Journal articles: 'Protoplast fusion technology'

1

Gleddie, Stephen, and Wilfred A. Keller. "Protoplast fusion technology." Journal of Tissue Culture Methods 12, no. 4 (1989): 157–61. http://dx.doi.org/10.1007/bf01404443.

Full text

APA, Harvard, Vancouver, ISO, and other styles

2

S., V. S. S. S. L. N. Hima Bindu, Samatha B, and A. Singara Charya M. "A study on strain improvement of Fomitopsis feei by protoplast fusion technology." International journal of Microbiology and Mycology (IJMM) 6, no. 1 (2017): 9–15. https://doi.org/10.5281/zenodo.8394142.

Full text

Abstract:

Protoplast fusion technology was used for the formation of improved strain from a brown rot fungus, Fomitopsis feei and a white rot fungus, Pycnoporous sps. for the enhanced production of exopolysaccharide. Self-fusion of Fomitopsis feei and intergeneric hybridization of Fomitopsis feei with Pycnoporus sp. were researched. A combination of chitinase and lysing enzymes were used for the release of protoplasts. Fusion was successful with the formation of visible stable morphological regenerated colonies after several subcultures. Mycelial growth and hyphal size of wild species and fusants were significantly different from those of the parental strains. Although, Self and intergeneric protoplast fusants were produced successfully but were not best producers of exopolysaccharide compared to wild fungi. Even though, strain improvement towards highest production of exopolysaccharide is not successful, this study shows the possibility of fusion between these two mushrooms and could be useful in other research areas. Published by the  International journal of Microbiology and Mycology (IJMM)

APA, Harvard, Vancouver, ISO, and other styles

3

Li, Jintao, Linling Liu, Lin Xu, et al. "Interspecific Hybridization between Ganoderma lingzhi and G. resinaceum by PEG-Induced Double-Inactivated Protoplast Fusion." Horticulturae 9, no. 10 (2023): 1129. http://dx.doi.org/10.3390/horticulturae9101129.

Full text

Abstract:

Ganoderma lingzhi is an important medicinal fungus, and it is particularly important to select strains with high yields and active substance contents. In this study, protoplasts of G. lingzhi were thermally inactivated to destroy intracellular enzyme proteins and preserve DNA. The DNA of G. resinaceum was damaged by ultraviolet (UV) radiation, and other components of the protoplasm except DNA were preserved. Then, the protoplast was induced using polyethylene glycol (PEG) for fusion. The results showed that the optimal thermal inactivation conditions for G. lingzhi were 30 min in a 45 °C water bath, and the optimal UV inactivation conditions for G. resinaceum were 70 s of irradiation using a 20 W UV lamp at a vertical distance of 15 cm. Antagonistic tests, internal transcribed space (ITS) and mitochondrial DNA identification, intersimple sequence repeat (ISSR) molecular markers and morphology were used to distinguish the parents from the fusants. Four true fusants were obtained, and the yield was 2.5%. The fruiting body yield of the fusants was significantly higher than that of G. lingzhi, and the polysaccharide and triterpene contents of the RAD-64 fusant were significantly higher than those of G. lingzhi. The results presented in this paper show that protoplast fusion technology can effectively improve G. lingzhi varieties and support the breeding of new varieties.

APA, Harvard, Vancouver, ISO, and other styles

4

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.

Full text

Abstract:

Protoplast fusion or somatic hybridization technology is an alternative technology for production hybrids of plants that are difficult to be produced by conventional methods due to their sexual incompatibility. An experiment was conducted to develop techniques for isolation, purification, and culture of rice protoplasts of cultivar IR64 and a wild rice species (Oryza officinalis). Optimization of protoplast isolation and purification methods from both rice genotypes were successfully done. The highest protoplast density was obtained by digesting embryonic callus or stems of young seedling in an enzyme solution containing of 2% cellulose, 0.1% pectolyase, 0.5% macerozyme, 0.5% driselase, 5 mM ES, and 13% mannitol in CPW solution. The protoplast digestion was done for three hours by soaking in the enzyme solution followed by shaking at 50 rpm under a room temperature. Purification of the protoplasts were done by separating them from plant debris using a 25% sucrose solution. Protoplast regeneration was not successful using although different media compositions and conditions. Growth process from cell division to cell aggregate was only successful on IR64 protoplast culture on a medium that contained AgNO3.

APA, Harvard, Vancouver, ISO, and other styles

5

Polivanova, O. B., A. S. Egorova, A. B. Sivolapova, and S. V. Goryunova. "Current state and prospects of protoplast technology and potato somatic hybridization (review)." Agricultural Science Euro-North-East 24, no. 1 (2023): 7–19. http://dx.doi.org/10.30766/2072-9081.2023.24.1.7-19.

