Friable callus was induced when immature seeds of floribunda rose were inoculated on MS medium supplemented with 2,4-D 3.0 mg.L^-1. When transfered onto subculture media, friable callus developed into embryogenic call...Friable callus was induced when immature seeds of floribunda rose were inoculated on MS medium supplemented with 2,4-D 3.0 mg.L^-1. When transfered onto subculture media, friable callus developed into embryogenic callus, which was used to establish cell suspension lines. Cell suspensions had to be subcultured at a interval of 4-5 days at the first several culture cycles. The best subculturing cycle for the stable cell suspensions was 8-10 days. The best inoculum quantity was 1 mL PCV（Packed Cell Volume） per 40 mL culture fluid.展开更多
Korean pine is an important afforestation tree species in Northeast China,which has a high ecological and economic value.Although regeneration of somatic embryogenesis using immature zygotic embryos of Korean pine as ...Korean pine is an important afforestation tree species in Northeast China,which has a high ecological and economic value.Although regeneration of somatic embryogenesis using immature zygotic embryos of Korean pine as explants has been successful,it cannot be applied to automation and large-scale production.Therefore,we urgently need a method that can increase the output of somatic embryos(SEs)to meet the needs of large-scale production.We used Korean pine 1-1 and 1-100 cell lines as research materials to evaluate the effects of inoculum-density,culture time,orbiting speed,vessel volume,plant growth regulator(PGR)concentration,and carbon source on the proliferation of embryogenic tissue(ET).The somatic embryogenesis ability of ET cultured in different liquid suspension media was also evaluated.We found that during liquid suspension culture of Korean pine ET,the sedimented cell volume(SCV),fresh weight(FW)and dry weight(DW)were affected by inoculumdensity,culture time,orbiting speed,2,4-D concentration,6-BA concentration and carbon source type.Fourty mg⋅mL^(−1)ET were transferred to a 200 mL Erlenmeyer flask containing 20 mL liquid medium,and cultured at 100 rpm/min for 14 days to obtain the maximum proliferation.In addition,we also found that SCV,FW and DW were higher when PGRs were reduced in the liquid suspension medium.The substitution of maltose for sucrose resulted in slow growth of cultures and limited SE yield(13 SEs g^(−1)FW).Although culture proliferation was high at 50 rpm,SE yield was inhibited by 48%compared with 100 rpm(50 rpm=33 SEs g^(−1)FW;100 rpm/min=70 SEs g^(−1)FW).Cultivation in low-concentration PGR(1.15μM⋅L^(−1)2,4-D,0.25μM⋅L^(−1)6-BA)and sucrose liquid medium at 100 rpm/min(80 SEs g^(−1)FW)could not only promote culture proliferation but also increase SE yield.The determination of the suspension culture scheme of Korean pine ET provides a reference for further expansion to bioreactor culture in the future and lays a foundation for the automation and scale of somatic embryogenesis of Korean pine.展开更多
Wild cotton species can contribute a valuable gene pool for agronomically desirable cultivated tetraploid cultivars. In order to exploit diploid cotton a regeneration system is required to achieve transformation based...Wild cotton species can contribute a valuable gene pool for agronomically desirable cultivated tetraploid cultivars. In order to exploit diploid cotton a regeneration system is required to achieve transformation based goals. The present studies aimed at optimizing the conditions for regeneration of local varieties as well as wild species of cotton. Different callus induction media were tested with varying concentrations of hormones in which sucrose was used as nutritional source. Different explants (hypo-cotyls, cotyledon, root) were used to check the regeneration of both local cotton plants and wild relatives using T & G medium, BAP medium, CIM medium, EMMS medium, and cell suspension medium. Different stages of embryogenicity such as early torpedo stage, late torpedo stage, heart stage, globular stage and cotyledonary stage were observed in wild relatives of cotton. The results of this study pave the way for establishing future transformation methods.展开更多
CRYOPRESERVATION by vitrification is a new ultralow-temperature-storage technique developed in the 1980s. Operationally, procedures for vitrification of biological specimens consist of the following basic steps: treat...CRYOPRESERVATION by vitrification is a new ultralow-temperature-storage technique developed in the 1980s. Operationally, procedures for vitrification of biological specimens consist of the following basic steps: treatment of the samples in a highly concentrated vitrification solution and then plunging the specimens into liquid nitrogen. The word 'vitrification' is sometimes used to describe the high moisture of the in vitro cultures or regenerants. However, in this note, this word means a biophysical process in which the cells and the solution are transformed into a homogeneous and amorphous state. The water in cells does not change into ice, but in展开更多
文摘Friable callus was induced when immature seeds of floribunda rose were inoculated on MS medium supplemented with 2,4-D 3.0 mg.L^-1. When transfered onto subculture media, friable callus developed into embryogenic callus, which was used to establish cell suspension lines. Cell suspensions had to be subcultured at a interval of 4-5 days at the first several culture cycles. The best subculturing cycle for the stable cell suspensions was 8-10 days. The best inoculum quantity was 1 mL PCV（Packed Cell Volume） per 40 mL culture fluid.
