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Protoplast

About: Protoplast is a research topic. Over the lifetime, 5474 publications have been published within this topic receiving 122468 citations.


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Journal ArticleDOI
TL;DR: Protoplasts from a total of thirty-six genotypes of Brassica species – B. napus, B. campestris and Xinjiang wild rape – were analysed for shoot regeneration using a feeder culture system, finding several genotypes with high regeneration ability were elite breeding lines.
Abstract: Protoplasts from a total of thirty-six genotypes of Brassica species – B. napus, B. campestris (syn. B. rapa), B. juncea, and three distant relatives, Orychophragmus violaceus, Isatis indigotica and Xinjiang wild rape – were analysed for shoot regeneration using a feeder culture system. With the exception of B. campestris and Xinjiang wild rape, some genotypes of all the species could regenerate plants with high efficiency (above 20% of isolated calli initiating shoots). Several genotypes with high regeneration ability were elite breeding lines. Culture conditions as well as genotype had a significant impact on shoot regeneration frequency. In particular, silver nitrate added to the regeneration medium at doses of 6 and 30 μM improved shoot regeneration frequency to 25.4% and 52.2% of isolated calli, respectively, compared to 7.3% percent shoot regeneration without silver nitrate in seven responsive genotypes. Addition of silver nitrate to the regeneration medium also induced shoot regeneration in non-responsive genotypes. Intact plants could be obtained within three months from protoplast isolation in the regenerative genotypes using the current culture system. Advantages of mesophyll protoplasts as compared to protoplasts isolated from hypocotyls for genetic manipulation in Brassica species are discussed.

44 citations

Book
01 Oct 1994
TL;DR: Somatic Hybridization is a Rich Source of Genetic Variability in Medicinal Plants - Including Tobacco, and the Selection of a New Strain, 694-L.
Abstract: Section I Protoplast Fusion, Somatic Hybrids, Asymmetric Hybrids, Cybrids - Transfer of Chloroplast Traits.- I.1 Somatic Hybridization - A Rich Source of Genetic Variability.- I.2 Fluorescence Microscope Study of Protoplast Fusion.- I.3 Somatic Hybridization by Microfusion of Protoplasts.- I.4 Asymmetric Somatic Hybrids.- I.5 Cybrids - Transfer of Chloroplast Traits Through Protoplast Fusion Between Sexually Incompatible Solanaceae Species.- Section II Somatic Hybridization in Cereals, Grasses, and Legumes.- II. 1 Somatic Hybridization in the Family Gramineae.- II.2 Somatic Hybridization Between Zea mays and Triticum sect, trititrigia.- II.3 Somatic Hybridization in Festuca and Lolium.- II.4 Somatic Hybridization Between Birdsfoot Trefoil (Lotus corniculatus L.) and Soybean (Glycine max L.).- II.5 Somatic Hybridization in the Genus Medicago.- Section III Somatic Hybridization in Potato, Tomato, Eggplant, and Lettuce.- III. 1 Cybridization in Potato.- III.2 Somatic Hybridization in Solanum Tuberosum x S. chacoense.- III.3 Somatic Hybridization Between Solanum tuberosum and Nicotiana plumbaginifolia.- III.4 Pomato: Potato Protoplast System and Somatic Hybridization Between Potato and a Wild Tomato.- III.5 Somatic Hybridization Between Lycopersicon esculentum Mill. and Lycopersicon peruvianum var. dentatum Dun.- III.6 Somatic Hybridization Between Tomato (Lycopersicon esculentum) and Pepino (Solanum muricatum).- III.7 Somatic Hybridization of Eggplant (Solanum melongena L.) with Its Close and Wild Relatives.- III.8 Somatic Hybridization in Lettuce (Lactuca Species).- Section IV Somatic Hybridization in Brassicaceae.- IV.1 Resynthesis of Brassica napus Through Protoplast Fusion Between B. oleracea and B. rapa.- IV.2 Analysis of Somatic Hybrids and Cybrids Obtained by Fusion of Brassica rapa and B. oleracea.- IV. 3 Somatic Hybridization Between Radish (Raphanus sativus) and Rapeseed (Brassica napus).- IV.4 Somatic Hybridization Between Brassica and Sinapi.- Section V Somatic Hybridization in Medicinal Plants - Including Tobacco.- V.I Somatic Hybridization of Medicinal Plants in the Family Solanaceae.- V.2 Somatic Hybridization in Datura.- V.3 Somatic Hybrids Between Nicotiana repanda and N. tabacum Show Resistance to Tobacco Mosaic Virus and Meloidogyne arenaria.- V.4 Somatic Hybridization Between Tobacco (Nicotiana tabacum L.) and Black Nightshade (Solanum nigrum L.), and the Selection of a New Strain, 694-L.- V.5 Transfer of Lincomycin Resistance Through Somatic and Sexual Cybridization in Nicotiana A. CSEPL? (With 4 Figures).- V.6 Somatic Hybridization in the Family Apocynaceae (Catharanthus, Rauwolfia, Rhazya, and Vinca Species).- Section VI Somatic Hybridization in Trees (Citrus, Poncinus, Prunus, Pyrus, and Populus Species).- VI. 1 Somatic Hybridization of Citrus with Sexually Incompatible Wild Relatives.- VI.2 Somatic Hybridization Between Citrus sinensis and Poncirus trifoliata.- VI.3 Somatic Hybridization Between Pyrus x Prunus Species.- VI.4 Somatic Hybridization in Populus Species (Poplars).- Section VII Somatic Hybridization in Algae, Bryophytes, and Ferns.- VII. 1 Somatic Hybridization in Algae.- VII.2 Somatic Hybridization in Bryophytes.- VII.3 Somatic Hybridization in Ferns.

