Non-embryogenic calli （NEC） was inevitably and heavily produced when grape embryogenic calli （EC） was induced from explants or during the subculture of EC.A stable and highly efficient NEC transformation platform ...Non-embryogenic calli （NEC） was inevitably and heavily produced when grape embryogenic calli （EC） was induced from explants or during the subculture of EC.A stable and highly efficient NEC transformation platform is required to further sort out and verify key genes which determine/switch the identity of NEC and EC.In this research,a vector pA5 containing a chitinase signal sequence fused to gfp （green fluorescent protein） and an HDEL motive was used to target and immobilize into Agrobacterium strain EHA105 to establish a transformation platform for Vitis vinifera L.cv.Chardonnay NEC.It was determined that NEC 10 d after subculture was the best target tissue;30 min for inoculation followed by 3 d co-cultivation with the addition of 200 μmol L-1 acetosyringone （AS） was optimized as protocol.The use of bacterial densities as 1.0 at OD600 did not result in serious tissue hypersensitive reaction and it had higher efficiency.Kanamycin at 200 mg L-1 was picked for positive expression selection.The stable transformation of NEC was proved by reverse transcription-polymerase chain reaction techniques （RT-PCR） and fluorescent microscopy after three sub-cultures of the selected cell line.Highly efficient genetic transformation protocol of grape NEC was achieved and some of the optimized parameters were different from that reported for EC.This transformation platform could facilitate the verification of candidate somatic embryogenesis （SE） decisive genes,and the successfully transformed NEC with certain genes can also be used as bioreactors for the production of functional products,as NEC not only proliferates fast,but also keeps in a rather stable condition.展开更多
A description of a successful direct somatic embryogenesis induction from immature zygotic embryos of a camphor tree （Cinnamomum camphora L.） is presented. After a subculture of 2-3 years, embryogenic calli could be...A description of a successful direct somatic embryogenesis induction from immature zygotic embryos of a camphor tree （Cinnamomum camphora L.） is presented. After a subculture of 2-3 years, embryogenic calli could be derived from primary somatic embryos. Immature zygotic embryos were cultured on a Murashige and Skoog （MS） basal medium supplemented with a range of combinations of cytokinins （BA） and auxins （2,4-D or NAA） for somatic embryo induction. Primary somatic embryos could be induced directly in almost all PGR combinations. A positive effect of 2,4-D on somatic embryogenesis from immature zygotic embryos of camphor tree was obtained. BA at appropriate concentrations （〈 5 mg-L-1） had an effect similar to 2,4-D, whereas high concentrations （〉 5 mg·L^-1） of BA had the effect of restraining somatic embryo induction. NAA had a less positive effect on somatic embryogenesis than 2,4-D.展开更多
Anliucheng (Citrus sinensis Osbeck), a very seedy and widely spread acidless sweet orange cul-tivar in south of China, was transformed by the strain of Agrobacterium Tumefaciens EHA105 carrying pTA29-barnase gene, whi...Anliucheng (Citrus sinensis Osbeck), a very seedy and widely spread acidless sweet orange cul-tivar in south of China, was transformed by the strain of Agrobacterium Tumefaciens EHA105 carrying pTA29-barnase gene, which will induce pollen sterility in transgenic plants. The embryogenic calli of Anliucheng were co-cultivated with Agrobacterium tumefaciens for 3 days, and then transferred to selective medium containing 50 mg I/1 basta (a kind of herbicide) for 5 weeks. The resistant calli were recovered and regenerated 118 embryoids. A total of 13 entire plants were obtained after micro-grafted on trifoliate orange. These regenerated plants were verified by PCR amplification and confirmed by PCR-Southern blotting analysis.展开更多
