• Title/Summary/Keyword: Non-Isolation

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Detection of Respiratoiry Tract Viruses in Busan, 1997-2000 (1997-2000년 부산지역 호흡기계 바이러스의 탐색)

  • 조경순;김영희
    • Korean Journal of Microbiology
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    • v.37 no.4
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    • pp.284-288
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    • 2001
  • Respiratory viruses are one of the most infectious agent in human. Six different respiratory tract viruses were detected from Busan while working on the preventive surveillance in 1997-2000. The isolation rate from suspected specimens were 8.4%. Influenza virus A, B type, parainfluenza virus, adenovirus, mumps virus, and measles virus were examined from throat swabs, serum, and secretions of patients. Influenza A/Sydney/05/97(H3N2)-like, A/Johanesburg/33/94(H3N2)-like, A/Beijing/262/95(H1N1)-like and Influenza B/Beijing/262/95-like, B/Harbin/07/94-like, B/Guangdong/08/93-like were found. Adenovirus serotype 1, 2, 3 and 5 were detected, antibody of mumps both IgM and IgG were shown and outbreaks of measles were confirmed. Different antigenic types of influenza virus were detected every year, one outbreak of parainfluenza in 1999, mumps outbreak in 1999 and 2000, and incidence of measles in 2000 were noticeable. Monthly outbreaks were November through following March with influenza virus, January through June with adenovirus, February through May and December with mumps, April through August and November, December with measles, respectively. The size of isolated viruses were 130 nm with influenza virus B type, non-enveloped, icosahedron with 70 nm with adenovirus, 170 nm with mumps virus and 180 nm with parainfluenza virus in diameter, respectively.

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Longitudinal Study of the Subgingival Microbial Change after Tetracycline Topical Application (Tetracycline 국소도포가 치은연하 세균분포에 미치는 영향)

  • Choi, Kwang-Choon;Lee, Young-Hee;Lee, Jin-Yong;Chung, Chong-Pyoung;Son, Seong-Heui
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.4
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    • pp.503-513
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    • 1986
  • Previous studies have developed the technique of topical application of tetracycline(TC) into the periodontal pockets and examined the change of clinical parameters and subgingival microbial morphotypes. The purpose of this study was to longitudinally examine the clinical and microbiological effects of topically applied TC in a double-blind and split-mouth design. Thirteen patients with moderate periodontitis, who were treated with or without TC application and scaling treatment, were examined. TC gel(3%) was used to apply into the selected periodontal pockets twice a week for 2 weeks. During the experiment, clinical parameters and subgingival microbial morphotypes were examined, and for isolation of black-pigmented Bacteroides(BPB) and streptococci, an anaerobic sample culturing was done at week 0, 2, and 7. In clinical observation the TC-scaled group exhibited a significant decrease of Gingival Inflammatory Index, Plaque Index, Sulcus Bleeding Index, pocket depth, and gingival crevicular fluid when compared to the TC-unsealed, placebo-scaled, and placebo-unsealed groups. The result of microbial morphotype observation showed a significant increase of coccal form and a decrease of spirochetes in the TC-scaled, TC-unscaled, and placebo-scaled groups. The culture study of streptococci revealed that TC with scaling treatment resulted in a significant increase of S. sanguis I at week 2, but its proportion had returned to the base line level. The anaerobic culture study showed that BPB was significantly reduced in the TC-scaled and TC-unsealed groups at week 7. Among BPB species, B. intermedius declined significantly with time treatment(week 2 and 7) in the TC-scaled and TC-unsealed groups. These results suggest that the settled pathogenic microflora can be succeeded by nonpathogenic microflora in periodontal pockets after TC treatment.

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Analysis of Potential Toxigenicity and Phylogeny using Target Genes in Aphanizomenon flos-aquae (Cyanophyceae) strains isolated from the Nakdong River (낙동강에서 분리된 Aphanizomenon flos-aquae (Cyanophyceae) 균주의 목표 유전자를 이용한 잠재적 독소 생성능 및 계통학적 분석)

