Radiation induced mutagenesis followed by in vitro selection was employed for salt tolerance in popular Indian sugarcane (Saccharum officinarum L.) cv. CoC-671. Embryogenic calli were gamma irradiated and exposed to...Radiation induced mutagenesis followed by in vitro selection was employed for salt tolerance in popular Indian sugarcane (Saccharum officinarum L.) cv. CoC-671. Embryogenic calli were gamma irradiated and exposed to different levels of NaCl (42.8, 85.6, 128.3, 171.1,213.9, 256.7, 299.5, or 342.2 mM). The relative growth rate (RGR) decreased progressively with increasing salt stress and was the least with a salt stress of 256.7 mM (0.25±0.009), almost 10 fold lesser than the control. The RGR was significantly lower in 85.6 mM and higher salt stressed calli than the control. The survival percent also decreased, with an increase in NaCl concentration. In case of 10 and 20 Gy irradiated calli, regeneration was observed up to 85.6 mM NaCl selection, medium, whereas, higher treatments (128.3 mM and beyond) exhibited browning initially. However, in the subsequent subcultures, regeneration was obtained in the case of 10 and 20 Gy irradiated calli on 128.3 and 171.1 mM NaCl selections. Higher dose of gamma irradiation (40 Gy) also showed regeneration, but only with 85.6 mM NaCI selection. The unirradiated calli regenerated the highest number of plantlets followed by 10 and 20 Gy irradiated calli on salt selection. A total of 147 plantlets were selected from different salt levels. The salt selected plants are being tested for their field performance.展开更多
In vitro selection of drought-tolerant mutants in sweetpotato cv. Lizixiang was studied by using PEG6000 as selection stress. Embryogenic suspension cultures were cultured in MS medium containing 035% PEG6000 and 2 mg...In vitro selection of drought-tolerant mutants in sweetpotato cv. Lizixiang was studied by using PEG6000 as selection stress. Embryogenic suspension cultures were cultured in MS medium containing 035% PEG6000 and 2 mg L 1 2,4-dichlorophenoxyacetic acid (2,4-D). The results indicated that 30% PEG6000 can be used for the optimal selection stress of drought-tolerance. Embryogenic suspension cultures irradiated with 80 Gy gamma-ray were cultured in MS medium containing 30% PEG6000 and 2 mg L 1 2,4-D and 20 drought-tolerant cell aggregates were obtained. These cell aggregates were transferred to solid MS me- dium supplemented with 2 mg L 1 2,4-D and formed embryogenic callus with somatic embryos. The embryogenic callus with somatic embryos was further transferred to MS medium supplemented with 1 mg L 1 abscisic acid (ABA), resulting in the germination of somatic embryos. In this study a total of 18 regenerated plants were obtained. The regenerated plants were transplanted in a greenhouse and 11 lines were formed. The analysis on drought treatment of seedlings, water retaining capacity of leaves and coefficient of drought-tolerance showed that 3 lines had significant drought-tolerance in comparison with the control plants.展开更多
The present investigation was undertaken to study the effect of gamma irradiation (dose from 10 to 100 Gy) and in vitro selection with fungus filtrate as selecting agent (concentration from 20% to 100%) on the suscept...The present investigation was undertaken to study the effect of gamma irradiation (dose from 10 to 100 Gy) and in vitro selection with fungus filtrate as selecting agent (concentration from 20% to 100%) on the susceptibility of the common bean to Rhizoctonia solani. The best results were found with a dose of 20 Gy or a concentration of 20% of fungus filtrate applied separately. These conditions were used to evaluate the combined effect of both approaches in a second experiment. The combined effect of irradiation and then selection adversely affected growth (height and roots) and survival of the in vitro plants. It may not be necessary to combine the variation generated by irradiation with the selection technique. For future assays we propose the application of: 1) gamma radiation, thereby inducing not only mutants with pathogen resistance, but also with other agronomic traits of interest. Later in the subculture MV4 potential fungus-resistant mutants will be evaluated in the field;or 2) selection pressure using fungus filtrate during three subcultures, which may be sufficient to induce the variation necessary to obtain in vitro plants resistant to fungus.展开更多
