in July, 2005. In case 2, all 19 embryos (100.0%) were successfully diagnosed and 4 out of 11 unaffected embryos were transferred in 2 cycles. Pregnancy was achieved in the second cycle and the healthy baby was delivered in March, 2008. The causative locus was confirmed as a normal by amniocentesis and postnatal diagnosis. Conclusions: To our knowledge, these two cases are the first successful PGD for OI in Korea. Our experience provides a further demonstration that PGD is a reliable and effective clinical techniques and a useful option for many couples with a high risk of transmitting a genetic disease.

• Horticultural Science & Technology
• /
• v.29 no.4
• /
• pp.366-373
• /
• 2011

To analyze the genetic relationships among Korean potato cultivars and to develop cultivar identification method using DNA markers, we carried out genotyping using simple sequence repeats (SSR) analysis and developed multiplex-SSR set. Initially, we designed 92 SSR primer combinations reported previously and applied them to twenty four Korean potato cultivars. Among the 92 SSR markers, we selected 14 SSR markers based on polymorphism information contents (PIC) values. PIC values of the selected 14 markers ranged from 0.48 to 0.89 with an average of 0.76. PIC value of PSSR-29 was the lowest with 0.48 and PSSR-191 was the highest with 0.89. UPGMA clustering analysis based on genetic distances using 14 SSR markers classified 21 potato cultivars into 2 clusters. Cluster I and II included 16 and 5 cultivars, respectively. And 3 cultivars were not classified into major cluster group I and II. These 14 SSR markers generated a total of 121 alleles and the average number of alleles per SSR marker was 10.8 with a range from 3 to 34. Among the selected markers, we combined three SSR markers, PSSR-17, PSSR-24 and PSSR-24, as a multiplex-SSR set. This multiplex-SSR set used in the study can distinguish all the cultivars with one time PCR and PAGE (Polyacrylamide gel electrophoresis) analysis and PIC value of multiplex-SSR set was 0.95.

• Microbiology and Biotechnology Letters
• /
• v.40 no.2
• /
• pp.111-116
• /
• 2012

In this study, we investigated the microbial flora changes in Gugija-Liriope tuber Makgeolli during fermentation and storage periods. We brewed Gugija-Liriope tuber Makgeolli for a week through twostage fermentations and stored the fermentation broth for a month at $4^{\circ}C$ or $20^{\circ}C$. We collected the samples periodically and analyzed microbial flora changes using viable cell counts and PCR-denaturing gradient gel electrophoresis (DGGE). Yeast viable cells were seen to have decreased to 13% of pre-storage levels after storage for 15 days at $20^{\circ}C$; however significant changes were not observed during storage at $4^{\circ}C$. Prolongation of storage time dramatically decreased the availability of viable cells. Yeast viable cell numbers had decreased to 38% of pre-storage levels at $4^{\circ}C$ and 4.8% at $20^{\circ}C$ after storage for 30 days. The results of the DGGE profile for yeast showed that Saccharomyces cerevisiae and Saccharomyces sp. were the predominant strains at the beginning of fermentation and throughout the whole period of storage. Viable cell counts for total bacteria had decreased to 36% of pre-storage levels after storage for 15 days but did not significantly change for the full 30 days of storage at $4^{\circ}C$. Similarly, viable cell counts for bacteria had decreased to 5% while viable cell numbers did not significantly change for the full 30 days at $20^{\circ}C$. Viable cell counts for lactic acid bacteria were performed and the results were similar to those for total bacteria. The results of the DGGE profile for bacteria showed that Weissella cibaria was the predominant strain at the beginning of fermentation. However it had disappeared by the end of fermentation, and Lactobacillus fermentum and Pediococcus acidilactici became the predominant species during storage.

• Journal of Life Science
• /
• v.20 no.2
• /
• pp.237-244
• /
• 2010

Five mold strains - Mucor racemousus, Rhizopus oryzae, Aspergillus oryzae, Aspergillus kawachii, and Monascus purpureus - were used for the fermentation of both freeze-dried and air-dried silkworm powders. The concentrations of proteins and minerals, electrophoretical protein patterns, fatty acid composition and the activities of fibrinolytic and antioxidation with freeze-dried silkworm (FDSW) or air-dried silkworm powders (ADSW) fermented by the five molds were investigated. The concentrations of major minerals in fermented FDSW and ADSW powders were K by 72.0-76.3 and 77.1-78.9 ppm, Mg by 29.6-49.7 and 44.3-58.7 ppm, Ca by 1.9-14.9 and 9.8-21.6 ppm and Zn by 0.64-0.70 and 4.17-4.52 ppm, respectively. Major fatty acids in fermented FDSW and ADSW powders were linolenic acid, oleic acid, and palmitic acid. There were slightly varietal differences in electrophoretical protein patterns when total protein patterns of fermented FDSW and ADSW powders were analyzed by native-and SDS-polyacrylamide gel electrophoresis (PAGE). DPPH radical scavenging activity was slightly stronger in fermented ADSW powders than that in fermented FDSW. Fibriolytic activity was only detected in the FDSW fermented by Aspergillus kawachi and Monascus purpureus. These results may provide the basic data to understand the biological activities and chemical characteristics of silkworm fermented by mold, which can be used for the development of functional foods.