In this study, we developed a reliable and efficient Agrobacterium-mediated genetic transformation system by applying sonication and vacuum infiltration to six chickpea cultivars (ICCV2, ICCV10, ICCV92944, ICCV37, JAKI9218, and JG11) using embryo axis explants. Wounded explants were precultured for 3 days in shoot induction medium (SIM) before sonication and vacuum infiltration with an Agrobacterium suspension and co-cultivated for 3 days in co-cultivation medium containing 100 µM/l of acetosyringone and 200 mg/l of L-cysteine. Responsive explants with putatively transformed shoots were selected using a gradual increase in kanamycin from 25 mg/l to 100 mg/l in selection medium to eliminate escapes. Results showed optimal transformation efficiency at a bacterial density of 1.0, an optical density at 600 nm wavelength (OD600), and an infection duration of 30 min. The presence and stable integration of the β-glucuronidase (gusA) gene into the chickpea genome were confirmed using GUS histochemical assay and polymerase chain reaction. A high transformation efficiency was achieved among the different factors tested using embryo axis explants of cv. JAKI 9218. Of the six chickpea cultivars tested, JAKI9218 showed the highest transformation efficiency of 8.6%, followed by JG11 (7.2%), ICCV92944 (6.8%), ICCV37 (5.4%), ICCV2 (4.8%), and ICCV10 (4.6%). These findings showed that the Agrobacterium-mediated genetic transformation system will help transfer novel candidate genes into chickpea.
Journal of the Korea Society of Computer and Information
/
v.28
no.4
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pp.113-120
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2023
Smart farms are steadily increasing in research to minimize labor, energy, and quantity put into crops as IoT technology and artificial intelligence technology are combined. However, research on efficiently managing crop growth information in smart farms has been insufficient to date. In this paper, we propose a management technique that can efficiently monitor crop growth information by applying autonomous sensors to smart farms. The proposed technique focuses on collecting crop growth information through autonomous sensors and then recycling the growth information to crop cultivation. In particular, the proposed technique allocates crop growth information to one slot and then weights each crop to perform load balancing, minimizing interference between crop growth information. In addition, when processing crop growth information in four stages (sensing detection stage, sensing transmission stage, application processing stage, data management stage, etc.), the proposed technique computerizes important crop management points in real time, so an immediate warning system works outside of the management criteria. As a result of the performance evaluation, the accuracy of the autonomous sensor was improved by 22.9% on average compared to the existing technique, and the efficiency was improved by 16.4% on average compared to the existing technique.
Jo, Hye-Mi;Kim, Ye-Jin;Yoo, Sang-Ho;Kim, Chang-Mu;Kim, KyeWon;Park, Cheon-Seok
Korean Journal of Food Science and Technology
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v.53
no.2
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pp.195-203
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2021
Aureobasidium pullulans, a black yeast, produces pullulan, a linear α-glucan composed of maltotriose repeating units linked by α(1→6)-glycosidic linkages. Pullulan can be widely used in food, cosmetic, and biotechnology industries. In this study, we isolated eight strains of A. pullulans from Forsythia koreana, Magnolia kobus DC., Spiraea prunifolia var. simpliciflora, Cornus officinalis, Cerasus, and Hippophae rhamnoides. Among them, A. pullulans MR was selected as the best pullulan producer. The effects of a carbon source, a nitrogen source, and pH on pullulan production were examined. The optimal cultivation conditions for pullulan production by A. pullulans MR were determined by response surface methodology as 15% sucrose, 0.4% soy peptone, and an initial pH of 7 at 26℃. Under these conditions, the predicted pullulan production was 47.6 g/L, which was very close to the experimental data (48.9 g/L).
