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Journal articles on the topic 'Protoplast'

Author: Grafiati

Published: 4 June 2021

Last updated: 25 July 2025

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1

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 (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 a

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2

Sinha, Anupam, Andrew C. Wetten, and P. D. S. Caligari. "Effect of biotic factors on the isolation of Lupinus albus protoplasts." Australian Journal of Botany 51, no. 1 (2003): 103. http://dx.doi.org/10.1071/bt01104.

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Several tissue types of Lupinus albus L. were investigated as sources for the isolation of protoplasts. Cotyledons from in vitro seedlings were found to yield the highest number of protoplasts compared with leaves, hypocotyls and roots. A combination of the protoplast isolation enzymes, cellulase and Pectolyase Y23, was capable of releasing the highest number of protoplasts compared with a combination of cellulase and Macerase. Protoplast yield increased with increasing cotyledon age but was accompanied by a progressive decline in protoplast viability. The optimal combination of protoplast yie

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3

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 (1998): 530–41. http://dx.doi.org/10.1139/b98-022.

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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

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4

Struck, C., R. Rohringer, and R. Heitefuss. "Isolation and lectin-binding properties of barley epidermal and mesophyll protoplasts." Canadian Journal of Botany 72, no. 11 (1994): 1688–91. http://dx.doi.org/10.1139/b94-207.

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Protoplasts from primary leaves of barley (Hordeum vulgare L.) were obtained by enzymatic digestion and fractionated by discontinuous density gradient centrifugation to yield highly enriched fractions of mesophyll and epidermal protoplasts. A characterization of both protoplast types resulted in a clear differentiation of the outer protoplast surfaces. The protoplasts were examined for affinity to various lectins by agglutination tests and by labeling with lectin – fluorescein isothiocyanate conjugates. Both types of protoplasts agglutinated with soybean lectin. Fluorescein isothiocyanate-labe

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5

Liu, Donglong, and Nancy A. Reichert. "PROTOPLAST ISOLATION AND CULTURE OF KENAF (HIBISCUS CANNABINUS L.)." HortScience 29, no. 7 (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 div

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6

Han, Jong-Eun, Han-Sol Lee, Hyoshin Lee, Hyunwoo Cho, and So-Young Park. "Embryogenic Stem Cell Identity after Protoplast Isolation from Daucus carota and Recovery of Regeneration Ability through Protoplast Culture." International Journal of Molecular Sciences 23, no. 19 (2022): 11556. http://dx.doi.org/10.3390/ijms231911556.

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Protoplasts are single cells isolated from tissues or organs and are considered a suitable system for cell studies in plants. Embryogenic cells are totipotent stem cells, but their regeneration ability decreases or becomes lost altogether with extension of the culture period. In this study, we isolated and cultured EC-derived protoplasts (EC-pts) from carrots and compared them with non-EC-derived protoplasts (NEC-pts) with respect to their totipotency. The protoplast isolation conditions were optimized, and the EC-pts and NEC-pts were characterized by their cell size and types. Both types of p

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7

Ahmed, Mohamed A. A., Miao Miao, Emmanouil D. Pratsinakis, et al. "Protoplast Isolation, Fusion, Culture and Transformation in the Woody Plant Jasminum spp." Agriculture 11, no. 8 (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 t

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8

Li, Xiaobin, Ying Qin, Yufei Kong, Samantha Chandranath Karunarathna, Yunjiang Liang, and Jize Xu. "Optimization of Protoplast Preparation Conditions in Lyophyllum decastes and Transcriptomic Analysis Throughout the Process." Journal of Fungi 10, no. 12 (2024): 886. https://doi.org/10.3390/jof10120886.

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Protoplasts are essential tools for genetic manipulation and functional genomics research in fungi. This study systematically optimized protoplast preparation conditions and examined transcriptional changes throughout the preparation and regeneration processes to elucidate the molecular mechanisms underlying the formation and regeneration of protoplasts in Lyophyllum decastes. The results indicated an optimal protoplast yield of 5.475 × 106 cells/mL under conditions of fungal age at 10 days, digestion time of 2.25 h, enzyme concentration of 2%, and digestion temperature of 28 °C. The Z5 medium

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9

Kurita-Tashiro, Asami, Noriko Hayashi, Tomoya Oyanagi, and Hamako Sasamoto. "New Factors for Protoplast-Callose-Fiber Formation in Salt-Tolerant Mangrove Plants, Avicennia alba and Bruguiera sexangula and Analysis of Fiber Substructures." Journal of Plant Studies 9, no. 2 (2020): 1. http://dx.doi.org/10.5539/jps.v9n2p1.

