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Studies on the Amylase Production by Bacteria (세균(細菌)에 의(依)한 Amylase생산(生産)에 관한 연구(硏究))

  • Park, Yoon-Joong
    • Applied Biological Chemistry
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    • v.13 no.2
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    • pp.153-170
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    • 1970
  • 1. Isolation and identification of amylase-producing bacteria. The powerful strain A-12 and S-8 were respectively isolated from air and soil after screening a large number of amylase-producing bacteria. Their bacterial characteristics have been investigated and it has been found that all characteristics of strain A-12 and S-8 are similar to Bac. subtilis of Bergey's manual except for the acid formation from a few carbohydrates and the citrate utilization, i.e., the strain A-12 shows negative in the citrate utilization, and the acid formation from arabinose and xylose, S-8 shows negative in the acid formation from xylose. 2. Amylase production by Liquid cultures with solid materials. Several conditions for amylase production by strain A-12 in stationary cultures have been studied. The results obtained are as follows. (1) The optimum conditions are:temperature $35^{\circ}C$, initial pH 6.5 to 7.0 and incubation time 3 to 4 days. (2) The amylase production is not affected by the preservation period of the stock cultures. (3) Among the various solid material, the defatted soy bean is found to be the best for t1e amylase production. However, the alkali treatment of the defatted soy bean gives no effect contrary to the cage of defatted rape seed. The addition of soluble starch to the alkali extract of defatted soy bean shows the increased amylase production. (4) Up to 1% addition of ethanol to carbon dificient media gives the improved amylase production, whereas the above effect is not found in the case of carbon rich media. (5) The amylase production can be increased 2.5 times when 10% of defatted soy bean is admixed to cheaply available wheat bran. (6) The excellent effect is found for amylase production when 20% of wheat bran is admixed to defatted dry milk which is a poor medium. The activity is found to be $D^{40^{\circ}}_{30'}$ 7,000(L.S.V. 1,800) in 10% medium. (7) No significant effect is observed due to the addition of various inorganic salts. 3. Amylase production by solid cultures. Several conditions for amylase production by strain A-12 in wheat bran cultures have been studied and the results obtained are as follows. (1) The optimum conditions: are temperature $33^{\circ}C$, incubation lime 2 days, water content added 150 to 175% and the thickness of the medium 1.5cm, The activity is found to be $D^{40^{\circ}}_{30'}$ 36,000(L.S.V. 15,000) (2) No significant effect is found in the case of the additions of various organic and inorganic substances.

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Preparation of Powdered Smoked-Dried Mackerel Soup and Its Taste Compounds (고등어분말수우프의 제조 및 정미성분에 관한 연구)

  • LEE Eung-Ho;OH Kwang-Soo;AHN Chang-Bum;CHUNG Bu-Gil;BAE You-Kyung;HA Jin-Hwan
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.20 no.1
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    • pp.41-51
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    • 1987
  • This study was carried out to prepare powdered smoked-dried mackerel which can be used as a soup base, and to examine storage stability and the taste compounds of Products. Raw mackerel are filleted, toiled for 10 minutes and pressed to remove lipids, and then soaked in extract solution of skipjack meat. This soaked mackerel are smoked 3 times to $10-12\%$ moisture content at $80^{\circ}C$ for 8 hours. And the smoked-dried mackerel were pulverized to 50 mesh. Finally, the powdered smoked-dried mackerel were packed in a laminated film $bag(PET/Al\;foil/CPP:\;5{\mu}m/15{\mu}m/70{\mu}m,\;15\times17cm)$ with air(product C), nitrogen(product N) and oxygen absorber(product O), and then stored at room temperature for 100 days. The moisture and crude lipid content of powdered smoked-dried mackerel was $11.3-12.3\%,\;12\%$, respectively, and water activity is 0.52-0.56. And these values showed little changes during storage. The pH, VBN and amino nitrogen content increased slowly during storage. Hydrophilic and lipophilic brown pigment formation showed a tendency of increase in product(C) and showed little change in product(N) and (O). The TBA value, peroxide value and carbonyl value of product(N) and (O) were lower than those of product (C). The major fatty acids of products were 16:0, 18:1, 22:6, 18:0 and 20:5, and polyenoic acids decreased, while saturated and monoenoic acids increased during processing and storage of products. The IMP content in products were 420.2-454.2 mg/100 g and decreased slightly with storage period. And major non-volatile organic acids in products were lactic acid, succinic acid and $\alpha-ketoglutaric$ acid. In free amino acids and related compounds, major ones are histidine, alanine, hydroxyproline, lysine, glutamic acid and anserine, which occupied $80.8\%$ of total free amino acids. The taste compounds of powdered smoked-dried mackerel were free amino acids and related compounds (1,279.4 mg/100 g), non-volatile organic acids(948.1 mg/100 g), nucleotides and their related compounds (672.8 mg/100 g), total creatinine(430.4 ntg/100 g), tetaine(86.6 mg/100 g) and small amount of TMAO. The extraction condition of powdered smoked-dried mackerel in preparing soup stock is appropriate at $100^{\circ}C$ for 1 minute. Judging from the results of taste and sensory evaluation, it is concluded that the powdered smoked-dried mackerel can be used as natural flavoring substance in preparing soups and broth.