Preformed albumin corona of albumin-nonselective nanoparticles(NPs)is widely exploited to inhibit the unavoidable protein adsorption upon intravenous administration.However,very few studies have concerned the preforme...Preformed albumin corona of albumin-nonselective nanoparticles(NPs)is widely exploited to inhibit the unavoidable protein adsorption upon intravenous administration.However,very few studies have concerned the preformed albumin corona of albumin-selective NPs.Herein,we report a novel type of albumin-selective NPs by decorating 6-maleimidocaproyl polyethylene glycol stearate(SA)onto PLGA NPs(SP NPs)surface,taking albuminnonselective PLGA NPs as control.PLGA NPs and SP NPs were prepared by emulsion-solvent evaporation method and the resultant NPs were in spherical shape with an average diameter around 180 nm.The corresponding albumin-coating PLGA NPs(PLGA@BSA NPs)and albumin-coating SP NPs(SP@BSA NPs)were formulated by incubating SP NPs or PLGA NPs with bovine serum albumin solution,respectively.The impact of albumin corona on particle characteristics,stability,photothermal effect,cytotoxicity,cell uptake,spheroid penetration and pharmacokinetics was investigated.In line with previous findings of preformed albumin coating,PLGA@BSA NPs exhibited higher stability,cytotoxicity,cell internalization and spheroid penetration performances in vitro,and longer blood circulation time in vivo than those of albumin-nonselective PLGA NPs,but albumin-selective SP NPs is capable of achieving a comparable in vitro and in vivo performances with both SP@BSA NPs and PLGA@BSA NPs.Our results demonstrate that SA decorated albumin-selective NPs pave a versatile avenue for optimizing nanoparticulate delivery without preformed albumin corona.展开更多
The selective toxicity of the mitochondria-specific cationic fluorescent dye rhodamine 123 (Rh-123) on He La cells in culture was studied. In this report, we demonstrate that with continuous exposure, Rh-123 markedly ...The selective toxicity of the mitochondria-specific cationic fluorescent dye rhodamine 123 (Rh-123) on He La cells in culture was studied. In this report, we demonstrate that with continuous exposure, Rh-123 markedly inhibited the growth of HeLa cells but had little effect on normal human kidney fibroblasts. With continuous exposure to Rh-123, the growth rate, colony forming ability, anl mitotic index of HeLa cells were decreased. The mechanism of toxicity of Rh-123 on HeLa cells was investigated by EM and enzyme cytochemistry stain. The mitochondria of carcinoma cells were the main targets for the inhibitory action of Rh-123, since they selectively accumulated the dye. At the dosage of Rh-123 which was toxic to HeLa cells, the structure and function of mitochondria were disrupted, as the mitochondria-related enzymes, i.e., ATPase, LDH and SDH were inhibited. The possible mechanism of the action of Rh-123 on HeLa cells is briefly discussed.展开更多
DNA-encoded chemical library(DEL)links the power of amplifiable genetics and the nonself-replicating chemical phenotypes,generating a diverse chemical world.In analogy with the biological world,the DEL world can evolv...DNA-encoded chemical library(DEL)links the power of amplifiable genetics and the nonself-replicating chemical phenotypes,generating a diverse chemical world.In analogy with the biological world,the DEL world can evolve by using a chemical central dogma,wherein DNA replicates using the PCR reactions to amplify the genetic codes,DNA sequencing transcripts the genetic information,and DNA-compatible synthesis translates into chemical phenotypes.Importantly,DNA-compatible synthesis is the key to expanding the DEL chemical space.Besides,the evolution-driven selection system pushes the chemicals to evolve under the selective pressure,i.e.,desired selection strategies.In this perspective,we summarized recent advances in expanding DEL synthetic toolbox and panning strategies,which will shed light on the drug discovery harnessing in vitro evolution of chemicals via DEL.展开更多
