• Title/Summary/Keyword: Mass culture

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Immunogenicity of Synthetic Peptide Specific for Major Immunogenic Determinat of Hepatitis B Surface Antigen (B형간염(型肝炎) 표면항원(表面抗原)의 주면역원(主免疫原) 결정기(決定基)에 특이(特異)한 합성(合成) Peptide의 면역원성(免疫原性)에 관한 연구(硏究))

  • Shin, Kwang-soon;Han, Su-nam
    • Korean Journal of Veterinary Research
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    • v.25 no.1
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    • pp.7-17
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    • 1985
  • Many investigators have been pursuing various attempts so far to produce hepatitis B surface antigen(HBsAg) vaccines using the techniques such as isolation from plasma of chronic HBsAg carrier, recombinant DNA technique or preparation of synthetic peptides specific for immunogenic determinants. Hepatitis B virus can not grow on any cell lines by the tissue culture technique at the present time. The plasma of chronic HBsAg carrier is expensive and its source is limited. The HBsAg from the recombinant DNA technique gave still very low yield. Another approach, therefore, has been initiated to develop a synthetic hepatitis B virus vaccine. The possible use of several distinct synthetic vaccines in prophylaxis can be facilitated by availability of full synthetic immunogens. Peptides synthesized for potential application as antiviral vaccines have been mostly tested in the form of conjugates with carrier proteins, although the free synthetic peptide can be immunogenic. To understand basic knowledges on the antigenicity and immunogenicity of a synthetic peptide specific for major immunogenic determinant of HBsAg, a nonapeptide, $H_2N^{139}Cys-Thr-Lys-Pro-Thr-Asp-Gly-^{146}Asn-Aba$ COOH, which corresponds to HBsAg amino acid residues 139 to 147, was synthesized by the Merrifield's solid-phase method with a slight modification. The antigenicity and immunogenicity of this specific synthetic peptide were examined comparing with purified plasma-derived natural HBsAg. The results obtained are as follows; 1. The peptide synthesized showed the identical amino acid composition to the theoretical value. The degree of purification and molecular weight were acertained by methods of high performance liquid chromatography and mass spectrometry. 2. Using m-maleimidobenzoyl-N-hydroxysuccinimide ester as a conjugating agent, the synthetic peptide was conjugated to rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin. Their conjugation yields were 8.3, 9.5, 15.8, 13.5, and 11.2%, respectively. 3. The natural HBsAg was purified from plasma of chronic HBsAg carrier. By the electron microscopic observation of the purified natural HBsAg preparation, no Dane particles were observed and the preparation showed negative DNA polymerase activity. 4. Antigenicity of the synthetic peptide and the plasma-derived natural HBsAg was determined by competition radioimmunoassay using $^{125}I$-natural HBsAg. Their 50% inhibitions appeared as $90{\mu}g/ml$ and $0.12{\mu}g/ml$ for the synthetic peptide and the natural HBsAg, respectively. This indicates that the former was about 750-fold less antigenic than the latter. 5. Immunogenicity of the synthetic peptide was determined by administering the peptide-carrier conjugates into rabbits with and without Freund's complete adjuvant. Regardless the carrier proteins and adjuvant, positive immune responses to the synthetic peptide were observed. The higher antibody titers, however, were shown in the groups administered with Freund's complete adjuvant. 6. Immunizing dose 50% in mice of the various peptide-carrier conjugates was 5.47, 6.00, 65.16, 31.25 and $13.03{\mu}g/dose$ for rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin, respectively, while the natural HBsAg showed $0.65{\mu}g/dose$. 7. It was postulated that homologous proteins prefer to heterologous ones as the carriers.

