Plant Regeneration from Protoplasts of Potato (Solarium tuberosum cv. Bintje)

Bokelmann, G. S.; Roest, S.

doi:10.1016/s0044-328x(83)80228-1

Cited by 115 publications

(47 citation statements)

References 14 publications

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“…After enzyme treatment, the protoplast suspension containing a mixture of micronucleate and mononucleate protoplasts, was filtered through 297-~tm and 88-1am nylon mesh, and repeatedly washed with half-strength V-KM medium (Bokelmann and Roest 1983) with macro-and microelements and NaC1 0.24 M (pH 5.6). Protoplasts were further purified as described previously .…”

Section: Genotypementioning

confidence: 99%

Isolation of sub-diploid microprotoplasts for partial genome transfer in plants: Enhancement of micronucleation and enrichment of microprotoplasts with one or a few chromosomes

Ramulu

Dijkhuis

Famelaer

et al. 1993

Planta

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Abstract.Results on the enhancement of the frequency of protoplasts with micronuclei, and on the isolation and enrichment of smaller sub-diploid microprotoplasts in transformed Nicotiana plumbaginifolia Viv. are reported. Suspension cells were treated with the spindle toxin amiprophos-methyl (APM) for 48 h, and subsequently incubated in a mixture of cell-wall-digesting enzymes in the presence of APM and cytochalasin-B. During enzyme incubation, the frequency of micronucleated protoplasts increased by a factor of 2-6. A shorter period (3 h) of incubation with a higher concentration of enzymes as well as a longer period (16 h) of incubation with a lower concentration of enzymes gave similar frequencies of micronucleated protoplasts and yields of micronuclei. Further, synchronization by sequential treatment with the DNA-synthesis inhibitor hydroxy urea, or aphidicolin, followed by APM and enzyme incubation, significantly increased the frequency of micronucleated protoplasts and the number of micronuclei. The suspension of protoplasts (mono-and micronucleated) obtained after enzyme incubation was fractionated throuh a continuous iso-osmotic gradient of Percoll, using high-speed centrifugation. This resulted in one large and a few small bands, which contained a heterogeneous population of microprotoplasts, protoplasts and cytoplasts. In contrast to the large band, the small bands contained a relatively higher frequency of small sub-diploid microprotoplasts. To separate the small sub-diploid microprotoplasts from the large microprotoplasts and protoplasts of the bands, discontinuous Percoll gradients and sequential filtration through nylon sieves of decreasing pore size (48--20-15-10-5 [am) were investigated. Compared with the former method, the latter gave a highly enriched fraction containing predominantly (~80%) small subdiploid microprotoplasts with DNA contents equivalent to that of one to four chromosomes, as revealed by miAbbreviations: APH =aphidicolin; APM = amiprophos-methyl; Au = arbitrary units; CB = cytochalasin B; DAPI = 4,6-diamidino-2-phenylindole; FDA = fluorescein diacetate; HU = hydroxy urea Correspondence to: K.S. Ramulu; FAX: 31 (8370) 18094 crodensitometric and flow-cytometric analyses. The application of this technique for partial genome and limited gene transfer is discussed.

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Section: Genotypementioning

confidence: 99%

Isolation of sub-diploid microprotoplasts for partial genome transfer in plants: Enhancement of micronucleation and enrichment of microprotoplasts with one or a few chromosomes

Ramulu

Dijkhuis

Famelaer

et al. 1993

Planta

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“…One possible reason for this behaviour could be the use of the modified V-KM medium since, in comparable assays, formation of the first cell colonies did not occur when using the original V-KM microelement solution (results not shown). Also the elimination of NH4NO 3 from the macroelements according to the culture medium of Boekelmann & Roest (1983) seemed to favour cellular divisions. Upadhya (1975) as well as Shepard & Totten (1977) observed that ammonia had a negative effect on the survival of potato protoplasts.…”

Section: Discussionmentioning

confidence: 99%

“…Immediately afterwards, 6 ml of modified V-KM medium (Boekelmann & Roest, 1983) was added to the fused protoplast suspension. In this medium the original composition of the microelements had been substituted with those of the MS medium.…”