Peanut (Arachis hypogaea L.) is an important oil crop globally and high oil content is one of the major targets in peanut breeding programs. Previous studies indicated that the osmotic pressure (OP) of the leaves of p...Peanut (Arachis hypogaea L.) is an important oil crop globally and high oil content is one of the major targets in peanut breeding programs. Previous studies indicated that the osmotic pressure (OP) of the leaves of peanut plants subjected to drought stress was negatively correlated with kernel oil content. Based on this knowledge, we established a practical and reliable method for creating new peanut varieties with high oil content using in vitro mutagenesis and directional OP-based selection. Using embryonic leaflets of peanut variety Huayu 20 as explants, pingyangmycin (PYM) as the mutagen, and hydroxyproline (HYP) as the OP regulator, we developed 15 HYP-tolerant regenerated plants. For each regenerated plant, we selected offspring with oil content>55% (relative to 49.5% for Huayu 20). We developed and released three new peanut varieties with high yield and high oil content from the offspring of the HYP-tolerant regenerated plants. The three new varieties were named as Yuhua 4, Yuhua 9 and Yuhua 14 and their oil contents were 57.7, 61.1 and 59.3%, respectively. The results indicate that in vitro mutagenesis with PYM followed by directed screening with HYP is a useful approach for breeding peanut varieties with high oil contents.展开更多
Cormels pieces of four Fusarium susceptible Gladiolus cultivars (Friendship, Peter Pears, Victor Borge and Novalux) formed friable calli when cultured in vitro on Murashige and Skoog basal medium containing various ...Cormels pieces of four Fusarium susceptible Gladiolus cultivars (Friendship, Peter Pears, Victor Borge and Novalux) formed friable calli when cultured in vitro on Murashige and Skoog basal medium containing various concentrations of auxin and cytokinin. The friable calli established cell suspensions. Plantlet regeneration was obtained from the control callus, control cell suspension derived callus and in vitro selected Fusarium oxysporum Schlecht. resistant cell-lines of Friendship. The in vitro cormlets showed 85-95% germination after breaking dormancy of 8 weeks at 4℃. Cell suspensions of all four Gladiolus cultivars were found to be highly sensitive to fusaric acid. Gradual increase in fusaric acid concentrations to the cell-suspension cultures decreased cell growth considerably. One albino plant was found from the second generation of the in vitro selected cell line of Friendship. The albino plant was found to be highly susceptible to F. oxysporum. The cormlets of all in vitro selected cell lines of Friendship were inoculated with a conidial suspension of the F. oxysporum before planting and were also sprayed with the same spore suspension for further characterization when the height of plants was about 6cm. The four selected cell lines showed the same response whether or not they were inoculated with conidia of the F. oxysporum. Plantlets of all of the selected cell lines exhibited significant growth as compared with the control after application of conidia of the F. oxysporum.展开更多
基金ASPEE Agricultural Research and Development Foundation,Malad,Mumbai,India,for research fellowship during the PG course
文摘Radiation induced mutagenesis followed by in vitro selection was employed for salt tolerance in popular Indian sugarcane (Saccharum officinarum L.) cv. CoC-671. Embryogenic calli were gamma irradiated and exposed to different levels of NaCl (42.8, 85.6, 128.3, 171.1,213.9, 256.7, 299.5, or 342.2 mM). The relative growth rate (RGR) decreased progressively with increasing salt stress and was the least with a salt stress of 256.7 mM (0.25±0.009), almost 10 fold lesser than the control. The RGR was significantly lower in 85.6 mM and higher salt stressed calli than the control. The survival percent also decreased, with an increase in NaCl concentration. In case of 10 and 20 Gy irradiated calli, regeneration was observed up to 85.6 mM NaCl selection, medium, whereas, higher treatments (128.3 mM and beyond) exhibited browning initially. However, in the subsequent subcultures, regeneration was obtained in the case of 10 and 20 Gy irradiated calli on 128.3 and 171.1 mM NaCl selections. Higher dose of gamma irradiation (40 Gy) also showed regeneration, but only with 85.6 mM NaCI selection. The unirradiated calli regenerated the highest number of plantlets followed by 10 and 20 Gy irradiated calli on salt selection. A total of 147 plantlets were selected from different salt levels. The salt selected plants are being tested for their field performance.
基金This research was supported by Outstanding Younger Science Foundation of China(30225028)the National High Tech R&D Program,China(863 Program,2002AA241031)+1 种基金the National Science-Technology Prograin(2001BA511B)the Teaching and Research Award Program for Outstanding Young Teachers in Higher Education In stitutions of Ministry of Education,China.