基金This work was supported by the Fundamental Research Funds for the Central Universities(2572020AW15)the National Key R&D Program of China(2017YFD0600600).
文摘Korean pine is an important afforestation tree species in Northeast China,which has a high ecological and economic value.Although regeneration of somatic embryogenesis using immature zygotic embryos of Korean pine as explants has been successful,it cannot be applied to automation and large-scale production.Therefore,we urgently need a method that can increase the output of somatic embryos(SEs)to meet the needs of large-scale production.We used Korean pine 1-1 and 1-100 cell lines as research materials to evaluate the effects of inoculum-density,culture time,orbiting speed,vessel volume,plant growth regulator(PGR)concentration,and carbon source on the proliferation of embryogenic tissue(ET).The somatic embryogenesis ability of ET cultured in different liquid suspension media was also evaluated.We found that during liquid suspension culture of Korean pine ET,the sedimented cell volume(SCV),fresh weight(FW)and dry weight(DW)were affected by inoculumdensity,culture time,orbiting speed,2,4-D concentration,6-BA concentration and carbon source type.Fourty mg⋅mL^(−1)ET were transferred to a 200 mL Erlenmeyer flask containing 20 mL liquid medium,and cultured at 100 rpm/min for 14 days to obtain the maximum proliferation.In addition,we also found that SCV,FW and DW were higher when PGRs were reduced in the liquid suspension medium.The substitution of maltose for sucrose resulted in slow growth of cultures and limited SE yield(13 SEs g^(−1)FW).Although culture proliferation was high at 50 rpm,SE yield was inhibited by 48%compared with 100 rpm(50 rpm=33 SEs g^(−1)FW;100 rpm/min=70 SEs g^(−1)FW).Cultivation in low-concentration PGR(1.15μM⋅L^(−1)2,4-D,0.25μM⋅L^(−1)6-BA)and sucrose liquid medium at 100 rpm/min(80 SEs g^(−1)FW)could not only promote culture proliferation but also increase SE yield.The determination of the suspension culture scheme of Korean pine ET provides a reference for further expansion to bioreactor culture in the future and lays a foundation for the automation and scale of somatic embryogenesis of Korean pine.
文摘Wild cotton species can contribute a valuable gene pool for agronomically desirable cultivated tetraploid cultivars. In order to exploit diploid cotton a regeneration system is required to achieve transformation based goals. The present studies aimed at optimizing the conditions for regeneration of local varieties as well as wild species of cotton. Different callus induction media were tested with varying concentrations of hormones in which sucrose was used as nutritional source. Different explants (hypo-cotyls, cotyledon, root) were used to check the regeneration of both local cotton plants and wild relatives using T & G medium, BAP medium, CIM medium, EMMS medium, and cell suspension medium. Different stages of embryogenicity such as early torpedo stage, late torpedo stage, heart stage, globular stage and cotyledonary stage were observed in wild relatives of cotton. The results of this study pave the way for establishing future transformation methods.
文摘CRYOPRESERVATION by vitrification is a new ultralow-temperature-storage technique developed in the 1980s. Operationally, procedures for vitrification of biological specimens consist of the following basic steps: treatment of the samples in a highly concentrated vitrification solution and then plunging the specimens into liquid nitrogen. The word 'vitrification' is sometimes used to describe the high moisture of the in vitro cultures or regenerants. However, in this note, this word means a biophysical process in which the cells and the solution are transformed into a homogeneous and amorphous state. The water in cells does not change into ice, but in
