인삼종자의 발아과정에 있어서 미세구조의 변화

김우갑,박홍덕,김은수,한성식,Kim, Woo-Kap,Park, Hong-Duok,Kim, Eun-Soo,Han, Sung-Sik 한국현미경학회 1979 Applied microscopy Vol.9 No.1

과피가 녹색이고 배의 길이가 $250{\mu}$인 미성숙 과정을 거쳐 발아시까지 배 및 배유세포의 미세구조 변화를 관찰한 결과를 요약하면 다음과 같다. 1. 배의 길이가 $250{\mu}$인 미성숙 종자의 배세포는 세포질이 비교적 충실하나 mitochondria의 Cristae 및 plastid는 미분화상태이고 가끔 dictyosome이 관찰되었다. 액포안에는 전자밀도가 높은 globoid 를 함유하는 것도 있었다. 소포체는 대부분 한면소포체이고 외형질에는 spherosome을 많이 함유하고 있었다. 배유세포는 spherosome과 이들로 둘러싸인 전자밀도가 높고 균질성인 protein body로 충만되었고 이들로 인하여 세포기관은 관찰되지 않았다. 2. 과피가 홍색인 종자의 배세포는 cristae가 발달한 mitochondria가 관찰되었으며, 전자밀도가 높은 globoid가 증가하였고 액포는 확장되었다. 배유세포에는 spherosome이 증가하고 있었으며 protein body는 확장되어. 그 내부에는 전자밀도가 높은 globoidal crystal이 분산되어 있었다. 3. 종피가 열개한 종자의 배축의 전형성층과 유아세포에서는 Golgi vacuole과 vesicle이 발달되었으며 mitochondria의 cristae는 매우 분화되었다. spherosome은 매우 많이 나타났고 유근세포, 배축의 주변세포 및 자엽세포의 액포는 매우 분화되었다. 배유세포는 spherosome으로 충만되어 있고 그 내질에는 전자밀도가 높고 작은 입자를 지니고 있었다. Protein body는 단일막으로 둘러싸여 있음이 확인되었다. 세포질내에는 구형체와 spherosome 주변의 세포질에 acid phosphatase의 활성을 나타냈다. 4. 파종시 자엽의 외면부 세포는 spherosome이 급격히 증가하였고 protein body도 관찰되었다. 세포기관들은 분화되어 있었고 전분을 함유하는 색소체가 출현하였다. 5. 파종시킨 후 발아할 때까지 자엽의 외측 $2{\sim}3$열세포, 배축의 주변세포, 유근부 세포에서는 spherosome이 증가하였고 전분립을 함유하는 plastid 도 증가하였다. 배를 구성하는 세포의 내형질의 액포부근에는 mitochondria와 microbody 가 혼재하고 있음이 관찰되었다. 발아시기가 가까워 질수록 배와 연접하는 배유세포에는 액포가 매우 분화되고 spherosome은 감소되는 반면 액포내에는 무정형의 전자밀도가 높은 물질이 증가하였다. 그밖의 배유세포에는 spherosome이 감소됨에 따라 mitochondria는 증가되고 전자밀도가 높은 소구체가 형성되어 증가되었다. Protein body의 외질은 감소되고 전자밀도가 낮은 부위는 확장되며 이는 서로 융합하여 외연부는 없어지고 기질속에 소입자들이 형성되었다. 6. 발아기의 배유세포안에는 spherosome이 감소되고, protein body는 서로 융합되어 한 세포내에 $2{\sim}3$개로 충만되어 있었다. The ultrastructural changes of embryo and endosperm cells were observed during the green fruit with embryo about $250{\mu}$ long to germination. 1. In the embryo cells of green fruit with embryo about $250{\mu}$ long, mitochondrial cristae and plastid are undifferentiated and dictyosome are occasionally observed. There are electron-opaque globoids in the vacuole and a lot of spherosomes in the outer layer of smooth endoplasmic reticulum. Endosperm is filled with spherosomes and electron-opaque protein bodies surrounded by spherosomes, and due to these, other organelle are not observed. 2. In the embryo cells of seeds with red seed coat, mitochondrial cristae are well developed, electron-opaque globoids increased, and vacuoles are enlarged. In the endosperm, however, spherosomes increased, protein bodies are enlarged, and electron-opaque globoidal crystals are dispersed within them. 3. In the procambium and epicotyl cells of dehiscent seed, Golgi vacuoles and vesicles are well developed, and mitochondrial cristae are also well differentiated. Spherosomes are numerously present and radicle cells, peripheral cells of hypocotyl, and vacuoles of cotyledon are well differentiated. Endosperm is filled with spherosomes containing electron-opaque granules and protein bodies are surrounded by a single membrane. There are acid phosphatase around globoids and spherosomes. 4. At the time of seeding, spherosomes markedly increased in the outer layer of cotyledon and protein bodies are also observed. Cell organelles are differentiated and plastids containing starch are also present. 5. In the outer $2{\sim}3$ layers of cotyledons, radicle cells, and peripheral cells of hypocotyl during post-seeding to germination, spherosomes and plastids with starch increased, and mitochondria and microbodies are also found around the nucleus of embryo cells. With approaching, the germination stage, in the endosperm contacting with embryo, vacuoles are well differentiated but spherosomes decreased. There increased electron-opaque materials within vacuoles. In other endosperm, with the decrease of spherosome, mitochondria increased and electro n-opaque globular bodies are formed and gradually increased. The outer layer of protein bodies are reduced while electron-transparent portions are enlarged and fused together to occupy the outer layer where small particles are formed. 6. In the endosperm of germination stage, spherosomes decreased while protein bodies, are fused together to form 2 or 3 within a cell.

