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야생 흰 제비꽃(viola patrinii DC.)callus의 현탁배양 방법이 세포 활성에 미치는 영향
김두현,정용모,정정한,이인,권오창,Kim, Du-Hyun,Chung, Yong-Mo,Chung, Chung-Han,Yeeh, Yeehn,Kwon, Oh-Chang 한국생명과학회 1996 생명과학회지 Vol.6 No.2
To understand effect of inoculum size, cell density, sucrose concentration and concentrations of MS basal on suspension culture and protoplast isolation of wild viola(Viola patrinii DC.) callus from petiole segments this experiment was conducted. In the lot of 30 mesh inoculum size, two observations were; One was that a considerable increase in the fresh and dry weight of callus was determined. Another was that the callus mass was relatively compact compared with others. A recommendable cell density was 0.4g for 20ml culture medium and the higher sucrose concentration, the higher fresh and dry weight were obtained. The dilution of MS basal salt was differently affected on fresh and dry weight; the highest fresh weight was found in 1x MS salt, while the higest dry weight was in 1/3x dilution.The addition of casein hydrolysate(3g/L) was more effective to increase of both fresh and dry weight. THe contents of protein was great in the inoculum lots with larger inoculum sizeand higher concentration of MS basal salts contenting 3g/L of casein hydrolysate and higher sucrose compared with others. The greatest protoplasts were isolated from the lot of 10 mesh size treated with 1%pectinase SE-150 and 2% cellulase YC. In general, for optimal protoplast isolation the followingconditions were recommended; 1) Use of smaller cell size cultured for 2-5 weeks, and 2) more than 5 hours incubation using the combined mixture of the enzymes with proper concentrations.
Kyung Soon Lee(이경순),Chung Han Chung(정정한),Yong Mo Chung(정용모),Doh Hoon Kim(김도훈),Soon Jae Jeong(정순재),Jae Sung Nam(남재성),Gyung Tae Kim(김경태),Young Byung Yi(이영병) 한국생명과학회 2007 생명과학회지 Vol.17 No.10
히아신스 Pink Pearl 품종의 인편배양에서 자구의 재생과 생장은 1.0 ㎎/l와 3.0 ㎎/l의 농도에서 IAA 보다 IBA가 더 효과적이었으며, 절편체의 기부가 배지에 삽식되는 정상적인 치상 방향이 역방향의 치상 방법에 비해 자구의 재생과 뿌리의 생장에 더 효율적인 것으로 나타났다. 계대배양에서 재생된 자구와 뿌리의 생장은 한천배지보다 펄라이트가 첨가된 액체배지에서 증가되었다. 이와 같은 히아신스 Pink Pearl 품종의 기내배양의 경우 자구 재생을 위한 초대배양은 한천 고체배지를, 재생된 자구의 생장은 펄라이트의 액체배지를 사용하는 2 단계의 교대배양 시스템이 더 유용함을 시사한다. The regeneration and growth of bulblets from the bulb scale segments of Hyacinthus orientalis L. cv. Pink Pearl were more efficient in IBA than IAA at the same concentrations (1.0 and 3.0 ㎎/l).. The normal (base-down) orientation of explants was more effective for bulblet regeneration and root growth than the inverted (base-up) orientation. The growth of bulblets and roots was increased higher in the perlite than the agar medium. These results suggested that the alternate culture system, first cultured in the agar medium for bulblet regeneration, and then in the perlite medium for bulblet growth, may be more useful for efficient in vitro culture of hyacinth (H. orientalils) cv. Pink Pearl.
Viola속 식물의 원형질체 및 융합세포의 전자현미경 관찰
정용모,임현희,손병구,서정해,정정한,권오창,Chung, Yong-Mo,Im, Hyun-Hee,Son, Beung-Gu,Suh, Jung-Hae,Chung, Chung-Han,Kwon, Oh-Chang 한국생명과학회 1997 생명과학회지 Vol.7 No.4
To obtain a basic information on the development of Genus Viola, ultrastructure and electrofusion process between the two protoplasts from wild Viola callus cells and pansy mesophyll cells were observed with a scanning electron microscopy(SEM) and transmission electron microscopy(TEM). In the ultrastructural observation of wild viola callus protoplasts and pansy mesophyll protoplasts using SEM, their cell walls were removed completely. A knob-like formation was observed on the enlarge surface of viola callus protoplasts. On the surface of pansy mesophyll protoplasts net-like chloroplasts were observed. In SEM observation of pansy mesophyll protoplasts, chloroplasts devoid of membrane were observed on the surface the protoplasts. Pearl chain was formed by applying AC field of 200 V/cm at 1.0 MHz for 43 sec. The lysis of plasma membranes and fusion process occurred by applying a 1,600 V/cm DC pulse twice for 1 sec. After 1-2 hours of a DC pulse application, it was observed that the two protoplasts were fused completely into one cell. In TEM observation of the fused cell, many small vacuoles were located in the fusion area of the two protoplasts. Indeed, two distinct regions were observed during fusing process; in one region, a nucleus was found, while in the other region, both nucleus and nucleous were found.