  • Ryu, Hui-Seong;An, Sung-Min;Lim, Chang-Kun;Shin, Ra-Young;Park, Jong-Guen;Lee, Jung-Ho
    • Korean Journal of Ecology and Environment
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    • v.50 no.1
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    • pp.137-147
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    • 2017
  • The identity of toxin producers remains only hypothesis unless there were identified by strain isolation and analytical confirmation of both the cyanotoxin production and the genetic identity of the monoculture. The purposes of this study were to identify a morphologic and phylogenetic classification in Aphanizomenon flos-aquae strains isolated from the Nakdong River and to investigate the potential ability of the strains to produce toxins such as saxitoxin and cylindrospermopsin using target genes. The 16S rRNA and sxtA, sxtI, cyrA, cyrJ genes were analyzed on two strains (DGUC001, DGUC003) isolated from the Nakdong River. Morphological features of the strains were observed a shape of aggregated trichomes in parallel fascicles which can reach up to macroscopic size and a hyaline terminal cell without aerotope. In addition, the 16S rRNA phylogenetic analyses showed that the strains were identified as the same species with high genetic similarity of 98.4% and grouped within a monospecific andsupported cluster I of Aphanizomenon flos-aquae selected from GenBank of the NCBI. The cyrA and cyrJ genes encoding for the cylindrospermopsin-biosynthesis were not detected in the present study. The sxtA gene was in detected both the two strains, whereas the sxtI gene which had been suggested as a suitable molecular marker to detect saxitoxin-producing cyanobacteria was not found both the strains. Thus, the two strains isolated from Nakdong River were identified as the same species of Aphanizomenon flos-aquae Ralfs ex Bornet et Flahault 1888, the two strains were confirmed as potential non-producing strains of the saxitoxin and cylindrospermopsin.

Isolation and Characteristics of a Phenol-degrading Bacterium, Rhodococcus pyridinovorans P21 (페놀분해세균 Rhodococcus pyridinovorans P21의 분리 및 페놀분해 특성)

  • Cho, Kwang-Sik;Lee, Sang-Mee;Shin, Myung-Jae;Park, Soo-Yun;Lee, Ye-Ram;Jang, Eun-Young;Son, Hong-Joo
    • Journal of Life Science
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    • v.24 no.9
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    • pp.988-994
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    • 2014
  • The effluents of chemical and petroleum industries often contain non-biodegradable aromatic compounds, with phenol being one of the major organic pollutants present among a wide variety of highly toxic organic chemicals. Phenol is toxic upon ingestion, contact, or inhalation, and it is lethal to fish even at concentrations as low as 0.005 ppm. Phenol biodegradation has been studied in detail using bacterial strains. However, these microorganisms suffer from substrate inhibition at high concentrations of phenol, whereby growth is inhibited. A phenol-degrading bacterium, P21, was isolated from oil-contaminated soil. The phenotypic characteristics and a phylogenetic analysis indicated the close relationship of strain P21 to Rhodococcus pyridinovorans. Phenol biodegradation by strain P21 was studied under shaking condition. The optimal conditions for phenol biodegradation by strain P21 were 0.09% $KNO_3$, 0.1% $K_2HPO_4$, 0.3% $NaH_2PO_4$, 0.015% $MgSO_4{\cdot}7H_2O$, 0.001% $FeSO_4{\cdot}7H_2O$, initial pH 9, and $20-30^{\circ}C$, respectively. When 1,000 ppm of phenol was added to the optimal medium, the strain P21 completely degraded it within two days. Rhodococcus pyridinovorans P21 could grow in up to 1,500 ppm of phenol as the sole carbon source in a batch culture, but it could not grow in a medium containing above 2,000 ppm. Moreover, strain P21 could utilize toxic compounds, such as toluene, xylene, and hexane, as a sole carbon source. However, no growth was detected on chloroform.

Isolation and Evaluation of an Antiviral Producing Serratia spp. Strain Gsm01 against Cucumber mosaic virus in Korea (한국에서 CMV에 항바이러스 효과를 나타내는 Serratia spp. Gsm01 균주의 분리 동정 및 효과 검정)

  • Ipper, Nagesh S.;Lee, Seon-Hwa;Suk, Jung-Ki;Shrestha, Anupama;Seo, Dong-Uk;Park, Duck-Hwan;Cho, Jun-Mo;Park, Dong-Sik;Hur, Jang-Hyun;Lim, Chun-Keun
    • The Korean Journal of Pesticide Science
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    • v.10 no.4
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    • pp.344-350
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    • 2006
  • An Antiviral producing bacterial strain was isolated from ginseng root environment in Hongcheon, Kangwon province of Republic of Korea. Identification of this bacterial strain was performed by physiological and biochemical tests along with 16S rRNA analyses. The results revealed that the bacterium was closer to genus Serratia, which was named as Gsm01. The strain was grown in Mannitol-Glutamate-Yeast (MGY) broth for 48 h. The culture was centrifuged and the filtrate obtained was tested for its ability to control Cucumber mosaic virus strain Y (CMV-Y) in greenhouse and field experiments. In the green house experiments, CF was evaluated for its ability to protect local host, Chenopodium amaranticolor and systemic host of CMV, Nicotiana tabacum cv. Xanthi-nc. It was found that, CF treatment reduced viral infection by 98% in local host; C. amaranticolor. The N. tabacum cv. Xanthi-nc plants treated with CF did not show visible viral symptoms 15 days post inoculation (dpi) and remained symptomless throughout the periods of the study. To evaluate effectiveness of CF under field conditions, experiment was carried out in a polyvinyl house. It was observed that, 52% plants were protected from viral diseases compared to non-treated plants, increasing the crop yield. This is the first report showing antiviral activity of a Serratia spp. against CMV.