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Isotopic Evidence of Marine Yeast to Artificial Culture of Moina macrocopa (물벼룩(Moina macrocopa)배양을 위한 해양효모의 유효성에 대한 안전 동위원소의 증거)

  • Kim Mu-Chan;Kang Chang-Keun;Park Hye-Young;Lee Dae-Seong;Kim Yun-Sook;Lee Won-Jae
    • Korean Journal of Microbiology
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    • v.42 no.2
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    • pp.111-115
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    • 2006
  • A feeding trial was conducted to test the use of marine yeasts isolated from seawaters and sediments as a dietary source in cultivating a Cladocera, Moina macrocopa which is available as an alternative live food for fish larvae. The marine yeast-fed M. macrocopa had similar essential amino acid profiles to the documented values for Rotifers and Artemia enriched in microalgae and commercial diets. Erythrobacter sp. $S{\pi}-1$ lacked ${\omega}-3$ high unsaturated fatty acids (HUFAs), $20:5{\omega}-3$ (EPA) and $22:6{\omega}-3$ (DHA), which were also poor but detected in both the marine yeasts. An increase in the $20:5{\omega}-3$ and $22:6{\omega}-3$ levels, compared with the levels in marine yeast strains themselves, was more pronounced in the $22:6{\omega}-3$ level of Moina fed the Candida sp. Y-16, resulting in a high DHA:EPA ratio. When the Moina diets were switched, their ${\delta}^{13}C$ values shifted gradually toward the values of the switched diets. Diet switch from Erythrobacter sp. $S{\pi}-1$to Candide sp. Y.16 resulted in a more rapid turnover of Moina tissue carbon than that in the inverse case. When fed a mixed diet, the ${\delta}^{13}C$ values of Moina tissue approached the value of marine yeasts immediately. These temporal changes in the ${\delta}^{13}C$ values of Moina tissue indicate the preferential ingestion of marine yeasts and a selective assimilation of the carbon originated from marine yeasts. These findings suggest that marine yeasts, particularly Candida sp. Y-16, are highly available to mass cultures of M. macrocopa, providing better nutritional and dietaty values than the commercial diet (Erythrobacter sp. $S{\pi}-1$).

Effect of an Acid pH Shock on Physiological Changes of Chlamydomonas acidophila (Chlorophyta), UTCC 122 (pH 충격에 의한 Chlamydomonas acidophila (Chlorophyta), UTCC 122의 생리적 변화에 관한 연구)

  • Lee, Kyung;Ki, Jang-Seu;Kim, Say-Wa;Han, Myung-Soo;Choi, Young-Kil;Yoo, Kwang-Il
    • Korean Journal of Ecology and Environment
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    • v.35 no.3 s.99
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    • pp.145-151
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    • 2002
  • The effect of low pH on physiological changes was studied with the acidophilic green alga, Clamydomonas acidophila, UTCC 122. The growthrates (${\mu}$) were identical, $0.5{\sim}0.7\;day^{-1}$, at pH 3.7${\sim}$6.7 and no significantly different (ANOVA, p =0.134), showing cell volume reduced gradually as they were growing, whereas that at pH 2.7 was falling to zero and cell volume increased dramatically. Chlorophyll a concentration of the cultures incubated for one day was $191{\sim}255\;pg\;cell^{-1}$, after then it declined from $60{\sim}103\;pg\;cell^{-1}$ at pH 3.7${\sim}$6.7 except $210\;pg\;cell^{-1}$ at pH 2.7, which was directly related with cell volume. External carbonic anhydrase (CA) activity was varied from1.1 to$3.7{\times}10^{-4}\;E.U.\;mm^{-2}$, showing the gradualincrease during culture, except at 2.7 and pH 5.7. However there was not found any relationship among the pH gradient cultures. CA molecular mass of C. acidophila was 29 kBa, and concentration of that was identical in all cultures. The proteins of 41 kDa and 63 were not or very faintly expressed in low pH cultures, in contrast that of 17 kDa more expressed. In this work, we found that C. acidophila could live optimally within a wide range of acidic pH, and 17 kDa of unidentified protein might be concerned with tolerating in low acid environment.