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Studies on the Factors Affecting Barley Injury Caused by Herbicides in Drained Paddy Field (제초제에 의한 답리작맥 약해발생 요인구명에 관한 연구)

  • Whan-Seung Ryang
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.14
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    • pp.147-157
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    • 1973
  • I. The effect of excessive soil moisture(at the time of germination) on germination of barley and crop damage of herbicides was investigated. Machete(Butachlor) and TOK(Nitrofen) were treated, respectively, at the rate of 150g ai/10a on each pot whose different soil moisture content was controlled by suppling 30, 40, 50 and 60ml of water per 100gr of air-dried soil, respectively. The results are summarized as follows: 1. Excessive soil moisture beyond field moisture capacity caused great inhibition, from 20 to 100%, of the germination of barley even at untreated pots(check pots). Also, further development of root and growth of barley were greatly inhibited even though the seeds germinated. 2. The same tendency in inhibition of germination and growth as at untreated pots was observed at treated pots, too. As a whole, however, the damage were heavier at treated pots. II. Wanju naked spring barley was seeded on four different soils and covered with soil to a depth of 1 em, and then Machete, TOK, Saturn and HE-314 were treated at the rate of 180, 150 and 200, 150, and 250g ai/10a, respectively, and the effect of soil texture on crop damage of the herbicides was investigated. The results are summarized as follows: 1. Machete(emulsion and granule, at 180g ai/10a) The degree of crop damage was quite different from one soil texture to another: while almost no crop damage was observed on a clay loam soil regardless of the type of formulation, the damage became heavier as the soil texture became sandier as sandy clay loam, volcanic ash loam and sandy loam, and great inhibition of growth was observed on sandy loam soil. In general heavier damage was caused by the application of emulsion than by granular formulation. 2. TOK(Wettable powder, at 150, 250g ai/l0a) Almost the same tendency as in the application of Machete was observed, and the damage became heavier as the application rate increased. 3. Saturn(at l50g ai/l0a) No great difference in crop damage among soil textures was observed. 4. HE-3l4(at 250g ai/l0a) Almost no difference in crop damage among soil textures was observed at this rate of 250g ai/l0a. III. To study a difference of crop damage on soil covering depth(4 levels), 9 herbicides(TOK, MO, HE-3l4, Machete, Saturn, Simetryne, Simazine, Gesaran, Lorox) were treated on the pots with two different soils, and the effect of soil covering depth on crop damage of the herbicides was investigated. The results obtained in this experiment are summarized as follows: Light Clay Soil 1. The growth of barley in relation to depth of soil covering at check pots followed the order vigorous to weak; lcm>1.5cm>0.5cm>0cm. And in case of 0 and 0.5cm covering the growth of barley was very poor. 2. The damage at 0 and 0.5cm covering at treated pots was very severe, but Saturn, Machete, MO and TOK at 100 to l50g ai/l0a, respectively and He-3l4 at 250 to 375g ai/l0a were relatively safe to barley at the depths of lcm and above. 3. Simazine, Lorox and Simetryne caused slight damage even at 1.5cm covering. Sandy Loam Soil The growth of barley in relation to depth of soil covering at untreated pots followed the order, from vigorous to weak; 1.5cm 0.5cm 3cm 5cm. While MO was safe to barley at 1.5cm covering, for other chemicals more than 3cm covering was require for safe use. Machete and Saturn at 100g ai/l0a, and HE-3l4 at 250g ai/l0a was relatively safe at more than 3cm covering. Simazine, Lorox, Simetryne and Gesaran were unsafe on sandy soil regardless of covering depth.

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Wind and Flooding Damages of Rice Plants in Korea (한국의 도작과 풍수해)

