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Relevant bibliographies by topics / Protoplast fusion

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Academic literature on the topic 'Protoplast fusion'

Author: Grafiati

Published: 4 June 2021

Last updated: 8 February 2022

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Journal articles on the topic "Protoplast fusion"

1

Navrátilová, B. "Protoplast cultures and protoplast fusion focused on Brassicaceae: A review." Horticultural Science 31, No. 4 (November 25, 2011): 140–57. http://dx.doi.org/10.17221/3809-hortsci.

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The subjects of this article are protoplast isolations and protoplast fusions, in particular their history, a review of factors influencing the protoplasts isolation and fusion, selection of hybrid plants and utilization of somatic hybrids in plant breed-ing. Somatic hybridization through protoplast fusion can overcome sexual incompatibility among plant species or genera; transfer genes of resistance to diseases (viral, bacterial, fungal), pests, herbicides and others stress factors; obtain cybrid plants; transfer cytoplasmic male sterility or incease content of secondary metabolites in hybrid plants. The article is focussed mainly on the family Brassicaceae because among representatives are significant crops for the human population. Various successful combination of intraspecific, interspecific and intergeneric protoplast fusion were reported between representatives of the family Brassicaceae with the genus Brassica which belonged to the first agricultural crops used for the isolation of protoplast.  

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2

Perera, Srini C., and Peggy Ozias-Akins. "Regeneration from Sweetpotato Protoplasts and Assessment of Growth Conditions for Flow-sorting of Fusion Mixtures." Journal of the American Society for Horticultural Science 116, no. 5 (September 1991): 917–22. http://dx.doi.org/10.21273/jashs.116.5.917.

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Petiole protoplasts of the sweetpotato [Ipomoea batatas (L.) Lam.] cultivars Red Jewel and Georgia Jet formed cell walls within 24 hours and divided in 2 to 3 days. Pretreating enzyme solutions with activated charcoal increased the viability and division frequency of protoplasts. Culture of protoplast-donor plants in a medium containing STS did not affect plant growth, protoplasm yield, or viability, but did increase the division frequency. Culture of protoplasts for 24 hours in a medium containing DB, a cell wall synthesis inhibitor, or staining of protoplasts with FDA did not significantly affect division frequency. The division frequency of protoplasts cultured in liquid medium was significantly higher than that of protoplasts cultured in agarose-solidified medium. Cell cycle analysis of petioles and freshly isolated protoplasts showed that the latter has a significantly higher proportion of nuclei in G1 phase. Protoplasts did not initiate DNA synthesis or mitosis within the first 24 hours of culture. Low-frequency regeneration of shoots from protoplast-derived callus was accomplished on MS medium containing 1.0 mg ldnetin/liter when preceded by MS medium modified to contain only (in mg·liter-1) 800 NH4NO3, 1400 KNO3, 0.5 2,4-D, 0.5 kinetin, and 1.0 ABA. Roots produced from protoplast-derived callus formed adventitious shoots after 4 weeks on MS medium containing 2% sucrose, 0.02 mg kinetin/liter and 0.2% Gelrite. Secondary shoot formation from regenerated roots will be a more effective means of obtaining plants from protoplasts than direct shoot regeneration from callus. Chemical names used: silver thiosulfate (STS): 2.6-dichlorobenzonitrile (DB); fluorescein diacetate (FDA): 2.4-diacetate (FDA); 2.4 dichlorophenoxyacetic acid (2,4-D); abscisic acid (ABA).

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Armita, Devi. "Plant Breeding Through Protoplast Fusion." Jurnal Biologi UNAND 8, no. 2 (December 31, 2020): 42. http://dx.doi.org/10.25077/jbioua.8.2.42-47.2020.

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Protoplast culture (protoplast fusion) is one method of tissue culture that is widely used in plant breeding programs in a relatively short time. This method is used to overcome the problem of plants that are difficult or impossible to cross conventionally as well as used for species improvement by transferring the desired gene from the donor plant to the target plant via protoplast fusion. Protoplast fusion makes it possible to produce plants that are resistant to a disease and various abiotic stresses, rapid growth rates and have a better quantity and quality of metabolites than their parents. Various factors affect the success of fusion and regeneration of protoplasts into whole plants, including the source of explants, the composition of the enzyme solution and the duration of incubation, fusagen type and culture media for regeneration.