文摘In vitro selection of drought-tolerant mutants in sweetpotato cv. Lizixiang was studied by using PEG6000 as selection stress. Embryogenic suspension cultures were cultured in MS medium containing 035% PEG6000 and 2 mg L 1 2,4-dichlorophenoxyacetic acid (2,4-D). The results indicated that 30% PEG6000 can be used for the optimal selection stress of drought-tolerance. Embryogenic suspension cultures irradiated with 80 Gy gamma-ray were cultured in MS medium containing 30% PEG6000 and 2 mg L 1 2,4-D and 20 drought-tolerant cell aggregates were obtained. These cell aggregates were transferred to solid MS me- dium supplemented with 2 mg L 1 2,4-D and formed embryogenic callus with somatic embryos. The embryogenic callus with somatic embryos was further transferred to MS medium supplemented with 1 mg L 1 abscisic acid (ABA), resulting in the germination of somatic embryos. In this study a total of 18 regenerated plants were obtained. The regenerated plants were transplanted in a greenhouse and 11 lines were formed. The analysis on drought treatment of seedlings, water retaining capacity of leaves and coefficient of drought-tolerance showed that 3 lines had significant drought-tolerance in comparison with the control plants.
文摘The present investigation was undertaken to study the effect of gamma irradiation (dose from 10 to 100 Gy) and in vitro selection with fungus filtrate as selecting agent (concentration from 20% to 100%) on the susceptibility of the common bean to Rhizoctonia solani. The best results were found with a dose of 20 Gy or a concentration of 20% of fungus filtrate applied separately. These conditions were used to evaluate the combined effect of both approaches in a second experiment. The combined effect of irradiation and then selection adversely affected growth (height and roots) and survival of the in vitro plants. It may not be necessary to combine the variation generated by irradiation with the selection technique. For future assays we propose the application of: 1) gamma radiation, thereby inducing not only mutants with pathogen resistance, but also with other agronomic traits of interest. Later in the subculture MV4 potential fungus-resistant mutants will be evaluated in the field;or 2) selection pressure using fungus filtrate during three subcultures, which may be sufficient to induce the variation necessary to obtain in vitro plants resistant to fungus.
基金financially supported by the National Basic Research Program of China(973 Program,No.2015CB932100)National Natural Science Foundation of China(No.81703451,81573371,81473164)。
文摘Preformed albumin corona of albumin-nonselective nanoparticles(NPs)is widely exploited to inhibit the unavoidable protein adsorption upon intravenous administration.However,very few studies have concerned the preformed albumin corona of albumin-selective NPs.Herein,we report a novel type of albumin-selective NPs by decorating 6-maleimidocaproyl polyethylene glycol stearate(SA)onto PLGA NPs(SP NPs)surface,taking albuminnonselective PLGA NPs as control.PLGA NPs and SP NPs were prepared by emulsion-solvent evaporation method and the resultant NPs were in spherical shape with an average diameter around 180 nm.The corresponding albumin-coating PLGA NPs(PLGA@BSA NPs)and albumin-coating SP NPs(SP@BSA NPs)were formulated by incubating SP NPs or PLGA NPs with bovine serum albumin solution,respectively.The impact of albumin corona on particle characteristics,stability,photothermal effect,cytotoxicity,cell uptake,spheroid penetration and pharmacokinetics was investigated.In line with previous findings of preformed albumin coating,PLGA@BSA NPs exhibited higher stability,cytotoxicity,cell internalization and spheroid penetration performances in vitro,and longer blood circulation time in vivo than those of albumin-nonselective PLGA NPs,but albumin-selective SP NPs is capable of achieving a comparable in vitro and in vivo performances with both SP@BSA NPs and PLGA@BSA NPs.Our results demonstrate that SA decorated albumin-selective NPs pave a versatile avenue for optimizing nanoparticulate delivery without preformed albumin corona.
文摘The selective toxicity of the mitochondria-specific cationic fluorescent dye rhodamine 123 (Rh-123) on He La cells in culture was studied. In this report, we demonstrate that with continuous exposure, Rh-123 markedly inhibited the growth of HeLa cells but had little effect on normal human kidney fibroblasts. With continuous exposure to Rh-123, the growth rate, colony forming ability, anl mitotic index of HeLa cells were decreased. The mechanism of toxicity of Rh-123 on HeLa cells was investigated by EM and enzyme cytochemistry stain. The mitochondria of carcinoma cells were the main targets for the inhibitory action of Rh-123, since they selectively accumulated the dye. At the dosage of Rh-123 which was toxic to HeLa cells, the structure and function of mitochondria were disrupted, as the mitochondria-related enzymes, i.e., ATPase, LDH and SDH were inhibited. The possible mechanism of the action of Rh-123 on HeLa cells is briefly discussed.
