A simple procedure for the isolation of streptomycin resistant plants in Solanaceae.

A system has been developed for rapid selection of streptomycin resistant mutants, as adventitious shoots arising from explants of several Solanaceous species. Efficient mutagenesis was achieved by incubating shoot culturederived leaf strips with 1 or 5 mM nitroso-methylurea, for 90 or 120 min. In Nicotiana tabacum and Lycopersicon peruvianum these treatments resulted in white or variegated adventitious shoots from up to 3.5% of explants placed on medium promoting shoot regeneration. Chlorophyll deficiencies were only observed very rarely in Solanum nigrum. Streptomycin resistant shoots were obtained from leaf explants placed on medium containing 500 mg 1- 1 streptomycin sulphate, under which conditions ex plants are bleached and adventitious shoot development suppressed. Green adventitious shoots appeared at a frequency dependent both on the mutagenic treatment and on the species. The best response was with S. nigrwn where > 70% of the ex plants produced streptomycin resistant shoots, most of which retained their resistance on subsequent testing. Maternal inheritance of streptomycin resistance has been confirmed for several N. tabacum and S. nigrum mutants, and there is also evidence for paternal transmission in the latter species. The procedure h3.s been successfully extended to other species, including N. s_vlvestris and N. plumbagin (fOlia, and also to obtain spectinomycin resistant mutants

A simple procedure for the isolation of streptomycin resistant plants in Solanaceae.

A system has been developed for rapid selection of streptomycin resistant mutants, as adventitious shoots arising from explants of several Solanaceous species. Efficient mutagenesis was achieved by incubating shoot culturederived leaf strips with 1 or 5 mM nitroso-methylurea, for 90 or 120 min. In Nicotiana tabacum and Lycopersicon peruvianum these treatments resulted in white or variegated adventitious shoots from up to 3.5% of explants placed on medium promoting shoot regeneration. Chlorophyll deficiencies were only observed very rarely in Solanum nigrum. Streptomycin resistant shoots were obtained from leaf explants placed on medium containing 500 mg 1- 1 streptomycin sulphate, under which conditions ex plants are bleached and adventitious shoot development suppressed. Green adventitious shoots appeared at a frequency dependent both on the mutagenic treatment and on the species. The best response was with S. nigrwn where > 70% of the ex plants produced streptomycin resistant shoots, most of which retained their resistance on subsequent testing. Maternal inheritance of streptomycin resistance has been confirmed for several N. tabacum and S. nigrum mutants, and there is also evidence for paternal transmission in the latter species. The procedure h3.s been successfully extended to other species, including N. s_vlvestris and N. plumbagin (fOlia, and also to obtain spectinomycin resistant mutants

Use of cell morphology to evaluate the effect of a peroxidase gene on cell death induction thresholds in tobacco

Tobacco suspension cultures were subjected to a range of heat stresses and used to compare morphological aspects of programmed cell death (PCD) and necrosis. Cells undergoing PCD were found to display characteristic death morphology, caused by cytoplasmic retraction of the protoplast, and to have cleaved DNA. We evaluated if the morphological characteristics of PCD could be used to monitor changes in cell death induction thresholds in transgenic cell cultures with high levels of peroxidase activity. Again, using a heat shock assay, we show that tobacco cell cultures with elevated levels of peroxidase have higher cell death induction threshold levels than wild type tobacco cell cultures. Thus, assessing PCD associated morphological changes can report on the effect of altering peroxidase genes on cell death activation in tobacco. This study demonstrates that PCD morphology could routinely be used to monitor the effects of introduced genes on programmed cell death induction thresholds in plants

High-throughput screening of monoclonal antibodies against plant cell wall glycans by hierarchical clustering of their carbohydrate microarray binding profiles

Antibody-producing hybridoma cell lines were created following immunisation with a crude extract of cell wall polymers from the plant Arabidopsis thaliana. In order to rapidly screen the specificities of individual monoclonal antibodies (mAbs), their binding to microarrays containing 50 cell wall glycans immobilized on nitrocellulose was assessed. Hierarchical clustering of microarray binding profiles from newly produced mAbs, together with the profiles for mAbs with previously defined specificities allowed the rapid assignments of mAb binding to antigen classes. mAb specificities were further investigated using subsequent immunochemical and biochemical analyses and two novel mAbs are described in detail. mAb LM13 binds to an arabinanase-sensitive pectic epitope and mAb LM14, binds to an epitope occurring on arabinogalactan-proteins. Both mAbs display novel patterns of recognition of cell walls in plant materials

US Cosmic Visions: New Ideas in Dark Matter 2017: Community Report

This white paper summarizes the workshop "U.S. Cosmic Visions: New Ideas in Dark Matter" held at University of Maryland on March 23-25, 2017

A simple procedure for the isolation of streptomycin resistant plants in Solanaceae.