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Ahmed, Mohamed A. A., Miao Miao, Emmanouil D. Pratsinakis, Hongliang Zhang, Wei Wang, Yuan Yuan, Meiling Lyu, et al. "Protoplast Isolation, Fusion, Culture and Transformation in the Woody Plant Jasminum spp." Agriculture 11, no. 8 (July 26, 2021): 699. http://dx.doi.org/10.3390/agriculture11080699.

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Plant protoplasts are significant for plant cell culture, somatic cell fusion, genetics, and breeding studies. In addition, in vitro plant regeneration has great importance for developmental biology, manifesting potential applications in agriculture and biotechnology. In this regard, we present a well-established protocol regarding protoplast isolation, cell culture and protoplast fusion of Jasminum spp. In particular, different tissues of Jasminum samab L. and Jasminum mesnyi were employed for protoplast isolation, and stem explants provided a high callus induction rate in a short period of time. The best source for protoplast isolation was calli tissues. The optimized isolation protocol consisted of digesting callus in an enzyme solution containing 0.4 M mannitol, 0.2 M MES, 1 M CaCl2, 0.2 M KCL and 1 M NaH2PO4, 1.5% Cellulases onozuka R-10, 0.4% Macerozyme R-10 and 0.8% Pectinase for 4 h at 26 °C in the dark, providing a yield of 23.8 × 106 Protoplast/gFW with 88% viability. Protoplasts were cultured both in liquid and agarose medium under optimum conditions, leading to microcalli formation after eight weeks. A 5% protoplast-fusion rate can be achieved when cultured in 40% (w/v) PEG-MW6000 supplemented with 0.1 M CaCl2, 0.1 M sorbitol and 1 M Tris for 20 min. Furthermore, we developed an efficient PEG-mediated transformation protocol for jasmine protoplasts. The best results regarding protoplast transformation were obtained when the protoplast concentration was 4 × 105 cells/mL and the exogenous plasmid DNA added had a concentration of 10 µg DNA/100 µL protoplast solution, followed by the application of 40% PEG-4000 for 10 min.

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Liu, Donglong, and Nancy A. Reichert. "PROTOPLAST ISOLATION AND CULTURE OF KENAF (HIBISCUS CANNABINUS L.)." HortScience 29, no. 7 (July 1994): 729e—729. http://dx.doi.org/10.21273/hortsci.29.7.729e.

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Protoplast isolation and culture protocols were developed for leaf tissue from 6 kenaf cultivars [Everglades 41 (E41), E71, Guatemala 4 (G4), G45, G51, and Tainung 1]. For protoplast isolation, the best combination of hydrolytic enzymes was cellulysin (1% w/v; Calbiochem) plus macerase (0.5% w/v; Calbiochem), with a 24 hour digestion at 30°C in the dark. Yields reached 7.2 (10)6 protoplasts/g leaf tissue. Protoplast viabilities ranged from 65% to 96%. Minor cultivar differences were observed related to protoplast yield, but all viability estimates were in an acceptable range. Greatest cell division frequencies and plating efficiencies were obtained when protoplasts were initially cultured in liquid medium at a density of 1.0 (10)5 protoplasts/ml. Electrofusion protocols were developed for kenaf protoplasts testing the range from 1200 to 3000 V/cm. A fusion voltage of 2000 V/cm yielded the highest fusion frequency and retained viability above 80%.

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Djajanegara, Ira, and Agus Masduki. "PROTOPLAST FUSION BETWEEN WHITE AND BROWN OYSTER MUSHROOMS." Indonesian Journal of Agricultural Science 11, no. 1 (July 8, 2013): 16. http://dx.doi.org/10.21082/ijas.v11n1.2010.p16-23.