In this study, an embryogenic callus induction and proliferation system for Hippeastrum vittatum was established, with the tender bulbs as explants. And then the sensitivity of the explants and calli to kanamycin and ...In this study, an embryogenic callus induction and proliferation system for Hippeastrum vittatum was established, with the tender bulbs as explants. And then the sensitivity of the explants and calli to kanamycin and hygromycin was evaluated. The results suggested that the embryogenic calli were induced from tender bulbs and cultured in Murashige and Skoog （MS） medium supplemented with 0.5 mg/L N-phenyl-N＇-1,2,3-thiadiazol-5-ylurea （TDZ）, 1.0 mg/L 2,4-diehloro- phenoxyacetic acid （2,4-D）, and 30 g/L sucrose （ pH5. 8） in the dark at 25 ±1℃. Further study of the influence of kanamyein and hygromycin on callus induction and multiplication showed that, the lethal doses of kanamycin and hygromycin to bulb explants were 100 and 30 mg/L, respectively. All explants of H. vittatum died on the medium supplemented with 100 mg/L kanamycin or 30 mg/L hygromycin at the induction stage, and callus proliferation was completely inhibited by 100 mg/L kanamycin or 25 mg,/L hygromyein, and all the calli died at last. These results will provide important reference for further studies of trausgenic H. vittaturn.展开更多
The plant regeneration frequencies of ealli fromplant tissue and cell culture,especially that of thecalli from rice tissue culture and rice anther cul-ture,and that of the foreign-DNA-transfor-mation-derived rice call...The plant regeneration frequencies of ealli fromplant tissue and cell culture,especially that of thecalli from rice tissue culture and rice anther cul-ture,and that of the foreign-DNA-transfor-mation-derived rice calli is very low(usually 10-15%).It is therefor very important to improve theplant regeneration frequency of rice calli.A1-展开更多
Wild rice is an important resource of usefulgenes to rice breeders. However, low regener-ation frequency is an obstacle to use the valu-able genes. We used desiccation to improve theregeneration frequency and studied ...Wild rice is an important resource of usefulgenes to rice breeders. However, low regener-ation frequency is an obstacle to use the valu-able genes. We used desiccation to improve theregeneration frequency and studied the bio-chemical changes of calli of wild rice after des-iccation.Materials used in this experiment werewild species O. rufipogon, O. meyeriana, O.alta, and O. brachyantha. Young panicles(0.1-0. 5 cm in length of the inflorescence)展开更多
Enrichment of copper to the culture mediumcould enhance the plant regeneration from cal-lus of indica rice variety Qiugui’ai 11. Westudied the effect of copper on plant regenera-tion of other rice varieties.Calli of ...Enrichment of copper to the culture mediumcould enhance the plant regeneration from cal-lus of indica rice variety Qiugui’ai 11. Westudied the effect of copper on plant regenera-tion of other rice varieties.Calli of 14 indica and 2 japonica varietieswere induced from disinfected mature embryoson an agar-gelled medium containing Nbasal展开更多
针对社区生活圈老年人步行友好出行环境理论缺乏与实践不足的问题,从社区建成环境对老年人活力出行影响的空间异质性问题展开了研究.结合老年人出行特征选取了人口密度、土地利用多样性、公交站点密度、绿视率等8个指标刻画建成环境,考...针对社区生活圈老年人步行友好出行环境理论缺乏与实践不足的问题,从社区建成环境对老年人活力出行影响的空间异质性问题展开了研究.结合老年人出行特征选取了人口密度、土地利用多样性、公交站点密度、绿视率等8个指标刻画建成环境,考虑路网非直线系数对社区生活圈尺度差异化界定,利用兴趣点(Point of Interest,POI)数据、道路网络、街景图像数据等地理空间数据测度建成环境.提取每日步行时间作为活力出行的测度,以重庆市居民出行行为调查数据作为实证研究数据来源,构建了考虑自变量尺度变异的多尺度地理加权回归模型(Multi-scale Geographic Weighted Regression,MGWR).研究结果表明:对比最小二乘回归(Ordinary Least Squares,OLS)模型与传统的地理加权回归(Geographically Weighted Regression,GWR)模型,纳入变量尺度异质性的MGWR模型降低了残差的自相关性,且调整后R2最高,相比于GWR模型与OLS模型分别提高了1.5倍与3.1倍;从标准化系数来看,社区建成环境指标中公交服务水平对老年人步行时间影响最大;社区建成环境指标中街道绿视率接近全局尺度,空间异质性较弱,其他建成环境变量均具有较强的空间异质性,需要进行差异化的空间设计,其中人口密度对老年人步行时间在不同区域呈现相反的影响,土地利用混合度、公交服务水平、蔬菜市场密度、公园广场密度与街道绿视率对老年人活力出行的影响呈现正效应.展开更多
