[ Objective] The aim was to study appropriate fusion conditions of protoplasts from 2 species Cosmos bipinnatus and Cosmos sulphureus. [ Method ] The protoplasts were separated from the blossom petals of Cosmos bipinn...[ Objective] The aim was to study appropriate fusion conditions of protoplasts from 2 species Cosmos bipinnatus and Cosmos sulphureus. [ Method ] The protoplasts were separated from the blossom petals of Cosmos bipinnatus and Cosmos sulphureus availed with the function of pectinase and cellulase, and the fusion of two pretoplasts was performed with PEG. [ Results] A great deal of free pretoplasts were obtained from Cosmos bipinnatus by using 6% mannitol + 1.3% cellu- lase + 1.2% pectinase, and from Cosmos sulphureus by using 10% mannitol + 1.3% cellulase + 1.2% pectinase. Five hours after of enzymolysis treatment, large amount of dissociated protoplasts were observed in the solution. Pretoplast fusion was be obtained by using 30% PEG fusion liquid and 0. lmol/1 CaC12 solution (pH value = 9.5). [ Conclusion] Our study established an optimal separation and fusion system of protoplasts from different coreopsis species, laying foundation for the breeding of novel coreopsis varieties.展开更多
This paper discussed the technique of algal protoplast fusion,including the method of protoplast-induced fusion,the screening and identification of hybrid cells and so on.In this paper,the research progress of protopl...This paper discussed the technique of algal protoplast fusion,including the method of protoplast-induced fusion,the screening and identification of hybrid cells and so on.In this paper,the research progress of protoplast-induced fusion of microalgae was reviewed,and the problems existing in the process of protoplast-induced fusion were discussed in order to provide a theoretical basis for the development of algae genetics and breeding.展开更多
This study generated two fused protoplasts of Antrodia cinnamomea and Cordyceps militaris in two ways.The protoplasts of A.cinnamomea were inactivated by heat to inactivate biochemical processes and enzymatic activiti...This study generated two fused protoplasts of Antrodia cinnamomea and Cordyceps militaris in two ways.The protoplasts of A.cinnamomea were inactivated by heat to inactivate biochemical processes and enzymatic activities in the cytoplasm,and the protoplasts of C.militaris were inactivated by UV radiation to invalidate their genome function,then they were fused under optimal conditions to get a fusion rate as(7.42±0.8)×10^(-6) fusants/mL;the new fusants were abbreviated as Ac-Cm.On the other hand,when A.cinnamomea and C.militaris were treated with heat and UV oppositely using similar experiments,the fusion rate was(9.70±0.68)×10^(-5) fusants/mL,and the new fusants were abbreviated as Cm-Ac.We selected each of two best-growing fused colonies Ac-Cm-1,Ac-Cm-2,Cm-Ac-1,and Cm-Ac-2,together with parental A.cinnamomea and C.militaris,and studied their morphology,growth antagonism tests,and genetic relationships by 18 S rRNA sequencing.In comparison with the initial cultures of 4 fusants,the yields of adenosine,biomass,cordycepic acid,cordycepin,total polysaccharide,and total triterpenoids were increased up 1.305-50.1563 times in the optimal medium conditions.For gene stability tests,those of the four fusants and their outputs were stabilized within 10 generations.展开更多
Among eight components of avermectin, B1 fractions have the most effective antiparasitic activities and the lowest level of toxic side-effects and are used widely in veterinary and agricultural fields. In-traspecific ...Among eight components of avermectin, B1 fractions have the most effective antiparasitic activities and the lowest level of toxic side-effects and are used widely in veterinary and agricultural fields. In-traspecific protoplast fusion between two strains of Streptomyces avermitilis, one an avermectin high producer (strain 76-05) and the other a genetically engineered strain containing the mutations aveDˉ and olmAˉ (strain 73-12) was performed for enhancement and selective production of avermectin B in the absence of oligomycin. Two recombinant strains (F23 and F29) were isolated and characterized with regards to the parental merits. F23 and F29 produced only the four avermectin B components with high yield and produced no oligomycin. The avermectin production of F23 and F29 was about 84.20% and 103.45% of the parental strain 76-05, respectively, and increased about 2.66-fold and 3.50-fold, re-spectively, compared to that of parental strain 73-12. F23 and F29 were genetically stable prototrophic recombinants and F29 was quite tolerant of fermentation conditions compared to avermectin high producer parental strain 76-05. The ability to produce avermectin B with high yield without the produc-tion of other avermectin components and oligomycin will make F23 and F29 useful strains for aver-mectin production. Strain F29's tolerance of fermentation conditions will also make it suitable for in-dustrial applications.展开更多