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Elongated and spiral &beta;-1,3-glucan (callose) fibers were obtained by new factors from protoplasts cultured in liquid medium from suspension cultured cells of two salt-tolerant mangrove species; Avicennia alba and Bruguiera sexangula. Differences in salt factor for protoplast-fiber formation were compared with those of the callose fibers developed from protoplasts of non-mangrove tree plants, Larix leptolepis and Betula platyphylla, which high concentrations of divalent cations, Mg2+ (50 mM) or Ca2+ (100 mM), were stimulatory. In the halophilic A. alba protoplasts, whose cell division w

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10

Shao, Yingying, Detian Mu, Limei Pan, et al. "Optimization of Isolation and Transformation of Protoplasts from Uncaria rhynchophylla and Its Application to Transient Gene Expression Analysis." International Journal of Molecular Sciences 24, no. 4 (2023): 3633. http://dx.doi.org/10.3390/ijms24043633.

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Protoplast-based engineering has become an important tool for basic plant molecular biology research and developing genome-edited crops. Uncaria rhynchophylla is a traditional Chinese medicinal plant with a variety of pharmaceutically important indole alkaloids. In this study, an optimized protocol for U. rhynchophylla protoplast isolation, purification, and transient gene expression was developed. The best protoplast separation protocol was found to be 0.8 M D-mannitol, 1.25% Cellulase R-10, and 0.6% Macerozyme R-10 enzymolysis for 5 h at 26 °C in the dark with constant oscillation at 40 rpm/

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11

Suryati, Emma, Andi Tenriulo, and Sri Rejeki Hesti Mulyaningrum. "ISOLASI DAN KULTUR PROTOPLAS RUMPUT LAUT Kappaphycus alvarezii DI LABORATORIUM." Jurnal Riset Akuakultur 2, no. 3 (2007): 399. http://dx.doi.org/10.15578/jra.2.3.2007.399-405.

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Isolasi protoplas rumput laut K. alvarezii, telah dilakukan dalam rangka penyiapan protoplas untuk penyilangan melalui fusi protoplas. Metode yang digunakan antara lain melalui cara kimia dengan melisis tallus rumput laut dengan campuran enzim komersial, kemudian enzim yang berasal dari viscera keong mas baik yang segar maupun yang beku, dengan media kultur yang digunakan pada pemeliharaan makro algae antara lain Conwy, PES, dan air laut steril. Tallus rumput laut yang digunakan berasal dari bagian pangkal, tengah dan ujung. Protoplas yang hidup diuji menggunakan evans blue 0,1%, hormon perang

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12

Sinha, Anupam, and Peter D. S. Caligari. "Aspects of isolation underpinning mitotic behaviour in lupin protoplasts." Australian Journal of Botany 52, no. 5 (2004): 669. http://dx.doi.org/10.1071/bt03153.

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This study reports on the influence of critical isolation factors on the subsequent culture of protoplasts of Lupinus albus L. Protoplasts were isolated from in vitro seedling cotyledons of five early maturing accessions in which protoplast yields and division frequencies appeared to be correlated as a high protoplast yield corresponded with a high division frequency. The overall difference among the accessions for mitosis was non-significant, although the highest yield and division frequency were observed in accession LA132, with Alban giving a significantly lower level. Accession Lucrop prod

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13

Monthony, Adrian S., and Andrew Maxwell P. Jones. "Enhancing Protoplast Isolation and Early Cell Division from Cannabis sativa Callus Cultures via Phenylpropanoid Inhibition." Plants 13, no. 1 (2024): 130. http://dx.doi.org/10.3390/plants13010130.

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De novo regeneration of Cannabis sativa L. (cannabis) using tissue culture techniques remains unreliable and infrequent. Conventional methods for the regeneration and transformation of cannabis have not achieved the reliability and replicability that need to be integrated into research and breeding programs. Protoplast systems are effective for gene expression studies and transformation and genome-editing technologies and open the possibility of somatic hybridization to create interspecific hybrids. To date, leaf-derived protoplasts have been isolated for transient gene expression studies, but

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14

Leinhos, Volker, and Rodney Arthur Savidge. "Isolation of protoplasts from developing xylem of Pinusbanksiana and Pinusstrobus." Canadian Journal of Forest Research 23, no. 3 (1993): 343–48. http://dx.doi.org/10.1139/x93-050.

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Protoplasts were isolated from developing xylem of Pinusbanksiana Lamb, and Pinusstrobus L. by incubating freshly harvested tissue in a cellulose–pectinase mixture having mannitol as osmoticum. Protoplasts were then purified using a discontinuous sucrose–mannitol gradient. More than 70% of the isolated protoplasts were of small diameter (12–27 μm) and had dense cytoplasm and many small vacuoles, suggesting that they originated from ray cells. Larger protoplasts constituted about 25% of the protoplast population; these contained single large vacuoles and only parietal cytoplasm, suggesting that

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15

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 (2018): 106–16. http://dx.doi.org/10.32508/stdjns.v1i6.621.