44 citations

Journal ArticleDOI
TL;DR: An efficient method for the transformation and regeneration of fertile transgenic rice (Oryza sativa L.) plants is presented and two-thirds of transgenic plants grown to maturity in the greenhouse bore viable seeds.
Abstract: An efficient method for the transformation and regeneration of fertile transgenic rice (Oryza sativa L.) plants is presented. In this protocol seed calli from the varietiesNipponbare andTaipei 309 were used to produce rice suspension cultures in General Medium. Protoplasts were isolated from suspension cells (8 × 106 protoplasts perg fresh weight), then were incubated with sterile DNA in the presence of MaMg solution, followed by addition of PEG to a final concentration of 25%. A hygromycin phosphotransferase (hph) gene under the plant transcriptional regulatory signals was used as a selectable marker gene. Hygromycin-resistant colonies were selected in the presence of 95 μM hygromycin B with apparent frequencies of 2×10−4 and 5×10−4 forNipponbare andTaipei 309, respectively. Plantlets were regenerated from resistant colonies in Murashige and Skoog plant regeneration medium. Among 628 transgenic plants grown to maturity in the greenhouse, two-thirds bore viable seeds.

44 citations

Journal ArticleDOI
TL;DR: Three kinds of enzymes, agarase, β‐1,4‐mannanase, and β-1,3‐xylanase, required for isolation of protoplasts from the red alga Bangia atropurpurea (Roth) C. atropolpurea were examined and resulted in successful protoplast isolation.
Abstract: Three kinds of enzymes, agarase, β-1,4-mannanase, and β-1,3-xylanase, required for isolation of protoplasts from the red alga Bangia atropurpurea (Roth) C. Ag. were prepared from bacterial culture fluids of Vibrio sp. PO-303, Vibrio sp. MA-138, and Alcaligenes sp. XY-234, respectively, isolated from the sea environment. The optimal pH of all enzymes was around 7.5. Suitable conditions for protoplast isolation from B. atropurpurea were examined. The pretreatment of the fronds with pa-pain solution (20 mM Mes buffer, pH 7.5, containing 2% papain and 0.5 M mannitol) contributed to successful protoplast isolation. When razor-cut fragments of the fronds (about 200 mg in fresh weight) immersed in 20 mM Mes buffer, 7.5, containing 0.5 M mannitol and one unit each of agarase, β-1,4-mannanase, and β-1,3-xylanase were incubated at 22°C for 90 min with gentle agitation, 5.7 × 106 protoplasts were released from them. Many protoplasts regenerated into fronds of regular or irregular shape.

44 citations

Journal ArticleDOI
TL;DR: An efficient system for protoplast isolation and subcellular localization of desired proteins using pineapple plants derived from tissue culture is developed and showed that the system is also suitable for protein–protein interaction studies.
Abstract: An efficient transformation protocol is a primary requisite to study and utilize the genetic potential of any plant species. A quick transformation system is also crucial for the functional analysis of genes along with the study of proteins and their interactions in vivo. Presently, however, quick and effective transformation systems are still lacking for many plant species including pineapple. This has limited the full exploration of the genetic repository of pineapple as well as the study of its genes, protein localization and protein interactions. To address the above limitations, we have developed an efficient system for protoplast isolation and subcellular localization of desired proteins using pineapple plants derived from tissue culture. A cocktail of 1.5% (W/V) Cellulase R-10 and 0.5% (W/V) Macerozyme R-10 resulted in 51% viable protoplasts with 3 h digestion. Compared to previously reported protocols, our protoplast isolation method is markedly faster (saving 4.5 h), requires only a small quantity of tissue sample (1 g of leaves) and has high yield (6.5 × 105). The quality of the isolated protoplasts was verified using organelle localization in protoplasts with different organelle markers. Additionally, colocalization analysis of two pineapple Mg2+ transporter genes in pineapple protoplasts was consistent with the results in a tobacco transient expression system, confirming that the protoplast isolation method can be used to study subcellular localization. Further findings showed that the system is also suitable for protein–protein interaction studies. Based on our findings, the presently described method is an efficient and effective strategy for pineapple protoplast isolation and transformation; it is convenient and time saving and provides a greater platform for transformation studies.

44 citations


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Performance
Metrics
No. of papers in the topic in previous years
YearPapers
202358
2022153
202160
202060
201978
201855