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Genetic crossing of white oyster mushroom (Pleurotus floridae) to introduce longer storage life trait can only be done within individuals in this particular species. However, longer storage life trait is possessed by brown oyster mushroom (Pleurotus cystidiosus). Therefore, a protoplast fusion experiment between white and brown oyster mushrooms was conducted to obtain an oyster mushroom strain showing high productivity and long storage life. The experiment was done at the biology laboratory of the University of Al Azhar Indonesia from May 2008 to August 2009. Protoplast fusion was done by isolating protoplast from 5-day old monokaryotic mycelia grown in potato dextrose broth (PDB). Around 3.15 x 105 protoplasts ml-1 were harvested using mixture of cellulase Onozuka R-10 (1%) and macerozyme R-10 (1%) from brown oyster mushroom with 80.61% viability. Similarly, 3.71 x 105 protoplasts ml-1 were harvested using lysing enzyme (2%) from white oyster mushroom with 83.68% viability. Protoplast fusions were conducted using 40% PEG6000 for 10 minutes. The candidate fusants were then screened using minimum regeneration media (MRM). There were 22 colonies grew on MRM media and four colonies (FS1, FS2, FS3, and FS4) showed clamp connection as well as primordia formation to be chosen as candidate fusants. However, isozyme studies using malate dehydrogenase and acid phosphatase as marker enzymes confirmed that only FS1 and FS2 were the hybridized products. The two colonies showed different mycelia growth patterns and hyphae sizes, fruit body morphology and productivity compared to their parents. These two fusants, however, did not indicate the presence of longer storage life trait as expected despite a higher productivity achieved by FS1. In this study, the protoplast fusion only yielded higher productivity strain of mushroom with different colors without any changes in storage life.

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Nolan, Richard A. "Stage-specific changes in cytoplasmic protein synthesis in Entomophaga aulicae protoplasts." Canadian Journal of Microbiology 35, no. 3 (March 1, 1989): 373–78. http://dx.doi.org/10.1139/m89-057.

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The patterns of protein synthesis associated with three sequential stages in protoplast morphogenesis (spindle-shaped, early fusion sphere, and late fusion sphere protoplasts) of the fungus Entomophaga aulicae were studied using both one-dimensional gels with general protein staining and two-dimensional gels with [35S]methionine protein labelling and fluorography. A total of 332 proteins were observed with 63.5% (211) common to all three developmental stages. Of the individual totals, 3.3% (8 out of 245), 7.3% (22 out of 301), and 4.5% (13 out of 286) of the proteins were unique to the spindle-shaped, early fusion sphere, and late fusion sphere protoplasts, respectively. The molecular mass and pI distribution profiles for early fusion sphere protoplast proteins are discussed.Key words: protein synthesis, stage-specific proteins, fungal protoplasts, Entomophaga aulicae.

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Pauls, K. P., and P. V. Chuong. "Flow cytometric identification of Brassica napus protoplast fusion products." Canadian Journal of Botany 65, no. 5 (May 1, 1987): 834–38. http://dx.doi.org/10.1139/b87-113.

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A flow cytometric method to identify Brassica somatic hybrids has been developed. The procedure is based on the use of fluorescein isothiocyanate stained hypocotyl protoplasts and unstained mesophyll protoplasts as partners for polyethylene glycol induced protoplast fusion. The fluorescein isothiocyanate stained hypocotyl protoplasts and unstained mesophyll protoplasts were passed through the flow cytometer – cell sorter separately to maximize the sensitivity of the red and green detectors to chlorophyll and fluorescein isothiocyanate fluorescence, respectively, and to ensure that there was no spillover of chlorophyll fluorescence into the green detector or fluorescein isothiocyanate fluorescence into the red detector. When adjusted properly, suspensions of a single protoplast type gave populations that fell on either the green or red axis of a two-dimensional green versus red fluorescence plot. With the same machine settings three populations of protoplasts could be identified in fusion mixtures of the two types of protoplasts: namely, the parental protoplasts that fell on the green or red axis as well as a third that had significant green and red fluorescence.

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Chen, Qin, H. Y. Li, Y. Z. Shi, D. Beasley, B. Bizimungu, and M. S. Goettel. "Development of an effective protoplast fusion system for production of new potatoes with disease and insect resistance using Mexican wild potato species as gene pools." Canadian Journal of Plant Science 88, no. 4 (July 1, 2008): 611–19. http://dx.doi.org/10.4141/cjps07045.

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Somatic hybridization through protoplast fusion is an important alternative approach for overcoming sexual incompatibility between diploid Solanum species and cultivated potatoes. However, compared with other potato species, methods for protoplast isolation and plant generation for several Mexican wild diploid potato species are not well established. In this study, a systematic procedure was designed for the isolation of a large number of high-quality protoplasts from various Mexican wild species that carry high levels of disease (late blight) and insect [Colorado potato beetle (CPB)] resistance. Using this procedure, an effective potato protoplast fusion system was developed to produce new somatic hybrids between two Mexican, one Argentina wild species, and cultivated potato clones. Regenerated plants and somatic hybrids were obtained at a high frequency from the protoplasts of the diploid wild species and their fused cells with S. tuberosum. Morphological, cytological and molecular marker analyses demonstrated that somatic hybrids were successfully obtained from the cell fusion of S. tuberosum and the diploid species S. pinnatisectum, S. cardiophyllum, and S. chacoense. Assessment of disease and insect reactions demonstrated that several of the protoplast-derived clones and somatic hybrids showed a higher level of resistance to both late blight and CPB than was found in S. tuberosum, confirming that the protoplast system is a powerful tool in potato breeding programs for the development of disease and insect resistance. This new fusion system provides breeders with opportunities to transfer disease and insect resistance genes from Mexican wild species into cultivated potato. Key words: Somatic hybrid, protoplast, fusion, potato, Solanum, late blight, disease resistance, Colorado potato beetle insect resistance