Full text

Abstract:

Wild Solanum species have often been used as sources of important agricultural traits, including resistance to various diseases, pests, and abiotic factors. However, their large-scale use in potato breeding is limited by complex barriers of sexual incompatibility with Solanum tuberosum. Fusion of protoplasts enzymatically isolated from somatic cells is one of the approaches to overcoming sexual incompatibility. The diverse nuclear and cytoplasmic traits exhibited by potato somatic hybrids provide new genetic material for breeding programs, which is confirmed by the creation of a large number of somatic hybrids of cultivated potatoes with wild Solanum species. The research in development of somatic potato hybrids by means of protoplast fusion has been carried out for more than 40 years already. In this review, the prospects for the use of this technology in modern potato breeding are considered. Genomic, transcriptomic, and proteomic studies provide further insight into the fundamental processes underlying the somatic hybrids formation, such as cell wall formation, chromosomal rearrangements in fusion products, regeneration, and also make a significant contribution to understanding the processes of genome stabilization. Improvement in the methods of molecular screening of both genome and cytoplasm also contributes to the expansion of the field of application of somatic hybrids in breeding. Finally, it has been shown that somatic hybridization promotes the introgression of important agricultural traits, primarily resistance to pathogens.

APA, Harvard, Vancouver, ISO, and other styles

6

Raikar, S. V., M. C. Christey, A. J. Conner Braun, and C. Bryant. "Protoplast isolation, colony formation and shoot regeneration from Lolium perenne." NZGA: Research and Practice Series 12 (January 1, 2006): 41–44. http://dx.doi.org/10.33584/rps.12.2006.3045.

Full text

Abstract:

Lolium perenne (ryegrass) is the most widely sown pasture grass in New Zealand providing forage for millions of sheep and cattle. Genetic improvement of Lolium species through conventional breeding has been particularly difficult because of its outbreeding and heterozygous nature. Biotechnology techniques that are not based on genetic modification technology, such as protoplast fusion, could help in the breeding of L. perenne as it enables the introgression of DNA (and associated desirable traits) from other species of plants that are otherwise unobtainable by conventional means. We intend to adapt this technology for L. perenne and report on our progress with the isolation of protoplasts, micro colony and callus formation, and shoot regeneration. Friable callus was induced from L. perenne seeds and used for isolation of protoplasts. After testing several combinations of enzyme mixtures at different molarities, a high yield of protoplasts (1.1 Ã— 107 g-1 FW) with a viability of 82% has been obtained. Callus colonies were formed from protoplasts cultured on a nitrocellulose membrane over a feeder layer. Plating efficiencies of 0.1-0.2% were obtained. Regeneration of plantlets from these colonies were obtained on Linsmaier and Skoog medium supplemented with growth hormones. All the plants regenerated to date have an albino appearance.

APA, Harvard, Vancouver, ISO, and other styles

7

Chen, Sheng, Yu Li, Wen Bin Jin, Yan Chen, Xiao Guang Liu, and Fu Ping Lu. "Selective Breeding of Oxygen-Tolerant and Oxalate-Degrading Lactic Acid Bacteria by Protoplast Fusion." Advanced Materials Research 750-752 (August 2013): 1489–94. http://dx.doi.org/10.4028/www.scientific.net/amr.750-752.1489.

Full text

Abstract:

Biﬁdobacterium lactiswith oxalate-degrading capacity can efficiently reduce the oxalate in vivo, and it can be used to prevent and treat kidney stone diseases. WhileBiﬁdobacterium lactisis poorly oxygen-tolerant, which hinders it from being as microbial ecological agents. To obtain oxygen-tolerant and oxalate-degrading lactic acid bacteria, protoplast fusion technology was used betweenB. lactisandL. acidophilus.Under the optimum conditions of protoplast fusion with PEG 6000 concentration 50%, the fusion time 7 min, the fusion temperature 30°C, the concentration of CaCl20. 02mol/ L and the concentration of MgCl20.5mol/ L, the fusion rate reached 7.6%, and three oxygen-tolerant fusant showing that the level of oxalate degradation were similar withB. lactiswas obtained. The fusants of SZY1-7 and SZY2-1 could tolerance to pH 2.5 and 0.5% (w/v) bile salt.

APA, Harvard, Vancouver, ISO, and other styles

8

Liu, Shuping, Xiaojie Li, Jiani Zhu, et al. "Modern Technologies Provide New Opportunities for Somatic Hybridization in the Breeding of Woody Plants." Plants 13, no. 18 (2024): 2539. http://dx.doi.org/10.3390/plants13182539.