The adoption of carbon foot print system is being activated mostly in the developed countries as one of the long-term response towards tightened up regulations and standards on carbon emission in the agricultural sector. The Korean Ministry of Environment excluded the primary agricultural products from the carbon foot print system due to lack of LCI (life cycle inventory) database in agriculture. Therefore, the research on and establishment of LCI database in the agriculture for adoption of carbon foot print system is urgent. Development of LCA (life cycle assessment) methodology for application of LCA to agricultural environment in Korea is also very important. Application of LCA methodology to agricultural environment in Korea is an early stage. Therefore, this study was carried out to find out the effect of lettuce cultivation on agricultural environment by establishing LCA methodology. Data collection of agricultural input and output for establishing LCI was carried out by collecting statistical data and documents on income from agro and livestock products prepared by RDA. LCA methodology for agriculture was reviewed by investigating LCA methodology and LCA applications of foreign countries. Results based on 1 kg of lettuce production showed that inputs including N, P, organic fertilizers, compound fertilizers and crop protectants were the main sources of major emission factor during lettuce cropping process. The amount of inputs considering the amount of active ingredients was required to estimate the actual quantity of the inputs used. Major emissions due to agricultural activities were $N_2O$ (emission to air) and ${NO_3}^-$/${PO_4}^-$ (emission to water) from fertilizers, organic compounds from pesticides and air pollutants from fossil fuel combustion in using agricultural machines. The softwares for LCIA (life cycle impact assessment) and LCA used in Korea are 'PASS' and 'TOTAL' which have been developed by the Ministry of Knowledge Economy and the Ministry of Environment. However, the models used for the softwares are the ones developed in foreign countries. In the future, development of models and optimization of factors for characterization, normalization and weighting suitable to Korean agricultural environment need to be done for more precise LCA analysis in the agricultural area.
Journal of the Korean Society of Food Science and Nutrition
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v.42
no.7
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pp.1125-1132
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2013
In the kimchi manufacturing process, the starter is cultured on a large-scale and needs to be supplied at a low price to kimchi factories. However, current high costs associated with the culture of lactic acid bacteria for the starter, have led to rising kimchi prices. To solve this problem, the development of a new medium for culturing lactic acid bacteria was studied. The base materials of a this novel medium consisted of Chinese cabbage extract, a carbon source, a nitrogen source, and inorganic salts. The optimal composition of this medium was determined to be 30% Chinese cabbage extract, 2% maltose, 0.25% yeast extract, and $2{\times}$ salt stock (2% sodium acetate trihydrate, 0.8% disodium hydrogen phosphate, 0.8% sodium citrate, 0.8% ammonium sulfate, 0.04% magnesium sulfate, 0.02% manganese sulfate). The newly developed medium was named MFL (medium for lactic acid bacteria). After culture for 24 hr at $30^{\circ}C$, the CFU/mL of Leuconostoc (Leuc.) citreum GR1 in MRS and MFL was $3.41{\times}10^9$ and $7.49{\times}10^9$, respectively. The number of cells in the MFL medium was 2.2 times higher than their number in the MRS media. In a scale-up process using this optimized medium, the fermentation conditions for Leuc. citreum GR1 were tested in a 2 L working volume using a 5 L jar fermentor at $30^{\circ}C$. At an impeller speed of 50 rpm (without pH control), the viable cell count was $8.60{\times}10^9$ CFU/mL. From studies on pH-stat control fermentation, the optimal pH and regulating agent was determined to be 6.8 and NaOH, respectively. At an impeller speed of 50 rpm with pH control, the viable cell count was $11.42{\times}10^9(1.14{\times}10^{10})$ CFU/mL after cultivation for 20 hr - a value was 3.34 times higher than that obtained using the MRS media in biomass production. This MFL media is expected to have economic advantages for the cultivation of Leuc. citreum GR1 as a starter for kimchi production.
Ryu, Myeong Seon;Yang, Hee-Jong;Jeong, Su-Ji;Seo, Ji Won;Ha, Gwangsu;Jeong, Seong-Yeop;Jeong, Do-Youn
Korean Journal of Microbiology
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v.54
no.4
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pp.384-397
/
2018
The aim of this study is to screen the strains of Bacillus spp. possessing safety, probiotic activity, and so on, which can be utilized as probiotic resource for using the feed and supplement food of companion animal. About 300 isolates were isolated from traditional Korean sauces, four isolates that did not have or produce the six kinds of B. cereus type vomiting and diarrhea toxin genes, ${\beta}$-hemolytic, and three kinds of carcinogenic enzymes were selected. Antibiotic gene retention, cell surface hydrophobicity, antibiotic sensitivity, and glucose utilization were analyzed for four isolates, and finally SRCM 100731 was selected. SRCM 100731 was named as Bacillus amyloliquefaciens SRCM 100731 16S rRNA sequencing analysis, and carried out optimization of cell growth for industrial applications such as pet food and feed. The effects of 14 different components on cell growth were investigated and three significant positive factors, molasses, sodium chloride, and potassium chloride were selected as the main factors based on a Plackett-Burman design. In order to find out optimal concentration on each constituent, we carried out central composite design. The predicted optimized concentrations were 7% molasses, 1.1% sodium chloride, 0.5% potassium chloride. Finally, an overall about 7-fold increase in dry cell weight yield ($12.6625{\pm}0.0658g/L$) was achieved using the optimized medium compared with the non-optimized medium ($1.8273{\pm}0.0214g/L$). This research is expected to be highly utilized in the growing pet industry by establishing optimal cultivation conditions for industrial application as well as screening Bacillus amyloliquefaciens SRCM 100731 as probiotic resource for companion animal.
