Academic literature on the topic 'Protoplast proliferation'
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Journal articles on the topic "Protoplast proliferation"
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Sun, M., H. Kieft, and AAM van Lammeren. "Cotyledon-derived diploid and haploid protoplast culture and diploid plant regeneration in Brassica napus cv. ' Topas '." Canadian Journal of Botany 76, no. 3 (March 1, 1998): 530–41. http://dx.doi.org/10.1139/b98-022.
Full textAbstract:
The present paper describes a simple and reliable protocol for the successful isolation, purification, culture, and regeneration of diploid cotyledon-derived protoplasts of Brassica napus L. cv. 'Topas'. Various protoplast isolation media, nutrient media, subculture procedures, and protoplast sources were tested under two culture temperatures. Protoplast viability, cell wall regeneration, and cell division were monitored. Single cotyledon-derived protoplasts formed calli in liquid protoplast medium, and when these were subcultured on solid proliferation medium and solid regeneration medium of appropriate composition, plants regenerated either by shoot formation or embryogenesis. Continuous culture at 32°C instead of 25°C favoured the initiation of cell division and cell proliferation but prevented regeneration, although calli maintained regeneration capacity. Viable haploid protoplasts were isolated from cotyledons of heat-shock-induced, microspore-derived haploid embryos and from young leaves of secondary embryos that were formed on microspore-derived embryos. Cell divisions were triggered in the two types of haploid protoplast cultures, and microcalli were formed at high frequencies. Differences between haploid and diploid protoplast cultures are discussed.Key words: cotyledon protoplast culture, haploid culture, plant regeneration.
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Kang, Hyun Hee, Aung Htay Naing, and Chang Kil Kim. "Protoplast Isolation and Shoot Regeneration from Protoplast-Derived Callus of Petunia hybrida Cv. Mirage Rose." Biology 9, no. 8 (August 16, 2020): 228. http://dx.doi.org/10.3390/biology9080228.
Full textAbstract:
Despite the increasing use of protoplasts in plant biotechnology research, shoot regeneration from protoplasts remains challenging. In this study, we investigated the factors involved in protoplast isolation, callus induction, and shoot regeneration in Petunia hybrida cv. Mirage Rose. The following conditions were found to be most optimal for protoplast yield and viability: 0.6 M mannitol, 2.0% cellulase, and 6 h digestion time. A plating density of 10 × 104 protoplasts/mL under osmoticum condition (0.58 M mannitol) showed high microcolony viability in liquid culture. The Kao and Michayluk medium was found to be appropriate for callus proliferation from microcalli under a 16-h light photoperiod. Calli cultured in Murashige and Skoog medium containing 1.0 mg/L 6-benzylaminopurine and 0.2 mg/L 3-indole butyric acid showed the highest shoot regeneration frequency and number of shoots obtained per explant. Random amplification of polymorphic DNA analysis showed that the protoplast-derived shoots exhibited the same banding patterns as those of donor plants. Collectively, these findings can contribute to solving problems encountered in protoplast isolation and shoot regeneration in other petunia cultivars and related species. As the protocol developed by us is highly reproducible, it can be applied in biotechnology research on P. hybrida cv. Mirage Rose.
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Godel-Jędrychowska, Maćkowska, Kurczyńska, and Grzebelus. "Composition of the Reconstituted Cell Wall in Protoplast-Derived Cells of Daucus is Affected by Phytosulfokine (PSK)." International Journal of Molecular Sciences 20, no. 21 (November 4, 2019): 5490. http://dx.doi.org/10.3390/ijms20215490.