Full text

Abstract:

Advances in cell fusion technology have propelled breeding into the realm of somatic hybridization, enabling the transfer of genetic material independent of sexual reproduction. This has facilitated genome recombination both within and between species. Despite its use in plant breeding for over fifty years, somatic hybridization has been limited by cumbersome procedures, such as protoplast isolation, hybridized-cell selection and cultivation, and regeneration, particularly in woody perennial species that are difficult to regenerate. This review summarizes the development of somatic hybridization, explores the challenges and solutions associated with cell fusion technology in woody perennials, and outlines the process of protoplast regeneration. Recent advancements in genome editing and plant cell regeneration present new opportunities for applying somatic hybridization in breeding. We offer a perspective on integrating these emerging technologies to enhance somatic hybridization in woody perennial plants.

APA, Harvard, Vancouver, ISO, and other styles

9

Christiansen, C., H. Kryvi, P. Sontum, and T. Skotland. "Isolation of intergeneric hybrids between Bacillus subtilis and Zymomonas mobilis and the production of thermostable amylase by hybrids." Biotechnology and Applied Biochemistry 20, no. 1 (1994): 109–16. http://dx.doi.org/10.1111/j.1470-8744.1994.tb00309.x.

Full text

Abstract:

Stable hybrids were obtained by protoplast fusion between Bacillus subtilis and Zymomonas mobilis. All the hybrids were able to hydrolyse starch and possessed ampicillin‐ and tetracycline‐resistant phenotypes. Two of the hybrids, BZ‐1 and BZ‐2, were hyperproducers of alpha‐amylase. The enzyme produced by these hybrids exhibited increased thermostability. The results show that stable intergeneric gene transfer can be achieved through poly(ethylene glycol)‐mediated protoplast fusion between two industrially important genera of bacteria.

APA, Harvard, Vancouver, ISO, and other styles

10

Prajwalitha, Kanala, Vijay Bahadur, and Saket Mishra. "Somatic Hybrids and Cybrids: Innovations in Vegetable Improvement." International Journal of Plant & Soil Science 37, no. 1 (2025): 331–46. https://doi.org/10.9734/ijpss/2025/v37i15275.

Full text

Abstract:

Somatic hybridization through protoplast fusion is indeed a promising method for producing asymmetrical and unbalanced polyploid somatic hybrids in various plant species. This technique involves merging protoplasts from different plant species to create hybrids that possess desirable traits for both scion and rootstock improvements. By circumventing the limitations of sexual hybridization, such as male/female sterility and sexual incompatibility, somatic hybridization enables the incorporation of beneficial genes from closely related or even distantly related species. The success of somatic hybridization in horticulture is evident in various crops such as citrus, potato, brinjal (eggplant), tomato, mango, avocado, banana, strawberry, pear, and cherry. It facilitates the transfer of numerous uncloned genes that confer resistance to biotic and abiotic stresses, thereby enhancing crop resilience and productivity. Unlike transgenic technology, which is often subject to regulatory constraints, somatic hybridization allows for the exchange of genetic material without the same legal formalities. Despite its potential benefits, somatic hybridization faces challenges and constraints compared to sexual hybridization. These include technical difficulties in protoplast isolation and fusion, as well as limitations in generating fertile hybrids and maintaining genome stability. However, advancements in genomic technologies provide optimism for overcoming these challenges. Improved understanding of plant genomes enables more precise manipulation and selection of desired traits through somatic hybridization. In conclusion, somatic hybridization is a crucial tool in modern plant breeding and crop improvement efforts. It expands the gene pool available for breeding programs by incorporating genetic variability from diverse sources, thus offering new opportunities to enhance agricultural productivity and sustainability. As research and technology continue to advance, somatic hybridization holds promise for addressing current and future challenges in global agriculture.

APA, Harvard, Vancouver, ISO, and other styles

11

Gupta, H. S., B. Bhattacharjee, and A. Pattanayak. "Transfer of cytoplasmic male sterility in indica rice through protoplast fusion." International Rice Research Notes 21, no. 2-3 (1996): 33–34. https://doi.org/10.5281/zenodo.7247089.

Full text

Abstract:

This article 'Transfer of cytoplasmic male sterility in indica rice through protoplast fusion' appeared in the International Rice Research Notes series, created by the International Rice Research Institute (IRRI) to expedite communication among scientists concerned with the development of improved technology for rice and rice-based systems. The series is a mechanism to help scientists keep each other informed of current rice research findings. The concise scientific notes are meant to encourage rice scientists to communicate with one another to obtain details on the research reported.

APA, Harvard, Vancouver, ISO, and other styles

12

Ozojiofor, Ugochukwu Okechukwu. "Protoplast fusion technology and its application in genetic improvement of plants: A review." International Journal of Advanced Biochemistry Research 1, no. 2 (2017): 45–52. http://dx.doi.org/10.33545/26174693.2017.v1.i2a.116.