Plant Growth Promoting Effect and Antifungal Activity of Bacillus subtilis S37-2 (Bacillus subtilis S37-2 균주의 항진균활성 및 식물생육촉진 효과)

  • Kwon, Jang-Sik;Weon, Hang-Yeon;Suh, Jang-Sun;Kim, Wan-Gyu;Jang, Kab-Yeul;Noh, Hyung-Jun
    • Korean Journal of Soil Science and Fertilizer
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    • v.40 no.6
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    • pp.447-453
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    • 2007
  • With a broad objective for the development of microbial based fertilizers, a total of 373 strains were isolated from rhizoplane and rhizosphere of pepper, tomato, lettuce, pasture, and grass. The efficacy of the isolates to augument overall plant growth was evaluated. After screening for their plant growth promotion and antagonistic properties in vitro efficient strains were further selected. The most efficient strains was characterized by 16S rRNA gene sequences and biochemical techniques and was designated as Bacillus subtilis S37-2. The strains facilitated plant growth and inhibited the plant phathogenic fungi such as Fusarium oxysporum (KACC 40037, Rhizoctonia solani (KACC 40140), and Sclerotinia sclerotiorum (KACC 40457). Pot based bioassay using lettuce as test plant was conducted by inoculating suspension ($10^5$ to $10^8cells\;mL^{-1}$) of B. subtilis S37-2 to the rhizosphere of lettuce cultivated in soil pots. Compared with non-inoculated pots, marked increase in leaf (42.3%) and root mass (48.7%) was observed in the inoculation group where the 50ml of cell mixture ($8.7{\times}10^8cells\;ml^{-1}$) was applied to the rhizosphere of letuce either once or twice. Antagonistic effects of B. subtilis S37-2 strain on S. sclerotiorum (KACC 40457) were tested. All the tested lettuce plants perished after 9 days in treatment containing only S. sclerotiorum, but only 17% of lettuce was perished in the inoculation plot. B. subtilis grew well in the TSB culture medium. The isolates grew better in yeast extracts than peptone and tryptone as nitrogen source. The growth rate was 2~4 times greater at $37^{\circ}C$ as compared with $30^{\circ}C$ incubation temperature. B. subitlis S37-2 produced $0.1{\mu}g\;ml^{-1}$ of IAA (indole 3-acetic acid) in the TSB medium containing L-tryptophan($20mg\;L^{-1}$) in 24 hours.

A study of Artificial Intelligence (AI) Speaker's Development Process in Terms of Social Constructivism: Focused on the Products and Periodic Co-revolution Process (인공지능(AI) 스피커에 대한 사회구성 차원의 발달과정 연구: 제품과 시기별 공진화 과정을 중심으로)

  • Cha, Hyeon-ju;Kweon, Sang-hee
    • Journal of Internet Computing and Services
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    • v.22 no.1
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    • pp.109-135
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    • 2021
  • his study classified the development process of artificial intelligence (AI) speakers through analysis of the news text of artificial intelligence (AI) speakers shown in traditional news reports, and identified the characteristics of each product by period. The theoretical background used in the analysis are news frames and topic frames. As analysis methods, topic modeling and semantic network analysis using the LDA method were used. The research method was a content analysis method. From 2014 to 2019, 2710 news related to AI speakers were first collected, and secondly, topic frames were analyzed using Nodexl algorithm. The result of this study is that, first, the trend of topic frames by AI speaker provider type was different according to the characteristics of the four operators (communication service provider, online platform, OS provider, and IT device manufacturer). Specifically, online platform operators (Google, Naver, Amazon, Kakao) appeared as a frame that uses AI speakers as'search or input devices'. On the other hand, telecommunications operators (SKT, KT) showed prominent frames for IPTV, which is the parent company's flagship business, and 'auxiliary device' of the telecommunication business. Furthermore, the frame of "personalization of products and voice service" was remarkable for OS operators (MS, Apple), and the frame for IT device manufacturers (Samsung) was "Internet of Things (IoT) Integrated Intelligence System". The econd, result id that the trend of the topic frame by AI speaker development period (by year) showed a tendency to develop around AI technology in the first phase (2014-2016), and in the second phase (2017-2018), the social relationship between AI technology and users It was related to interaction, and in the third phase (2019), there was a trend of shifting from AI technology-centered to user-centered. As a result of QAP analysis, it was found that news frames by business operator and development period in AI speaker development are socially constituted by determinants of media discourse. The implication of this study was that the evolution of AI speakers was found by the characteristics of the parent company and the process of co-evolution due to interactions between users by business operator and development period. The implications of this study are that the results of this study are important indicators for predicting the future prospects of AI speakers and presenting directions accordingly.