A system has been developed for rapid selection of streptomycin resistant mutants, as adventitious shoots arising from explants of several Solanaceous species. Efficient mutagenesis was achieved by incubating shoot culturederived leaf strips with 1 or 5 mM nitroso-methylurea, for 90 or 120 min. In Nicotiana tabacum and Lycopersicon peruvianum these treatments resulted in white or variegated adventitious shoots from up to 3.5% of explants placed on medium promoting shoot regeneration. Chlorophyll deficiencies were only observed very rarely in Solanum nigrum. Streptomycin resistant shoots were obtained from leaf explants placed on medium containing 500 mg 1- 1 streptomycin sulphate, under which conditions ex plants are bleached and adventitious shoot development suppressed. Green adventitious shoots appeared at a frequency dependent both on the mutagenic treatment and on the species. The best response was with S. nigrwn where > 70% of the ex plants produced streptomycin resistant shoots, most of which retained their resistance on subsequent testing. Maternal inheritance of streptomycin resistance has been confirmed for several N. tabacum and S. nigrum mutants, and there is also evidence for paternal transmission in the latter species. The procedure h3.s been successfully extended to other species, including N. s_vlvestris and N. plumbagin (fOlia, and also to obtain spectinomycin resistant mutants

A simple procedure for the isolation of streptomycin resistant plants in Solanaceae.

A system has been developed for rapid selection of streptomycin resistant mutants, as adventitious shoots arising from explants of several Solanaceous species. Efficient mutagenesis was achieved by incubating shoot culturederived leaf strips with 1 or 5 mM nitroso-methylurea, for 90 or 120 min. In Nicotiana tabacum and Lycopersicon peruvianum these treatments resulted in white or variegated adventitious shoots from up to 3.5% of explants placed on medium promoting shoot regeneration. Chlorophyll deficiencies were only observed very rarely in Solanum nigrum. Streptomycin resistant shoots were obtained from leaf explants placed on medium containing 500 mg 1- 1 streptomycin sulphate, under which conditions ex plants are bleached and adventitious shoot development suppressed. Green adventitious shoots appeared at a frequency dependent both on the mutagenic treatment and on the species. The best response was with S. nigrwn where > 70% of the ex plants produced streptomycin resistant shoots, most of which retained their resistance on subsequent testing. Maternal inheritance of streptomycin resistance has been confirmed for several N. tabacum and S. nigrum mutants, and there is also evidence for paternal transmission in the latter species. The procedure h3.s been successfully extended to other species, including N. s_vlvestris and N. plumbagin (fOlia, and also to obtain spectinomycin resistant mutants

Use of cell morphology to evaluate the effect of a peroxidase gene on cell death induction thresholds in tobacco

Tobacco suspension cultures were subjected to a range of heat stresses and used to compare morphological aspects of programmed cell death (PCD) and necrosis. Cells undergoing PCD were found to display characteristic death morphology, caused by cytoplasmic retraction of the protoplast, and to have cleaved DNA. We evaluated if the morphological characteristics of PCD could be used to monitor changes in cell death induction thresholds in transgenic cell cultures with high levels of peroxidase activity. Again, using a heat shock assay, we show that tobacco cell cultures with elevated levels of peroxidase have higher cell death induction threshold levels than wild type tobacco cell cultures. Thus, assessing PCD associated morphological changes can report on the effect of altering peroxidase genes on cell death activation in tobacco. This study demonstrates that PCD morphology could routinely be used to monitor the effects of introduced genes on programmed cell death induction thresholds in plants

Use of cell morphology to evaluate the effect of a peroxidase gene on cell death induction thresholds in tobacco

Tobacco suspension cultures were subjected to a range of heat stresses and used to compare morphological aspects of programmed cell death (PCD) and necrosis. Cells undergoing PCD were found to display characteristic death morphology, caused by cytoplasmic retraction of the protoplast, and to have cleaved DNA. We evaluated if the morphological characteristics of PCD could be used to monitor changes in cell death induction thresholds in transgenic cell cultures with high levels of peroxidase activity. Again, using a heat shock assay, we show that tobacco cell cultures with elevated levels of peroxidase have higher cell death induction threshold levels than wild type tobacco cell cultures. Thus, assessing PCD associated morphological changes can report on the effect of altering peroxidase genes on cell death activation in tobacco. This study demonstrates that PCD morphology could routinely be used to monitor the effects of introduced genes on programmed cell death induction thresholds in plants

Use of cell morphology to evaluate the effect of a peroxidase gene on cell death induction thresholds in tobacco

Tobacco suspension cultures were subjected to a range of heat stresses and used to compare morphological aspects of programmed cell death (PCD) and necrosis. Cells undergoing PCD were found to display characteristic death morphology, caused by cytoplasmic retraction of the protoplast, and to have cleaved DNA. We evaluated if the morphological characteristics of PCD could be used to monitor changes in cell death induction thresholds in transgenic cell cultures with high levels of peroxidase activity. Again, using a heat shock assay, we show that tobacco cell cultures with elevated levels of peroxidase have higher cell death induction threshold levels than wild type tobacco cell cultures. Thus, assessing PCD associated morphological changes can report on the effect of altering peroxidase genes on cell death activation in tobacco. This study demonstrates that PCD morphology could routinely be used to monitor the effects of introduced genes on programmed cell death induction thresholds in plants