Full text

APA, Harvard, Vancouver, ISO, and other styles

13

Geerts, P., A. Henneguez, P. Druart, and B. Watillon. "PROTOPLAST ELECTRO-FUSION TECHNOLOGY AS A TOOL FOR SOMATIC HYBRIDISATION BETWEEN STRAWBERRIES AND RASPBERRIES." Acta Horticulturae, no. 842 (August 2009): 495–98. http://dx.doi.org/10.17660/actahortic.2009.842.101.

Full text

APA, Harvard, Vancouver, ISO, and other styles

14

Chen, Honglei, Huaiyu Zhan, Yuancai Chen, and Shiyu Fu. "Construction of engineering microorganism degrading chlorophenol efficiently by protoplast fusion technique." Environmental Progress & Sustainable Energy 32, no. 3 (2012): 443–48. http://dx.doi.org/10.1002/ep.11626.

Full text

APA, Harvard, Vancouver, ISO, and other styles

15

Ge, Jingping, Jingwen Zhao, Luyan Zhang, Mengyun Zhang, and Wenxiang Ping. "Construction and Analysis of High-Ethanol-Producing Fusants with Co-Fermentation Ability through Protoplast Fusion and Double Labeling Technology." PLoS ONE 9, no. 9 (2014): e108311. http://dx.doi.org/10.1371/journal.pone.0108311.

Full text

APA, Harvard, Vancouver, ISO, and other styles

16

Rathod, L. B. Laxmikanth, Shaik Muzammil Pasha, Anju Rajan, Madhavi ., and Chand Pasha. "Sequential Strain Improvement of Cordyceps militaris." Journal of Advances in Microbiology 23, no. 3 (2023): 13–20. http://dx.doi.org/10.9734/jamb/2023/v23i3711.

Full text

Abstract:

Aim: As a highly valued fungus, Cordyceps militaris has been widely used all over the world. Although the wild resources of C. militaris are limited, the fruiting bodies of C. militaris have been successfully cultivated on a large-scale with limited Cordycepin production. However, the high-frequency degeneration of C. militaris during subculture and preservation seriously limits the development of the C. militaris industry. Methods and Results: In this study, three different techniques for strain improvement viz., protoplast fusion, UV irradiation and chemical mutagenesis were assessed for improved biomass and cordycepin production. Conclusion: It was observed that the fusant subsequently mutated were stable for many generation and yielding improved biomass and Cordycepin. It was found that fusants after mutagenesis will be genetically stable and this methodology can be used for strain improvement of genetically unstable microorganisms.

APA, Harvard, Vancouver, ISO, and other styles

17

Dai, Wen, Dongchen Zhang, Chuanzhen Wang, and Fanglue Wang. "Construction of engineering bacteria degrading residual polyacrylamide in coal slime water by protoplast fusion technique." International Journal of Global Energy Issues 43, no. 4 (2021): 340. http://dx.doi.org/10.1504/ijgei.2021.10040098.

Full text

APA, Harvard, Vancouver, ISO, and other styles

18

Zhang, Dongchen, Fanglue Wang, Chuanzhen Wang, and Wen Dai. "Construction of engineering bacteria degrading residual polyacrylamide in coal slime water by protoplast fusion technique." International Journal of Global Energy Issues 43, no. 4 (2021): 340. http://dx.doi.org/10.1504/ijgei.2021.117025.

Full text

APA, Harvard, Vancouver, ISO, and other styles

19

Jianyou, Long, Gao Xiaoning, and Xia Jianrong. "Enhancement production of qinlingmycin by a soil-derived Streptomyces No. 24 using protoplast fusion technology and assessment of antibacterial activity." African Journal of Biotechnology 10, no. 54 (2011): 11217–23. http://dx.doi.org/10.5897/ajb10.2665.

Full text

APA, Harvard, Vancouver, ISO, and other styles

20

Keskitalo, Marjo Kristiina. "Application of Protoplast Fusion Technology to Tansy (Tanacetum vulgareL.): Biodiversity as a Source to Enhance Biological Activity of Secondary Compounds." Journal of Herbs, Spices & Medicinal Plants 9, no. 2-3 (2002): 197–203. http://dx.doi.org/10.1300/j044v09n02_28.

Full text

APA, Harvard, Vancouver, ISO, and other styles

21

Keskitalo, M. K. "APPLICATION OF PROTOPLAST FUSION TECHNOLOGY TO TANSY (Tanacetum vulgare L.) BIODIVERSITY AS A SOURCE TO ENHANCE BIOLOGICAL ACTIVITY OF SECONDARY COMPOUNDS." Acta Horticulturae, no. 560 (October 2001): 263–67. http://dx.doi.org/10.17660/actahortic.2001.560.50.