Full textAbstract:
Phytosulfokine-α (PSK), a peptidyl plant growth factor, has been recognized as a promising intercellular signaling molecule involved in cellular proliferation and dedifferentiation. It was shown that PSK stimulated and enhanced cell divisions in protoplast cultures of several species leading to callus and proembryogenic mass formation. Since PSK had been shown to cause an increase in efficiency of somatic embryogenesis, it was reasonable to check the distribution of selected chemical components of the cell walls during the protoplast regeneration process. So far, especially for the carrot, a model species for in vitro cultures, it has not been specified what pectic, arabinogalactan protein (AGP) and extensin epitopes are involved in the reconstruction of the wall in protoplast-derived cells. Even less is known about the correlation between wall regeneration and the presence of PSK during the protoplast culture. Three Daucus taxa, including the cultivated carrot, were analyzed during protoplast regeneration. Several antibodies directed against wall components (anti-pectin: LM19, LM20, anti-AGP: JIM4, JIM8, JIM13 and anti-extensin: JIM12) were used. The obtained results indicate a diverse response of the used Daucus taxa to PSK in terms of protoplast-derived cell development, and diversity in the chemical composition of the cell walls in the control and the PSK-treated cultures.
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Zhang, Demei, Rui Wang, Shijian Han, Zhigang Li, Jiming Xiao, Yangrui Li, Lingqiang Wang, and Suli Li. "Transcriptome Analysis of between Sugarcane Young Leaves and Protoplasts after Enzymatic Digestion." Life 12, no. 8 (August 9, 2022): 1210. http://dx.doi.org/10.3390/life12081210.
Full textAbstract:
Sugarcane somatic cell hybridization can break through the barrier of genetic incompatibility between distantly related species in traditional breeding. However, the molecular mechanisms of sugarcane protoplast regeneration and the conditions for protoplast preparation remain largely unknown. In this study, young sugarcane (ROC22) leaves were enzymatically digested, and the viability of protoplasts reached more than 90% after enzymatic digestion (Enzymatic combination: 2% cellulase + 0.5% pectinase + 0.1% dissociative enzyme + 0.3% hemicellulase, pH = 5.8). Transcriptome sequencing was performed on young sugarcane leaves and protoplasts after enzymatic digestion to analyze the differences in gene expression in somatic cells before and after enzymatic digestion. A total of 117,411 unigenes and 43,460 differentially expressed genes were obtained, of which 21,123 were up-regulated and 22,337 down-regulated. The GO terms for the 43,460 differentially expressed genes (DEGs) were classified into three main categories: biological process, cellular component and molecular function, which related to developmental process, growth, cell proliferation, transcription regulator activity, signal transducer activity, antioxidant activity, oxidative stress, kinase activity, cell cycle, cell differentiation, plant hormone signal transduction, and so on. After enzymatic digestion of young sugarcane leaves, the expressions of GAUT, CESA, PSK, CyclinB, CyclinA, CyclinD3 and cdc2 genes associated with plant regeneration were significantly down-regulated to 65%, 47%, 2%, 18.60%, 21.32%, 52% and 45% of young leaves, respectively. After enzymatic digestion, Aux/IAA expression was up-regulated compared with young leaves, and Aux/IAA expression was 3.53 times higher than that of young leaves. Compared with young leaves, these key genes were significantly changed after enzymatic digestion. These results indicate that the process of somatic enzymatic digestion process may affect the regeneration of heterozygous cells to a certain extent.
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Goverse, Aska, Jeroen Rouppe van der Voort, Charlotte Rouppe van der Voort, Annemieke Kavelaars, Geert Smant, Arjen Schots, Jaap Bakker, and Johannes Helder. "Naturally Induced Secretions of the Potato Cyst Nematode Co-stimulate the Proliferation of Both Tobacco Leaf Protoplasts and Human Peripheral Blood Mononuclear Cells." Molecular Plant-Microbe Interactions® 12, no. 10 (October 1999): 872–81. http://dx.doi.org/10.1094/mpmi.1999.12.10.872.