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Nolan, Richard A. "Influence of a negatively charged surface (Teflon disk) on Entomophaga aulicae protoplast morphogenesis under mass fermentation conditions." Canadian Journal of Botany 69, no. 11 (November 1, 1991): 2578–81. http://dx.doi.org/10.1139/b91-321.

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The effects of a negatively charged surface (Teflon disk) on protoplast morphogenesis for the fungus Entomophaga aulicae under mass fermentation conditions were determined. The control consisted of a vessel lacking such a disk. In the presence of the disk the initial three and sequentially produced protoplast stages (spindle shaped, early fusion sphere, and late fusion sphere protoplasts) recycled with the early fusion sphere predominating. The production of the subsequent and walled stage (i.e., hyphal body) was suppressed. The results are in contrast with those obtained in a previous study using a neutral (Mylar) and a positively charged (polypropylene) disk in which hyphal body production was enhanced. This technique provides a new and subtle approach for altering protoplast developmental patterns which avoids the use of mutagens or added chemical metabolic inhibitors. Key words: Entomophaga aulicae, fungal protoplast morphogenesis, negatively charged surface, mass fermentation.

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Dissertations / Theses on the topic "Protoplast fusion"

1

Illing, G. T. "Protoplast fusion and regeneration in Streptomyces clavuligerus." Thesis, University of Nottingham, 1987. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.378980.

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Mandegaran, Zohreh. "Genetic improvement of roses by protoplast fusion." Thesis, University of East London, 1996. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.339583.

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Brites, Anny Stella Monteiro. "Seleção de linhagens de Saccharomyces cerevisiae potencializadas pelo fator Killer, H2S- e o carater floculante." Universidade de São Paulo, 2003. http://www.teses.usp.br/teses/disponiveis/11/11141/tde-19052003-144728/.

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Dentre as características desejáveis em leveduras fermentadoras alcóolicas estão a capacidade de floculação, a não produção de H2S e o caráter "killer". Neste trabalho foram selecionadas sete linhagens de Saccharomyces cerevisiae com algumas destas características, que passaram por testes confirmativos e pela cariotipagem eletroforética resultando na escolha de duas linhagens: ATCC 26602 (altamente floculante) e K1 (H2S - e possuidoras do caráter "killer"). Estas linhagens foram utilizadas em um cruzamento via fusão de protoplasto para se obter um produto de fusão estável com as características de interesse tecnológico. Na seleção das linhagens híbridas com base em caracteres naturais foram isolados 1291 híbridos em meio seletivo e entre essas colônias somente 1,5% foram inicialmente consideradas híbriadas. Após três subcultivos em YEPD líquido, estes produtos de fusão não se mostraram estáveis.
Flocculative and "killer" skills and lack in production of H2S are desirable characteristics of the ethanolic fermentative yeasts. Seven selected strains of Saccharomyces cerevisiae with some of these characteristics were evaluated for confirmation of these habilities and their genetic characterization was undertaken by eletrophoretic kariotyping. The strain ATCC 26602 had flocculant hability and the strain K1 was H2S - and "killer". The strains were selected for protoplast fusion aiming to obtain a stable fusion strain with these desirable technologyc characteristics. The selection of the hybrid strains were based on natural characters and have shown 1291 hybrids (frequency of 1,5%) in the medium for the isolation of the fusionants (protoplasts). The protoplast stability were monitored by three continuous growth in the YEPD liquid midium and the stable fusion products were not obtained.

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Maren, Nathan Allen. "Symmetric Protoplast Fusion in Interserial Syringa (Oleaceae) Hybridization." Thesis, North Dakota State University, 2016. https://hdl.handle.net/10365/28026.