  • 강양순
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.34 no.s02
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    • pp.45-65
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    • 1989
  • The Korean peninsular having the complexity of the photography and variability of climate is located within passing area of a lots of typhoon occurring from the southern islands of Philippines. So, there are various patterns of wind and flooding damages in paddy field occuring by the strong wind and the heavy rain concentrated during the summer season of rice growing period in Korea. The wind damages to rice plants in Korea were mainly caused by saline wind, dry wind and strong wind when typhoon occurred. The saline wind damage having symptom of white head or dried leaves occurred by 1.1 to 17.2 mg of salt per dry weight stuck on the plant which was located at 2. 5km away from seashore of southern coastal area during the period(from 27th to 29th, August, 1986) of typhoon &Vera& accompanying 62-96% of relative humidity, more than 6 m per second of wind velocity and 22.5 to 26.4$^{\circ}C$ of air temperature without rain. Most of the typhoons accompanying 4.0 to 8. 5m per second of wind and low humidity (lesp an 60%) with high temperature in the east coastal area and southen area of Korea. were changed to dry and hot wind by the foehn phenomenon. The dry wind damages with the symptom of the white head or the discolored brownish grain occurred at the rice heading stage. The strong wind caused the severe damages such as the broken leaves, cut-leaves and dried leaves before heading stage, lodging and shattering of grain at ripening stage mechanically during typhoon. To reduce the wind damages to rice plant, cultivation of resistant varieties to wind damages such as Sangpoongbyeo and Cheongcheongbyeo and the escape of heading stage during period of typhoon by accelerating of heading within 15th, August are effective. Though the flood disasters to rice plant such as earring away of field, burying of field, submerging and lodging damage are getting low by the construction of dam for multiple purpose and river bank, they are occasionally occurred by the regional heavy rain and water filled out in bank around the river. Paddy field were submerged for 2 to 4 days when typhoon and heavy rain occurred about the end of August. At this time, the rice plants that was in younger growing stage in the late transplanting field of southern area of Korea had the severe damages. Although panicles of rice plant which was in the meiotic growing stage and heading stage were died when flooded, they had 66% of yield compensating ability by the upper tilling panicle produced from tiller with dead panicle in ordinary transplanting paddy field. It is effective for reduction of flooding damages to cultivate the resistant variety to flooding having the resistance to bacterial leaf blight, lodging and small brown planthopper simultaneously. Especially, Tongil type rice varieties are relatively resistant to flooding, compared to Japonica rice varieties. Tongil type rice varieties had high survivals, low elongation ability of leaf sheath and blade, high recovering ability by the high root activity and photosynthesis and high yield compensating ability by the upper tillering panicle when flooded. To minimize the flooding and wind damage to rice plants in future, following research have to be carried out; 1. Data analysis by telemetering and computerization of climate, actual conditions and growing diagnosis of crops damaged by disasters. 2. Development of tolerant varieties to poor natural conditions related to flooding and wind damages. 3. Improvement of the reasonable cropping system by introduction of other crops compensating the loss of the damaged rice. 4. Increament of utilization of rice plant which was damaged.

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The Analysis of Dose in a Rectum by Multipurpose Brachytherapy Phantom (근접방사선치료용 다목적 팬톰을 이용한 직장 내 선량분석)

  • Huh, Hyun-Do;Kim, Seong-Hoon;Cho, Sam-Ju;Lee, Suk;Shin, Dong-Oh;Kwon, Soo-Il;Kim, Hun-Jung;Kim, Woo-Chul;K. Loh John-J.
    • Radiation Oncology Journal
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    • v.23 no.4
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    • pp.223-229
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    • 2005
  • Purpose: In this work we designed and made MPBP(Multi Purpose Brachytherapy Phantom). The MPBP enables one to reproduce the same patient set-up in MPBP as the treatment of the patient and we tried to get an exact analysis of rectal doses in the phantom without need of in-vivo dosimetry. Materials and Methods: Dose measurements were tried at a point of rectum 1, the reference point of rectum, with a diode detector for 4 patients treated with tandem and ovoid for a brachytherapy of a cervix cancer. Total 20 times of rectal dose measurements were made with 5 times a patient. The set-up variation of the diode detector was analyzed. The same patient set-ups were reproduced in self-made MPBP and then rectal doses were measured with TLD. Results: The measurement results of the diode detector showed that the set-up variation of the diode detector was the maximum $11.25{\pm}0.95mm$ in the y-direction for Patient 1 and the maximum $9.90{\pm}4.50mm,\;20.85{\pm}4.50mm,\;and\;19.15{\pm}3.33mm$ in the z-direction for Patient 2, 3, and 4, respectively. Un analyzing the degree of variation in 3 directions the more variation was showed in the z-direction than x- and y-direction except Patient 1. The results of TLD measurements in MPBP showed the relative maximum error of 8.6% and 7.7% at a point of rectum 1 for Patient 1 and 4, respectively and 1.7% and 1.2% for Patient 2 and 3, respectively. The doses measured at R1 and R2 were higher than those calculated except R point of Patient 2. this can be thought to related to the algorithm of dose calculation, whcih corrects for air and water but is guessed not to consider the correction for the scattered rays, but by considering the self-error (${\pm}5%$) TLD has the relative error of values measured and calculated was analyzed to be in a good agreement within 15%. Conclusion: The reproducibility of dose measurements under the same condition as the treatment could be achieved owing to the self-made MPMP and the dose at the point of interest could be analyzed accurately. If a treatment is peformed after achieving dose optimization using the data obtained in the phantom, dose will be able to be minimized to important organs.

