• 농업과학연구
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• 제39권1호
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• pp.23-28
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• 2012

In order to cloning of PPO gene as a melanin formation related genes involved in hardening and pigmentation of insect integument or wing, we cloned cDNA and analyzed the sequence of PPO gene of H. axyridis. PPO2 primer were designed based on the sequences of PPO genes of Tribolium castaneum and Drosophila melanogaster, and then plasmid DNA were cloned from PCR products obtained from different two color patterns. When the plasmid DNA band pattern were digested by restriction enzymes, BamH1, Xba1, and EcoR1, we found same size band pattern. However, this sequence was not homologous to sequence of T. castaneum PPO gene. Using the primer designed based on the sequence of D. melanogaster, 209 bp PCR product was observed.

• Applied Microscopy
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• 제18권2호
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• pp.91-101
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• 1988

The ultrastructure of the sac and tail portion of the large ampullate gland and production of the silk materials in the orb web spider, Nephila clavata L. Koch, are studied with electron microscope. Large ampullate glands, the largest glands among the seven kinds of silk glands in this species, are composed of three parts which are the excretory duct, the storage sac and the convoluted tail. The wall of the sac is composed of a single layer of columnar epithelial cells. In the cytoplasm of these cells several kinds of the secretory granules, which are commonly associated with the rough endoplasmic reticula and had characteristic crystalloid, are seen. According to the morphology and internal textures of these granules, the sac portion is subdivided into proximal(connected with the convoluted tail) and distal(connected with excretory duct) portion. Between these two portions, the proximal portion is longer than the distal by four times. Silk materials, being synthesized to the shape of secretory granules, within the glandular epithelial cell of the tail portion are released to the inner cavity by the mechanism of the eccrine secretion. These secretory granules are originated from the rough endoplasmic reticula of the glandular epithelial cells, whereas no Golgi complexes has been found in any of the cells which have been examined.

• Applied Microscopy
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• 제19권1호
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• pp.59-69
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• 1989

Ultrastructure of the secretory portion of the small ampullate gland in the orb web spider, Nephila clavata, has been investigated using the electron microscope. The secretory portion of this gland comprise two different regions which are a S-shaped storage sac and a long, convoluted tail. By the internal textures of the secretory granules, the sac is subdivided into two regions ; the proximal region near the tail and the distal region near the duct. Commonly single layered connectives cover the basal portion of the sac epithelium, and apical portion of the epithelial cells is occupied by the thick cuticles. Within the epithelial cells of both the proximal and distal region, several types of the secretory granules surrounded by a limiting membrane and had characteristic crystalloid are scattered throughout the cytoplasm. The granular size and its electron densities are not coincide with each other according to the maturation level of the granules. The wall of the tail is composed of single layer of columnar epithelial cells, and their nuclei are found at the basal portion of the cells. Dissimilar to the epithelial cells of the sac, apical cuticles are not found at this portion. In the cytoplasm of these cells, numerous secretory granules, synthesized from the rough endoplasmic reticula commonly and had fine fibrous materials, are found. At the cell surface bordering the lumen, microvilli are seen, arid along the cellular boundaries specialized septate junctions and desmosomes appeared.

• Applied Microscopy
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• 제19권1호
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• pp.49-58
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• 1989

Ultrastructure of the excretory duct of the small ampullate gland in the orb web spider, Nephila clavata L. Koch are studied with light and electron microscopes. The small ampullate glands, located near the midline portion of the abdominal cavity, are connected with the spigots(large spinning tubes) on the middle spinnerets and composed of three parts which are the excretory duct, the storage sac and the convoluted tail. The excretory duct of this gland is enclosed by a thin layer of the outer connective tissues. By the morphology of the apical cuticles and internal textures of the epithelial cells, the duct is subdivided into two regions which are proximal duct region near the sac and distal duct region near the spinnerets. At the distal region of the ducts, the subcuticle which had the function of water removal form the progenetive silk material is well developed, whereas at the proximal region this cuticle disappeared and instead of these, endocuticle is developed. Moreover the epithelium of the distal duct region is composed of columnar epithelial cells, but at the proximal region the epithelium is changed to squamous or cuboidal forms. In the cytoplasm of the epithelial cells, rough endoplasmic reticula, Golgi comlexes and large secretory vesicles related to the production of the cuticular materials are well developed. And between the adjacent epithelial cells, specialized septate junction and desmosomes are formed along the plasma membrane.