식물세포(植物細胞)의 Cytolysomes에 관(關)한 전자현미경적(電子顯微鏡的) 연구(硏究)

김우갑,Kim, Woo-Kap 한국현미경학회 1974 Applied microscopy Vol.4 No.1

Glycine max Merr., Zea mays L.의 엽육세포와 근단세포에서 cytolysome의 기원과 기능에 관하여 추구하였다. Cytolysomes은 원형질막과 세포질의 소포체에서 유래되는데, 원형질막의 함입으로 다소포체인 것과 환상인 다층막성구조로 분화되는 두가지 기본형이 있고, 또 세포질에서는 무과입성 소포체가 세포질의 일부를 분리시키으로서 세포질분리체로 분화된다. 원형질막과 세포질에서 유래된 이들 구조는 각각 막성구조가 증가되면서 액포내로 유리되고 자식성소화과정을 밟아 퇴화되는것 같다. The origin and the function of cytolysomes were studied in the mesophyll cells and the root-tip cells of Glycine max Merr. and Zea mays L. fixed by paraformaldehyde-glutaraldehyde-$OsO_4$. The cytolysome-like structures were found of three main types of configurations: multivesicular, myelin like (multilamllar) and multitubular. More complex and mixed ones were also observed. The origin of these structures seems to be initiated by invaginations or in holdings of the plasmalemma into the cell interior, and that by aggregation and convolution of endoplasmic reticulum in the cytoplasm. Invagination of the plasmalemma were found of two main types of configurations: concentric whorls of lamellar and multivesicular. The structures were also observed within vacuoles and cytoplasm. Since the structurers are widely distributed in the cells and are greatly varied in sizes and shapes. These structures originate from the plasmalemma and the cytoplasm subsequently protrudes into the vacuole, and that seem to play an important role on the formation of the autophagic vacuoles. The possible function and fate of these structures are discussed.