A Study of Local Festival for the China Hebeisheng (중국 하북성 마을제 연구 - 하북성조현범장이월이룡패회중룡신적여인(河北省趙縣范庄二月二龍牌會中龍神的與人) -)

  • Park, Kwang-Jun
    • Korean Journal of Heritage: History & Science
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    • v.36
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    • pp.347-377
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    • 2003
  • China is a country with large agricultural areas and subject to frequent calamities. Drought is the top of them. It has been a key problem for development of agriculture in the country. In the long struggle against drought, Chinese have accumulated many rational and irrational experiences. The Dragon Kings Belief, which is popular in North China and discussed in a thesis, is one of their irrational experiences. The belief was passed together with Buddhism from India to China in the Tang Dynasty. After it settled down, it was incorporated with the local five dragons belief and a set of beliefs in dragon kings came into existence. The emergence of the dragon kings belief ended the history that the title of rain got was not clear in China and Dragon kings finally got the status. Irrigation is the lifeblood of agriculture in China. In a Chinese mind, Dragon kings are the most important gods who take charge of rain and thus offer the lifeblood. In understanding the nature and characteristics of Chinese traditional culture, it is important for us to make clear the origin and evolution of the belief, find out its nature, function and operation. In the every year beginning of February of the Fanzhuang calendar in the people of Hebeisheng Zhaoxian, would all hold a festival to offer sacrifices to the $^{{\circ}TM}^{\prime}longpai$. Longpai was regarded as the core of the temple fair, thus the native sons came to call this festival; "longpaihui". In this region the'Fanzhuang longpaihui'developed into a well knownand grand temple fair. It was able to attract numerous pilgrims with its special magic power, occupying a place in $China^{{\circ}TM}$ 'eryueer'festival with festive dragon activities. The dragon is a common totem among Chinese nationals. The belief worship of the dragon dates from the start time of primitive societies. Dragon oneself the ancients worship's thunder lightning. In the worship of the great universe, at first afterwards this belief with the tribe's totem worships to combine to become the animal spirit. In ancient myths legends, along with folk religion and beliefs all hold a very important position. The longpaihui is a temple fair without a temple; this characteristic is a distinction between longpaihui and other temple fairs. As for longpaihui must of the early historical records are unclear. The originator of a huitou system has a kind of organized form of the special features rather, originator of a huitou not fix constant, everything follows voluntarily principle, can become member with the freedom, also can back at any time the meeting. There is a longpaihui for 'dangjiaren', is total representative director in the originator of a huitou will. 'banghui' scope particularly for extensive, come apparently every kind of buildup that help can return into the banghui, where is the person of this village or outside village of, the general cent in banghui work is clear and definite, for longpaihui would various businesses open smoothly the exhibition provides to guarantees powerfully. Fanzhuang longpaihui from the beginning of February to beginning six proceed six days totally. The longpai is used as the ancestry absolute being to exsits with the community absolute being at the same time in fanzhuang first took civil faith, in reality is a kind of method to support social machine in native folks realize together that local community that important function, it provided a space, a kind of a view to take with a relation, rising contact, communication, solidify the community contents small village, formation with fanzhuang. The fanzhuang is used as supplies for gathering town, by luck too is this local community trade exchanges center at the same time therefore can say the faith of the longpai, in addition to its people's custom, religious meaning, still have got the important and social function. Moreover matter worthy of mentioning, Longpai would in organize process, from prepare and plan the producing of meeting every kind of meeting a longpeng of the matter do, all letting person feeling is to adjust the popular support of, get the mass approbation with positive participate. Apart from the originator of a huitou excluding, those although not originator of a huitou, however enthusiasm participate the banghui of its business, also is too much for the number.

