Cell walls of facultative alkaliphilic Bacillus sp. C-125 consist of three polymers (peptidoglycan, teichuronopeptide and teichuronic acid). Protoplasts prepared from the strain with egg-white lysozyme regenerated cell walls at neutral pH, but not at pH above 8.5. The protoplasts were susceptible to lysis at alkaline pH. The protoplasts exposed to alkaline pH rapidly burst and lost ability to r...

Sucrose (Suc):Suc 1-fructosyltransferase (1-SST) is the key enzyme in plant fructan biosynthesis, since it catalyzes de novo fructan synthesis from Suc. We have cloned 1-SST from onion (Allium cepa) by screening a cDNA library using acid invertase from tulip (Tulipa gesneriana) as a probe. Expression assays in tobacco (Nicotiana plumbaginifolia) protoplasts showed the formation of 1-kestose fro...

Proximity-dependent biotin identification (BioID), which detects physiologically relevant proteins based on the proximity-dependent biotinylation process, has been successfully used in different organisms. In this report, we established the BioID system in rice protoplasts. Biotin ligase BirAG was obtained by removing a cryptic intron site in the BirA∗ gene when expressed in rice protoplasts. W...

We have developed a general method for electrically introducing DNA into plant cells. Gene transfer occurs when a high-voltage electric pulse is applied to a solution containing protoplasts and DNA. Carrot protoplasts were used as a model system to optimize gene-transfer efficiency, which was measured 24-48 hr after electroporation by the amount of chloramphenicol acetyltransferase activity res...

Neutral sugar, free amino acid, and anthocyanin levels and vacuole/extravacuole distribution were determined for Hippeastrum and Tulipa petal and Tulipa leaf protoplasts. Glucose and fructose, the predominant neutral monosaccharides observed, were primarily vacuolar in location. Glutamine, the predominant free amino acid found, was primarily extravacuolar. gamma-Methyleneglutamate was identifie...

A method is described for the isolation of protoplasts (Pisum sativum, Phaseolus vulgaris, Avena sativa, Arabidopsis thaliana) in preparation for ion flux studies using patch clamp electrophysiology. Protoplasts that have been exposed to hydrolytic, cell wall degrading, enzymes for as little as 5 minutes form gigaseals (seal resistance higher than 10 giga Ohm) with the patch pipette with succes...

An efficient system has been developed for the reproducible regeneration of plants from cell suspension-derived protoplasts of the two commercial Iranian Japonica rice cultivars Tarom and Khazer. Friable embryogenic calli were used for cell suspension initiation and protoplasts were isolated from both varieties. When embedded in agarose, protoplasts failed to divide. However, sustained division...

Responses to auxin of Lotus corniculatus root tips or protoplasts transformed by Agrobacterium rhizogenes strains 15834 and 8196 were compared to those of their normal counterparts. Three different types of experiments were performed, involving long-term, medium-term, or short-term responses to a synthetic auxin, 1-naphthaleneacetic acid. Root tip elongation, proton excretion by root tips, and ...

in this study techniques for laboratory identification of dermatophyte fungi through protoplast hybridization were established. firstly, auxotrophic mutants of different species of microsporum and trichophyton were induced and identified. secondly, protoplasts from these mutants were isolated by digestion of their mycelium with novozyme 234 using caci, (o.4m) as an osmotic stabilizer and glycin...

Kinsky, Stephen C. (Washington University, St. Louis, Mo.). Effect of polyene antibiotics on protoplasts of Neurospora crassa. J. Bacteriol. 83:351-358. 1962.-The polyene antibiotics nystatin, amphotericin B, and filipin induced shrinkage of Neurospora crassa protoplasts stabilized in sucrose. At low concentrations of filipin (5 mug per ml), or higher concentrations of nystatin and amphotericin...