인삼 분비관의 구조와 기능

김우갑,김은수,Kim, Woo-Kap,Kim, Eun-Soo 한국현미경학회 1980 Applied microscopy Vol.10 No.1

인삼의 근피층에서 확인된 분비관의 분포, 분비상피세포의 미세구조, 후형질성 함유물의 성분등을 광학 및 전자현미경을 이용하여 추구하였다. 1) 분비관은 배축, 근피층, 엽맥 유관속의 배축면과 향축면, 줄기의 유관속을 사이에 두고 피층과 수층 및 수내(髓內)에서 관찰되었다. 2) 근피층에 존재하는 분비관은 형성층에서 분화함이 확인되었고, 분비관의 환상배열층은 근령(根令)을 추정하는 구조적인 지표가 된다고 믿어진다. 3) 분비상피세포는 인(仁), 미토콘드리아, 소포체가 매우 발달되었으며, ribosome, chromoplast, leucoplast, dictyosome과 저장과립인 전분립이 관찰되었다. 4) 상피세포로는 단층이고 세포질과 분비관내에는 sudanophyl, osmophyl inclusion이 확인되었지만 Pyridin 및 alcohol로 lipid를 추출하였을 때에는 이들 함유물이 관찰되지 않았다. 5) Nile blue반응에서 상피세포내에는 oxazin에 반응하는 산성지질이, 분비관에는 oxazone에 반응하는 중성지질이 검출되었다. 6) 세포내 분비과립은 $OsO_4$단일고정시 전자밀도가 매우 높았다. 따라서 이 물질의 주성분은 불포화성지질인것 같으며 인삼의 분비관은 지질대사와 밀접한 관련성이 있는 것으로 믿어지므로 인삼이 지니는 분비관은 지질분비관(lipid canal 또는 lipid duct)으로 명명됨이 바람직하다. The distribution of the secretory ducts, fine structures of the secretory epithelial cells, and the ingredients of the metaplasmic inclusions were studied at light and electron microscopical levels in seeds, stems, leaves, and roots of ginseng. The secretory ducts occurred in the hypocotyl of the embryo, in the cortex of the roots, and also both inside and outside of each vascular bundle in the stems and leaves. Especially, it is considered that the circular layers of the secretory ducts in roots may represent their ages. The epithelial cell has well developed nucleolus, mitochondria and smooth endoplasmic reticulum. Sudanophyl and osmiophilic inclusions were found in the epithelial cytoplasm and duct lumen. But these inclusions were not observed when extracted with pyridin or alcohol. In contrast to the lumen with red color, the epithelial cells were blue in color as stained with nile blue, suggesting that the former inclusions are neutral lipid while the latter are acidic lipid. The electron density of the cell inclusions was quite high as fixed with osmium tetroxide, indicating that most of these secretory materials seem to be unsaturated lipid. Therefore, since ginseng secretory ducts are closely associated with the lipid metabolism, it should be called lipid canal or lipid duct.

배추흰나비 식도하신경절(食道下神經節)의 신경세포(神經細胞)에 관(關)한 미세구조적(微細構造的) 연구(硏究)

김우갑,이봉희,Kim, Woo-Kap,Lee, Bong-Hee 한국현미경학회 1981 Applied microscopy Vol.11 No.1

The study on the nerve cells in the subesophageal ganglion of 5-day-old cabbage butterfly, Pieris rapae L., was performed to observe their ultrastructures and classify them on the basis of the differences in size, shape and relative distribution of cell organelles. 1. Type I neurons: These cells are neurosecretory granules ranging 100 to 300 nm in size. 2. Type II neurons: As giant neurons averaging 25 to $30{\mu}m$ in size, such as mitochondria and Golgi apparatus. 3. Type III neurons: These spindle-shaped cells range 9 to $15{\mu}m$ in width. 4. Type IV neurons: These cells have a range of diameter from 12 to $16 {\mu}m$. The cells are abundantly observed in the subesophageal ganglion. 5. Type V neurons: These cells are very small nerve cells with 4.5 to $8.0{\mu}m$ in size and have a prominent nucleus.

