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Studies on Ichthyophthirius multifiliis Fouquet, 1876 in Freshwater Fishes II. Experimental Infection and Development of I. multifiliis (담수산 백점충(Ichthyophthirius multifiliis)에 관한 연구 II. 백점충의 인위 감염 및 어체내 충체 발달상)

  • Ji, Bo-Young;Kim, Ki-Hong;Park, Soo-Il;Kim, Yi-Cheong
    • Journal of fish pathology
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    • v.11 no.1
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    • pp.51-60
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    • 1998
  • Concerned to the lyfe cycle of Ichthyophthirius multifiliis, the experimental infection and development of the parasites were studied in the several freshwater cultured fishes. Opitimum conditions for the propagation of the parasite by serial passage with the rainbow trout fry was observed. Visiable white spots were examined in the body surface, fins and gills of the healthy fries, and a stable infection has been maintained for 2 months in the experimental system (Temperature: $18{\pm}1^{\circ}C$ DO: 7-7.5 ppm; pH: $7{\pm}0.2$). Induction conditions for artificial infection of the parasite by interms of the host fishes, stages of the parasites, and rearing temperature regimes was investigated. Rainbow trout fries showed a positive infection which was resulted from exposure of theront at $18^{\circ}C$. The rainbow trout fries induced white spots on the body surface at 3-7 days exposure to the theronts at $18^{\circ}C$. It was found that exposure of the rainbow trout fries exposed to 1,000 theronts per fish (10 theront/ml) for 45-60 minutes at $18^{\circ}C$ would consistently produce infection. Perfect infection (100%) was induced when the fries were exposed to 1,500 theront per fish (15 theront/ml) under laboratory condition. Development of I. multifiliis in the rainbow trout was observed for 7 days postexposure (PE). The parasite increased in average diameter from $54{\mu}m$ on the 1st day PE to $426{\mu}m$ on the 7th day PE. In the initial infestation period, the parasites were found on the gill epithelium, and on the 3rd day PE they invaded into the basal part of the gill filament adjacent to the major blood vessels, particularly the afferent vessels. Morphological change of buccal apparatus were observed on the 2nd day PE. Contractile vacuoles were more prominent on the 4th day PE, and they had notable changes on the 7th day PE.

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Seasonal Changes of Species Composition and Standing Crop of Phytoplankton in the Ark Shell Scapharca broughtonii Farming Areas of Jinhae Bay (진해만 피조개 Scapharca broughtonii 양식장 식물플랑크톤의 종조성과 현존량의 월별변화)

  • Kim, Byoung-Hak;Min, Byeong-Hee;Choi, Nak-Joong;Oh, Bong-Se;Park, Ki-Yeol;Min, Kwang-Sik
    • Journal of Aquaculture
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    • v.21 no.3
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    • pp.157-166
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    • 2008
  • Species composition and standing crop of phytoplankton were investigated in the ark shell Scapharca broughtonii farming areas from March, 2006 to February, 2007 in Jinhae Bay. Water temperature ranged from 7.56 to $25.90^{\circ}C$, salinity from 13.74 to 34.78 psu, dissolved oxygen from 4.13 to 13.20 mg/L, chlorophyll $\alpha$ from 2.77 to 104.98 $mg/m^2$ and pH from 7.83 to 8.65. Dominant species was Nitzschia and Rhizosolenia from March to May, Skeletonama costatum and Prorocentrum from June to August, Skeletonama costatum, Thalassiosira, Chaetoceros from September to November and Thalassiosira, Chaetoceros from December to February. Colonial diatoms were more dominant than the single cell diatoms. Standing crop was the highest in July at three stations. Standing crop of Skeletonama costatum was the highest as 1,760.0 cells/mL at St. 1, 1,075.2 cells/mL at St. 2 and 698.4 cells/mL at St. 3 in July.