基金Financial support was provided by the National Natural Science Foundation of China(grant numbers 22177073,21977070,21907085,and U19A2011)the Natural Science Foundation of Shanghai,China(grant numbers 21ZR1442900 and 23ZR1437600)+2 种基金the Natural Science Foundation of Zhejiang Province,China(grant number LY22H300001)Shanghai Frontiers Science Center of Degeneration and Regeneration in Skeletal SystemShanghai Key Laboratory of Orthopedic Implants(grant number KFKT202207,China)for financial support.
文摘DNA-encoded chemical library(DEL)links the power of amplifiable genetics and the nonself-replicating chemical phenotypes,generating a diverse chemical world.In analogy with the biological world,the DEL world can evolve by using a chemical central dogma,wherein DNA replicates using the PCR reactions to amplify the genetic codes,DNA sequencing transcripts the genetic information,and DNA-compatible synthesis translates into chemical phenotypes.Importantly,DNA-compatible synthesis is the key to expanding the DEL chemical space.Besides,the evolution-driven selection system pushes the chemicals to evolve under the selective pressure,i.e.,desired selection strategies.In this perspective,we summarized recent advances in expanding DEL synthetic toolbox and panning strategies,which will shed light on the drug discovery harnessing in vitro evolution of chemicals via DEL.
基金This research was funded by the National Natural Science Foundation of China(31571705 and 31872875)the Key R&D Program of Shandong Province,China(2018GNC111014)the Open Oroject of the Key Lab of Oil Crop Biology and Genetic Breeding,Ministry of Agriculture and Rural Affairs of China(KF2018008).
文摘Peanut (Arachis hypogaea L.) is an important oil crop globally and high oil content is one of the major targets in peanut breeding programs. Previous studies indicated that the osmotic pressure (OP) of the leaves of peanut plants subjected to drought stress was negatively correlated with kernel oil content. Based on this knowledge, we established a practical and reliable method for creating new peanut varieties with high oil content using in vitro mutagenesis and directional OP-based selection. Using embryonic leaflets of peanut variety Huayu 20 as explants, pingyangmycin (PYM) as the mutagen, and hydroxyproline (HYP) as the OP regulator, we developed 15 HYP-tolerant regenerated plants. For each regenerated plant, we selected offspring with oil content>55% (relative to 49.5% for Huayu 20). We developed and released three new peanut varieties with high yield and high oil content from the offspring of the HYP-tolerant regenerated plants. The three new varieties were named as Yuhua 4, Yuhua 9 and Yuhua 14 and their oil contents were 57.7, 61.1 and 59.3%, respectively. The results indicate that in vitro mutagenesis with PYM followed by directed screening with HYP is a useful approach for breeding peanut varieties with high oil contents.
文摘Cormels pieces of four Fusarium susceptible Gladiolus cultivars (Friendship, Peter Pears, Victor Borge and Novalux) formed friable calli when cultured in vitro on Murashige and Skoog basal medium containing various concentrations of auxin and cytokinin. The friable calli established cell suspensions. Plantlet regeneration was obtained from the control callus, control cell suspension derived callus and in vitro selected Fusarium oxysporum Schlecht. resistant cell-lines of Friendship. The in vitro cormlets showed 85-95% germination after breaking dormancy of 8 weeks at 4℃. Cell suspensions of all four Gladiolus cultivars were found to be highly sensitive to fusaric acid. Gradual increase in fusaric acid concentrations to the cell-suspension cultures decreased cell growth considerably. One albino plant was found from the second generation of the in vitro selected cell line of Friendship. The albino plant was found to be highly susceptible to F. oxysporum. The cormlets of all in vitro selected cell lines of Friendship were inoculated with a conidial suspension of the F. oxysporum before planting and were also sprayed with the same spore suspension for further characterization when the height of plants was about 6cm. The four selected cell lines showed the same response whether or not they were inoculated with conidia of the F. oxysporum. Plantlets of all of the selected cell lines exhibited significant growth as compared with the control after application of conidia of the F. oxysporum.