Topic
Protoplast
About: Protoplast is a research topic. Over the lifetime, 5474 publications have been published within this topic receiving 122468 citations.
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TL;DR: Since rooting was almost impossible in these thidiazuron-induced shoots, shoots were grafted onto young pea seedlings and regenerated to fertile plants.
Abstract: Protoplasts isolated from lateral shoot buds of cotyledon-free pea embryo axes were regenerated to callus. Protoplast derived calluses with a diameter of about 1cm were transferred to shoot induction media, containing different concentrations (1–50µM) of thidiazuron. Shoot formation was observed after 16 weeks up to 12% efficiency. Thidiazuron (10µM) was the most effective concentration in all experiments. Shoot buds elongated in medium supplemented with N-isopentenyl adenine and indole-3-butyric acid. Since rooting was almost impossible in these thidiazuron-induced shoots, shoots were grafted onto young pea seedlings and regenerated to fertile plants.
53 citations
TL;DR: Ghosts were prepared by attaching freshly isolated protoplasts to glass coverslips or formvar/carbon-coated grids with poly-L-lysine and then bursting them either osmotically or by detergent treatment in the presence of a microtubule stabilizing buffer, and proved very useful for measuring the density and length of cortical microtubules.
Abstract: The arrangement of cortical microtubules in tobacco protoplasts is described using the following techniques: 1. Transmission electron microscopy (TEM) of thin sections of whole protoplasts, 2. TEM of negatively stained protoplast ghosts, and 3. Indirect immunofluorescence microscopy of protoplast ghosts. Ghosts were prepared by attaching freshly isolated protoplasts to glass coverslips or formvar/carbon-coated grids with poly-L-lysine and then bursting them either osmotically or by detergent treatment in the presence of a microtubule stabilizing buffer. Osmotic bursting of protoplasts yielded large pieces of plasma membrane with attached microtubules. These preparations proved very useful for measuring the density and length of cortical microtubules. Detergent treatment dissolved the plasma membrane and altered the distribution of cortical microtubules.
53 citations
TL;DR: A simplified protoplast regeneration system for Vigna aconitifolia was developed and the plant cultivar used was an important factor in determining transformation frequencies since one of the cultivars had an 85 fold higher transformation rate than the other.
Abstract: A simplified protoplast regeneration system for Vigna aconitifolia was developed. A plating efficiency of 60% was obtained using mesophyll protoplasts from 10-day-old seedlings. By co-cultivation of protoplasts with Agrobacterium tumefaciens containing the Ti plasmid derivative pGV 3850∶1103 neo kanamycin-resistant colonies were obtained; 23% of the transformed lines showed expression of the nonselected co-transferred nopaline synthase gene. Transformation was confirmed by Southern blot analysis using a nonradioactive detection system. The plant cultivar used was an important factor in determining transformation frequencies since one of the cultivars had an 85 fold higher transformation rate than the other.
53 citations
TL;DR: The first protocol to isolate large numbers of viable protoplasts from both leaves and roots of two switchgrass genotypes is reported, indicating an efficient protoplast isolation and transient assay system that can be used to facilitate gene expression studies in switchgrass.
Abstract: Transient assay systems using protoplasts have been utilized in several plant species and are a powerful tool for rapid functional gene analysis and biochemical manipulations. A protoplast system has not been used in switchgrass (Panicum virgatum L.), even though it is a bioenergy crop that has received considerable attention. Here we report the first protocol to isolate large numbers of viable protoplasts from both leaves and roots of two switchgrass genotypes. Furthermore, we demonstrate transient expression of PEG-mediated DNA uptake in the isolated protoplasts by measuring the activity of β-glucuronidase (GUS) reporter gene driven by either the Cauliflower mosaic virus (CaMV) 35S promoter or the maize ubiquitin 1 promoter. Protoplast transformation with either the 35S or the ubiquitin promoter resulted in an increase in GUS activity compared to the untransformed controls; however, the extent of GUS activity was considerably higher for the ubiquitin promoter than for the 35S promoter. Collectively, our results indicate an efficient protoplast isolation and transient assay system that can be used to facilitate gene expression studies in switchgrass.
53 citations
Patent•
09 Oct 1996TL;DR: In this paper, a method of producing a cereal plant cell or protoplast useful for regeneration of a water stress or salt stress tolerant cereal plant by transforming the rice plant cell with a nucleic acid encoding a late embryogenesis abundant protein was presented.
Abstract: The present invention is directed to a method of producing a cereal plant cell or protoplast useful for regeneration of a water stress or salt stress tolerant cereal plant by transforming the cereal plant cell or protoplast with a nucleic acid encoding a late embryogenesis abundant protein. A transgenic cereal plant or cereal plant cell or protoplast transformed with a nucleic acid encoding a late embryogenesis abundant protein is also provided. An LEA protein gene, HVA1, from barley (Hordeum vulgare L.) was transformed into rice (Oryza sativa L.) plants. The resulting transgenic rice plants accumulate the HVA1 protein in both leaves and roots. Transgenic rice plants showed significantly increased tolerance to water stress (drought) and salt stress.
53 citations