Characteristic study and optimization of culture conditions for Bacillus amyloliquefaciens SRCM 100731 as probiotic resource for companion animal (Bacillus amyloliquefaciens SRCM 100731의 반려 동물용 프로바이오틱스 소재로서의 특성 규명 및 배양 조건 최적화)

  • 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
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    • 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 Effects of Self-regulatory Resources and Construal Levels on the Choices of Zero-cost Products (자아조절자원 및 해석수준이 공짜대안 선택에 미치는 영향)

  • Lee, Jinyong;Im, Seoung Ah
    • Asia Marketing Journal
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    • v.13 no.4
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    • pp.55-76
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    • 2012
  • Most people prefer to choose zero-cost products they may get without paying any money. The 'zero-cost effect' can be explained with a 'zero-cost model' where consumers attach special values to zero-cost products in a different way from general economic models (Shampanier, Mazar and Ariely 2007). If 2 different products at the regular prices of ₩200 and ₩400 simultaneously offer ₩200 discounts, the prices will be changed to ₩0 and ₩200, respectively. In spite of the same price gap of the two products after the ₩200 discounts, people are much more likely to select the free alternative than the same product at the price of ₩200. Although prior studies have focused on the 'zero-cost effect' in isolation of other factors, this study investigates the moderating effects of a self-regulatory resource and a construal level on the selection of free products. Self-regulatory resources induce people to control or regulate their behavior. However, since self-regulatory resources are limited, they are to be easily depleted when exerted (Muraven, Tice, and Baumeister 1998). Without the resources, consumers tend to become less sensitive to price changes and to spend money more extravagantly (Vohs and Faber 2007). Under this condition, they are also likely to invest less effort on their information processing and to make more intuitive decisions (Pocheptsova, Amir, Dhar, and Baumeister 2009). Therefore, context effects such as price changes and zero cost effects are less likely in the circumstances of resource depletion. In addition, construal levels have profound effects on the ways of information processing (Trope and Liberman 2003, 2010). In a high construal level, people tend to attune their minds to core features and desirability aspects, whereas, in a low construal level, they are more likely to process information based on secondary features and feasibility aspects (Khan, Zhu, and Kalra 2010). A perceived value of a product is more related to desirability whereas a zero cost or a price level is more associated with feasibility. Thus, context effects or reliance on feasibility (for instance, the zero cost effect) will be diminished in a high level construal while those effects may remain in a low level construal. When people make decisions, these 2 factors can influence the magnitude of the 'zero-cost effect'. This study ran two experiments to investigate the effects of self-regulatory resources and construal levels on the selection of a free product. Kisses and Ferrero-Rocher, which were adopted in the prior study (Shampanier et al. 2007) were also used as alternatives in Experiments 1 and 2. We designed Experiment 1 in order to test whether self-regulatory resource depletion will moderate the zero-cost effect. The level of self-regulatory resources was manipulated with two different tasks, a Sudoku task in the depletion condition and a task of drawing diagrams in the non-depletion condition. Upon completion of the manipulation task, subjects were randomly assigned to one of a decision set with a zero-cost option (i.e., Kisses ₩0, and Ferrero-Rocher ₩200) or a set without a zero-cost option (i.e., Kisses ₩200, and Ferrero-Rocher ₩400). A pair of alternatives in the two decision sets have the same price gap of ₩200 between a low-priced Kisses and a high-priced Ferrero-Rocher. Subjects in the no-depletion condition selected Kisses more often (71.88%) over Ferrero-Rocher when Kisses was free than when it was priced at ₩200 (34.88%). However, the zero-cost effect disappeared when people do not have self-regulatory resources. Experiment 2 was conducted to investigate whether constual levels influence the magnitude of the 'zero-cost effect'. To manipulate construal levels, 4 different 'why (in the high construal level condition)' or 'how (in the low construal level condition)' questions about health management were asked. They were presented with 4 boxes connected with downward arrows. In a box at the top, there was one question, 'Why do I maintain good physical health?' or 'How do I maintain good physical health?' Subjects inserted a response to the question of why or how they would maintain good physical health. Similar tasks were repeated for the 2nd, 3rd, and 4th responses. After the manipulation task, subjects were randomly assigned either to a decision set with a zero-cost option, or to a set without it, as in Experiment 1. When a low construal level is primed with 'how', subjects chose free Kisses (60.66%) more often over Ferrero-Rocher than they chose ₩200 Kisses (42.19%) over ₩400 FerreroRocher. On contrast, the zero-cost effect could not be observed any longer when a high construal level is primed with 'why'.