The discharge of waste nutrient solution from greenhouse to natural ecosystem leads to the accumulation of excess nutrients that results in contamination or eutrophication. There is a need to recycle the waste nutrient solution in order to prevent the environmental hazards. The amount and kind of nutrients in waste nutrient solution might be enough to grow photosynthetic microorganisms. Hence in the present study, we examined the growth and mass cultivation of cyanobacteria in the waste nutrient solution with an objective of removing N and P and concomitantly, its mass cultivation. Four photosynthetic filamentous cyanobacteria (Anabaena HA101, HA701 and Nostoc HN601, HN701) isolated from composts and soils of the Chungnam province were used as culture strains. Among the isolates, Nostoc HN601 performed faster growth rate and higher N and P uptake in the BG-II ($NO_3{^-}$) medium when compared to those of other cyanobacterial strains. Finally, the selected isolate was tested under optimum conditions (airflow at the rate of $1L\;min^{-1}$. in 15 L reactor, initial pH 8) in waste nutrient solution from tomato hydroponic in green house condition. Results showed to remove 100% phosphate from the waste nutrient solution in the tomato hydroponics recorded over a period of 7 days. The growth rate of Nostoc HN601 was $16mg\;Chl-a\;L^{-1}$ in the waste nutrient solution from tomato hydroponics with optimum condition, whereas growth rate of Nostoc HN601 was only $9.8mg\;Chl-a\;L^{-1}$ in BG-11 media. Nitrogen fixing capacity of Nostoc HN601 was $20.9nmol\;C_2H_4\;mg^{-1}\;Chl-a\;h^{-1}$ in N-free BG-11. The total nitrogen and total phosphate concentration of Nostoc HN601 were 63.3 mg N gram dry weight $(GDW)^{-1}$ and $19.1mg\;P\;GDW^{-1}$ respectively. Collectively, cyanobacterial mass production using waste nutrient solution under green house condition might be suitable for recycling and cleaning of waste nutrient solution from hydroponic culture system. Biomass of cyanobacteria, cultivated in waste nutrient solution, could be used as biofertilizer.
Kim, Sun-Hee;Kim, Sung-Hoon;Lee, Jo-No;An, Sang-Wook;Kim, Kwang-Soo;Hwnag, Baik;Lee, Hyeong-Yong
Microbiology and Biotechnology Letters
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v.26
no.5
/
pp.435-441
/
1998
Optimal conditions for the production of natural color, betacyanin were investigated by varying light intensity, C/N ratio, concentrations of phosphate and kinds of elicitors. Batch cultivation was employed to characterize cell growth and betacyanin production of 32 days. The maximum specific growth rate, ${\mu}$$\sub$max/, was 0.3 (1/day) for batch cultivation. The maximum specific production rate, q$\^$max/$\sub$p/, was enhanced 0.11 (mg/g-cell/day) at 3 klux. A light intensity of 3 klux was shown to the best for both cell growth and betacyanin production. The maximum specific production rate was 0.125 (mg/g-cell/day) at 0.242 (1/day), the maximum specific growth rate. The dependence of specific growth rate on the light lintensity is fit to the photoinhibition model. The correlation between ${\mu}$ and q$\sub$p/ showed that the product formation parameters, ${\alpha}$ and ${\beta}$$\sub$p/ were 0.3756 (mg/cell) and 0.001 (mg/g-cell/day), respectively. The betacyanin production was partially cell growth related process, which is different from the production of a typical product in plant cell cultures. In C/N ratio experiment, high carbon concentration, 42.1 (w/w) improved cell growth rate while lower concentration, 31.6 (w/w) increased the betacyanin production rate. The ${\mu}$$\sub$max/ and q$\^$max/$\sub$p/ were 0.26 (1/day) and 0.075 (mg/g-cell/day), respectively. Beta vulgaris L. cells under 1.25 mM phosphate concentration produced 10.15 mg/L betacyanin with 13.46 (g-dry wt./L) of maximum cell density. The production of betacyanin was elongated by adding 0.1 ${\mu}$M of kinetin. This also increased the cell growth. Optimum culture conditions of light intensity, C/N, phosphate concentration were obtained as 5.5 klux, 27 (w/w), 1.25 mM, respectively by the response surface methodology. The maximum cell density, X$\sub$max/, and maximum production, P$\sub$max/, in optimized conditions were 16 (g-dry wt./L), 12.5 (mg/L) which were higher than 8 (g-dry wt./L), 4.48 (mg/L) in normal conditions. The ${\mu}$$\sub$max/ and q$\^$max/$\sub$p/ were 0.376 (1/day) and 0.134 (mg/g-cell/day) at the optimal condition. The overall results may be useful in scaling up hairy root cell culture system for commercial production of betacyanin.