Multivesicular bodies 및 Cytolysomes에 관(關)한 전자현미경적(電子顯微鏡的) 연구(硏究)

김우갑,김창환,박홍덕,양희영,Kim, Woo-Kap,Kim, Chang-Whan,Park, Hong-Duok,Yang, He-Young 한국현미경학회 1976 Applied microscopy Vol.6 No.1

Rhizopus nigricans, Aspergillus niger, Aspergillus ochraceus의 균계 Agricus bisporus, Rhizopogon rubescens의 자실층균계 및 담자기세포, Marchantia polymorpha, Pogonatum inflexum, Pteridium aqiulium 의 동화조직, Pinus densiflora, Ginkgo biloba, Panax ginseng 의 엽육조직의 소편을 glutaralehyde-paraformalehyde 및 $OsO_4$에 고정하여 다소포체 및 세포질용해체와 관련된 다양상인 막성구조의 기원과 기능에 관하여 전 자현미경적으로 추구하였다. Rhizopus nigricans, Aspergillus niger, A. ochraceus, Agricus bisporus, Rhizopogon rubescens 의 동심원상인 다층막, 미에린一소포-관상구조의 복합체, 평행다층막구조등은 원형질막에서 유래된다. Marchantia polymorpha, Pogonatum inflexum, Pteridium aquilinum, Pinus densiflora, Ginkgo biloba, Panax ginseng 에 있어서 pinocytotic vesicles 외에 다양상안 막성구조는 원형질막과 세포질안의 무과립성 소포체에서 유래된다. 원형질막에서 유래되는 것은 다소포체인 것과 미에린 양구조인 것으로 구분되고, 이것은 2차액포를 형성하거나 또는 1차액포안으로 유리되어 퇴화한다. 세포질안의 소포체가 세포질의 일부를 포위하고 그 내부에 막증식과 더불어 세포질이 감소되면서 다층막구조로 변하고, 이것은 액포안으로 유리되어 퇴화한다. 세포질의 소포체에서 유래되는 것은 액포분화와 밀접한 관계가 있는 세포질분리체 또는 세포용해체인 것으로 믿어진다. The origins and the functions of the multi vesicular bodies and the various structures of the membranes related to the cytolysomes were studied in the mycelium cells of Rhizopus nigricans, Aspergillus niger and A. ochraceus, in the hymenium and basidium cells of Agricus bisporus sand Rhizopogon rubesecens, in the cells of assimilation tissue of Marchtantia polymorpha and Pogonalum inflexum and in the mesophyll cells of Pteridium aqiulinum, Pinus densiflora, Ginkgo biloba and Panax ginseng fixed with glutaraldehyde-paraformaldehyde-$ OsO_4$. In Rhizopus nigricans, Aspergillus niger, A. ochraceus, Agricus bisporus sand Rhizopogon rubescens, the concentric multilamellar, multivesicular, myelin-vesicle-tubular and concentric parallel-lamellar complexes were originated from the plasmalemma, while in Marehantia polymorpha, Pogonatum inflexum, Pteridium aquilinum, Pinus densiflora, Ginkgo biloba and Panax ginseng, they were originated from plasmalemma and the cytoplasm. The structures originated from the plasmalemma may be grouped into multi vesicular body and myelin-like structure, both forming the secondary vacuoles or protruding into the central vacuoles and finally degrading, In some cases, endoplasmic reticulum within the cytoplasm encloses some part of the cytoplasm to form a circle where the membranous lamellae increase in number, while the enclosed cytoplasm decrease to be eventually replaced by the multilamellar structure which is released into the vacuoles and subsquently degraded. The structures originated from the cytoplasm are believed to be the cytosegresomes or cytolysomes closely related to the differentiation of the vacuoles. The possible fate of these structures are also discussed.