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Ecological Changes of Insect-damaged Pinus densiflora Stands in the Southern Temperate Forest Zone of Korea (I) (솔잎혹파리 피해적송림(被害赤松林)의 생태학적(生態学的) 연구(研究) (I))

  • Yim, Kyong Bin;Lee, Kyong Jae;Kim, Yong Shik
    • Journal of Korean Society of Forest Science
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    • v.52 no.1
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    • pp.58-71
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    • 1981
  • Thecodiplosis japonesis is sweeping the Pinus densiflora forests from south-west to north-east direction, destroying almost all the aged large trees as well as even the young ones. The front line of infestation is moving slowly but ceaselessly norhwards as a long bottle front. Estimation is that more than 40 percent of the area of P. densiflora forest has been damaged already, however some individuals could escapes from the damage and contribute to restore the site to the previous vegetation composition. When the stands were attacked by this insect, the drastic openings of the upper story of tree canopy formed by exclusively P. densiflora are usually resulted and some environmental factors such as light, temperature, litter accumulation, soil moisture and offers were naturally modified. With these changes after insect invasion, as the time passes, phytosociologic changes of the vegetation are gradually proceeding. If we select the forest according to four categories concerning the history of the insect outbreak, namely, non-attacked (healthy forest), recently damaged (the outbreak occured about 1-2 years ago), severely damaged (occured 5-6 years ago), damage prolonged (occured 10 years ago) and restored (occured about 20 years ago), any directional changes of vegetation composition could be traced these in line with four progressive stages. To elucidate these changes, three survey districts; (1) "Gongju" where the damage was severe and it was outbroken in 1977, (2) "Buyeo" where damage prolonged and (3) "Gochang" as restored, were set, (See Tab. 1). All these were located in the south temperate forest zone which was delimited mainly due to the temporature factor and generally accepted without any opposition at present. In view of temperature, the amount and distribution of precipitation and various soil factor, the overall homogeneity of environmental conditions between survey districts might be accepted. However this did not mean that small changes of edaphic and topographic conditions and microclimates can induce any alteration of vegetation patterns. Again four survey plots were set in each district and inter plot distance was 3 to 4 km. And again four subplots were set within a survey plot. The size of a subplot was $10m{\times}10m$ for woody vegetation and $5m{\times}5m$ for ground cover vegetation which was less than 2 m high. The nested quadrat method was adopted. In sampling survey plots, the followings were taken into account: (1) Natural growth having more than 80 percent of crown density of upper canopy and more than 5 hectares of area. (2) Was not affected by both natural and artificial disturbances such as fire and thinning operation for the past three decades. (3) Lower than 500 m of altitude (4) Less than 20 degrees of slope, and (5) Northerly sited aspect. An intensive vegetation survey was undertaken during the summer of 1980. The vegetation was devided into 3 categories for sampling; the upper layer (dominated mainly by the pine trees), the middle layer composed by oak species and other broad-leaved trees as well as the pine, and the ground layer or the lower layer (shrubby form of woody plants). In this study our survey was concentrated on woody species only. For the vegetation analysis, calculated were values of intensity, frequency, covers, relative importance, species diversity, dominance and similarity and dissimilasity index when importance values were calculated, different relative weights as score were arbitrarily given to each layer, i.e., 3 points for the upper layer, 2 for the middle layer and 1 for the ground layer. Then the formula becomes as follows; $$R.I.V.=\frac{3(IV\;upper\;L.)+2(IV.\;middle\;L.)+1(IV.\;ground\;L.)}{6}$$ The values of Similarity Index were calculated on the basis of the Relative Importance Value of trees (sum of relative density, frequency and cover). The formula used is; $$S.I.