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Few other woody plants embody the preeminence of temperate woody plants in garden cultivation like the lilacs. In spite of their relationship, the trees lack the diversity of cultivated floral forms observed within the shrub lineages. Typical selection and cross-pollination schemes within the tree lilacs or between trees and shrubs have failed to yield the diversity of colors and fragrances on a tree form. With somatic fusion in Citrus spp. as a guideline for Syringa spp. protoplast isolation and culture, experiments were designed to optimize the conditions through somatic fusion. Protoplast isolation experiments revealed yield increases with increased exposure to cell wall degrading enzymes as well as losses in viability with increased exposure. Electrofusion experiments yielded somatic hybrids, yet further investigation is necessary to optimize the fusion electroporation settings and beyond.

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Hansen, Christine S. "Construction of galactose assimilating, carotenoid producing yeasts by protoplast fusion." Thesis, University of British Columbia, 1988. http://hdl.handle.net/2429/27935.

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Protoplasts were prepared from two yeast strains P. rhodozyma (ATCC 24202) and K. fragilis (ATCC 8455). Protoplasts prepared from P. rhodozyma were facilitated by prior growth of the cells in a media containing S-(2-aminoethyl)-L-cysteine. Protoplasts from these two yeast genera were fused either by the use of electrofusion or polyethylene glycol treatment. Stable carotenoid producing cell lines were selected by growth at 30°C on yeast nitrogen base plus galactose. Selected single fusants display taxonomic characteristics common to both genera with a cellular morphology and a carotenoid composition similar to that of P. rhodozyma.
Land and Food Systems, Faculty of
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Raikar, Sanjeev Vencu. "Protoplast fusion of Lolium perenne and Lotus corniculatus for gene introgression." Phd thesis, Lincoln University. Bio-Protection and Ecology Division, 2007. http://theses.lincoln.ac.nz/public/adt-NZLIU20080214.105406/.

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Protoplast fusion of Lolium perenne and Lotus corniculatus for gene introgression by Sanjeev V. Raikar Lolium perenne is one of the most important forage crops globally and in New Zealand. Lotus corniculatus is a dicotyledonous forage that contains valuable traits such as high levels of condensed tannins, increased digestibility, and high nitrogen fixing abilities. However, conventional breeding between these two forage crops is impossible due to their markedly different taxonomic origin. Protoplast fusion (somatic hybridisation) provides an opportunity for gene introgression between these two species. This thesis describes the somatic hybridisation, the regeneration and the molecular analysis of the putative somatic hybrid plants obtained between L. perenne and L. corniculatus. Callus and cell suspensions of different cultivars of L. perenne were established from immature embryos and plants were regenerated from the callus. Of the 10 cultivars screened, cultivars Bronsyn and Canon had the highest percentage of callus induction at 36% each on 5 mg/L 2,4-D. Removal of the palea and lemma which form the seed coat was found to increase callus induction ability of the embryos. Plant regeneration from the callus was achieved when the callus was plated on LS medium supplemented with plant growth regulators at different concentrations. Variable responses to shoot regeneration was observed between the different cultivars with the cv Kingston having the lowest frequency of shoot formation (12%). Different factors affecting the protoplast isolation of L. perenne were investigated. The highest protoplast yield of 10×106 g-1FW was obtained when cell suspensions were used as the tissue source, with enzyme combination A (Cellulase Onozuka RS 2%, Macerozyme R-10 1%, Driselase 0.5%, Pectolyase 0.2%), for 6 h incubation period in 0.6 M mannitol. Development of microcolonies was only achieved when protoplasts were plated on nitrocellulose membrane with a L. perenne feeder layer on PEL medium. All the shoots regenerated from the protoplast-derived calli were albino shoots. The highest protoplast yield (7×106 g-1FW) of L. corniculatus was achieved from cotyledons also with enzyme combination A (Cellulase Onozuka RS 2%, Macerozyme R-10 1%, Driselase 0.5%, Pectolyase 0.2%), for 6 h incubation period in 0.6 M mannitol. The highest plating efficiency for L. corniculatus of 1.57 % was achieved when protoplasts were plated on nitrocellulose membrane with a L. perenne feeder layer on PEL medium. The highest frequency of shoot regeneration (46%) was achieved when calli were plated on LS medium with NAA (0.1 mg/L) and BA (0.1 mg/L). Protoplast fusion between L. perenne and L. corniculatus was performed using the asymmetric somatic hybridisation technique using PEG as the fusogen. L. perenne protoplasts were treated with 0.1 mM IOA for 15 min and L. corniculatus protoplasts were treated with UV at 0.15 J/cm2 for 10 min. Various parameters affecting the fusion percentage were investigated. Successful fusions were obtained when the fusions were conducted on a plastic surface with 35% PEG (3350 MW) for 25 min duration, followed by 100 mM calcium chloride treatment for 25 min. A total of 14 putative fusion colonies were recovered. Shoots were regenerated from 8 fusion colonies. Unexpectedly, the regenerated putative hybrid plants resembled L. corniculatus plants. The flow cytometric profile of the putative somatic hybrids resembled that of L. corniculatus. Molecular analysis using SD-AFLP, SCARs and Lolium specific chloroplast microsatellite markers suggest that the putative somatic hybrids could be L. corniculatus escapes from the asymmetric protoplast fusion process. This thesis details a novel Whole Genome Amplification technique for plants using Strand Displacement Amplification technique.