Full text

APA, Harvard, Vancouver, ISO, and other styles

22

Evans, David A. "FOOD QUALITY GENES AND GENETIC MODIFICATION OF FRUIT AND VEGETABLE PRODUCE." HortScience 25, no. 12 (1990): 1531E—1531. http://dx.doi.org/10.21273/hortsci.25.12.1531.

Full text

Abstract:

For many branded food products, the raw material is purchased as a commodity. Food companies provide added-value based on processing technology or marketing. The tools of plant biotechnology, which shorten the time for crop improvement and permit development of novel germplasm, offer the food industry the opportunity to modify raw materials and develop proprietary branded products. Such modifications will permit development of plant cultivars specifically selected for traits with added-value for the processor or the consumer. Biotechnology-developed cultivars offer the opportunity to develop produce that can be branded. The cell genetics tools of clonal propagation, somaclonal variation, gametoclonal variation, and protoplast fusion permit new cultivar development in an intermediate time scale, making them attractive for market introduction of proprietary products. When integrated with conventional breeding, these intermediate-term technologies will permit modification of fruits and vegetables over the course of the next several years. The longer-term technologies of plant genetic engineering will continue to have an impact on manipulation of specific traits, resulting in second-generation products specifically designed for consumers.

APA, Harvard, Vancouver, ISO, and other styles

23

Singh, A. K., N. K. Mishra, Sanjeev Kumar, and Sudhakar Pandey. "Application of somatic hybridization for the improvement of horticultural crops." Horticultural Biotechnology Research 1 (December 5, 2015): 39. http://dx.doi.org/10.19071/jhcbt.2015.v1.2897.

Full text

Abstract:

Somatic hybridization (SH) using protoplast fusion is a promising tool to produce symmetrical and asymmetrical polyploidy somatic hybrids in many agricultural crops. The technique of SH could facilitate conventional breeding by providing of novel lines so as to use them as elite breeding materials in conventional crosses for both scion and rootstock improvement. Further, SH can overcome those problems associated with sexual hybridization viz., sexual incompatibility, nucellar embryogenesis, and male/female sterility. Successful exploitation of SH in horticultural crops mainly comes from transfer of resistance genes for biotic and abiotic stresses from related species in several horticultural crops, viz., citrus, potato, brinjal, tomato, mango, avocado, banana, strawberry, pear, cherry etc. Unlike transgenic technology, SH is not affected by legal formalities and able to transfer uncloned multiple genes. However, certain boundaries and limitations of SH restricts its use over sexual hybridization but, envisage of new genomic technologies providing better insight into the plant genomes will increase the potentiality of SH in betterment of agriculture.

APA, Harvard, Vancouver, ISO, and other styles

24

Omere, E. A., C. N. C. Nwaoguala, and T. O. Emede. "Polyploidy and its relevance in crop improvement." Nigerian Journal of Biotechnology 39, no. 2 (2023): 9–19. http://dx.doi.org/10.4314/njb.v39i2.2.

Full text

Abstract:

Many areas of research in crop production have been geared towards crop improvement and increased yield. Crop improvement include but not restricted to; plant introduction and acclimatization, domestication, ploidy manipulation (polyploidization), recombinant DNA technology, crossing for superior selection (cultivar development), molecular genetics, etc. Polyploidy is a condition where the genome of an organism has more than the usual number of complete sets of chromosomes and the product of this phenomenon is called a Polyploid. Polyploidy occurs naturally, and can be induced chemically using antimitotic agents or physically using protoplast fusion and temperature shock. It is mostly artificially induced through a process called polyploidization. Polyploids are more advantageous in important plant attributes than the regular diploid. Relative success has been reported in the application of polyploidization for crop improvement which resulted chiefly in increased amount of beneficial secondary metabolites (phytochemicals), larger stomata and leaves,improved adaptation to stress and unfavourable conditions, to mention but a few. Therefore, it is imperative to state that polyploidy is an area of research that has been and will continually be deployed in crop improvement .

APA, Harvard, Vancouver, ISO, and other styles

25

Surya Krishna, Sakthivel, S. R. Harish Chandar, Maruthachalam Ravi, et al. "Transgene-Free Genome Editing for Biotic and Abiotic Stress Resistance in Sugarcane: Prospects and Challenges." Agronomy 13, no. 4 (2023): 1000. http://dx.doi.org/10.3390/agronomy13041000.