Density Effects on the Size of 2-1 Korean Pine and 1-1 Jack Pine Nursery Stock (잣나무 2-1 묘(苗)와 방크스소나무 1-1 묘(苗)의 생장(生長)에 미치는 밀도효과(密度効果))

  • Ma, Sang Kyu
    • Journal of Korean Society of Forest Science
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    • v.32 no.1
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    • pp.1-8
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    • 1976
  • One of the most common needle leaf species used in planting in Korea is korean pine (Pinus koraiensis S. et Z.), and jack pine (Pinus banksiana Lamb.) is one of the test species for suitability. The relation of nursery bed density of 2-1 korean pine and 1-1 jack pine was studied at the Kwang Nung Nursery, Central Branch Station of Forest Research Institute, and about 40km north of Seoul. Nursery bed density of 2-1 korean pine, which ranged from 36 to 324 trees per square meters and of 1-1 jack pine, which ranged from 25 to 169 trees per square meters, had a marked effect on caliper, height, dry weight and percent and amount of plantable stock. The soil physical and chemical properties is silt plus clay, 50.55 percent; organic matter, 2.09 percent; total nitrogen, 0.13 percent; available phosphorus, 253.25 ppm; exchangeable potash, 0.46 m.e/100g; and pH, 5.58. As the density of the nursery seedling stand of 2-1 korean pine increases, the average tree height increases (Fig. 1A), but in 1-1 jack pine density do not affect to increase or decrease the average tree height. As the density of nursery bed increases, the average stem caliper (at 2cm above ground line) and dry weight decrease (Fig. 1B), but the decreasing rate is more seriously in 1-1 jack pine than 2-1 korean pine (Fig.5). As increasing the density of nursery bed, the T/R ratio of trees of the test species increase. Also the dry weight of leaf, stem and root parts are decreasing in proportion to the increase of stand density, but the drop rate of jack pine is more rapid than korean pine (table. 1) The patent facts of difference of growth characteristics between 2-1 korean pine and 1-1 jack pine were studied. These facts should be used to select the scale of stand density at the nursery bed or the plantable site. Korean pine is demanded high density, on the other hand in jack pine low density are more suitable to manage the stand density. Stands of comparatively low density had the greatest percentage of high-quality stock, and the stands of high density had less than the high quality trees of low density. An important criterion of the best density is percent and number of high-quality trees produced per square meter of bed area. Stem caliper and stem height of seedling is used in most public nurseries to sort seedling into plantable grades. The stock grade standard has set at 4.5mm caliper and 16cm height of 2-1 korean pine as the minimum desired stem caliper and height. By the result studies, the plantable stock grade standards of 2-1 korean pine used at stem height 16cm and stem caliper 4.5mm from public nurseries should be reformed to stem height 18cm and stem caliper 4.0mm by the growth characteristics and the tree distribution of stem height and caliper of relation to density. For the 2-1 korean pine, best density should be about 160 to 200 trees per square meter according to soil fertility. For the 1-1 jack pine, the suitable standard of plantable stock should be at stem height 25cm and caliper 6mm (at 2cm above ground line) and best density was about 100 to 120 trees.

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Developmental Abnormality in Agricultural Region and Toxicity of the Fungicide Benomyl on Korea salamander, Hynobius leechii (한국산 도롱뇽(Hynobius leechii)의 농경지에서의 배 발생 이상과 살균제 Benomyl의 독성효과)

  • Choi, Yeoung-Ju;Yoon, Chun-Sik;Park, Joo-Hung;Jin, Jung-Hyo;Cheong, Seon-Woo
    • Korean Journal of Ecology and Environment
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    • v.35 no.3 s.99
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    • pp.198-212
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    • 2002
  • A numerical variation and abnormalities were studied on egg bags and embryos of Korean salamander, Hynobius leechii from agricultural habitat. The teratogenic and toxic effects of fungicide benomyl were also investigated with early embryos from non-agricultural habitat. We collected 144 egg bags from agricultural region, and 3418 of early embryos were contained. The lengths of egg bags were varied from 10 to 23 cm and the most frequent length was 19 cm. The number of embryos was varied from 7 to 43, and the most frequent range was 22 to 26. Spontaneous abnormalities were occurred in 406 embryos among 116 egg bags, and 24 kinds of external abnormalities were found. Individuals showing severe external defect were histologically studied and they showed optic dyspalsia, thyroid carcinoma, somatic muscular dysplasia, partial biaxial structure, decrease of red blood cells in the heart, cephalic degeneration and intestinal dysplasia. 385 embryos from non-agricultural region were exposed to 200 nM${\sim}$ 1 ${\mu}$M of benomyl at blastula or gastrula for 12 days. All embryo were dead in the concentration of 1 ${\mu}$M (LD$_{100}$) and 75% of embryos were dead in 800nM of benomyl. Speciflc effect due to benomyl was acrania or cephalic dysplasia and this restult suggests that the benomyl inhibit stongly to the development of neural tissue. These abnormal developments may be caused by antimitotic action, inhibition of tubulin complex, destruction of microtubule, inhibitions of neurulation and closing of neural fold, and by the inhibition of the movement of neural crest cells.