A study on dermatologic diseases of workers exposed to cutting oil (절삭유 취급 근로자의 피부질환에 관한 연구)

  • Chun, Byung-Chul;Kim, Hee-Ok;Kim, Soon-Duck;Oh, Chil-Hwan;Yum, Yong-Tae
    • Journal of Preventive Medicine and Public Health
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    • v.29 no.4 s.55
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    • pp.785-799
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    • 1996
  • We investigated the 1,004 workers who worked in a automobile factory to study the epidemiologic characterists of dermatoses due to cutting oils. Among the workers, 667(66.4%) answered the questionaire. They are belong to 5 departments of the factory-the Engine-Work(258 workers), Gasoline engine Assembly(210), Diesel engine Assembly(96), Power train Work(86), Power train Assembly(17). We measured the oil mist concentration in air of the departments and examined the workers who had dermatologic symptoms. The results were follows; 1) Oil mist concentration ; Of all measured points(52),9 points(17.2%) exeeded $5mg/m^3$- the time-weighed PEL-and one department had a upper confidence limit(95%) higher than $5mg/m^3$. 2) Dermatologists examined 213 workers. 172 of them complained any skin symptoms at that time - itching(32.5%), papule(21.6%), scale(15.7%), vesicle(12.5%) in order. The abnormal skin site found by dermatologist were palm(29.3%), finger & nail(24.6%), forearm(16.2%), back of hand(8.4%) in order. 3) As the result of physical examination, we found that 160 workers had skin diseases. Contact dermatitis was the most common; 69 workers had contact dermatitis alone(43.1%), 11 had contact dermatitis with acne(6.9%), 10 had contact dermatitis with folliculitis(6.3%), 1 had contact dermatitis with acne & folliculitis, and 1 had contact dermatitis with abnormal pigmentation. Others were folliculitis(9 workers, 5.6%), acne(8, 5.0%), folliculitis & acne (2, 1.2%), keratosis(1, 0.6%), abnormal pigmentation (1, 0.6%), and non-specific hand eczema (47, 29.3%). 4) The prevalence of any skin diseases was 34.0 pet 100 in cutting oil users, and 13.3 per 100 in non- users. Especially, the prevalence of contact dermatitis was 23.0 per 100 in cutting oil users and 23.0 per 100 in non-users. 5) We tried patch test(standard serise, oil serise, organic solvents) on 49 patients to differentiate allergic contact dermatitis from irritant contact dermatitis and found 20 were positive. 6) In a multivariate analysis(independant=age, tenure, kinds of cutting oil), the risk of skin diseases was higher in the water-based cutting oil user and both oil user than non-user or neat oil user(odds ratio were 2.16 and 2.78, respectively). And the risk of contact dermatitis was much higher at the same groups(odds ratio were 5.16 and 6.82, respectively).

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Evaluation of the Usefulness of MapPHAN for the Verification of Volumetric Modulated Arc Therapy Planning (용적세기조절회전치료 치료계획 확인에 사용되는 MapPHAN의 유용성 평가)

  • Woo, Heon;Park, Jang Pil;Min, Jae Soon;Lee, Jae Hee;Yoo, Suk Hyun
    • The Journal of Korean Society for Radiation Therapy
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    • v.25 no.2
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    • pp.115-121
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    • 2013
  • Purpose: Latest linear accelerator and the introduction of new measurement equipment to the agency that the introduction of this equipment in the future, by analyzing the process of confirming the usefulness of the preparation process for applying it in the clinical causes some problems, should be helpful. Materials and Methods: All measurements TrueBEAM STX (Varian, USA) was used, and a file specific to each energy, irradiation conditions, the dose distribution was calculated using a computerized treatment planning equipment (Eclipse ver 10.0.39, Varian, USA). Measuring performance and cause errors in MapCHECK 2 were analyzed and measured against. In order to verify the performance of the MapCHECK 2, 6X, 6X-FFF, 10X, 10X-FFF, 15X field size $10{\times}10$ cm, gantry $0^{\circ}$, $180^{\circ}$ direction was measured by the energy. IGRT couch of the CT values affect the measurements in order to confirm, CT number values : -800 (Carbon) & -950 (COUCH in the air), -100 & 6X-950 in the state for FFF, 15X of the energy field sizes $10{\times}10$, gantry $180^{\circ}$, $135^{\circ}$, $275^{\circ}$ directionwas measured at, MapPHAN allocated to confirm the value of HU were compared, using the treatment planning computer for, Measurement error problem by the sharp edges MapPHAN Learn gantry direction MapPHAN of dependence was measured in three ways. GANTRY $90^{\circ}$, $270^{\circ}$ in the direction of the vertically erected settings 6X-FFF, 15X respectively, and Setting the state established as a horizontal field sizes $10{\times}10$, $90^{\circ}$, $45^{\circ}$, $315^{\circ}$, $270^{\circ}$ of in the direction of the energy-6X-FFF, 15X, respectively, were measured. Without intensity modulated beam of the third open arc were investigated. Results: Of basic performance MapCHECK confirm the attenuation measured by Couch, measured from the measured HU values that are assigned to the MAP-PHAN, check for calculation accuracy for the angled edge of the MapPHAN all come in a range of valid measurement errors do not affect the could see. three ways for the Gantry direction dependence, the first of the meter built into the value of the Gantry $270^{\circ}$ (relative $0^{\circ}$), $90^{\circ}$ (relative $180^{\circ}$), 6X-FFF, 15X from each -1.51, 0.83% and -0.63, -0.22% was not affected by the AP/PA direction represented. Setting the meter horizontally Gantry $90^{\circ}$, $270^{\circ}$ from the couch, Energy 6X-FFF 4.37, 2.84%, 15X, -9.63, -13.32% the difference. By-side direction measurements MapPHAN in value is not within the valid range can not, because that could be confirmed as gamma pass rate 3% of the value is greater than the value shown. You can check the Open Arc 6X-FFF, 15X energy, field size $10{\times}10$ cm $360^{\circ}$ rotation of the dose distribution in the state to look at nearly 90% pass rate to emerge. Conclusion: Based on the above results, the MapPHAN gantry direction dependence by side in the direction of the beam relative dose distribution suitable for measuring the gamma value, but accurate measurement of the absolute dose can not be considered is. this paper, a more accurate treatment plan in order to confirm, Reduce the tolerance for VMAT, such as lateral rotation investigation in order to measure accurate absolute isodose using a combination of IMF (Isocentric Mounting Fixture) MapCHEK 2, will be able to minimize the impact due to the angular dependence.