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Raikar, S. V. "Protoplast fusion of Lolium perenne and Lotus corniculatus for gene introgression." Diss., Lincoln University, 2007. http://hdl.handle.net/10182/301.

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Lolium perenne is one of the most important forage crops globally and in New Zealand. Lotus corniculatus is a dicotyledonous forage that contains valuable traits such as high levels of condensed tannins, increased digestibility, and high nitrogen fixing abilities. However, conventional breeding between these two forage crops is impossible due to their markedly different taxonomic origin. Protoplast fusion (somatic hybridisation) provides an opportunity for gene introgression between these two species. This thesis describes the somatic hybridisation, the regeneration and the molecular analysis of the putative somatic hybrid plants obtained between L. perenne and L. corniculatus. Callus and cell suspensions of different cultivars of L. perenne were established from immature embryos and plants were regenerated from the callus. Of the 10 cultivars screened, cultivars Bronsyn and Canon had the highest percentage of callus induction at 36% each on 5 mg/L 2,4-D. Removal of the palea and lemma which form the seed coat was found to increase callus induction ability of the embryos. Plant regeneration from the callus was achieved when the callus was plated on LS medium supplemented with plant growth regulators at different concentrations. Variable responses to shoot regeneration was observed between the different cultivars with the cv Kingston having the lowest frequency of shoot formation (12%). Different factors affecting the protoplast isolation of L. perenne were investigated. The highest protoplast yield of 10×10⁶ g⁻¹FW was obtained when cell suspensions were used as the tissue source, with enzyme combination 'A' (Cellulase Onozuka RS 2%, Macerozyme R-10 1%, Driselase 0.5%, Pectolyase 0.2%), for 6 h incubation period in 0.6 M mannitol. Development of microcolonies was only achieved when protoplasts were plated on nitrocellulose membrane with a L. perenne feeder layer on PEL medium. All the shoots regenerated from the protoplast-derived calli were albino shoots. The highest protoplast yield (7×10⁶ g⁻¹FW) of L. corniculatus was achieved from cotyledons also with enzyme combination 'A' (Cellulase Onozuka RS 2%, Macerozyme R-10 1%, Driselase 0.5%, Pectolyase 0.2%), for 6 h incubation period in 0.6 M mannitol. The highest plating efficiency for L. corniculatus of 1.57 % was achieved when protoplasts were plated on nitrocellulose membrane with a L. perenne feeder layer on PEL medium. The highest frequency of shoot regeneration (46%) was achieved when calli were plated on LS medium with NAA (0.1 mg/L) and BA (0.1 mg/L). Protoplast fusion between L. perenne and L. corniculatus was performed using the asymmetric somatic hybridisation technique using PEG as the fusogen. L. perenne protoplasts were treated with 0.1 mM IOA for 15 min and L. corniculatus protoplasts were treated with UV at 0.15 J/cm² for 10 min. Various parameters affecting the fusion percentage were investigated. Successful fusions were obtained when the fusions were conducted on a plastic surface with 35% PEG (3350 MW) for 25 min duration, followed by 100 mM calcium chloride treatment for 25 min. A total of 14 putative fusion colonies were recovered. Shoots were regenerated from 8 fusion colonies. Unexpectedly, the regenerated putative hybrid plants resembled L. corniculatus plants. The flow cytometric profile of the putative somatic hybrids resembled that of L. corniculatus. Molecular analysis using SD-AFLP, SCARs and Lolium specific chloroplast microsatellite markers suggest that the putative somatic hybrids could be L. corniculatus escapes from the asymmetric protoplast fusion process. This thesis details a novel Whole Genome Amplification technique for plants using Strand Displacement Amplification technique.