• 환경생물
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• 제22권2호
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• pp.300-307
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• 2004

The chloroacetanilide herbicide alachlor (2-chloro-2', 6'-diethyl-N-(methoxymethyl)-acetanilide) and organochlorine insecticide endosulfan (6, 7, 8, 9, 10, 10-hexachloro-l, 5, 5a, 6, 9, 9a-hexahydro-6, 9-methano-2, 3, 4-benzodioxathiepin-3-oxide)are the highly toxic agricultural chemicals. Bombina orientalis is one of the most common amphibians in the world and comprises a large proportion of their total number. B. orientalis spawns in the farming regions at Spring when the massive application of agricultural chemicals occurs. These chemicals in farmland may threaten the reproduction of this frog. Therefore, we examined the embryotoxic and survival effects of alachlor and endosulfan at various concentrations in B. orientalis embryos. The survival rates of embryos at 312h post fertilization treated with alachlor and endosulfan were decreased with concentration dependent manner. Also, developmental malformations appeared by alachlor and endosulfan in B. orientalis embryos. The malformations showed in order of frequency with bent trunk, tail dysplasia, bent tail, thick-set body and ventral blister in alachlor treated embryos. The exposure of endosulfan produced 7 types of severe external malformations with tail dysplasia, pectoral blister, bent trunk, bent tail, cephalic dysplasia, ventral blister, and thick-set body. Following exposure to alachlor and endosulfan the types of malformations were diverse, suggesting these chemicals target multiple events in embryonic and larval development in this species. These results suggested that alachlor and endosulfan were detrimental for survival and development of B. orientalis embryos.

• 환경생물
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• 제22권2호
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• pp.329-335
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• 2004

To develop a biomarker for the monitoring of the contamination of estrogenic endocrine disrupters in the aquatic environment, reverse transcription -polymerase chain reaction (RT-PCR) analysis of vitellogenin (Vg) mRNA expression was optimized in Bombina orientalis, a Korean red bellied toad species. Based on partial cDNA sequences of both Vg and beta actin genes of B. orientalis, specific primers for RT-PCR of Vg and beta actin mRNAs were developed. Semiquantitative RT-PCR of the Vg mRNA in liver was optimized using a beta actin mRNA as an internal control in both sexes. In female RT-PCR using $1\;\mu{g}$ of the liver cDNA resulted in a linear increment in the PCR product of Vg from 18 to 34 cycles of amplification. In male, on the contrary, the RT- PCR product was first detected at 30 cycles of amplification and a linear increment was observed from 30 to 40 cycles of amplification, suggesting that male B. orientalis expresses minute amount of Vg mRNA which is a $2^{-12}$ equivalent of female. In conclusion, the optimized protocol for semiquantitative RT-PCR analysis of Vg mRNA level in B. orientalis male liver will be useful for the environmental monitoring the xenoestrogen contamination in the freshwater environment in Korea.

• Applied Microscopy
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• 제28권3호
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• pp.329-343
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• 1998

Early wound healing responses in the skin of the Korean fire bellied toad, Bombina orientalis, were examined using histochemical , scanning and transmission electron microscopical techniques. Cutaneous wounds were induced by sharp razor blade , and maintained in special cages for up to 3 days. By this treatment whole parts of epithelium and dermis were profoundly affected. After injury, retraction of damaged surface of the skin was observed. At 3 hours after injury, poly-band was form by mucous substance. The wound skin consisted of mucous gland secreted mucous substance which flows into wound cavity and the wound area was sealed by mucous substance which form thick layer at 6 hours. It appeared that natural disposition showed acid and neutral mucous substance by reaction of PAS and Alcian blue (pH 2.5) staining. After formation of the poly-band, epithelial cells were formed by migration of regenerating epithelial cells. These cells moved over the wound surface by migration at 12 hours after wounding. Poly-band was much increased in thickness and migrating cells were completely formed wound closure at 24 hours post-wounding. After 72 hours regenerated epithelial layer consists of four or five cell layers which are similar to clear, granular, and spinous layer.