Full text

Abstract:

Sugarcane (Saccharum spp.) is one of the most valuable food and industrial crops. Its production is constrained due to major biotic (fungi, bacteria, viruses and insect pests) and abiotic (drought, salt, cold/heat, water logging and heavy metals) stresses. The ever-increasing demand for sugar and biofuel and the rise of new pest and disease variants call for the use of innovative technologies to speed up the sugarcane genetic improvement process. Developing new cultivars through conventional breeding techniques requires much time and resources. The advent of CRISPR/Cas genome editing technology enables the creation of new cultivars with improved resistance/tolerance to various biotic and abiotic stresses. The presence of genome editing cassette inside the genome of genome-edited plants hinders commercial exploitation due to regulatory issues. However, this limitation can be overcome by using transgene-free genome editing techniques. Transgene-free genome editing approaches, such as delivery of the RNPs through biolistics or protoplast fusion, virus-induced genome editing (VIGE), transient expression of CRISPR/Cas reagents through Agrobacterium-mediated transformation and other approaches, are discussed. A well-established PCR-based assay and advanced screening systems such as visual marker system and Transgene killer CRISPR system (TKC) rapidly identify transgene-free genome edits. These advancements in CRISPR/Cas technology speed up the creation of genome-edited climate-smart cultivars that combat various biotic and abiotic stresses and produce good yields under ever-changing conditions.

APA, Harvard, Vancouver, ISO, and other styles

26

Li, Meng, Jiachang Li, Yufei Li, Xian Zhang, and Jianzhong Xu. "Synergistic Optimization of Bacillus subtilis for Efficiently Producing Menaquinone-7 (MK-7) by Atmospheric and Room Temperature Plasma (ARTP) Mutagenesis and Metabolic Engineering." Fermentation 11, no. 3 (2025): 137. https://doi.org/10.3390/fermentation11030137.

Full text

Abstract:

Menaquinone-7 (MK-7) plays a crucial role in preventing fractures and certain cardiovascular diseases and is one of the essential vitamins in the human body. In this study, a strain of Bacillus subtilis that produces MK-7 was isolated from commercially available natto fermentation agents, with an MK-7 titer of 75 mg/L. It was named L-5. Firstly, by employing Atmospheric and Room Temperature Plasma (ARTP) mutagenesis technology and protoplast fusion techniques, mutants resistant to 1-hydroxy-2-naphthoic acid (HNA) and diphenylamine (DPA) were obtained, with the titer of MK-7 reaching 196 mg/L. It was named R-8. Based on whole-genome sequencing technology, four mutants involved in the MK-7 synthesis pathway of strain L-5 were identified: 2-succinyl-5-enol-pyruvate-6-hydroxy-3-cyclohexen-1-carboxylic acid, MenD (S249L); (1,4)-dihydroxy-2-naphthalic acid-heptaisoprenyltransferase, MenA (S196L); 1-deoxy-D-xylose-5-phosphate synthetase, Dxs (N60D, Q185H); and hydroxy acid reductive isomerase, Dxr (Q351K). The overexpression of these mutants led to increases in MK-7 production of 19 mg/L, 20 mg/L, 17 mg/L, and 16 mg/L, respectively, compared to the unmutated genes. These mutations have been shown to be effective. To further enhance the production of MK-7, the mutants menD (S249L), menA (S196L), Dxs (N60D, Q185H), and Dxr (Q351K) were co-expressed. The final titer of MK-7 reached 239 mg/L. This study provides theoretical support for the future genetic modification of key enzymes in the MK-7 biosynthetic pathway.

APA, Harvard, Vancouver, ISO, and other styles

27

K. C., Bigyan, Rishav Pandit, Bishnu Prasad Kandel, Kanchan Kumar K. C., Arpana K. C., and Mukti Ram Poudel. "Scenario of Plant Breeding in Nepal and Its Application in Rice." International Journal of Agronomy 2021 (June 30, 2021): 1–9. http://dx.doi.org/10.1155/2021/5520741.

Full text

Abstract:

Rice, the number one staple food crop of Nepal, contributes nearly 20% to the agricultural gross domestic product, almost 7% to gross domestic product, and supplies with 40% of the food calorie consumption of Nepalese people. Despite of increasing production, the national demand of rice cannot be fulfilled, and billions of rupees are spent yearly for importing rice from India. This article reviews history, recent scenario, prospects, and importance of rice breeding research in Nepal for self-sufficiency. Though plant breeding inception point in Nepal was 1951, the systematic research in rice was started from 1972 after the establishment of the National Rice Research Program. Introduction and evaluation of high yielding exotic germplasm of rice was the initial rice breeding activities. After analysing several foreign materials, CH-45 variety of rice was first recommended in 1959. Nepal Agriculture Research Council (NARC), the leading organisation for rice varietal improvement to date, has been doing rice research activities such as germplasm conservation, exchange, varietal development, verification of technologies, and knowledge updates in collaboration with various organizations. International Rice Research Institute (IRRI), since 1966, has been playing an important role in Nepal’s rice research. Major plant breeding achievements are introduction of exotic materials and their evaluation, local landraces identification, collection and analysis, crossing of local landraces with exotic genetic materials, heterosis breeding, tissue culture technology, genetic marker technology, and National Genebank. RAPD and SSR, DNA-based markers, are mostly used for diversity appraisal and selecting desirable genotypes. Integration of advanced tools such as genetic engineering, bioinformatics, protoplast fusion, and SNP markers could not be still applied in rice research and development. Hence, emphasis should be given to improve the capacity and abilities of the rice scientists and should release more short duration, drought, and submergence-tolerant rice varieties by optimum utilisation of local landraces combining modern and conventional breeding.