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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

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16

Pauk, János, Sándor Fekete, Juha Vilkki, and Seppo Pulli. "Protoplast culture and plant regeneration of different agronomically important Brassica species and varieties." Agricultural and Food Science 63, no. 5 (1991): 371–78. http://dx.doi.org/10.23986/afsci.72416.

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Protoplast cultures were prepared from 6-day-old hypocotyls of six spring, seven winter cultivars of Brassica napus L. and one line of Brassica campestris L. The molarity of enzyme solution was raised to 0,714 M mannitol resulting in well manipulable, cytoplasm dense protoplasts. In the protoplast purification procedure density gradient centrifugation was used to minimize physical damage of protoplasts. Three different protoplast culture systems —(1) liquid, (2) 2nd day embedded, (3) directly embedded in low melting agarose were compared. The two different protoplast embedding techniques resul

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17

Xu, Mengxue, Qingwei Du, Caihuan Tian, Ying Wang, and Yuling Jiao. "Stochastic gene expression drives mesophyll protoplast regeneration." Science Advances 7, no. 33 (2021): eabg8466. http://dx.doi.org/10.1126/sciadv.abg8466.

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Cell pluripotency is fundamental to biology. It has long been known that differentiated somatic plant cells may reacquire pluripotency, but the underlying mechanism remains elusive. In many plant species, a single isolated mesophyll protoplast may regenerate into an entire plant, which is widely used in gene transformation. Here, we identified two transcription factors whose ectopic activation promotes protoplast regeneration. Furthermore, we found that their expression was induced by protoplast isolation but at a very low frequency. Using live imaging and single-cell transcriptomics, we show

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18

Adjei, Mark Owusu, Huan Zhao, Xiaoguang Tao, et al. "Using A Protoplast Transformation System to Enable Functional Studies in Mangifera indica L." International Journal of Molecular Sciences 24, no. 15 (2023): 11984. http://dx.doi.org/10.3390/ijms241511984.

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Mangoes (Mangifera indica L.) are an important kind of perennial fruit tree, but their biochemical testing method and transformation technology were insufficient and had not been rigorously explored. The protoplast technology is an excellent method for creating a rapid and effective tool for transient expression and transformation assays, particularly in plants that lack an Agrobacterium-mediated plant transformation system. This study optimized the conditions of the protoplast isolation and transformation system, which can provide a lot of help in the gene expression regulation study of mango

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19

Lee, Chang-Hoo, N. I. Hyung, and S. E. Kim. "SOME FACTORS AFFECTING ISOLATION OF MESOPHYLL PROTOPLASTS FROM APPLE (MALUS DOMESTICA BORKH CV. FUJI)." HortScience 27, no. 6 (1992): 693g—694. http://dx.doi.org/10.21273/hortsci.27.6.693g.

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Experiments were conducted to investigate the factors influencing mesophyll protoplast isolation in `Fuji' apple. Half an hour pretreatment in 0.6M mannitol gave the highest protoplast yield.The enzyme solution containing 2% Cellulase Onozuka R-10 and 0.5% macrozyme R-10 with CPW 0.6M mannitol at pH 5.5 was most effective for protoplast isolation from leaf. Effective incubation time for the enzyme treatment was found to be 15-20 hrs at 25°C in the dark. Use of 1.0-2.0% PVP and 0.5mM MES was essential for higher yield and viability of protoplast. Supplementation of BA and IBA to the shoot cultu

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20

Qiu, Quan-Sheng, Ze-Zhou Wang, Nang Zhang, Qi-Gui Cai, and Rong-Xi Jiang. "Aquaporins in the plasma membrane of leaf callus protoplasts of Actinidia deliciosa var. deliciosa cv. Hayward." Functional Plant Biology 27, no. 1 (2000): 71. http://dx.doi.org/10.1071/pp99033.

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The water transport activity of Actinidia deliciosa protoplasts was determined using a cell imaging system. Results showed that the protoplast volume increased swiftly when placed in a hypoton-ic medium, and also increased with an increase in medium osmotic gradients. The osmotic water permeability coefficient (Pf) values were 0.118 × 10–3, 0.121 × 10–3, and 0.133 × 10–3 cm s–1 when the osmotic gradients were 75, 100, and 125 mosmol, respectively. The water transport activity of protoplas-ts could be inhibited by HgCl2 and stimulated by amphotericin B. Moreover, ZnCl2 and ZnSO4 had a significa

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Meng, Ruirui, Chenchen Wang, Lihua Wang, et al. "An efficient sorghum protoplast assay for transient gene expression and gene editing by CRISPR/Cas9." PeerJ 8 (October 13, 2020): e10077. http://dx.doi.org/10.7717/peerj.10077.