Protoplast fusion was induced between sainfoin and alfalfa by an improved polyethyleneglycol (PEG) method. The intergeneric somatic calluses were selected based on complementation of hydroxyproline-resistance of sainf...Protoplast fusion was induced between sainfoin and alfalfa by an improved polyethyleneglycol (PEG) method. The intergeneric somatic calluses were selected based on complementation of hydroxyproline-resistance of sainfoin and hormone autonomy growth of alfalfa transformation cell line. 17 somatic hybrid plantlets were regenerat-ed. PEG could induce the tight agglutination of protoplasts. During diluting and washing process, cyclization of the linked membrane and formation of vesicle-like structures were observed, resulting in protoplast fusion. 5%-10% glycerol supplemented in the fusion inducing solution markedly increased the frequency of heterogeneous fusion. Better fusion results were obtained when mixed protoplast suspension was dripped in petri dishes in which PEG solution was previously placed. Chromosome number of regenerated hybrid buds varied from 30 to 60. The genome of hybrids in-cluded the small chromosome from sainfoin and two chromosomes with two clear constrictions from alfalfa. The展开更多
Laser tweezers can manipulate small particles, such as cells and organdies. When coupling them with laser microbeam selective fusion of two tobacco protoplasts containing some chloroplast was achieved. Physical and bi...Laser tweezers can manipulate small particles, such as cells and organdies. When coupling them with laser microbeam selective fusion of two tobacco protoplasts containing some chloroplast was achieved. Physical and biological variables that affect laser trapping and laser-induced fusion were also discussed. The results show that the effect of chloroplast content and distribution on the yield of cell fusion is remarkable.展开更多
[Objectives] This study was conducted to improve the genetic character of Epipremnum aureum and provide a new way for genetic breeding of E. aureum. [Methods] In the paper,protoplasts were isolated from tender leaves ...[Objectives] This study was conducted to improve the genetic character of Epipremnum aureum and provide a new way for genetic breeding of E. aureum. [Methods] In the paper,protoplasts were isolated from tender leaves of E. aureum and Scindapsus aureus var. wilcoxii by using 1% cellulase and 1% peetinase. Then the protoplasts were purified and dynamically identified,and fusion of protoplasts of E. aureum and S. aureus was induced by PEG. [Results]A large quantity of protoplasts were obtained from tender leaves of E. aureum and S. aureus successfully,and the activity of protoplasts was over 90%. Fusion product was obtained by PEG induction,and the fusion rate was 16. 3%. [Conclusions]Protoplasts were isolated from tender leaves of E. aureum and S. aureus successfully,and a stable fusion system was built by PEG induction.展开更多
The progress in protoplast technology and its application in fruit breeding were summarized,and the existing problems and application prospect were also discussed.
文摘[ Objective] The aim was to study appropriate fusion conditions of protoplasts from 2 species Cosmos bipinnatus and Cosmos sulphureus. [ Method ] The protoplasts were separated from the blossom petals of Cosmos bipinnatus and Cosmos sulphureus availed with the function of pectinase and cellulase, and the fusion of two pretoplasts was performed with PEG. [ Results] A great deal of free pretoplasts were obtained from Cosmos bipinnatus by using 6% mannitol + 1.3% cellu- lase + 1.2% pectinase, and from Cosmos sulphureus by using 10% mannitol + 1.3% cellulase + 1.2% pectinase. Five hours after of enzymolysis treatment, large amount of dissociated protoplasts were observed in the solution. Pretoplast fusion was be obtained by using 30% PEG fusion liquid and 0. lmol/1 CaC12 solution (pH value = 9.5). [ Conclusion] Our study established an optimal separation and fusion system of protoplasts from different coreopsis species, laying foundation for the breeding of novel coreopsis varieties.
文摘This paper discussed the technique of algal protoplast fusion,including the method of protoplast-induced fusion,the screening and identification of hybrid cells and so on.In this paper,the research progress of protoplast-induced fusion of microalgae was reviewed,and the problems existing in the process of protoplast-induced fusion were discussed in order to provide a theoretical basis for the development of algae genetics and breeding.