基金supported by the National Natural Science Foundation of China (30471212,30500347)the Earmarked Fund for Modern Agro-Industry Technology Research System,Ministry of Agriculture,China (NYCYTX-3-CY-04)
文摘Non-embryogenic calli （NEC） was inevitably and heavily produced when grape embryogenic calli （EC） was induced from explants or during the subculture of EC.A stable and highly efficient NEC transformation platform is required to further sort out and verify key genes which determine/switch the identity of NEC and EC.In this research,a vector pA5 containing a chitinase signal sequence fused to gfp （green fluorescent protein） and an HDEL motive was used to target and immobilize into Agrobacterium strain EHA105 to establish a transformation platform for Vitis vinifera L.cv.Chardonnay NEC.It was determined that NEC 10 d after subculture was the best target tissue;30 min for inoculation followed by 3 d co-cultivation with the addition of 200 μmol L-1 acetosyringone （AS） was optimized as protocol.The use of bacterial densities as 1.0 at OD600 did not result in serious tissue hypersensitive reaction and it had higher efficiency.Kanamycin at 200 mg L-1 was picked for positive expression selection.The stable transformation of NEC was proved by reverse transcription-polymerase chain reaction techniques （RT-PCR） and fluorescent microscopy after three sub-cultures of the selected cell line.Highly efficient genetic transformation protocol of grape NEC was achieved and some of the optimized parameters were different from that reported for EC.This transformation platform could facilitate the verification of candidate somatic embryogenesis （SE） decisive genes,and the successfully transformed NEC with certain genes can also be used as bioreactors for the production of functional products,as NEC not only proliferates fast,but also keeps in a rather stable condition.
基金supported by the Natural Science Foundation of Henan Province of China(0611033300).
文摘A description of a successful direct somatic embryogenesis induction from immature zygotic embryos of a camphor tree （Cinnamomum camphora L.） is presented. After a subculture of 2-3 years, embryogenic calli could be derived from primary somatic embryos. Immature zygotic embryos were cultured on a Murashige and Skoog （MS） basal medium supplemented with a range of combinations of cytokinins （BA） and auxins （2,4-D or NAA） for somatic embryo induction. Primary somatic embryos could be induced directly in almost all PGR combinations. A positive effect of 2,4-D on somatic embryogenesis from immature zygotic embryos of camphor tree was obtained. BA at appropriate concentrations （〈 5 mg-L-1） had an effect similar to 2,4-D, whereas high concentrations （〉 5 mg·L^-1） of BA had the effect of restraining somatic embryo induction. NAA had a less positive effect on somatic embryogenesis than 2,4-D.
文摘Anliucheng (Citrus sinensis Osbeck), a very seedy and widely spread acidless sweet orange cul-tivar in south of China, was transformed by the strain of Agrobacterium Tumefaciens EHA105 carrying pTA29-barnase gene, which will induce pollen sterility in transgenic plants. The embryogenic calli of Anliucheng were co-cultivated with Agrobacterium tumefaciens for 3 days, and then transferred to selective medium containing 50 mg I/1 basta (a kind of herbicide) for 5 weeks. The resistant calli were recovered and regenerated 118 embryoids. A total of 13 entire plants were obtained after micro-grafted on trifoliate orange. These regenerated plants were verified by PCR amplification and confirmed by PCR-Southern blotting analysis.