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Protoplasts are commonly used in genetic and breeding research. In this study, the isolation of sorghum protoplasts was optimized and applied to transient gene expression and editing by CRISPR/Cas9. The protoplast was most viable in 0.5 M mannitol, which was the highest of three concentrations after 48- and 72-hours treatments. Using this method we can derive an average of 1.6×106 cells which vary from 5 to 22 nm in size. The average transfection of the protoplasts was 68.5% using the PEG-mediated method. The subcellular assays located Sobic.002G279100-GFP and GFP proteins in the cell compartm

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22

Reni Mahmudah, Edy Setiti Wida Utami, Yosephine Sri Wulan Manuhara, Junairiah, and Sucipto Hariyanto. "Improvements in Protoplast Isolation Protocol of Dendrobium antennatum Lindl." Open Access Research Journal of Science and Technology 13, no. 2 (2025): 108–15. https://doi.org/10.53022/oarjst.2025.13.2.0049.

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This study aimed to determine the effect of a combination of enzyme concentrations, incubation time, and osmoticum concentrations on protoplast isolation mesophyll of leaves of Dendrobium antennatum Lindl. This was an experimental study using a 3×3×3 factorial design and three replications.The treatment variable was a combination of 3 levels of enzyme concentration (0.5% cellulase + 0.2% pectinase, 1% cellulase + 0.2% pectinase, and 1.5% cellulase + 0.2% pectinase), 3 levels of long incubation time (6 hours, 9 hours, and 12 hours), and 3 levels of osmoticum concentration (0.3 M sucrose, 0.4 M

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23

Moon, Ki-Beom, Ji-Sun Park, Su-Jin Park, et al. "A More Accessible, Time-Saving, and Efficient Method for In Vitro Plant Regeneration from Potato Protoplasts." Plants 10, no. 4 (2021): 781. http://dx.doi.org/10.3390/plants10040781.

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Both obtaining high-yielding, viable protoplasts and following reliable regeneration protocols are prerequisites for the continuous expansion and development of newly emerging systems involving protoplast utilization. This study determines an efficient process from protoplast isolation to shoot regeneration in vitro. The maximum yield of protoplast extraction, which was 6.36 ± 0.51 × 106 protoplasts/g fresh weight (FW), was approximately 3.7 times higher than that previously reported for potato protoplasts. To obtain data, wounded leaves were used by partially cutting both sides of the midrib,

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Stajič, Ester. "Improvements in Protoplast Isolation Protocol and Regeneration of Different Cabbage (Brassica oleracea var. capitata L.) Cultivars." Plants 12, no. 17 (2023): 3074. http://dx.doi.org/10.3390/plants12173074.

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Protoplasts are a versatile tool in plant biotechnology since they can be used for basic biological studies as well as for breeding strategies based on genome editing. An efficient protoplast isolation protocol is essential for conducting protoplast-based studies. To optimize the protoplast isolation protocol in cabbage (Brassica oleracea var. capitata L.), different enzyme solutions were tested for the isolation of leaf mesophyll protoplasts. In our experiments, the combination of 0.5% Cellulase Onozuka RS and 0.1% Macerozyme R-10 showed the best result. The optimized protocol proved suitable

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Rahayu Opi Anggoro, Decenly, Yosua Hambit, and Nanang Wahyu Prajaka. "Establishment of A Transient Expression Using PEG-Mediated Protoplast Transformation System in Black Rice Cempo Ireng." Journal of Biotropical Research and Nature Technology 2, no. 1 (2023): 43–49. https://doi.org/10.52850/borneo.v2i1.11019.

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Indonesia black rice is a potential crop which consider to be develop as functional food because of high nutritional values. However, some agronomical traits, such as high culm and long harvesting time need to be improved for high productivity. Genetic engineering based on protoplast system is one of tools that can be used for improving black rice agronomical traits. The purpose of this study was to establish an efficient method for obtaining protoplasts, and to get information on whether the PEG-mediated transformation method can be carried out on black rice ‘Cempo Ireng’ using GFP transient

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Ren, Rui, Jie Gao, Chuqiao Lu, et al. "Highly Efficient Protoplast Isolation and Transient Expression System for Functional Characterization of Flowering Related Genes in Cymbidium Orchids." International Journal of Molecular Sciences 21, no. 7 (2020): 2264. http://dx.doi.org/10.3390/ijms21072264.