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Johnson, Alexander Arthur Theodore. "Protoplast Fusion for the Production of Intermonoploid Somatic Hybrids in Cultivated Potato." Thesis, Virginia Tech, 1998. http://hdl.handle.net/10919/46514.

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Monoploid potato genotypes represent plant material that is free from the "genetic load" of lethal and severely deleterious alleles normally present in the highly heterozygous cultivated potato species. Field evaluations enabled the identification of agronomically superior monoploid potato genotypes from a population of more than 100 anther-derived monoploids. Chemical fusion and electrofusion between pairs selected from 31 superior monoploids resulted in the production of three different groups of intermonoploid somatic hybrids. The hybridity of somatic hybrid plants and calluses was confirmed through PCR-based amplification of simple sequence repeat (SSR) sequences in the potato genome. Polymorphic SSR loci between the monoploid parents of a particular group of somatic hybrids were used to separate true somatic hybrids (heterozygous at the loci) from parental somaclones regenerating from unfused protoplasts (homozygous for one parental band at the loci). One group of somatic hybrids (SH1, SH2 and SH2B) was of particular interest because it resulted from the fusion of a S. phureja monoploid to a high acetylleptinidine-producing monoploid derived from an F1 hybrid between S. chacoense and S. phureja. The leptine acetylleptinidine (ALD) is produced only by some accessions of S. chacoense Bitt. and provides resistance to feeding by the Colorado potato beetle (Leptinotarsa decemlineata Say) when present in sufficient concentrations. The somatic hybrids produced moderate levels of ALD in leaves and stems (roughly 60% that of a high ALD-producing S. chacoense clone). Pollinations of SH1, SH2 and SH2B by several diploid and tetraploid potato clones resulted in three fruit on SH2, one fruit on SH2B and no fruit on SH1. Two resulting progeny populations of SH2 [SH2A = SH2 × S. andigena 8-1 (4x); SH2P = SH2 × S. phureja 66AP11-53 (2x)] expressed higher fertility than the original somatic hybrids and were sexually crossed as both male and female parents to S. tuberosum cv. Atlantic. All of the SH2 progeny populations expressed acetylleptinidines, albeit at lower levels than the SH2 somatic hybrid, providing strong evidence that the genes controlling acetylleptinidine production are dominant. Variation for ALD expression in the SH2 progeny indicated one or a few genes with additive effect controlling the ALD trait. In addition, the choice of male parent in sexual crosses to SH2 affected subsequent ALD expression in progeny populations. The SH2 progeny represent an important first step towards transferring acetylleptinidines to cultivated potato. SH1, SH2 and SH2B appeared to be negatively affected by an unusually high ploidy (hexaploid, 6x). Field-grown plants produced many tubers (mean = 35) of low weight (mean = 10.4 g) and were stunted in appearance. Anther culture of SH2 yielded triploid regenerants (3x). These regenerants may be more phenotypically normal than the original somatic hybrids because of lower ploidy. Segregation of SSR alleles in the triploid anther culture regenerants provided evidence that the hexaploid somatic hybrid SH2 genome is comprised of four homologous genomes of CP2-103 (the high leptine-producing monoploid) and two homologous genomes of 13-14 203 (the S. phureja monoploid).
Master of Science

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Hothersall, Joanne. "Metabolite production and molecular characterisation of interspecific Aspergilli." Thesis, University of Nottingham, 1998. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.285775.

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Ravichandran, Vidya. "Application of molecular markers to characterize potato plants derived from anther culture and protoplast fusion." Thesis, This resource online, 1996. http://scholar.lib.vt.edu/theses/available/etd-11072008-063210/.

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Books on the topic "Protoplast fusion"

1

Mandegaran, Zohreh. Genetic improvement of roses by protoplast fusion. London: University of East London, 1996.

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Clugston, Denise B. Embryo culture and protoplast fusion for the introduction of Mexican wild species germplasm into the cultivated potato. Birmingham: University of Birmingham, 1988.

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Rice, Glenn Edward. Protoplast fusion in the genus Rhodotorula. 1987.

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Gleba, Yury Y., and Konstantin M. Sytnik. Protoplast Fusion: Genetic Engineering in Higher Plants. Springer, 2011.