For large and rapid screening of high-yielding mutants of lovastatin produced by filamentous fungal cells of Aspergillus terreus, one of the most important stage is to test as large amounts of mutated strains as possible. For this purpose, we intended to develop a miniaturized cultivation method using $7m{\ell}$ culture tube instead of traditional $250m{\ell}$ flask (working volume $50m{\ell}$). For obtaining large amounts of conidiospores to be used as inoculums for miniaturized cultures, 4 components i.e., glucose, sucrose, yeast extract and $KH_2PO_4$ were intensively investigated, which had been observed to show positive effect on enhancement of spore production through Plackett-Burman design experimet. When optimum concentrations of these components that were determined through application of response surface method (RSM) based on central composite design (CCD) were used, maximum spore numbers amounting to $1.9\times10^{10}$ spores/plate were obtained, resulting in approximately 190 fold increase as compared to the commonly used PDA sporulation medium. Using the miniaturized cultures, intensive strain development programs were carried out for screening of lovastatin high-yielding as well as highly reproducible mutants. It was observed that, for maximum production of lovastatin, the producers should be activated through 'PaB' adaptation process during the early solid culture stage. In addition, they should be proliferated in condensed filamentous forms in miniaturized growth cultures, so that optimum amounts of highly active cells could be transferred to the production culture-tube as reproducible inoculums. Under these highly controlled fermentation conditions, compact-pelleted morphology of optimum size (less than 1 mm in diameter) was successfully induced in the miniaturized production cultures, which proved essential for maximal utilization of the producers' physiology leading to significantly enhanced production of lovastatin. As a result of continuous screening in the miniaturized cultures, lovastatin production levels of the 81% of the daughter cells derived from the high-yielding producers turned out to be in the range of 80%$\sim$120% of the lovastatin production level of the parallel flask cultures. These results demonstrate that the miniaturized cultivation method developed in this study is efficient high throughput system for large and rapid screening of highly stable and productive strains.
As information technology fusion is accelerated, the researches to improve the quality and productivity of crops inside a plant factory actively progress. Advanced growth environment management technology that can provide thermal environment and air flow suited to the growth of crops and considering the characteristics inside a facility is necessary to maximize productivity inside a plant factory. Currently running plant factories are designed to rely on experience or personal judgment; hence, design and operation technology specific to plant factories are not established, inherently producing problems such as uneven crop production due to the deviation of temperature and air flow and additional increases in energy consumption after prolonged cultivation. The optimization process has to be set up in advance for the arrangement of air flow devices and operation technology using computational fluid dynamics (CFD) during the design stage of a facility for plant factories to resolve the problems. In this study, the optimum arrangement and air flow of air circulation fans were investigated to save energy while minimizing temperature deviation at each point inside a plant factory using CFD. The condition for simulation was categorized into a total of 12 types according to installation location, quantity, and air flow changes in air circulation fans. Also, the variables of boundary conditions for simulation were set in the same level. The analysis results for each case showed that an average temperature of 296.33K matching with a set temperature and average air flow velocity of 0.51m/s suiting plant growth were well-maintained under Case 4 condition wherein two sets of air circulation fans were installed at the upper part of plant cultivation beds. Further, control of air circulation fan set under Case D yielded the most excellent results from Case D-3 conditions wherein air velocity at the outlet was adjusted to 2.9m/s.
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