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Protoplast systems have been proven powerful tools in modern plant biology. However, successful preparation of abundant viable protoplasts remains a challenge for Cymbidium orchids. Herein, we established an efficient protoplast isolation protocol from orchid petals through optimization of enzymatic conditions. It requires optimal D-mannitol concentration (0.5 M), enzyme concentration (1.2 % (w/v) cellulose and 0.6 % (w/v) macerozyme) and digestion time (6 h). With this protocol, the highest yield (3.50 × 107/g fresh weight of orchid tissue) and viability (94.21%) of protoplasts were obtained

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Li, Cai Hong, De Feng Xu, Ri Ying Ye, Ya Ling Wang, and Li Jun Sun. "An Efficient Method for Monitoring and Improving the Regeneration Efficiency of Protoplasts from Aspergillus Oryzae HN3042." Advanced Materials Research 781-784 (September 2013): 808–11. http://dx.doi.org/10.4028/www.scientific.net/amr.781-784.808.

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Protoplast technique is a powerful tool for strain improvement and the preparation of protoplasts with high regeneration efficiency is the basis of subsequent manipulations. In this study, the effects of enzymatic hydrolysis time on release and regeneration of protoplasts from Aspergillus oryzae HN3042 were investigated by analyzing the time-series image. The results showed that there was a significant correlationship between regeneration efficiency and morphological size of protoplasts (P < 0.01). An efficient method for monitoring and improving the regeneration efficiency of protoplast wa

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CHAMANI, Esmaeil, Seyyed Karim TAHAMI, Nasser ZARE, Rasool Asghari-ZAKARIA, Mehdi MOHEBODINI, and Daryl JOYCE. "Effect of Different Cellulase and Pectinase Enzyme Treatments on Protoplast Isolation and Viability in Lilium ledebeourii Bioss." Notulae Botanicae Horti Agrobotanici Cluj-Napoca 40, no. 2 (2012): 123. http://dx.doi.org/10.15835/nbha4028055.

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For overcoming interspecific incompatibility, protoplast combination method is a proper procedure for making a new plant withdesired traits. For this purpose, protoplast preparation is a first and important step. Hence, experiments were conducted to evaluatevarious combinations of cellulose, pectinase and their treatment times on protoplast production and protoplast viability in Liliumledebeourii Bioss. The results of experiment revealed that the protoplast yield was significantly affected by different treatment levels.Cellulase at 4% gave the highest numbers of protoplasts at 3.71Ã—105 protop

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Attree, S. M., D. I. Dunstan, and L. C. Fowke. "Initiation of embryogenic callus and suspension cultures, and improved embryo regeneration from protoplasts, of white spruce (Picea glauca)." Canadian Journal of Botany 67, no. 6 (1989): 1790–95. http://dx.doi.org/10.1139/b89-227.

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Rapid and high frequency somatic embryo regeneration from protoplasts isolated from 10 embryogenic cell lines of white spruce (Picea glauca) is reported. Embryogenic callus was initiated from immature zygotic embryos as source material for protoplast isolation. Individual cell lines exhibited different capabilities for sustained growth. Protoplast plating efficiencies depended on the concentrations of macroelements included in the medium. Using a medium with reduced salts, individual protoplasts developed directly into embryos with no disorganized growth period. Protoplasts from newly establis

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Kantharajah, AS, and WA Dodd. "Factors That Influence the Yield and Viability of Cucumber (Cucumis sativus L) Cotyledon Protoplasts." Australian Journal of Botany 38, no. 2 (1990): 169. http://dx.doi.org/10.1071/bt9900169.

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Protoplasts isolated from cotyledons of aseptically germinated cucumber seedlings were divided into three size classes. The relationships between tissue age, isolation procedure, yield and protoplast size were investigated. During germination and up to an age of 13 days, the percentage of protoplasts in each size class underwent considerable change with a big reduction in percentage of the largest protoplasts in older cotyledons. Protoplast size and yield could also be manipulated by varying the isolation technique. In this context, temperature, incubation time and shaker speed were significan

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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 (2020): 228. http://dx.doi.org/10.3390/biology9080228.

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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 med

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Zeng, Xiaojian, Xiaolei Cao, Qiuyue Zhao, et al. "Isolation of Haustorium Protoplasts Optimized by Orthogonal Design for Transient Gene Expression in Phelipanche aegyptiaca." Plants 13, no. 15 (2024): 2163. http://dx.doi.org/10.3390/plants13152163.