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Studies on Neck Blast Infection of Rice Plant (벼 이삭목도열병(病)의 감염(感染)에 관(關)한 연구(硏究))

  • Kim, Hong Gi;Park, Jong Seong
    • Korean Journal of Agricultural Science
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    • v.12 no.2
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    • pp.206-241
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    • 1985
  • Attempts to search infection period, infection speed in the tissue of neck blast of rice plant, location of inoculum source and effects of several conditions about the leaf sheath of rice plants for neck blast incidence have been made. 1. The most infectious period for neck blast incidence was the booting stage just before heading date, and most of necks have been infected during the booting stage and on heading date. But $Indica{\times}Japonica$ hybrid varieties had shown always high possibility for infection after booting stage. 2. Incubation period for neck blast of rice plants under natural conditions had rather a long period ranging from 10 to 22 days. Under artificial inoculation condition incubation period in the young panicle was shorter than in the old panicle. Panicles that emerged from the sheath of flag leaf had long incubation period, with a low infection rate and they also shown slow infection speed in the tissue. 3. Considering the incubation period of neck blast of rice plant, we assumed that the most effective application periods of chemicals are 5-10 days for immediate effective chemicals and 10-15 days for slow effective chemicals before heading. 4. Infiltration of conidia into the leaf sheath of rice plant carried out by saturation effect with water through the suture of the upper three leaves. The number of conidia observed in the leaf sheath during the booting stage were higher than those in the leaf sheath during other stages. Ligule had protected to infiltrate of conidia into the leaf sheath. 5. When conidia were infiltrated into the leaf sheath, the highest number of attached conidia was observed on the panicle base and panicle axis with hairs and degenerated panicle, which seemed to promote the infection of neck blast. 6. The lowest spore concentration for neck blast incidence was variable with rice varietal groups. $Indica{\times}Japonica$ hybrid varieties were infected easily compared to the Japonica type varieties, especially. The number of spores for neck blast incidence in $Indica{\times}Japonica$ hybrid varieties was less than 100 and disease index was higher also in $Indica{\times}Japonica$ hybrid than in Japonica type varieties. 7. Nitrogen content and silicate content were related with blast incidence in necks of rice plants in the different growing stage changed during growing period. Nitrogen content increased from booting stage to heading date and then decreased gradually as time passes. Silicate content increased from booting stage after heading with time. Change of these content promoted to increase neck blast infection. 8. Conidia moved to rice plant by ascending and desending dispersal and then attached on the rice plant. Conidia transfered horizontally was found very negligible. So we presumed that infection rate of neck blast was very low after emergence of panicle base from the leaf sheath. Also ascending air current by temperature difference between upper and lower side of rice plant seemed to increase the liberation of spores. 9. Conidial number of the blast fungus collected just before and after heading date was closely related with neck blast incidence. Lesions on three leaves from the top were closely related with neck blast incidence, because they had high potential for conidia formation of rice blast fungus and they were direct inoculum sources for neck blast. 10. The condition inside the leaf sheath was very favorable for the incidence of neck blast and the neck blast incidence in the leaf sheath increased as the level of fertilizer applied increased. Therefore, the infection rate of neck blast on the all panicle parts such as panicle base, panicle branches, spikelets, nodes, and internodes inside the leaf sheath didn't show differences due to varietal resistance or fertilizers applied. 11. Except for others among dominant species of fungi in the leaf sheath, only Gerlachia oryzae appeared to promote incidence of neck blast. It was assumed that days for heading of varieties were related with neck blast incidence.

