Plant protoplast technology: Current status

Abstract

115 9516334; phone +44 115 9513057 Key words: Proto plasts, ge netic ma nip u la tion, nuclear and organelle trans for ma tion, phys i o log i cal in ves ti ga tions, protoplast-to-plant sys tems, so matic hy bridi sa tion ACTA PHYSIOLOGIAE PLANTARUM Vol. 27. No. 1. 2005: 117-129re view liv ing cell con tents into a hypertonic so lu tion. Several years later, what may now be con sid ered as early 'bio tech nol ogy' ex per i ments em a nated from in ves ti ga tions of Kuster (1909) andMichel (1937). These work ers dem on strated that the plasma membranes of iso lated proto plasts (wall-less 'na ked cells') could be in duced to fuse, per mit ting co alescence of their cytoplasms. How ever, such in ves tiga tions passed vir tu ally un no ticed for many years un til the com mer cial avail abil ity, dur ing the 1960s, of wall de grad ing pectinases and cellulases. Such en zymes, when used ei ther se quen tially or in combi na tion, fa cil i tated the re pro duc ible iso la tion of large, work able pop u la tions (>1 x 10 6 ) of viable protoplasts from primary tissues of a range of plant species.

Plant regeneration from protoplasts of Gentiana straminea Maxim

“…The calli derived from sLD and liquid culture systems were non-embryogenic, while those derived from aPL culture system showed both embryogenic and non-embryogenic characteristics. The effects of various culture systems on callus formation from protoplasts have also been reported in other plants [7,8,15,24]. In Gentiana kurroo, globular somatic embryos were detected one week after the microcolonies obtained in the thin layer culture and bead culture had been transferred to induction medium, but the cell aggregates obtained in the liquid culture died when transferred to the same medium [15].…”

“…In Ulmus minor, all the proliferating calli derived from the agarose droplet culture acquired the embryogenic characteristics 15 weeks after culture initiation [24]. Previous research suggested that the regeneration potential of the protoplasts in different culture systems might be influenced through stimulating cell wall formation, stabilizing the plasmalemma, supply of nutrients, the dilution of inhibitory substances, etc [7,8]. In this study, the aPL method gave compact colonies with small homogenous cells, which readily turned into embryogenic calli, once transplanted onto subculture medium.…”

Phytosulfokine stimulates cell divisions in sugar beet (Beta vulgaris L.) mesophyll protoplast cultures

“…nonionic surfactants or artificial oxygen carriers which enhance physiological status of the cultured cells. Such factors are able to increase plating efficiencies even a fewfold (Davey et al 2005b). In this report we show beneficiary effect of phytosulfokine (PSK) which represents a new class of supplements-the peptide growth factors.…”

“…For example, npropyl gallate (nPG) seems to target the first category of phenomena through its antioxidant activity (Saleem and Cutler 1987;Krens et al 1994). In turn, the stimulating effect of nurse cells results from the chemical signals they emit to cultured protoplasts (Hall et al 1993, Davey et al 2005b. It is obvious that phytosulfokine also falls into the second category of factors and once its action was even referred to as chemical nursing (Matsubayashi et al 2004).…”

Plant regeneration from leaf-derived protoplasts within the Daucus genus: effect of different conditions in alginate embedding and phytosulfokine application

“…Among them, incubation of protoplasts in liquid medium is the most simple to establish but results in unfavorable cell agglutination (Davey et al 2005a). Therefore, for many species, techniques ensuring physical separation of protoplasts based on their embedding in semi-solid media, with agar being first used as the gelling agent (Nagata and Takebe 1971), have been preferred (Davey et al 2005b).…”