Full textAbstract:
Naturally induced secretions from infective juveniles of the potato cyst nematode Globodera rostochiensis co-stimulate the proliferation of tobacco leaf protoplasts in the presence of the synthetic phytohormones α-naphtha-leneacetic acid (NAA) and 6-benzylaminopurine (BAP). With the use of a protoplast-based bioassay, a low-molecular-weight peptide(s) (<3 kDa) was shown to be responsible for the observed effect. This mitogenic oligopeptide(s) is functionally dissimilar to auxin and cytokinin and, in addition, it does not change the sensitivity of the protoplasts toward these phytohormones. In combination with the mitogen phytohemagglutinin (PHA), cyst nematode secretions also co-stimulated mitogenesis in human peripheral blood mononuclear cells (PBMC). The stimulation of plant cells isolated from nontarget tissue—these nematodes normally invade the roots of potato plants—suggests the activation of a general signal transduction mechanism(s) by an oligopeptide(s) secreted by the nematode. Whether a similar oligopeptide-induced mechanism underlies human PBMC activation remains to be investigated. Reactivation of the cell cycle is a crucial event in feeding cell formation by cyst nematodes. The secretion of a mitogenic low-molecular-weight peptide(s) by infective juveniles of the potato cyst nematode could contribute to the redifferentiation of plant cells into such a feeding cell.
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Lee, Cheol Hee, Kee Yoeup Paek, J. Brian Power, and Edward C. Cocking. "ASSESSMENT OF THE LIMITATIONS OF SOMATIC HYBIDIZATION." HortScience 25, no. 9 (September 1990): 1164c—1164. http://dx.doi.org/10.21273/hortsci.25.9.1164c.
Full textAbstract:
This study was designed to assess the general limitations of somatic hybridization as one of the key technologies for genetic manipulation in plants. The limits of somatic hybridization against different taxonomic backgrounds, intraspecific to interfamilial, were also assessed. Protoplast culture studies provided essential information relating to the species cultural and morphogenetic capacity. several #elect Ion strategies for the recovery of somatic hybrid colonies/plants were developed and assessed using various combinations of protoplast sources and species in the genera Petunia, Nicotiana, Salpiglossis and Chrysanthemum. Morphological, cytological and biochemical analyses were performed to confirm the hybridity of plants or cell lines recovered following protoplasm fusion (using 4-5 methods) and selection.The somatic hybrid callus/plants were obtained at intraspecific to interfamilial levels by complementation to chlorophyll proficiency, together with media selection or complementation of nitrate reductase deficient mutants as follows; P. Hybrida var. Monsanto (+) P. hybrida cv. Blue Lace (intraspecific), P. hybrida var. Monsanto (+) P. inflata and P. parviflora (interspecific), P. parviflora (+) N. tabacum (intergeneric), S. sinuata (+) P. hybrida var. Monsanto, P. parodii and N. tabacum (intertribal), and C. morifolium (+) S. sinuata.From this study, it appeared that there were no taxonomic limits to the production and proliferation of somatic hybrid cell lines. However, obtaining morphologically normal hybrid plants met with limited success as the taxonomic relationships became more distant. The regeneration capacity of somatic hybrids seemed to be controlled by both parental species. Somatic incompatibility mechanism was also shown to operate on chromosome elimination. Such chromosome elimination may well be advantageous in plant improvement.
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Tran, Huong Thanh, and Cuong Quoc Vo. "Protoplast isolation and culture from different explants of Musa spp. Cau man." Science and Technology Development Journal - Natural Sciences 1, no. 6 (December 7, 2018): 106–16. http://dx.doi.org/10.32508/stdjns.v1i6.621.
Full textAbstract:
In this paper, the roles of type and concentration of enzymes on protoplast isolation from in vitro leaves, multi-scalps (highly proliferating meristem culture), and young male flower of banana cv. cau man were studied. Respiration rate and content of plant hormones of these materials were analysed. Different techniques were used to culture these protoplasts. The development of protoplasts was observed under fluorescence microscope. The highest yield of protoplast (69.5 x 106 protoplasts / g fresh weight) was obtained from young male flowers after 16 hours treatment with 1.5 % cellulase, 0.25 % pectinase and 0.25 % hemicellulase. The combination of hanging drop cell technique (in 6 days), and carrot feeder layer cells in N6PKM medium supplemented with 0.2 mg/L 2,4- dichlorophenoxyacetic acid (2,4-D), 1 mg/L α- naphthalene acetic acid (NAA), and 0.5 mg/L zeatin are suitable for protoplast development. Protoplasts that were isolated from multi-scalps and young male flowers created the wall and divided when cultured by this method. The development of protoplasts from young male flower began with cell walls creation after 4 days, the cell division was after 6 days, and small colonies formation was after 28 days of culture. The differentiation and physiological activity of cells play an important role on quantity and quality of protoplasts, as on the well as protoplast development.