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Studies on the Internal Changes and Germinability during the Period of Seed Maturation of Pinus koraiensis Sieb. et Zucc. (잣나무 종자(種字) 성숙과정(成熟過程)에 있어서의 내적변화(內的變化)와 발아력(發芽力)에 대(對)한 연구(硏究))

  • Min, Kyung-Hyun
    • Journal of Korean Society of Forest Science
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    • v.21 no.1
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    • pp.1-34
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    • 1974
  • The author intended to investigate external and internal changes in the cone structure, changes in water content, sugar, fat and protein during the period of seed maturation which bears a proper germinability. The experimental results can be summarized as in the following. 1. Male flowers 1) Pollen-mother cells occur as a mass from late in April to early in May, and form pollen tetrads through meiosis early and middle of May. Pollen with simple nucleus reach maturity late in May. 2) Stamen number of a male flower is almost same as the scale number of cone and is 69-102 stamens. One stamen includes 5800-7300 pollen. 3) The shape is round and elliptical, both of a pollen has air-sac with $80-91{\mu}$ in length, and has cuticlar exine and cellulose intine. 4) Pollen germinate in 68 hours at $25^{\circ}C$ with distilled water of pH 6.0, 2% sugar and 0.8% agar. 2. Female flowers 1) Ovuliferous scales grow rapidly in late April, and differentiation of ovules begins early in May. Embryo-sac-mother cells produce pollen tetrads through meiosis in the middle of May, and flower in late May. 2) The pollinated female flowers show repeated divisions of embryo-sac nucleus, and a great number of free nuclei form a mass for overwintering. Morphogenesis of isolation in the mass structure takes place from the middle of March, and that forms albuminous bodies of aivealus in early May. 3. Formation of pollinators and embryos. 1) Archegonia produce archegonial initial cells in the middle and late April, and pollinators are produced in the late April and late in early May. 2) After pollination, Oespore nuclei are seen to divide in the late May forming a layer of suspensor from the diaphragm in early June and in the middle of June. Thus this happens to show 4 pro-embryos. The organ of embryos begins to differentiate 1 pro-embryo and reachs perfect maturation in late August. 4. The growth of cones 1) In the year of flowering, strobiles grow during the period from the middle of June to the middle of July, and do not grow after the middle of August. Strobiles grow 1.6 times more in length 3.3 times short in diameter and about 22 times more weight than those of female flower in the year of flowering. 2) The cones at the adult stage grow 7 times longer in diameter, 12-15 times shorter diameter than those of strobiles after flowering. 3) Cone has 96-133 scales with the ratio of scale to be 69-80% and the length of cone is 11-13cm. Diameter is 5-8cm with 160-190g weight, and the seed number of it is 90-150 having empty seed ratio of 8-15%. 5. Formation of seed-coats 1) The layers of outer seed-coat become most for the width of $703{\mu}$ in the middle of July. At the adult stage of seed, it becomes $550-580{\mu}$ in size by decreasing moisture content. Then a horny and the cortical tissue of outer coats become differentiated. 2) The outer seed-coat of mature seeds forms epidermal cells of 3-4 layers and the stone cells of 16-21 layers. The interior part of it becomes parenchyma layer of 1 or 2 rows. 3) Inner seed-coat is formed 2 months earlier than the outer seed-coat in the middle of May, having the most width of inner seed-coat $667{\mu}$. At the adult stage it loses to $80-90{\mu}$. 6. Change in moisture content After pollination moisture content becomes gradually increased at the top in the early June and becomes markedly decreased in the middle of August. At the adult stage it shows 43~48% in cone, 23~25% in the outer seed-coat, 32~37% in the inner seed-coat, 23~26% in the inner seed-coat and endosperm and embryo, 21~24% in the embryo and endosperm, 36~40% in the embryos. 7. The content compositions of seed 1) Fat contents become gradually increased after the early May, at the adult stage it occupies 65~85% more fat than walnut and palm. Embryo includes 78.8% fat, and 57.0% fat in endosperm. 2) Sugar content after pollination becomes greatly increased as in the case of reducing sugar, while non-reducing sugar becomes increased in the early June. 3) Crude protein content becomes gradually increased after the early May, and at the adult stage it becomes 48.8%. Endosperm is made up with more protein than embryo. 8. The test of germination The collected optimum period of Pinus koraiensis seeds at an adequate maturity was collected in the early September, and used for the germination test of reduction-method and embryo culture. Seeds were taken at the interval of 7 days from the middle of July to the middle of September for the germination test at germination apparatus.