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The efficient protoplast transient transformation system in plants is an important tool to study gene expression, metabolic pathways, and various mutagenic parameters, but it has not been established in Phelipanche aegyptiaca. As a root parasitic weed that endangers the growth of 29 species of plants in 12 families around the world, there is still no good control method for P. aegyptiaca. Even the parasitic mechanisms of P. aegyptiaca and the related genes regulating parasitism are not yet understood. In this study, by comparing the factors related to protoplast isolation and transfection, we

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Ishikawa, Francine Hiromi, Quélen de Lima Barcelos, Elaine Aparecida de Souza, and Eustáquio Souza Dias. "Factors affecting the production and regeneration of protoplasts from Colletotrichum lindemuthianum." Ciência e Agrotecnologia 34, no. 1 (2010): 74–79. http://dx.doi.org/10.1590/s1413-70542010000100009.

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The present work reports factors affecting the production and regeneration of protoplasts from Colletotrichum lindemuthianum. The usefulness of protoplast isolation is relevant for many different applications and has been principally used in procedures involving genetic manipulation. Osmotic stabilizers, lytic enzymes, incubation time and mycelial age were evaluated in terms of their effects on protoplast yield. The optimal condition for protoplast production included the incubation of young mycelia (48 h) in 0.6 mol l-1 NaCl as the osmotic stabilizer, with 30 mg ml-1 Lysing Enzymes from Trich

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Zhang, Huali, Qianqian Geng, Zhanghua Sun, et al. "A Dual-Channel Microfluidic Chip for Single Tobacco Protoplast Isolation and Dynamic Capture." Micromachines 13, no. 12 (2022): 2109. http://dx.doi.org/10.3390/mi13122109.

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Protoplasts are widely used in gene function verification, subcellular localization, and single-cell sequencing because of their complete physiological activities. The traditional methods based on tissues and organs cannot satisfy the requirement. Therefore, the isolation and capture of a single protoplast are most important to these studies. In this study, a dual-channel microfluidic chip based on PDMS with multi-capture cavities was designed. The design theory of the dual-channel microfluidic chip’s geometry was discussed. The capture mechanism of the single cell in a dual-channel microfluid

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35

Echeverria, Ed. "Preparation and Characterization of Protoplasts from Citris Juice Vesicles." Journal of the American Society for Horticultural Science 112, no. 2 (1987): 393–96. http://dx.doi.org/10.21273/jashs.112.2.393.

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Abstract A method for the preparation of protoplasts from juice vesicles of citrus fruits is described. Protoplasts were obtained after incubating juice vesicles for 16 hr at room temperature in a medium containing cellulase and pectinase. The solution containing the protoplast was filtered through a 200-μm nylon mesh and the filtrate layered on a Percoll density gradient of 4%, 3.5%, and 3%. After 1 to 2 hr at 4°C, protoplasts were collected from the 4% Percoll layer. Protoplast viability was assessed by different methods and estimated to be ≈ 85%. The use of several stains and observations w

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36

Navrátilová, B. "Protoplast cultures and protoplast fusion focused on Brassicaceae: A review." Horticultural Science 31, No. 4 (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

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37

Tamaru, Yutaka, Sadaharu Ui, Koichiro Murashima, et al. "Formation of Protoplasts from Cultured Tobacco Cells and Arabidopsis thaliana by the Action of Cellulosomes and Pectate Lyase from Clostridium cellulovorans." Applied and Environmental Microbiology 68, no. 5 (2002): 2614–18. http://dx.doi.org/10.1128/aem.68.5.2614-2618.2002.

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ABSTRACT The crude culture supernatants from Clostridium cellulovorans were tested for their ability to convert plant cells to protoplasts. The supernatants readily released protoplasts from cultured tobacco cells and Arabidopsis thaliana. The crude culture supernatant from pectin-grown cells was more active than supernatants from glucose-, cellobiose-, xylan-, and locust bean gum-grown cells. After removal of cellulosomes, the crude culture supernatant lost its protoplast formation activity. The protoplast formation activity of the crude culture supernatant from C. cellulovorans was more effe

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Mieth, Hannelore, Volker Speth, and Jürgen Ebel. "Phytoalexin Production by Isolated Soybean Protoplasts." Zeitschrift für Naturforschung C 41, no. 1-2 (1986): 193–201. http://dx.doi.org/10.1515/znc-1986-1-229.

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Abstract Protoplasts isolated enzymatically from suspension-cultured cells of soybean (Glycine max L. Merr., cv. Harosoy 63) were used to study the production of the isoflavonoid-derived phytoalexin, glyceollin. A large enhancement in the in vivo rates of synthesis and catalytic activities of two of the enzymes associated with glyceollin biosynthesis, phenylalanine ammonia-lyase and chalcone synthase, preceded phytoalexin accumulation during early stages of culture of isolated protoplasts while cell wall regeneration occurred. A glucan elicitor from cell walls of the fungus Phytophthora megas

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Saxena, Praveen K., and John King. "Optimal conditions for the isolation and culture of protoplasts from a cell suspension culture of an isoleucine–valine-requiring auxotroph of Datura innoxia." Canadian Journal of Botany 65, no. 8 (1987): 1736–40. http://dx.doi.org/10.1139/b87-237.