Method Development for Determination of Chlorogenic Acid and Arbutin Contents in Fruits by UHPLC-MS/MS (UHPLC-MS/MS를 이용한 과일류 중 클로로젠산 및 알부틴 동시분석법 개발)

  • Choi, Young-Ju;Jeon, Jong-Sup;Kim, Woon-Ho;Jung, You-Jung;Ryu, Ji-Eun;Choi, Jong-Chul;Chae, Kyung-Suk;Lee, Jin-Hee;Do, Young-Sook;Park, Young-Bae;Yoon, Mi-Hye
    • Journal of Food Hygiene and Safety
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    • v.34 no.5
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    • pp.413-420
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    • 2019
  • In this study, a sample preparation method and a simultaneous determination method by ultra-high performance liquid chromatography coupled with tandem mass spectrometry for 9 isomers of chlorogenic acid and arbutin in fruits were developed. The samples were extracted using 90% methanol (pH 3.0), with the solutions being shaken and then sonicated for 10 min each. After centrifugation at 4,000 rpm for 10 min, the extraction was concentrated under a vacuum at $40^{\circ}C$ using a vacuum evaporator. The residue was dissolved in 5 mL of 5% methanol and filtered through a $0.45{\mu}m$ membrane before UHPLC-MS/MS analysis. The separations were performed on a C18 column with gradient elution of water (containing 0.1% formic acid) and methanol (containing 0.1% formic acid). The specificity, linearity, limit of detection, limit of quantification, accuracy, and precision of the proposed methods were also evaluated.

Compaction Characteristics of Multi-cropping Paddy Soils in South-eastern Part of Korea (우리나라 동남부 다모작 논토양의 경반화 특성)

  • Yun, Eul-Soo;Jung, Ki-Yeul;Park, Ki-Do;Sonn, Yeon-Kyu;Park, Chang-Yeong;Hwang, Jae-Bog;Nam, Min-Hee
    • Korean Journal of Soil Science and Fertilizer
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    • v.44 no.5
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    • pp.688-695
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    • 2011
  • This study was carried out for some survey about soil compaction in the multi-cropping system of paddy field. Investigated sites were 90 farmer's fields in south-eastern part of Korea. The tillage practices season was different according to cropping system of paddy; in spring for mono rice cultivation and in autumn for the multi-cropping field. The average tillage depth in investigated sites was about 25 cm, however, it is different between the farmer's tillage practices and soil characteristics. It is high correlation to tillage deep and minimum resistance of penetration. The reaching soil deep to maximum resistance of penetration was about 27 cm, and average penetration resistance of the deep is 1.8~2.0 MPa for moderately fine-textured soils and more than 3.0 MPa for moderately coarse-textured soils. The difference of penetration resistance between cultivating and compacted layer was in order to sandy loam > clayey loam > clayey, and the difference was lesser in poorly drained soils than somewhat poorly ones. In the rice mono cropping field, the maximum resistance in no-tillage for 15 years was 1.18~1.25 Mpa at 20~25 cm in soil deep, however, the resistance of field with every year tillage practices was 2.03~2.21 Mpa. In the extremely compacted sandy loam textured soils, the penetration resistance at 30 cm in soil depth was drastically reduced by the subsoil from 5.2 Mpa to 3.2 Mpa, and the watermelon root in plastic film house was deep elongated.