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Studies on Sclerotium rolfsii Sacc. isolated from Magnolia kobus DC. in Korea (목련(Magnolia kobus DC.)에서 분리한 흰비단병균(Sclerotium rolfsii Sacc.)에 관한 연구)

  • Kim Kichung
    • Korean journal of applied entomology
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    • v.13 no.3 s.20
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    • pp.105-133
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    • 1974
  • The present study is an attempt to solve the basic problems involved in the control of the Sclerotium disease. The biologic stranis of Sclerotium rolfsii Sacc., pathogen of Sclerotium disease of Magnolia kobus, were differentiated, and the effects of vitamins, various nitrogen and carbon sources on its mycelial growth and sclerotial production have been investigated. In addition the relationship between the cultural filtrate of Penicillium sp. and the growth of Sclerotium rolfsii, the tolerance of its mycelia or sclerotia to moist heat or drought and to Benlate (methyl-(butylcarbamoy 1)-2-benzimidazole carbamate), Tachigaren (3-hydroxy-5-methylisoxazole) and other chemicals were also clarified. The results are summarizee as follows: 1. There were two biologic strains, Type-l and Type-2 among isolates. They differed from each other in the mode of growth and colonial appearance on the media, aversion phenomenon and in their pathogenicity. These two types had similar pathogenicity to the Magnolia kobus and Robinia pseudoacasia, but behaved somewhat differently to the soybaen and cucumber, the Type-l being more virulent. 2. Except potassium nitrite, sodium nitrite and glycine, all of the 12 nitrogen sources tested were utilized for the mycelial growth and sclerotial production of this fungus when 10r/l of thiamine hydrochloride was added in the culture solution. Considering the forms of nitrogen, ammonium nitrogen was more available than nitrate nitrogen for the growth of mycelia, but nitrate nitrogen was better for sclerotia formation. Organic nitrogen showed different availabilities according to compounds used. While nitrite nitrogen was unavailable for both mycelial growth and sclerotial formation whether thiamine hydrochlioride was added or not. 3. Seven kinds of carbon sources examined were not effective in general, as long as thiamine hydrochloride was not added. When thiamine hydrochloride was added, glucose and saccharose exhibited mycelial growth, while rnaltose and soluble starch gave lesser, and xylose, lactose, and glycine showed no effect at all,. In the sclerotial production, all the tested carbon sources, except lactose, were effective, and glucose, maltose, saccharose, and soluble starch gave better results. 4. At the same level of nitrogen, the amount of mycelial growth increased as more carbon Sources were applied but decreased with the increase of nitrogen above 0.5g/1. The amount of sclerotial production decreased wi th the increase of carbon sources. 5. Sclerotium rolfsii was thiamine-defficient and required thiamine 20r/l for maximun growth of mycelia. At a higher concentration of more than 20r/l, however, mycelial growth decreased as the concentration increased, and was inhibited at l50r/l to such a degree of thiamine-free. 6. The effect of the nitrogen sources on the mycelial growth under the presence of thiamine were recognized in the decreasing order of $NH_4NO_3,\;(NH_4)_2SO_4,\;asparagine,\;KNO_3$, and their effects on the sclerotial production in the order of $KNO_3,\;NH_4NO_3,\;asparagine,\;(NH_4)_2SO_4$. The optimum concentration of thiamine was about 12r/l in $KNO_3$ and about 16r/l in asparagine for the growth of mycelia; about 8r/l in $KNO_3$ and $NH_4NO_3$, and 16r/l in asparagine for the production of sclerotia. 7. After the fungus started to grow, the pH value of cultural filtrate rapidly dropped to about 3.5. Hereafter, its rate slowed down as the growth amount increased and did not depreciated below pH2.2. 8. The role of thiamine in the growth of the organism was vital. If thiamine was not added, the combination of biotin, pyridoxine, and inositol did not show any effects on the growth of the organism at all. Equivalent or better mycelial growth was recognized in the combination of thiamine+pyridoxine, thiamine+inositol, thiamine+biotin+pyridoxine, and thiamine+biotin+pyridoxine+inositol, as compared with thiamine alone. In the combinations of thiamine+biotin and thiamine+biotin+inositol, mycelial growth was inhibited. Sclerotial production in dry weight increased more in these combinations than in the medium of thiamine alone. 9. The stimulating effects of the Penicillium cultural filtrate on the mycelial growth was noticed. It increased linearly with the increase of filtrate concentration up to 6-15 ml/50ml basal medium solution. 10. $NH_4NO_3$. as a nitrogen source for mycelial growth was more effective than asparasine regardless of the concentration of cultural filtrate. 