Ginseng Research in Natural Products Research Institute (NPRI) and the Pharmaceutical Industry Complex in Gaesong (생약연구소의 인삼연구와 약도개성)

  • Park, Ju-young
    • Journal of Ginseng Culture
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    • v.3
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    • pp.54-73
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    • 2021
  • The Natural Products Research Institute (NPRI, 生藥硏究所), an institution affiliated with Keijo Imperial University (京城帝國大學), was the predecessor of the NPRI at Seoul National University and a comprehensive research institute that focused on ginseng research during the Japanese colonial era. It was established under the leadership of Noriyuki Sugihara (杉原德行), a professor of the second lecture in pharmacology at the College of Medicine in Keijo Imperial University. Prof. Sugihara concentrated on studying Korean ginseng and herbal medicine beginning in 1926 when the second lecture of pharmacology was established. In addition to Prof. Sugihara, who majored in medicine and pharmacology, Kaku Tenmin (加來天民), an assistant professor who majored in pharmacy; Tsutomu Ishidoya (石戶谷勉), a lecturer who majored in agriculture and forestry; and about 36 researchers actively worked in the laboratory before the establishment of the NPRI in 1939. Among these personnel, approximately 14 Korean researchers had basic medical knowledge, derived mostly from specialized schools, such as medical, dental, and pharmaceutical institutions. As part of the initiative to explore the medicinal herbs of Joseon, the number of Korean researchers increased beginning in 1930. This increase started with Min Byung-Ki (閔丙祺) and Kim Ha-sik (金夏植). The second lecture of pharmacology presented various research results in areas covering medicinal plants in Joseon as well as pharmacological actions and component analyses of herbal medicines. It also conducted joint research with variousinstitutions. Meanwhile, in Gaesong (開城), the largest ginseng-producing area in Korea, the plan for the Pharmaceutical Industry Complex was established in 1935. This was a large-scale project aimed at generating profits through research on and the mass production of drugs and the reformation of the ginseng industry under collaboration among the Gaesong Ministry, Kwandong (關東) military forces, Keijo Imperial University, and private organizations. In 1936 and 1938, the Gyeonggi Provincial Medicinal Plant Research Institute (京畿道立 藥用植物硏究所) and the Herb Garden of Keijo Imperial University (京城帝國大學 藥草園) and Pharmaceutical Factory were established, respectively. These institutions merged to become Keijo Imperial University's NPRI, which wasthen overseen by Prof. Sugihara as director. Aside from conducting pharmacological research on ginseng, the NPRI devoted efforts to the development and sale of ginseng-based drugs, such as Sunryosam (鮮麗蔘), and the cultivation of ginseng. In 1941, the Jeju Urban Test Center (濟州島試驗場) was established, and an insecticide called Pancy (パンシ) was produced using Jeju-do medicinal herbs. However, even before research results were published in earnest, Japanese researchers, including Prof. Sugihara, hurriedly returned to Japan in 1945 because of the surrender of Japanese forces and the liberation of Korea. The NPRI was handed over to Seoul National University and led by Prof. Oh Jin-Sup (吳鎭燮), a former medical student at Keijo Imperial University. Scholars such as Woo Lin-Keun (禹麟根) and Seok Joo-Myung (石宙明) worked diligently to deal with the Korean pharmaceutical industry.