APA, Harvard, Vancouver, ISO, and other styles

28

Guo, Jianan, Yipeng Zhao, Huihui Cheng, et al. "Enhancing Plant Stress Tolerance: The Role of LcWRKY40 Gene in Drought and Alkaline Salt Resistance in Tobacco and Yeast." International Journal of Molecular Sciences 25, no. 18 (2024): 10149. http://dx.doi.org/10.3390/ijms251810149.

Full text

Abstract:

Leymus chinensis, a halophytic perennial grass belonging to the Poaceae family, thrives in saline-alkali grasslands and harbors a rich repository of resistance-related genetic resources. This study focused on deciphering the stress-responsive mechanisms of L. chinensis by conducting transcriptomic sequencing under NaHCO3 stress, which resulted in the annotation of a segment corresponding to the 51WRKY gene. The alkali-induced gene LcWRKY40 (QIG37591) was identified by phylogenetic analysis. Real-time quantitative PCR analysis was performed on L. chinensis plants subjected to PEG6000 and alkaline salt (NaHCO3) stress, and the results indicated that the LcWRKY40 gene was upregulated in both the leaves and roots. The localization of the LcWRKY40 protein was confirmed by the use of green fluorescent protein (GFP) fusion technology in transformed rice protoplast cells. The GAL4-driven transformation of the LcWRKY40 gene in INVScI yeast cells, which exhibited enhanced tolerance upon overexpression of the LcWRKY40 gene under mannitol and alkaline salt (NaHCO3) stress conditions. Under drought stress using mannitol, the fresh weight of Nicotiana tabacum overexpressing the LcWRKY40 gene was significantly higher than that of wild-type(WT) tobacco. Through drought and salt alkali stress, we found that overexpressed tobacco at different stages always outperformed the wild type in terms of fresh weight, SOD, MDA, and Fv/Fm. This study provides preliminary insights into the involvement of the LcWRKY40 gene in responding to drought and alkaline salt stresses, highlighting its role in enhancing plant resistance to drought and saline-alkaline conditions. These findings lay the foundation for future molecular breeding strategies aimed at improving grass resistance from different aspects.

APA, Harvard, Vancouver, ISO, and other styles

29

Tachibana, Sanro, Miyuki Wada, Kazutaka Itoh, Tomohito Itoh, and Tae Oki. "Studies on Biological Treatment of Mechanical Pulps(VIII). Formation of a fusant having ability for depression of light-induced color reversion of mechanical pulps by protoplast fusion of Phanerochaete chrysosporium and Fusarium solani." JAPAN TAPPI JOURNAL 50, no. 10 (1996): 1446–55. http://dx.doi.org/10.2524/jtappij.50.1446.

Full text

APA, Harvard, Vancouver, ISO, and other styles

30

Bhattacharjee, B., H. S. Gupta, and A. P. Sane. "Transferring wild abortive cytoplasmic male sterility through asymmetric fusion of protoplasts." International Rice Research Notes 23, no. 1 (1998): 11. https://doi.org/10.5281/zenodo.6947623.

Full text

Abstract:

This article 'Transferring wild abortive cytoplasmic male sterility through asymmetric fusion of protoplasts' appeared in the International Rice Research Notes series, created by the International Rice Research Institute (IRRI) to expedite communication among scientists concerned with the development of improved technology for rice and rice-based systems. The series is a mechanism to help scientists keep each other informed of current rice research findings. The concise scientific notes are meant to encourage rice scientists to communicate with one another to obtain details on the research reported.

APA, Harvard, Vancouver, ISO, and other styles

31

Leaungthitikanchana, Sumana, Khachapohn Thongdonyod, and Nootjaree Singphan. "Effect of Enzyme Treatments on Protoplast Isolation from Leaves of Vetiver (Vetiveria spp.)." Chiang Mai University Journal of Natural Sciences 20, no. 3 (2021). http://dx.doi.org/10.12982/cmujns.2021.048.