인삼 종자의 저장단백질에 관한 면역 세포화학적 연구 - Tris 완충액 가용성 단백질 -

김우갑,Kim, Woo-Kap 한국현미경학회 1989 Applied microscopy Vol.19 No.2

인삼 종자의 배유조직에서 Tris 완충액 가용성 저장단백질을 추출한 후 전기 영동적 분석으로 분리하여 $SP_{1}$(MW=160,000)과 $SP_2$(MW=70,000)의 두가지 저장단백질을 정제하였다. 이 두가지 저장단백질을 항원으로 사용하여 토끼에 피하주사하여 항체를 얻었으며, 이 항체를 이용하여 면역 세포화학적 금입자표지법을 실시한 결과, $SP_1$과 $SP_2$ 모두 구형의 protein body내에 산재하여 있음을 확인되었으며, globoid에는 이러한 두가지 단백질중 어느 것도 함유되어 있지 않는 것으로 나타났다. 또한 각각의 protein body에 함유된 $SP_1$과 $SP_2$의 상대적 함량에는 서로 차이가 있음이 확인되었다. Buffer soluble storage proteins of ginseng seed have been localized by electron microscopy using post-embedding immunocytochemical gold labelling technique. Major components of the storage proteins were revealed to be storage protein-1($SP_{1}$, MW 160,000) and storage protein-2($SP_{2}$, MW 70,000). Both of the storage proteins are glycoproteins. Anti-$SP_{1}$ and anti-$SP_{2}$ from rabbit, against $SP_1$ and $SP_2$, respectively, reacted on sections of ginseng endosperm tissue embedded in Spurr's epoxy resin. The rabbit antibodies were visualized indirectly by reaction with protein A labelled with colloidal gold. Both storage proteins were found to be accumulated together in the same protein bodies, but their relative contents are not equal.

지질 및 전분성 종자에서 지질 및 지질가수분해효소의 분포

김우갑(Woo Kap Kim),유성철(Seong Cheol Yu) 한국식물학회 1992 Journal of Plant Biology Vol.35 No.3

N/A

식물생장소가 대두 엽육세포의 Cytolysome - like Organelle 에 미치는 영향

김우갑(Woo Kap Kim),박만규(Man Kyu Park),박홍덕(Hong Duok Park) 한국식물학회 1974 Journal of Plant Biology Vol.17 No.4

Leaf tissue of Glycine max Merr. was fixed in para-formaldehyde-glutarldehyde and postfixed in osmium tetroxide or postassium permanganate for electron microscopy. The origin of cytolysome-like organelles of mesophyll cell was studied and changes of fine structure of the organelles according to treating solutions such as gibberellin(GA), kinetin(KI), 2,4-dichlorophenoxy acetic acid(2,4-D) or 2,4-D+GA(1㎎/ℓ, respectively) were observed. The cytolysome-like organelles differentiate in endoplasmic reticulum and plasmalemma, and they drop into vacuoles being isolated from the formers. They seem to change into myelin-like structure and to be degenerated by autodigestion. Cytolysome-like organelles involved in cell walls and vacuoles showed activity of acid phosphatase. In the group of GA and KI treatment, cytolysome-like organelles were similar to that of the control group. But in the treatmental groups of 2,4-D and 2,4-D+GA, myelin-like structures increased in size and autodigestion of this organelles were similar to that of the control group. But in the treatmental groups of 2,4-D and 2,4-D+GA, myelin-like structures increased in size and autodigestion of this organelle seemed to be accelerated. In the treatmental group of 2,4-D+GA, myelin-like structures shown high electron density were observed in cytoplasm and vacuoles together.

인삼 ( Panax ginseng C. A. Meyer ) 종피의 (種皮) 구조 및 분화에 관한 연구

김우갑(Woo Kap Kim),김은수(Eun Soo Kim),정병갑(Byung Kap Jeong) 한국식물학회 1986 Journal of Plant Biology Vol.29 No.4