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Conditions have been standardized for obtaining high yields of viable protoplasts from cell suspension cultures of an isoleucine–valine-requiring auxotroph (IV-1) of Datura innoxia P. Mill. Isolation of protoplasts critically required the use of a salt solution, containing sodium chloride and potassium chloride (0.125 M each), as osmotic stabilizers. The yield, viability, and divisional activity of the protoplasts isolated with mannitol or sucrose were poor. Protoplast-releasing enzymes used to isolate IV-1 protoplasts could be used twice without any loss in the yield or viability of the proto

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Du, Jiageng, Huitao Zhang, Weilong Li, et al. "Optimization of Protoplast Preparation System from Leaves and Establishment of a Transient Transformation System in Apium graveolens." Agronomy 13, no. 8 (2023): 2154. http://dx.doi.org/10.3390/agronomy13082154.

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Protoplast culture and transformation technology offer a novel method for developing new plant varieties. Nonetheless, the effective preparation of protoplasts and transformation technology specific to celery has yet to be achieved. This study utilized celery seedling leaves as the primary materials to examine the key factors influencing protoplast isolation. The aim was to prepare leaf protoplasts with a high yield and of high quality and subsequently conduct transient gene transformation and expression. The findings indicated that the most effective procedure for isolating and purifying prot

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Inoue, Aya, Daisuke Mori, Reiko Minagawa, Yoshiharu Fujii, and Hamako Sasamoto. "Allelopathy in a Leguminous Mangrove Plant, Derris indica: Protoplast Co-culture Bioassay and Rotenone Effect." Natural Product Communications 10, no. 5 (2015): 1934578X1501000. http://dx.doi.org/10.1177/1934578x1501000512.

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To investigate allelopathic activity of a leguminous mangrove plant, Derris indica, the ‘Protoplasts Co-culture Method’ for bioassay of allelopathy was developed using suspension culture. A suspension culture was induced from immature seed and sub-cultured in Murashige and Skoog's (MS) basal medium containing 10 μM each of 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzyladenine (BA). The protoplasts were isolated using the separate wells method with 2% each of Cellulase RS, Driselase 20 and Macerozyme R10 in 0.4 M mannitol solution. Protoplast cultures of D. indica revealed that high concen

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Sasamoto, Hamako, Yosuke Kobayashi, Tomoya Oyanagi, and Yutaka Sasamoto. "In vitro Bioassay of Allelopathic Activities of Soybean, and Three Isoflavones, Using Protoplast Co-culture Method with Digital Image Analysis." Journal of Plant Studies 11, no. 1 (2022): 19. http://dx.doi.org/10.5539/jps.v11n1p19.

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Inhibitory allelopathic activities of dried leaves of potted plants and protoplasts of Glycine max (soybean) cv. Okuhara-wase were studied using two in vitro bioassay methods. Strong inhibition (90%) of growth of recipient lettuce root was obtained with 50 mg soybean leaves using the lettuce seedling growth test (sandwich method), and moderate inhibition (65%) of lettuce protoplast division was obtained with 100 &times; 103 mL-1 protoplasts of etiolated soybean seedlings using the protoplast co-culture method with digital image analysis (DIA-PP method). Three isoflavones, genistein, genist

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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 (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)] resistan

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Sukmadjaja, Deden, Novianti Sunarlim, Endang G. Lestari, Ika Roostika, and Tintin Suharlini. "Teknik Isolasi dan Kultur Protoplas Tanaman Padi." Jurnal AgroBiogen 3, no. 2 (2016): 60. http://dx.doi.org/10.21082/jbio.v3n2.2007.p60-65.

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<p>Protoplast<br />fusion or somatic hybridization technology is an alternative<br />technology for production hybrids of plants that are difficult<br />to be produced by conventional methods due to their sexual<br />incompatibility. An experiment was conducted to develop<br />techniques for isolation, purification, and culture of rice<br />protoplasts of cultivar IR64 and a wild rice species (Oryza<br />officinalis). Optimization of protoplast isolation and purification<br />methods from both rice genotypes were successfully<br />don

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Schulz, Margot, and Gottfried Weissenböck. "Isolation and Separation of Epidermal and Mesophyll Protoplasts from Rye Primary Leaves — Tissue-Specific Characteristics of Secondary Phenolic Product Accumulation." Zeitschrift für Naturforschung C 41, no. 1-2 (1986): 22–27. http://dx.doi.org/10.1515/znc-1986-1-205.