11. In the series of fractionations of the cultural filtrate, mycelial growth occured in unvolatile, ether insoluble cation-adsorbed or anion-unadsorbed substance fractions among the fractions of volatile, unvolatile acids, ether soluble organic acids, ether insoluble, cation-adsorbed, cation-unadsorbed, anion-adsorbed and anion-unadsorbed. and anion-un-adsorbed substance tested. Sclerotia were produced only in cation-adsorbed fraction. 12. According to the above results, it was assumed that substances for the mycelial growth and sclerotial formation and inhibitor of sclerotial formation were include::! in cultural filtrate and they were quite different from each other. I was further assumed that the former two substances are un volatile, ether insotuble, and adsorbed to cation-exchange resin, but not adsorbed to anion, whereas the latter is unvolatile, ether insoluble, and not adsorbed to cation or anion-exchange resin. 13. Seven amino acids-aspartic acid, cystine, glysine, histidine, Iycine, tyrosine and dinitroaniline-were detected in the fractions adsorbed to cation-exchange resin by applying the paper chromatography improved with DNP-amino acids. 14. Mycelial growth or sclerotial production was not stimulated significantly by separate or combined application of glutamic acid, aspartic acid, cystine, histidine, and glysine. Tyrosine gave the stimulating effect when applied .alone and when combined with other amino acids in some cases. 15. The tolerance of sclerotia to moist heat varied according to their water content, that was, the dried sclerotia are more tolerant than wet ones. The sclerotia harvested directly from the media, both Type-1 and Type-2, lost viability within 5 minutes at $52^{\circ}C$. Sclerotia dried for 155 days at$26^{\circ}C$ had more tolerance: sclerotia of Type-l were killed in 15 mins. at $52^{\circ}C$ and in 5 mins. at $57^{\circ}C$, and sclerotia of Type-2 were killed in 10 mins. both at $52^{\circ}C$ or $57^{\circ}C$. 16. Cultural sclerotia of both strains maintained good germinability for 132 days at$26^{\circ}C$. Natural sclerotia of them stored for 283 days under air dry condition still had good germinability, even for 443 days: type-l and type-2 maintained $20\%$ and $26.9\%$ germinability, respectively. 17. The tolerance to low temperature increased in the order of mycelia, felts and sclerotia. Mycelia completely lost the ability to grow within 1 week at $7-8^{\circ}C$> below zero, while mycelial felts still maintained the viability after .3 weeks at $7-20^{\circ}C$ below zero, and sclerotia were even more tolerant. 18. Sclerotia of type-l and type-2 were killed when dipped into the $0.05\%$ solution of mercury chloride for 180 mins. and 240 mins. respectively: and in the $0.1\%$ solution, Type-l for 60 mins. and Type-2 for 30 mins. In the $0.125\%$ uspulun solution, Type-l sclerotia were killed in 180 mins., and those of Type-2 were killed for 90 mins. in the$0.125\%$solution. Dipping into the $5\%$ copper sulphate solution or $0.2\%$ solution of Ceresan lime or Mercron for 240 mins. failed to kill sclerotia of either Type-l or Type-2. 19. Inhibitory effect on mycelial growth of Benlate or Tachi-garen in the liquid culture increased as the concentration increased. 6 days after application, obvious inhibitory effects were found in all treatments except Benlate 0.5ppm; but after 12 days, distingushed diflerences were shown among the different concentrations. As compared with the control, mycelial growth was inhibited by $66\%$ at 0.5ppm and by $92\%$ at 2.0ppm of Benlate, and by$54\%$ at 1ppm and about $77\%$ at 1.5ppm or 2.0ppm of Tachigaren. The mycelial growth was inhibited completely at 500ppm of both fungicides, and the formation of sclerotia was checked at 1,000ppm of Benlate ant at 500ppm or 1,000ppm of Tachigaren. 20. Consumptions of glucose or ammonium nitrogen in the culture solution usually increased with the increment of mycelial growth, but when Benlate or Tachigaren were applied, consumptions of glucose or ammonium nitrogen were inhibited with the increment of concentration of the fungicides. At the low concentrations of Benlate (0.5ppm or 1ppm), however, ammonium nitrogen consumption was higher than that of the ontrol. 21. The amount of mycelia produced by consuming 1mg of glucose or ammonium nitrogen in the culture solution was lowered markedly by Benlate or Tachigaren. Such effects were the severest on the third day after their treatment in all concentrations, and then gradually recovered with the progress of time. 22. In the sand culture, mycelial growth was not inhibited. It was indirectly estimated by the amount of $CO_2$ evolved at any concentrations, except in the Tachigaren 100mg/g sand in which mycelial growth was inhibited significantly. Sclerotial production was completely depressed in the 10mg/g sand of Benlate or Tachigaren. 23. There was no visible inhibitory effect on the germination of sclerotia when the sclerotia were dipped in the solution 0.1, 1.0, 100, 1.000ppm of Benlate or Tachigaren for 10 minutes or even 20 minutes.

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