基金supported by grants from the Ministry of Science and Technology of Taiwan(Grant number:MOST 106-2320-B-037008-MY2,MOST 108-2320-B-037-022-MY3,108-2811-B-037-511,and 109-2927-I-037-502)funded by the Drug Development and Value Creation Research Center,Kaohsiung Medical UniversityDepartment of Medical Research,Kaohsiung Medical University Hospital(Grant number:KMU-TC108A03-11)。
文摘This study generated two fused protoplasts of Antrodia cinnamomea and Cordyceps militaris in two ways.The protoplasts of A.cinnamomea were inactivated by heat to inactivate biochemical processes and enzymatic activities in the cytoplasm,and the protoplasts of C.militaris were inactivated by UV radiation to invalidate their genome function,then they were fused under optimal conditions to get a fusion rate as(7.42±0.8)×10^(-6) fusants/mL;the new fusants were abbreviated as Ac-Cm.On the other hand,when A.cinnamomea and C.militaris were treated with heat and UV oppositely using similar experiments,the fusion rate was(9.70±0.68)×10^(-5) fusants/mL,and the new fusants were abbreviated as Cm-Ac.We selected each of two best-growing fused colonies Ac-Cm-1,Ac-Cm-2,Cm-Ac-1,and Cm-Ac-2,together with parental A.cinnamomea and C.militaris,and studied their morphology,growth antagonism tests,and genetic relationships by 18 S rRNA sequencing.In comparison with the initial cultures of 4 fusants,the yields of adenosine,biomass,cordycepic acid,cordycepin,total polysaccharide,and total triterpenoids were increased up 1.305-50.1563 times in the optimal medium conditions.For gene stability tests,those of the four fusants and their outputs were stabilized within 10 generations.
基金Supported by National Basic Research Project (Grant No. 2003CB114205)Key Technologies R&D Programme (Grant No. 2004BA713B02-03)
文摘Among eight components of avermectin, B1 fractions have the most effective antiparasitic activities and the lowest level of toxic side-effects and are used widely in veterinary and agricultural fields. In-traspecific protoplast fusion between two strains of Streptomyces avermitilis, one an avermectin high producer (strain 76-05) and the other a genetically engineered strain containing the mutations aveDˉ and olmAˉ (strain 73-12) was performed for enhancement and selective production of avermectin B in the absence of oligomycin. Two recombinant strains (F23 and F29) were isolated and characterized with regards to the parental merits. F23 and F29 produced only the four avermectin B components with high yield and produced no oligomycin. The avermectin production of F23 and F29 was about 84.20% and 103.45% of the parental strain 76-05, respectively, and increased about 2.66-fold and 3.50-fold, re-spectively, compared to that of parental strain 73-12. F23 and F29 were genetically stable prototrophic recombinants and F29 was quite tolerant of fermentation conditions compared to avermectin high producer parental strain 76-05. The ability to produce avermectin B with high yield without the produc-tion of other avermectin components and oligomycin will make F23 and F29 useful strains for aver-mectin production. Strain F29's tolerance of fermentation conditions will also make it suitable for in-dustrial applications.
基金Project supported by the National Natural Science Foundation of China.
文摘Protoplast fusion was induced between sainfoin and alfalfa by an improved polyethyleneglycol (PEG) method. The intergeneric somatic calluses were selected based on complementation of hydroxyproline-resistance of sainfoin and hormone autonomy growth of alfalfa transformation cell line. 17 somatic hybrid plantlets were regenerat-ed. PEG could induce the tight agglutination of protoplasts. During diluting and washing process, cyclization of the linked membrane and formation of vesicle-like structures were observed, resulting in protoplast fusion. 5%-10% glycerol supplemented in the fusion inducing solution markedly increased the frequency of heterogeneous fusion. Better fusion results were obtained when mixed protoplast suspension was dripped in petri dishes in which PEG solution was previously placed. Chromosome number of regenerated hybrid buds varied from 30 to 60. The genome of hybrids in-cluded the small chromosome from sainfoin and two chromosomes with two clear constrictions from alfalfa. The
基金Project supported by the National Natural Science Foundation of China (Grant No. 39470205)State Science and Technology Commision, and Science and Technolgy Commision of Anhui Province
文摘Laser tweezers can manipulate small particles, such as cells and organdies. When coupling them with laser microbeam selective fusion of two tobacco protoplasts containing some chloroplast was achieved. Physical and biological variables that affect laser trapping and laser-induced fusion were also discussed. The results show that the effect of chloroplast content and distribution on the yield of cell fusion is remarkable.
文摘[Objectives] This study was conducted to improve the genetic character of Epipremnum aureum and provide a new way for genetic breeding of E. aureum. [Methods] In the paper,protoplasts were isolated from tender leaves of E. aureum and Scindapsus aureus var. wilcoxii by using 1% cellulase and 1% peetinase. Then the protoplasts were purified and dynamically identified,and fusion of protoplasts of E. aureum and S. aureus was induced by PEG. [Results]A large quantity of protoplasts were obtained from tender leaves of E. aureum and S. aureus successfully,and the activity of protoplasts was over 90%. Fusion product was obtained by PEG induction,and the fusion rate was 16. 3%. [Conclusions]Protoplasts were isolated from tender leaves of E. aureum and S. aureus successfully,and a stable fusion system was built by PEG induction.
基金Supported by National Agro-industry(Apple Industry)Technology Research System/China Agriculture Research System of Apple(nycytx-27)
文摘The progress in protoplast technology and its application in fruit breeding were summarized,and the existing problems and application prospect were also discussed.