Protoplast isolation

Refinement of techniques involved in protoplast isolation has enhaneed the success of isolation of vaeuoles and other intracellular structures... 

Rosen, A. Tal, M. (1981). Salt tolerance in the wild relatives of the cultivated tomato responses of naked protoplasts isolated from leaves of Lycopersicon esculentum and L. peruvianum to NaCl and proline. Zeitschrift fur Pflanzenphy-siologie, 102, 91-4. 

Characteristics of parental strain, mutants and conditions for protoplast isolation. 

The cultivated potato and many of its relatives are amenable to eell and tissue culture procedures, ineluding protoplast isolation and fusion. Somatic cell fusions have frequently been used to eombine the genomes of Solarium speeies that are sexually ineompatible because of pollen-stylar interactions or mismatched EBN numbers. Somatic fusion circumvents sexual reproduetion and results in novel eombinations of not only nuclear genomes, but also cytoplasmic genomes (Trabelsi et al., 2005 Bidani et al., 2007 Lovene et al., 2007). However, recalcitrant genotypes... 

Urao et al. (1994) [87] cloned two closely related Ca dependent protein kinases from Arabidopsis and showed that they are ABA inducible. Overexpression of these kinases in protoplasts isolated from maize leaves leads to induction of an ABA responsive promoter [75]. Coexpression of ABIl inhibits the induction. It is difficult to make this fit with the role of ABIl as a mediator of ABA induced gene expression. The mechanism of action of ABIl and the dominant effects of the abil mutation have not yet been clarified. It is possible that overexpression of ABIl has the same effect as the abil mutant protein. 

Several studies have examined the abilities of plant tissue cultures to efficiently produce Taxol. Suspension cell cnltnres of T. chinensis are capable of prodncing Taxol. Fnrthermore, protoplasts isolated from cultnred cells of T. cmpidita, either in static cnlture or immobilized in agarose gel, effectively prodnce Taxol. Thus, several systems exist for the potential economical production of Taxol. 

Aoyagi, H., DiCosmo, F., and Tanaka, H., Efficient paclitaxel production using protoplasts isolated from cultured cells of Taxus cuspidata, Planta Medica, 68,420,2002. 

After initially washing the fixed material, protoplast isolation is carried out in 1.5-mL microcentrifuge tubes at room temperature. If plenty of material is available, 10- to 15-mL polypropylene or glass centrifuge tubes can be used. If no metaphases are found, a squash preparation is required to check the material. 

Attree, S.M Bekkaoui, F. Dunstan, D.L. Fowke, L.C. (1987). Regeneration of somatic embryos from protoplasts isolated from an embryogenic suspension culture of white spruce (Pkea glauca). Plant Cell Reports, Vol. 6, pp. 480-483. 

Cocking EC (1972) Plant cell protoplasts - isolation and development. Annu Rev Plant Physiol 23 29-50... 

HCA may also be covalenty attached to aliphatic components of cutin and suberin. The amount of covalently bound phenolic compounds m-, /7-coumaric acids and flavonoids) in tomato fruit cutin increased during fruit development and accounted for as much as 6% of cutin membranes. Protoplasts isolated from immature tomato fruit secrete a wall that has been shown to contain suberin, in which phenolic compounds formed 25% of total monomers [3]. 

Behera BC, Sonone A, Makhija U (2009b) Protoplast isolation from cultured lichen Usnea ghattensis, their fusion with protoplasts of Aspergillus nidulans, fusant regeneration and production of usnic acid. Folia Microbiol 54 415-418 Behera BC, Mahadik N, Morey M (2012) Antioxidative and cardiovascular-protective activities of metabolite usnic acid and psoromic acid produced by lichen species Usnea complanata under submerged fermentation. Pharm Biol 50 968-979 Bezivin C, Tomasi S, Rouaud I et al (2004) Cytotoxic activity of compounds from the lichen Cladonia convoluta. Planta Med 70 874-877... 

Kinoshita Y, Hayase S, Higuchi M, Yamamoto Y, Ahmadjian V, Yoshimura I, Yamada Y (1991) Improvement of Protoplast Isolation from Lichen Mycobionts. Agric Biol Chem 55 1891... 

J.D. Mlkkelsen, Synthesis of lipids by epidermal and mesophyll protoplasts Isolated from barley leaf sheaths, in "Biogenesis and Function of Plant Lipids," P. Mazllak, P. Benvenlste, C. Costes, and R. Douce, eds., Elsevler/North-Holland Biomedical Press, Amsterdam (1980). 

Content of major lipid classes in pmol fatty acids per cell was determined. MGDG, 276 DGDG, 25 TG, 67 PC, 65 PG, 56 FFA, 23 PE, 21. The results showed no sign of extensive degradation of lipids which was reported to occur during protoplast isolation". ... 

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