• Applied Microscopy
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• 제28권3호
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• pp.379-390
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• 1998

Dermal wound healing responses in the skin of the toad, Bombina orientalis, were examined using transmission electron microscopy. At 12 hours after wounding, debridement and collagenolysis occurred in damaged dermis. Histocyte has a large nucleus and long cytoplasm process. Phagocytic vesicle and lysosome were observed in the cytoplasm. Damaged blood cells were transformed spindle to irregular shape. Autolysis was observed in their cytoplasm. Histocytes are found in poly-band. The irregularly shaped nucleus is located peripheral region in cytoplasm. At 2 days after wounding, partial aggregation of blood cells is observed. Phagocytic, activity is observed in histocyte and collagenolytic collagen fibers are scattered. Fibroblast is observed in the dermis at 3 days after wounding. Clusters of ribosomes and some short cisternae of rough-surfaced endoplasmic reticulum are found in the cytoplasm. In histocyte at 7 days post wounding, various size granules composed of moderately dense material are found the cytoplasm. In this period histocyte is round to rod in profile, with slender processes projecting from the surface. At 7 day after injury, it was observed that formation of connective tissue fibers and amorphous ground substance in regenerating skin.

• Parasites, Hosts and Diseases
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• 제33권2호
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• pp.75-84
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• 1995

Gorgoderidae에 속하는 흡충류는 어류, 양서류 및 파충류 등의 방광에 기생하며 우리 나라의 양서류에서는 Gonodera japonica 1종만이 기록되어 있다가 유와 이(1983)에 의해 Corgoderina bombinae 1종이 신종으로 보고된 바 있다. 본 연구에서는 1989년부터 1994년에 걸쳐 전국 각지에서 채집한 양서류를 대상으로 그들의 방광에 기생하는 윤충류를 조사한 결과 Gorgoder japonica 및 Conoderinc bombinae 2종을 동정하였으며. 분류된 2종에 대한 형태학적인 특징을 기술하고 이들의 분류학적 위치에 대해 상세히 논하였다 G. japonica가 G. cygnoides와 명확히 구별되는 점은 난황선이 깊은 열개에 의해 뚜렷한 여러 개의 엽을 형성하는 점이었다. G. bombinoe는 기존의 종들과 비교한 결과 뚜렷한 종으로서 확인되었으며, 이 종에 대한 국명을 "무당개구리방광흡충"으로 제안한다.

• 한국균학회지
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• 제19권3호
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• pp.175-185
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• 1991

1989년 3월부터 1990년 10월까지 지리산 국립공원 일대에서 한국산 고등균류를 약 70여종 채집하여 동정한 결과 미기록 1속과 미기록 12종을 확인 하였으며 이들에 대하여 한국 보통명을 신칭하였다. Tubaria (겨나팔버섯속), Collybia areruata (단풍애기버섯), Hygrocybe calyptraeformis (새벽벚꽃버섯), Tubaria fufuracea (겨나팔버 섯), Amanita hemibapha subsp. javanicea (노란띠쟈바버섯), A. alboflabescens (백황색광대버섯), A. neoovoidea (신알광대버섯), Leccinum holopus var. holopus (으뜸껄껄이그물버섯아재비), L. griseum (회색껄껄이그물버섯), Tylopilus nigropurpureus (흑자색쓴맛그물버섯), Lactarius zonarius var. zonarius (당귀젖버섯아재비), L. necator (독젖버섯), Russula alboareolata (흰둘레무당버섯).