Section: Methodsmentioning

confidence: 99%

Transfer of PLRV resistance fromSolanum verrucosum Schlechdt to potato (S. tuberosum L.) by protoplast electrofusion

et al. 2000

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SummarySomatic fusions between an accession of the diploid wild species Solanum verrucosum and a dihaploid S. tuberosum genotype were produced in order to incorporate resistance to potato leafroll virus (PLRV). In total 15 somatic hybrids out of 16 regenerants were Qbtained. Identification of hybrids was based on additive RAPD patterns, general morphological characteristics, chromosome numbers and chloroplast counts in stomata guard cells. A field trial was performed with the hybrids, their two parents and the control cultivar Kennebec to assess field performance and phenotypic variability. Yield parameters varied considerably among somatic hybrids. Some of the hybrids gave significantly higher yields, tuber numbers and tuber weights than both parents. Pollen fertility of hybrids ranged from 19 to 77%. Twelve hybrids were found to be resistant to PLRV.

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“…Consequently, modification has been made to MS macronutrient composition, mainly the (Takahata and Jomori 1989;Jomori et al 1995) or its complete withdrawal and replacement with glutamine (Fiuk and Rybczyński 2007;Tomiczak et al 2015). Other media used successfully for gentian protoplast culture include nutrient-rich KM8P medium developed by Kao and Michayluk (1975) for cells and protoplasts cultured at a very low densities (Meng et al 1996), V-KM medium reported by Bokelmann and Roest (1983) for potato protoplasts (O'Brien and Lindsay 1993), and B5 medium of Gamborg et al (1968), as used by Takahata and Jomori (1989) and Nakano et al (1995). Glucose and sucrose typically served as carbon sources, whereas mannitol ensured the correct osmotic pressure.…”

Section: Protoplast Culturementioning

confidence: 99%

Protoplast Culture and Somatic Cell Hybridization of Gentians

Tomiczak

Mikuła

Rybczyński

2015

The Gentianaceae - Volume 2: Biotechnology and Applications

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During the last three decades, less than fifteen papers have described the results of scientific investigations in the field of gentian protoplast technology and somatic hybridization. Despite rather limited research already done on this subject, several important goals have been achieved. Protoplast-to-plant systems have been developed either for leading ornamental species or for specific medicinal plants. Two major protoplast sources were evaluated in gentians, namely differentiated leaf mesophyll cells and undifferentiated callus/cell suspensions. Plant regeneration proceeded by the two different pathways of shoot organogenesis or somatic embryogenesis. Some examples of somaclonal variation at the ploidy level were demonstrated within the pool of protoplast-derived regenerants. Totipotency exhibited by gentian protoplasts was exploited to create three different somatic hybrid combinations: intergeneric Swertia mussotii (+) Bupleurum scorzonerifolium, and interspecific Gentiana kurroo (+) G. cruciata and G. cruciata (+) G. tibetica. IntroductionPlant cell totipotency as postulated by Schleiden and Schwann in the middle of the nineteenth century is the basis of modern plant biotechnology (Vasil 2008). Based on the ability of a living single plant cell to dedifferentiate and to convert into other cell types, it is possible to obtain a completely new plant from a cell and even from its protoplast. Consequently, plant regeneration from single protoplasts underlies the genetic manipulation technologies of somatic hybridization and direct genetic transformation by DNA uptake. However, the prerequisite for practical application of these technologies is the development of efficient and reproducible protoplast-to-plant systems for the species of interest.

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Plant Regeneration from Protoplasts of Potato (Solarium tuberosum cv. Bintje)

Cited by 115 publications

References 14 publications

Isolation of sub-diploid microprotoplasts for partial genome transfer in plants: Enhancement of micronucleation and enrichment of microprotoplasts with one or a few chromosomes

Isolation of sub-diploid microprotoplasts for partial genome transfer in plants: Enhancement of micronucleation and enrichment of microprotoplasts with one or a few chromosomes

Transfer of PLRV resistance fromSolanum verrucosum Schlechdt to potato (S. tuberosum L.) by protoplast electrofusion

Protoplast Culture and Somatic Cell Hybridization of Gentians

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