基金Supported by Science and Technology Program of Guangzhou City(2016201604030076)Science and Technology Program of Guangdong Province(2014A020208062)+2 种基金Cooperation Project of Industry,Education and Academy of Dongguan City(2015509105105)the Fund for Demonstration and Extension of Introduced Intellectual Achievements in Guangdong "Forcing Cultivation and Flowering Regulation Technology Introduction,Demonstration and Promotion in Hippeastrum vittatum" and "The Characteristics of Taiwan Varieties and Cultivation Technology Introduction and Promotion in Hippeastrum vittatum"Open Project of Key Laboratory of Urban Agriculture in South China,Ministry of Agriculture,China
文摘In this study, an embryogenic callus induction and proliferation system for Hippeastrum vittatum was established, with the tender bulbs as explants. And then the sensitivity of the explants and calli to kanamycin and hygromycin was evaluated. The results suggested that the embryogenic calli were induced from tender bulbs and cultured in Murashige and Skoog （MS） medium supplemented with 0.5 mg/L N-phenyl-N＇-1,2,3-thiadiazol-5-ylurea （TDZ）, 1.0 mg/L 2,4-diehloro- phenoxyacetic acid （2,4-D）, and 30 g/L sucrose （ pH5. 8） in the dark at 25 ±1℃. Further study of the influence of kanamyein and hygromycin on callus induction and multiplication showed that, the lethal doses of kanamycin and hygromycin to bulb explants were 100 and 30 mg/L, respectively. All explants of H. vittatum died on the medium supplemented with 100 mg/L kanamycin or 30 mg/L hygromycin at the induction stage, and callus proliferation was completely inhibited by 100 mg/L kanamycin or 25 mg,/L hygromyein, and all the calli died at last. These results will provide important reference for further studies of trausgenic H. vittaturn.
文摘The plant regeneration frequencies of ealli fromplant tissue and cell culture,especially that of thecalli from rice tissue culture and rice anther cul-ture,and that of the foreign-DNA-transfor-mation-derived rice calli is very low(usually 10-15%).It is therefor very important to improve theplant regeneration frequency of rice calli.A1-
文摘Wild rice is an important resource of usefulgenes to rice breeders. However, low regener-ation frequency is an obstacle to use the valu-able genes. We used desiccation to improve theregeneration frequency and studied the bio-chemical changes of calli of wild rice after des-iccation.Materials used in this experiment werewild species O. rufipogon, O. meyeriana, O.alta, and O. brachyantha. Young panicles(0.1-0. 5 cm in length of the inflorescence)
文摘Enrichment of copper to the culture mediumcould enhance the plant regeneration from cal-lus of indica rice variety Qiugui’ai 11. Westudied the effect of copper on plant regenera-tion of other rice varieties.Calli of 14 indica and 2 japonica varietieswere induced from disinfected mature embryoson an agar-gelled medium containing Nbasal
文摘针对社区生活圈老年人步行友好出行环境理论缺乏与实践不足的问题,从社区建成环境对老年人活力出行影响的空间异质性问题展开了研究.结合老年人出行特征选取了人口密度、土地利用多样性、公交站点密度、绿视率等8个指标刻画建成环境,考虑路网非直线系数对社区生活圈尺度差异化界定,利用兴趣点(Point of Interest,POI)数据、道路网络、街景图像数据等地理空间数据测度建成环境.提取每日步行时间作为活力出行的测度,以重庆市居民出行行为调查数据作为实证研究数据来源,构建了考虑自变量尺度变异的多尺度地理加权回归模型(Multi-scale Geographic Weighted Regression,MGWR).研究结果表明:对比最小二乘回归(Ordinary Least Squares,OLS)模型与传统的地理加权回归(Geographically Weighted Regression,GWR)模型,纳入变量尺度异质性的MGWR模型降低了残差的自相关性,且调整后R2最高,相比于GWR模型与OLS模型分别提高了1.5倍与3.1倍;从标准化系数来看,社区建成环境指标中公交服务水平对老年人步行时间影响最大;社区建成环境指标中街道绿视率接近全局尺度,空间异质性较弱,其他建成环境变量均具有较强的空间异质性,需要进行差异化的空间设计,其中人口密度对老年人步行时间在不同区域呈现相反的影响,土地利用混合度、公交服务水平、蔬菜市场密度、公园广场密度与街道绿视率对老年人活力出行的影响呈现正效应.