Analysis of an ancient textiles from the Xianbei period tombs of the Shiveet Khairkhan site, Mongolia (몽골 시베트 하이르한 유적 선비 시기(1~3세기) 고분 출토 직물의 섬유와 염료 분석)

  • YUN Eunyoung;YU Jia;PARK Serin;AN Boyeon
    • Korean Journal of Heritage: History & Science
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    • v.55 no.4
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    • pp.166-177
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    • 2022
  • The Shiveet Khairkhan is located on Tsengel Som in the middle of Bayan-ulgi Aimag in the Altai region. Various remains have been identified, and it has been found to be an important area of the Eurasian steppe. In this study, the characteristics of textile fibers and dyes excavated from the tombs of the 1st~3rd century Xianbei period in the sites of Shiveet Khairkhan, Mongolia were investigated. As a result of analysis using optical microscopic observation and attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR) for fiber identification, green and yellow fabrics were identified as silk fabrics. To investigate the properties of the dye, the surface reflectance of the dyed fabric was measured using an fiber optic reflectance spectrophotometer for non-destructive analysis. The green fabric appeared similar to the reflection spectrum of indigo dye. In addition, as a result of component analysis using gas chromatography-mass spectrometry, isatin and indigotine were detected. Isatin and indigotine are characteristic components of indigo dye, and it was found that the green fabric of the tombs of the Xianbei period was dyed using indigo dye. It was difficult to identify the type of dye in the yellow fabric as a result of reflectance spectrum and gas chromatography analysis. Indigo plants are a dye used for blue dyeing from thousands of years ago, and many species are distributed around the world. It was confirmed that the fabric was relatively well preserved and indigo dye was used for the green Jikryeongui (garment with a straight collar) in the ancient tomb of the Xianbei period about 1,800 years ago, even though it was buried for a long time. Scientific investigation of textile cultural heritage is an essential process for conservation treatment, restoration, exhibition, and the creation of a conservation environment. It is expected that related research will be activated in the future and will be helpful in interpreting the living culture at the time, preserving textiles, and a conservation environment.

Viability Test and Bulk Harvest of Marine Phytoplankton Communities to Verify the Efficacy of a Ship's Ballast Water Management System Based on USCG Phase II (USCG Phase II 선박평형수 성능 평가를 위한 해양 식물플랑크톤군집 대량 확보 및 생물사멸시험)

  • Hyun, Bonggil;Baek, Seung Ho;Lee, Woo Jin;Shin, Kyoungsoon
    • Journal of the Korean Society of Marine Environment & Safety
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    • v.22 no.5
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    • pp.483-489
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    • 2016
  • The type approval test for USCG Phase II must be satisfied such that living natural biota occupy more than 75 % of whole biota in a test tank. Thus, we harvested a community of natural organisms using a net at Masan Bay (eutrophic) and Jangmok Bay (mesotrophic) during winter season to meet this guideline. Furthermore, cell viability was measured to determine the mortality rate. Based on the organism concentration volume (1 ton) at Masan and Jangmok Bay, abundance of ${\geq}10$ and $<50{\mu}m$ sized organisms was observed to be $4.7{\times}10^4cells\;mL^{-1}$and $0.8{\times}10^4cells\;mL^{-1}$, and their survival rates were 90.4 % and 88.0 %, respectively. In particular, chain-forming small diatoms such as Skeletonema costatum-like species were abundant at Jangmok Bay, while small flagellate ($<10{\mu}m$) and non chain-forming large dinoflagellates, such as Akashiwo sanguinea and Heterocapsa triquetra, were abundant at Masan Bay. Due to the size-difference of the dominant species, concentration efficiency was higher at Jangmok Bay than at Masan Bay. The mortality rate in samples treated by Ballast Water Treatment System (BWMS) (Day 0) was a little lower for samples from Jangmok Bay than from Masan Bay, with values of 90.4% and 93%, respectively. After 5 days, the mortality rates in control and treatment group were found to be 6.7% and >99%, respectively. Consequently, the phytoplankton concentration method alone did not easily satisfy the type approval standards of USCG Phase II ($>1.0{\times}10^3cells\;mL^{-1}$ in 500-ton tank) during winter season, and alternative options such as mass culture and/or harvesting system using natural phytoplankton communities may be helpful in meeting USCG Phase II biological criteria.

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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