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Book chapters on the topic "Protoplast fusion"

1

Gooch, Jan W. "Protoplast Fusion." In Encyclopedic Dictionary of Polymers, 918. New York, NY: Springer New York, 2011. http://dx.doi.org/10.1007/978-1-4419-6247-8_14602.

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Bengochea, Teresa, and John H. Dodds. "Protoplast fusion." In Plant Protoplasts, 44–58. Dordrecht: Springer Netherlands, 1986. http://dx.doi.org/10.1007/978-94-009-4095-6_4.

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Gingold, Elliot B. "Yeast Protoplast Fusion." In Springer Protocols Handbooks, 247–53. Totowa, NJ: Humana Press, 1986. http://dx.doi.org/10.1007/978-1-60327-405-0_24.

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Spencer, John F. T., Dorothy M. Spencer, and I. J. Bruce. "Protoplast Formation and Fusion." In Yeast Genetics, 64–69. Berlin, Heidelberg: Springer Berlin Heidelberg, 1989. http://dx.doi.org/10.1007/978-3-642-73356-7_9.

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Zimmermann, Martin, and Matthias Sipiczki. "Protoplast Fusion of Yeasts." In Nonconventional Yeasts in Biotechnology, 83–99. Berlin, Heidelberg: Springer Berlin Heidelberg, 1996. http://dx.doi.org/10.1007/978-3-642-79856-6_2.

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Schieder, O., P. P. Gupta, G. Krumbiegel, and T. Hein. "Protoplast Fusion and Transformation." In Ciba Foundation Symposium 97 - Better Crops for Food, 213–36. Chichester, UK: John Wiley & Sons, Ltd., 2008. http://dx.doi.org/10.1002/9780470720783.ch15.

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Sink, K. C. "Protoplast Fusion and Somatic Hybridization." In Genetics and Breeding of Ornamental Species, 53–68. Dordrecht: Springer Netherlands, 1991. http://dx.doi.org/10.1007/978-94-011-3296-1_4.

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Gnanam, Annie Juliet. "Protoplast Fusion Techniques in Fungi." In Laboratory Protocols in Fungal Biology, 483–88. New York, NY: Springer New York, 2012. http://dx.doi.org/10.1007/978-1-4614-2356-0_45.

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Bravo, J. E., and D. A. Evans. "Protoplast Fusion for Crop Improvement." In Plant Breeding Reviews, 193–218. Hoboken, NJ, USA: John Wiley & Sons, Inc., 2011. http://dx.doi.org/10.1002/9781118061008.ch4.

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Grosser, Jude W., and Frederick G. Gmitter. "Protoplast Fusion and Citrus Improvement." In Plant Breeding Reviews, 339–74. Hoboken, NJ, USA: John Wiley & Sons, Inc., 2011. http://dx.doi.org/10.1002/9781118061053.ch10.

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Conference papers on the topic "Protoplast fusion"

1

Kantawang, Titirat, Sompid Samipak, Jumras Limtrakul, and Nattaporn Chattham. "Application of optical tweezers and excimer laser to study protoplast fusion." In International Conference on Photonics Solutions 2015, edited by Surasak Chiangga and Sarun Sumriddetchkajorn. SPIE, 2015. http://dx.doi.org/10.1117/12.2195700.

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Sang, Wen-Jiao, Bei-Ping Zhang, Zhen Liu, Lei Wang, Chun-Hua Zhu, and Lan Gao. "Experimental Study on Improving Nitrogen Removal Efficiency of Denitrify Bacteria by Protoplast Fusion." In 2008 2nd International Conference on Bioinformatics and Biomedical Engineering. IEEE, 2008. http://dx.doi.org/10.1109/icbbe.2008.394.

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Shiwei Wang, Qinghui Wang, Ying Zhai, and Min Wang. "Screening of NHase high-producing Rhodococcus ruber Strain TQD-58 by both parents inactivated protoplast fusion." In 2012 International Symposium on Information Technology in Medicine and Education (ITME 2012). IEEE, 2012. http://dx.doi.org/10.1109/itime.2012.6291408.

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Guosheng Chen, Zegen Wang, Min Wu, Wei Chen, and Min Wang. "Selection of Streptomyces roseosporus strain with high yield by the inactivated parental strain protoplasts fusion." In 2011 International Conference on Remote Sensing, Environment and Transportation Engineering (RSETE). IEEE, 2011. http://dx.doi.org/10.1109/rsete.2011.5965999.

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