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Grymaszewska, Grażyna, and Władysław Golinowski. "Structure of syncytia induced by Heterodera schachtii Schmidt in roots of susceptible and resistant radish (Raphanus sativus L., var. oleiformis)." Acta Societatis Botanicorum Poloniae 67, no. 3-4 (2014): 207–16. http://dx.doi.org/10.5586/asbp.1998.024.
Full textAbstract:
The structure of syncytia induced by <i>Heterodera schachtii</i> Schmidt in roots of susceptible <i>Raphanus sativus</i> L. cv. "Siletina" and resistant radish cv. "Pegletta" was investigated. In the radish cultivar "Siletina" the syncytia most often appeared in the elongation zone of lateral roots. They were initiated in the procambium and pericycle but also included the parenchyma cells of vascular cylinder. In the susceptible cultivar "Siletina" the cells forming the female's syncytia were subject to hypertrophy. Their cytoplasmic density increased. The cytoplasm contained numerous organella. The proliferation of the smooth endoplasmic reticulum took place. Branched cell wall ingrowths were formed next to the vessels. In the male's syncytia the cells were only slightly increased. Their protoplasts contained few organelles. The cell wall ingrowths were poorly developed. In the syncytia of the resistant cultivar "Pegletta" there was only a slight increase of the cell volume. A well developed system of rough endoplasmic reticulum was observed in the protoplast. Distended ER cisterns contained fine fibrillar material. Material of similar structure also appeared in numerous small vacuoles. In resistant plants only some, not numerous, syncytia spreading in procambium fully developed and functioned long enough for the parasite females to mature. At an advanced stage of infection a well developed system of a rough ER was observed also in those syncytia and numerous vacuoles appeared.
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Yang, Honglin, Yihua Yang, Qiang Wang, Jinyu He, Liyun Liang, Hui Qiu, Yue Wang, and Lijuan Zou. "Adventitious Shoot Regeneration from Leaf Explants in Sinningia Hybrida ‘Isa’s Murmur’." Plants 11, no. 9 (May 2, 2022): 1232. http://dx.doi.org/10.3390/plants11091232.
Full textAbstract:
As a valuable ornamental plant, Sinningia hybrida ‘Isa’s Murmur’ (S. hybrida) has genetic flower diversity, which has great potential to develop different flower characters in the horticultural market. The present study focuses on establishing a practical approach for the sustainable propagation of S. hybrida. Compared with aseptic seeding leaves explants, field-grown leaves explants are more suitable for adventitious shoot regeneration. Adding 0.1 mg L−1 NAA and 2.0 mg L−1 TDZ could obtain the highest adventitious shoot proliferation coefficient (24.5), and the induction rate was 91.7%. The shoot proliferation coefficient (20.7) and the greatest shoot length and induction rate (95.3%) were achieved in 0.1 mg L−1 NAA and 2.0 mg L−1 BA medium, accompanied by rooting formation. Adding 0.5 mg L−1 GA3, 1.0 mg L−1 BA, and 0.2 mg L−1 IBA to MS medium can effectively prolong the regenerated buds for rooting. The best for rooting was 1/2 MS medium containing 0.3 mg L−1 IBA, with the maximum number of roots (13.4 per shoot) and survival rate for transplanting (100%). This work aims to build an efficient, definitive, and scalable protocol for S. hybrida regeneration useful for large-scale cultivation and even more protoplast fusion and genetic transformation to develop more colorful or fragrant flowers.