Structure and differentiation mechanism of the seed coat of Panax ginseng are studied with light and electron microscopes to clarify the developmental processes of seed coat and the structural changes during the differentiation of the seed. The seed coat of ginseng is differentiated from the inner cell layers of ovary wall, which can be compared with the seed coat differentiated from integument(s) in other plants. The single integument is differentiated into endothelium, which is degenerated to one layer of 4 ㎛ in thickness, composed of remants of cell wall components in fully ripened seed. The ripened seed coat is composed of three layers; fringe layer, inner layer and palisade layer, and all of them are crossed at right angles with one another. This may be the cause of protection of the kernel from other mechanical injuries. The thickness of fully ripened seed coat is about 300∼600 ㎛, and arrangements of sclereids are irregular. However the raphe region of seed coat is thin about 200 ㎛ in thickness and sclereids in that region are arranged regularly. This is the important cause for the cleavage of the seed coat during post-maturation process. The vascular bundles on the raphe are still remaining after sarcocarps are removed, and one of the branches of vascular bundles entered into the seed coat through the hilum and extended to chalazal region. During post-maturation process, the supply of water being necessary for growth of embryo may be accompolished by the vascular bundles entered into the seed coat through the opened hilum.

마우스위장관(胃腸管) 점막(粘膜)의 점액분비세포(粘液分泌細胞)에 대(對)한 조직화학적(組織化學的) 및 전자현미경적(電子顯微鏡的) 연구(硏究)

김두규,김우갑,Kim, Doo-Kyu,Kim, Woo-Kap 한국현미경학회 1976 Applied microscopy Vol.6 No.1

The present study was performed to clarify the histochemical compositions and fine structure of the mucus secreting cells in the gastrointestinal mucosa of normal mice. The mucus cells in the surface epithelium of stomach body had neutral mucin and some quantity of weak acid mucin. And the mucus cells in gastric pits and mucus neck cells had neutral mucin. The goblet cells in villial epithelium of small intestine contain strong acid sulfated mucin as their main content and a little of neutral mucopolysaccharide. However, the goblet cells in intestinal glands-Liberkuhn crypt were confirmed to contain non-sulfated weak acid mucin. The goblet cells in the surface epithelium of colon had the same component as the small intestine did. But the cells in the crypts of colon contained neutral and weak acid mucin as their main contents. The majority of secretory granules of the surface epithelial cells of the stomach body had high electron density, and some granules with low electron density appeared too. While the mucin granules in the mucus neck cells were low in its electron density, and some of those granules were frequently found to have dense core in them. Secretory granules in goblet cells of small and large intestines had low electron density. The mode of secretion in mucin-containing cells in gastro-intestinal tract was found to be merocrine. 성숙한 정상마우스 ( 체중 약 20gr)의 위장관 점막 내에 출현하는 점액함유 세포의 점액질의 화학적 조성을 구명하고자 PAS, AB pH 2.5-PAS, AB pH 1.0-PAS 염색법을 이용하여 중성점액질, 황화점액질 및 산성점액질의 염색반응의 유무 내지 강약으로서 비교 검토하였고, 또 전자현미경을 이용하여 이들 세포의 미세구조를 관찰하였다. 위체부 표면상피에 출현하는 점액함유 세포는 주성분이 중성점액질이고, 약간의 약산성 점액질도 존재하였다. 위소와에 있는 점액세포와 점액경세포는 중성점액질을 함유하고 있었다. 소장의 점막상피에 출현하는 배상세포에서는 약산성, 강산성, 황화점액질이 주성분이었고, 약간의 중성점액질을 함유하고 있었다. 한편, 장선의 장상세포는 비황화 약산성 점액질을 갖고 있다. 대장의 점막상피에 출현하는 배상세포는 표면상피에서는 소장과 대동소이 하였으나 은와부에서는 중성 및 약산성점액질이 주성분이었다. 위체부 표면상피에 있는 점액세포의 과랍은 전자밀도가 높은 것이 대부분이었고, 전자밀도가 낮은 것이 소수 출현하였다. 점액경세포의 점액과립은 전자밀도가 낮았으나, 그 속에 dense core 가 있는 것이 출현하였다. 대 소장 점막상피에 출현하는 배상세포는 모두 전자밀도가 낮은 과립이 존재하였다. 위장관의 점액세포들은 모두 부분분필방식을 취하고 있었다.