Growth of Intestinal Bacteria and Intestinal Inflammation of Sprout Extract from Common Buckwheat and Tartary Buckwheat (일반메밀과 쓴메밀의 새싹 추출물의 장내 유익균 증식 및 염증조절 효능 평가)

  • Su Jeong Kim;Hwang Bae Sohn;Jong Won Kim;Sanghyun Lim;Jong Nam Lee;Su Hyoung Park;Jung Hwan Nam;Do Yeon Kim;Ye Jin Lee;Dong Chil Chang;Yul Ho Kim
    • Korean Journal of Plant Resources
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    • v.36 no.5
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    • pp.455-468
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    • 2023
  • We aimed to assess the potential growth-promoting effects of buckwheat sprout on intestinal bacteria and their anti-inflammation effects in a cellular model of intestinal inflammation. The growth of Bifidobacterium longum ssp. infantis BT1 was enhanced with the addition of the sprout extract of tartary buckwheat. Further, in the inflammatory model cells cultured with Raw 264.7 cells were treated with buckwheat sprout including each 10 probiotics before the addition of lipopolysaccharide (LPS) to induce inflammation in Raw 264.7 cells. Buckwheat sprout in both Bifidobacterium longum ssp. infantis BT1 and Lacticaseibacillus paracasei LPC5 significantly reduced the production of NO and PGE2. The above results indicate that buckwheat sprout extract which contains with various physiologically active substances such as rutin, quercetin, and choline is effective in suppressing NO and PGE2 production, which are inflammation-related indicators. The present study suggests that buckwheat sprout could induce positive effects on the intestinal beneficial bacteria and in anti-inflammation.

Expression of TIMP1, TIMP2 Genes by Ionizing Radiation (이온화 방사선에 의한 TIMP1, TIMP2 유전자 발현 측정)

  • Park Kun-Koo;Jin Jung Sun;Park Ki Yong;Lee Yun Hee;Kim Sang Yoon;Noh Young Ju;Ahn Seung Do;Kim Jong Hoon;Choi Eun Kyung;Chang Hyesook
    • Radiation Oncology Journal
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    • v.19 no.2
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    • pp.171-180
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    • 2001
  • Purpose : Expression of TIMP, intrinsic inhibitor of MMP, is regulated by signal transduction in response to genotoxins and is likely to be an important step in metastasis, angiogenesis and wound healing after ionizing radiation. Therefore, we studied radiation mediated TIMP expression and its mechanism in head and neck cancer cell lines. Materials and Methods : Human head and neck cancer cell lines established at Asan Medical Center were used and radiosensitivity $(D_0)$, radiation cytotoxicity and metastatic potential were measured by clonogenic assay, n assay and invasion assay, respectively. The conditioned medium was prepared at 24 hours and 48 hours after 2 Gy and 10 Gy irradiation and expression of TIMP protein was measured by Elisa assay with specific antibodies against human TIMP. hTIMP1 promoter region was cloned and TIMP1 luciferase reporter vector was constructed. The reporter vector was transfected to AMC-HN-1 and -HN-9 cells with or without expression vector Ras, then the cells were exposed to radiation or PMA, PKC activator. EMSA was peformed with oligonucleotide (-59/-53 element and SP1) of TIMP1 promoter. Results : $D_0$ of HN-1, -2, -3, -5 and -9 cell lines were 1.55 Gy, 1.8 Gy, 1.5 Gt, 1.55 Gy and 2.45 Gy respectively. n assay confirmed cell viability, over $94\%$ at 24hrs, 48hrs after 2 Gy irradiation and over 73% after 10 Gy irradiation. Elisa assay confirmed that cells secreted TIMP1, 2 proteins continuously. After 2 Gy irradiation, TIMP2 secretion was decreased at 24hrs in HN-1 and HN-9 cell lines but after 10 Gy irradiation, it was increased in all cell lines. At 48hrs after irradiation, it was increased in HN-1 but decreased in HN-9 cells. But the change in TIMP secretion by RT was mild. The transcription of TIMP1 gene in HN-1 was induced by PMA but in HN-9 cell lines, it was suppressed. Wild type Ras induced the TIMP-1 transcription by 20 fold and 4 fold in HN-1 and HN-9 respectively. The binding activity to -59/-53, AP1 motif was increased by RT, but not to SP1 motif in both cell lines. Conclusions : We observed the difference of expression and activity of TIMPs between radiosensitive and radioresistant cell line and the different signal transduction pathway between in these cell lines may contribute the different radiosensitivity. Further research to investigate the radiation response and its signal pathway of TIMPs is needed.

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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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