• 한국균학회소식:학술대회논문집
• /
• 한국균학회 2009년도 추계학술대회 및 정기총회
• /
• pp.14-16
• /
• 2009

Poria cocos is an importantant medical macrofungus,the sclerotium of Poria cocos has specific value as the drug material. There are few papers about its breeding and spawn rejuvenation. In this project, the protoplasts of cultivated strain T and wild strain L were prepared and treated separately by ultraviolet and heating, then fused with the PEG6000. The tural fusants were selected and identified by the affinity and ISSR analysis. 71 incompatibility strains between parents and reg regenerations were obtained from 118 regenerations by the affinity analysis. Five incompatibility strains were amplified with different primers, the results were showed that they had specific bands of both parents in the profile amplified with 3 primers, which proved these 5 strains were fusants by means of molecular biology marker. On the other hand, 25 strain were selected from 168 protoplast regenerations of cultivated strain T for cultivation experiment. The fresh sclerotium weight of these protoplast regenerations were better than the original strain.significantil 3 strains (T-1, T-4, T-7) increased respectively 118%, 73% and 73% than original strain. This method could be the effective in the rejuvenation Poria cocos.

• 한국균학회지
• /
• 제20권1호
• /
• pp.44-50
• /
• 1992

Rhizoctonia solani의 균사융합군(AG) 중에서 생장속도가 빠른 AG-1과 AG-4는 가장 많은 원형질체를 형성하였으며 속도가 느린 AG-3은 저조하였고 중간인 AG-2와 AG-5는 또한 중간이었다. 균핵의 형성은 AG에 따라 차이가 있었다. Cellulase "onozuka" R-l0, macerozyme R-10, ${\beta}-glucuronidase$의 효소 복합액에서 가장 많은 원형질체를 형성하였다. 5개 융합군은 30시간 배양한 3g의 균사를 0.6 M mannitol이 첨가된 효소 복합액에 $32^{\circ}C$에서 3-4시간 처리하면 가장 많은 원형질체를 형성하였다. 균핵을 형성하는 AG-1을 열처리로 불활성화시킨 후 균핵을 형성하지 않는 AG-5의 원형질체를 PEG와 ${Ca}^{2+}$로 융합시켰다. 재생배지상에서의 균총의 특징과 균핵의 형성에 의해 7 개의 원형질 융합체를 선발하고 esterase 동위효소의 zymogram 및 AG-1 과 융합체인 F150l 사이에서 일어나는 killing reaction으로 확인하였다.

• 한국식물생명공학회:학술대회논문집
• /
• 한국식물생명공학회 2005년도 추계학술대회 및 한일 식물생명공학 심포지엄
• /
• pp.106-111
• /
• 2005

For breeding of a new rootstock for eggplant production, somatic hybrids between two species, Solanum integrifolium and S. sanitwongsei were obtained through protoplast fusion. The former species has been commonly used for rootstock for eggplant production in Japan. Eggplants on these rootstocks are more productive than ungrafted plants, but are susceptible to bacterial wilt caused Ralstonia solanacearum. While the latter species is resistant, the growth of eggplants on this rootstock is rather slow and low yield. Protoplast of both species were isolated from cotyledons, and inactivated with iodoacetamide or UV-irradiation, then fused electrically. The fused products were then cultured. Regenerated plantlets were then transplanted on soil then maintained in a green house. The plants were classified into four groups. Those in the first group showed morphological characters intermediate of the parentalspecies. The plants bore fruit with viable seeds. The plants showed a chromosome number of 2n=48, the sum of those of the parental species, and are suggested to be symmetric fusion products. While plants in the other groupswas less vigorous and showed chromosome number 2n= 68 to 72 suggesting asymmetric fusion products by genomic in situ hybridization(GISH). Isozyme pattern of shikimate dehydrogenase (SKDH; EC 1.1.1.25), isocitrate dehydrogenase (IDH; EC 1.1.1.41) and phosphoglucomutase (PGM; EC 2.7.5.1) showed that 24 regenerated plants in three groups were somatic hybrids. Analysis of random amplified polymorphic DNA (RAPD) showed that 43 S. integrifolium-specific and 57 S. sanitwongsei-specific bands were all found in 24 plants. Both somatic hybrids and its S1 plants were found to be resistant to bacterial wilt, and eggplant grafted these plants using for rootstocks were more productive than grafted mother plants. Now, S1 progenies are used for commercial eggplant production in Osaka Prefecture.

• 미생물학회지
• /
• 제30권3호
• /
• pp.160-163
• /
• 1992

원형질체 융합을 통하여 육성한 killer 효모 융합주 FWKS 260 의 killer toxin 을 ammonium sulfate fractionation, Amicon PM 10 concentration, Sephadex G-200 및 Sephadex G-75 column chromatography 를 행하여 정제한 결과 단일 단백질 band 를 보여 순수하게 정제되었음을 알 수 있었고, 단백질 분해효소를 처리한 결과 killer 활성이 소실되어 killer toxin 의 단백질 부분이 killer 활성을 나타냄을 알 수 있었다. 그리고 이 toxin 은 20.deg.C 에서는 거의 안정하였으나, 온도가 증가함에 따라 점차 활성이 소실되었고, pH 2.0-5.0 에서 비교적 안정하였다. 한편, SDS-polyacrylamide gel electrophoresis 결과 분자량은 약 13.000 임을 알 수 있었고, SDS polyacrylamide gel electrophoresis 를 행한 후 Schiffs reagent 로 염색한 결과 붉은 단일 band 를 보여 정제된 killer toxin 은 glycoprotein 임을 확인 할 수 있었다.