=\frac{2C}{S_1+S_2}{\times}100=\frac{2C}{100+100}{\times}100=C(%)$$ Where: C = The sum of the lower of the two quantitative values for species shared by the two communities. $S_1$ = The sum of all values for the first community. $S_2$ = The sum of all values for the second community. In Tab. 3, the species composition of each plot by layer and by district is presented. Without exception, the species formed the upper layer of stands was Pinus densiflora. As seen from the table, the relative cover (%), density (number of tree per $500m^2$), the range of height and diameter at brest height and cone bearing tendency were given. For the middle layer, Quercus spp. (Q. aliena, serrata, mongolica, accutissina and variabilis) and Pinus densiflora were dominating ones. Genus Rhodedendron and Lespedeza were abundant in ground vegetation, but some oaks were involved also. (1) Gongju district The total of woody species appeared in this district was 26 and relative importance value of Pinus densiflora for the upper layer was 79.1%, but in the middle layer, the R.I.V. for Quercus acctissima, Pinus densiflora, and Quercus aliena, were 22.8%, 18.7% and 10.0%, respectively, and in ground vegetation Q. mongolica 17.0%, Q. serrata 16.8% Corylus heterophylla 11.8%, and Q. dentata 11.3% in order. (2) Buyeo district. The number of species enumerated in this district was 36 and the R.I.V. of Pinus densiflora for the uppper layer was 100%. In the middle layer, the R.I.V. of Q. variabilis and Q. serrata were 8.6% and 8.5% respectively. In the ground vegetative 24 species were counted which had no more than 5% of R.I.V. The mean R.I.V. of P.densiflora ( totaling three layers ) and averaging four plots was 57.7% in contrast to 46.9% for Gongju district. (3) Gochang-district The total number of woody species was 23 and the mean R.I.V. of Pinus densiflora was 66.0% showing greater value than those for two former districts. The next high value was 6.5% for Q. serrata. As the time passes since insect outbreak, the mean R.I.V. of P. densiflora increased as the following order, 46.9%, 57.7% and 66%. This implies that P. densiflora was getting back to its original dominat state again. The pooled importance of Genus Quercus was decreasing with the increase of that for Pinus densiflora. This trend was contradict to the facts which were surveyed at Kyonggi-do area (the central temperate forest zone) reported previously (Yim et al, 1980). Among Genus Quercus, Quercus acutissina, warm-loving species, was more abundant in the southern temperature zone to which the present research is concerned than the central temperate zone. But vice-versa was true with Q. mongolica, a cold-loving one. The species which are not common between the present survey and the previous report are Corpinus cordata, Beltala davurica, Wisturia floribunda, Weigela subsessilis, Gleditsia japonica var. koraiensis, Acer pseudosieboldianum, Euonymus japonica var. macrophylla, Ribes mandshuricum, Pyrus calleryana var. faruiei, Tilia amurensis and Pyrus pyrifolia. In Figure 4 and Table 5, Maximum species diversity (maximum H'), Species diversity (H') and Eveness (J') were presented. The Similarity indices between districts were shown in Tab. 5. Seeing Fig. 6, showing two-dimensional ordination of polts on the basis of X and Y coordinates, Ai plots aggregate at the left site, Bi plots at lower site, and Ci plots at upper-right site. The increasing and decreasing patterns as to Relative Density and Relative Importance Value by genus or species were given in Fig. 7. Some of the patterns presented here are not consistent with the previously reported ones (Yim, et al, 1980). The present authors would like to attribute this fact that two distinct types of the insect attack, one is the short war type occuring in the south temperate forest zone, which means that insect attack went for a few years only, the other one is a long-drawn was type observed at the temperate forest zone in which the insect damage went on continuously for several years. These different behaviours of infestation might have resulted the different ways of vegetational change. Analysing the similarity indices between districts, the very convincing results come out that the value of dissimilarity index between A and B was 30%, 27% between B and C and 35% between A and C (Table 6). The range of similarity index was obtained from the calculation of every possible combinations of plots between two districts. Longer time isolation between communities has brought the higher value of dissimilarity index. The main components of ground vegetation, 10 to 20 years after insect outbreak, become to be consisted of mainly Genus Lespedeza and Rhododendron. Genus Quercus which relate to the top dorminant state for a while after insect attack was giving its place to Pinus densiflora. It was implied that, provided that the soil fertility, soil moisture and soil depth were good enough, Genus Quercuss had never been so easily taken ever by the resistant speeies like Pinus densiflora which forms the edaphic climax at vast areas of forest land. Usually they refer Quercus to the representative component of the undisturbed natural forest in the central part of this country.

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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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