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Abstract We have develop ed a technique for the large-scale isolation of epidermal and mesophyll protoplasts, as well as the vascular strands, of rye primary leaf blades. Separation of the two types of protoplasts has been successful only from leaves harvested at the end of a 13-h light period, when chloroplasts were enriched in starch. The occurrence of different flavonoid compounds, and amounts, in epidermal and mesophyll protoplasts can be used as criteria for protoplast purity and viability since C-glucosylflavone O-glycosides are characteristic of epidermal protoplasts whereas flavone O-

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46

Armita, Devi. "Plant Breeding Through Protoplast Fusion." Jurnal Biologi UNAND 8, no. 2 (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 parent

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47

Kuźniak, Elżbieta, Marzena Wielanek, Urszula Małolepsza, and Henryk Urbaniak. "Effects of environmental preconditioning, donor tissue and isolation conditions on tomato (Lycopersicon esculentum Mill.) protoplast yield." Acta Agrobotanica 48, no. 2 (2013): 113–20. http://dx.doi.org/10.5586/aa.1995.022.

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The effects of soil or <em>in vitro</em> grown plants, pretreatment conditions, donor tissue and isolation procedure on protoplast yield from cotyledons and leaves of tomato cv. 'Perkoz' and 'Zorza' were studied. The highest protoplast yield of 1.5 x 10<sup>7</sup>/g FW was obtained from leaves of <em>in vitro</em> grown plants. Low light intensity during donor plants <em>in vitro</em> culture and dark pretreatment were essential for successful protoplast isolation while cold pretreatment was not. Tissue preplasmolysis prior to transfer to enzyme

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Krasova, Yuliya V., Vladimir V. Fadeev, Yelizaveta М. Moiseeva, Yury S. Gusev, and Mikhail I. Chumakov. "Optimization of the technique for maize protoplast isolation and their nativity after electroporation." Izvestiya of Saratov University. Chemistry. Biology. Ecology 22, no. 4 (2022): 445–54. http://dx.doi.org/10.18500/1816-9775-2022-22-4-445-454.

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We optimizes the composition and concentration of the enzymes, the time for enzymatic treatment and the volume of the enzyme mixture. We also optimized the concentration osmotic of agent, the centrifugation mode, and filter pore size for protoplasts isolating from epidermal cells of maize roots (Zea mays L.) of the Brown Marker (BM) line. It was found that 150 minutes is the optimal time for 150 mg root tissue maceration. The yield of intact protoplasts was ~ 4.4 ± 0.2 × 105 cells/mL at the following concentrations of enzymes and osmotic stabilizer: cellulase – 17,4, pectolase – 1.2, hemicellu

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Sasamoto, Hamako, and Shinso Yokota. "In vitro Bioassay of Allelopathic Activities of a Mangrove Tree, Kandelia obovata, and Fast-growing Trees, Betula platyphylla and Populus alba, Using Protoplast Co-culture Method." Journal of Plant Studies 10, no. 2 (2021): 8. http://dx.doi.org/10.5539/jps.v10n2p8.

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Allelopathic activities of a salt-tolerant and low-temperature tolerant mangrove tree, Kandelia obovata, which grows in brackish water regions of sub-tropical areas, and two fast-growing trees, Betula platyphylla and Populus alba, which grow in the temperate area, were examined by two in vitro bioassay methods, the sandwich method using dried leaves and the protoplast co-culture method using leaf protoplasts. Lettuce root growth examined by the sandwich method, was inhibited 50% by 50 mg dried mature leaves of K. obovata. In the protoplast co-culture method, inhibition rates of cell division o

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Tusa, N., J. W. Grosser, and F. G. Gmitter. "Plant Regeneration of `Valencia' Sweet Orange, `Femminello' Lemon, and the Interspecific Somatic Hybrid following Protoplasm Fusion." Journal of the American Society for Horticultural Science 115, no. 6 (1990): 1043–46. http://dx.doi.org/10.21273/jashs.115.6.1043.

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Protoplasm culture following the chemical fusion of `Valencia' sweet orange [Citrus sinensis (L.) Osb.] protoplasts, isolated from an embryogenic suspension culture, with `Femminello' lemon [Citrus limon (L.) Burro. f.] leaf protoplasts resulted in the regeneration of an interspecific allotetraploid somatic hybrid plant, two autotetraploid lemon plants, and diploid plants from both parents. The regeneration of plants from lemon leaf protoplasts is an example of protoplast-to-plant regeneration from non-nucellus-derived tissue for Citrus. Regenerated plants were classified according to leaf mor

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