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Chen, L., S. O. Park, S. Dhir, and A. S. Bhagsari. "331 Effects of Explant Type, Sucrose Level and Callus Development Time on In Vitro Plant Regeneration of Sweetpotato." HortScience 35, no. 3 (June 2000): 449A—449. http://dx.doi.org/10.21273/hortsci.35.3.449a.
Full textAbstract:
Several experiments were conducted to evaluate the influence of explant type, sucrose level, and callus development time on sweetpotato [Ipomoea batatas (L.) Lam] in vitro culture. Shoot tip, petiole, and leaf of Selection 75-96-1 was used as explants in Murashige and Skoog (MS) media with different plant growth regulators. Calli derived from shoot tip and petiole produced 42.1% and 10.3% somatic embryos, respectively, but the leaf failed to produce somatic embryos. The effect of sucrose level was determined using shoot tip as explants. Compared with 3% sucrose in the same plant growth regulators level medium during callus initiation and callus proliferation periods, 5% sucrose level suppressed root growth and improved shoot regeneration. The callus development time was measured by using shoot tips on callus initiation medium containing 1.5 mg/L alpha-Naphthaleneacetic acid (NAA) and 0.25 mg/L Kinetin (KIN) plus 5% sucrose. When explants were cultured for less than 6 weeks during callus initiation, then transferred onto plant regeneration medium, plant regeneration via organogenesis occurred; whereas, maintaining cultures for more than 12 weeks on the same callus initiation/proliferation medium, plant regeneration was favored via embryogenesis. Explant type and other factors affecting plant regeneration noted here could be applied to protoplast culture, somatic hybridization, and transformation in sweetpotato.
Dissertations / Theses on the topic "Protoplast proliferation"
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de, Jong Femke. "Analysis of protoplasts and somatic embryogenesis in Medicago truncatula." Phd thesis, 2006. http://hdl.handle.net/1885/7161.
Full textAbstract:
This thesis examined protoplast proliferation and somatic embryogenesis, by
comparing a highly with a poorly embryogenic Medicago truncatula line through microscopic, proteomic and in situ hybridization analysis. Proteome analysis of M. truncatula was used to identify proteins involved in protoplast proliferation and the
initiation of somatic embryogenesis. Furthermore, an in situ hybridization study was done to compare the expression of genes known to be involved in zygotic embryogenesis with the expression during somatic embryogenesis. A large number
of proteins were up-, and down-regulated during the first 5 days of protoplast culture
indicating that cellular reorganization took place. An up-regUlation of PR 1 O-like
proteins and flavonoid synthesis proteins and a down-regulation of energy
metabolism proteins were observed, indicating an initiation of a stress response. The observed stress response in protoplasts was down-regulated before the first cell divisions at 5-7 d. A stress-inducing bioassay on protoplasts showed that the ability of protoplasts to overcome stress and to proliferate under stress conditions depended on the level of stress and density of the protoplast culture, whereby more stress or a lower culture density resulted in higher levels of cell death. Proteomic analysis of the initiation of somatic embryogenesis showed that similar metabolic pathways were involved in the initiation of somatic embryogenesis and protoplast proliferation. By using a highly embryogenic (2HA) line, and a poorly embryogenic (A 17) line of M. truncatu!a, it was shown that particular proteins were specifically accumulated during the initiation of somatic embryogenesis. A high accumulation of a peroxidase was observed only in At7 tissue at the time of initiation of somatic embryogenesis and might be the reason why the initiation of somatic embryogenesis is inhibited in A 17 tissue. The specific accumulation of flavonoid synthesis proteins might also indicate that flavonoids are involved during the initiation of somatic embryogenesis.
In situ hybridization with probes to genes known to be involved in zygotic embryogenesis, showed that M. truncatula somatic and Arabidopsis thaliana zygotic
embryogenesis both followed similar developmental pathways. However, a few
genes showed distinct patterns of gene expression in M. truncatula somatic embryos.ARC Centre of Excellence for Integrative Legume Research for funding and scholarship