Full text

Abstract:

Protoplast isolation is a first and important step for establishing a new plant with desired traits through protoplast fusion technology. This experiments were conducted to evaluate various concentration of enzymes and incubation time on protoplast yield and viability in two vetiver ecotypes, Kamphaeng Phet 2 (Vetiveria zizanioides Nash) and Prachuap Khiri Khan (V. nemoralis A.Camus). The results revealed that protoplast yields were significantly affected by different enzyme treatments. The highest protoplast yield (6.12x105 protoplasts/ml) and high viability (98.61%) in Kamphaeng Phet 2 was obtained through the process of cell wall digestion when treated with enzyme solution containing 0.5% (w/v) cellulase onozuka R-10 and 0.5% (w/v) macerozyme R-10 in combination. While, the optimal enzyme solution for protoplast isolation from leaves of Prachuap Khiri Khan was the combination of 1.0% (w/v) cellulase onozuka R-10 and 0.4% (w/v) macerozyme R-10, resulting in the highest yield (6.80x105 protoplasts/ml) and viability (96.56%) of protoplasts. Meanwhile, incubation time of 24 h with the optimal enzyme solution resulted in the highest protoplast yields of both ecotypes. Our findings have the potential to generate an efficient protocol to isolate the protoplast from leaves of vetiver which can be used for further research studies in protoplast culture and fusion for vetiver improvement. Keywords: Cellulase onozuka R-10, Macerozyme R-10, Protoplast isolation, Vetiver

APA, Harvard, Vancouver, ISO, and other styles

32

Yue, Jin-Jun, Jin-Ling Yuan, Fu-Hui Wu, et al. "Protoplasts: From Isolation to CRISPR/Cas Genome Editing Application." Frontiers in Genome Editing 3 (August 11, 2021). http://dx.doi.org/10.3389/fgeed.2021.717017.

Full text

Abstract:

In the clustered regulatory interspaced short palindromic repeats (CRISPR)/CRISPR associated protein (Cas) system, protoplasts are not only useful for rapidly validating the mutagenesis efficiency of various RNA-guided endonucleases, promoters, sgRNA designs, or Cas proteins, but can also be a platform for DNA-free gene editing. To date, the latter approach has been applied to numerous crops, particularly those with complex genomes, a long juvenile period, a tendency for heterosis, and/or self-incompatibility. Protoplast regeneration is thus a key step in DNA-free gene editing. In this report, we review the history and some future prospects for protoplast technology, including protoplast transfection, transformation, fusion, regeneration, and current protoplast applications in CRISPR/Cas-based breeding.

APA, Harvard, Vancouver, ISO, and other styles

33

Zaranek, Magdalena, Reneé Pérez-Pérez, Anna Milewska-Hendel, Alexander Betekhtin, and Ewa Grzebelus. "Promotive effect of phytosulfokine - peptide growth factor - on protoplast cultures development in Fagopyrum tataricum (L.) Gaertn." BMC Plant Biology 23, no. 1 (2023). http://dx.doi.org/10.1186/s12870-023-04402-9.

Full text

Abstract:

Abstract Background Fagopyrum tataricum (Tartary buckwheat) is a valuable crop of great nutritional importance due to its high level of bioactive compounds. Excellent opportunities to obtain plants with the high level or the desired profile of valuable metabolites may be provided by in vitro cultures. Among known in vitro techniques, protoplast technology is an exciting tool for genetic manipulation to improve crop traits. In that context, protoplast fusion may be applied to generate hybrid cells between different species of Fagopyrum. To apply protoplast cultures to the aforementioned approaches in this research, we established the protoplast-to-plant system in Tartary buckwheat. Results In this work, cellulase and pectinase activity enabled protoplast isolation from non-morphogenic and morphogenic callus (MC), reaching, on average, 2.3 × 106 protoplasts per g of fresh weight. However, to release protoplasts from hypocotyls, the key step was the application of driselase in the enzyme mixture. We showed that colony formation could be induced after protoplast embedding in agarose compared to the alginate matrix. Protoplasts cultured in a medium based on Kao and Michayluk supplemented with phytosulfokine (PSK) rebuilt cell walls, underwent repeated mitotic division, formed aggregates, which consequently led to callus formation. Plating efficiency, expressing the number of cell aggregate formed, in 10-day-old protoplast cultures varied from 14% for morphogenic callus to 30% for hypocotyls used as a protoplast source. However plant regeneration via somatic embryogenesis and organogenesis occurred only during the cultivation of MC-derived protoplasts. Conclusions This study demonstrated that the applied protoplast isolation approach facilitated the recovery of viable protoplasts. Moreover, the embedding of protoplasts in an agarose matrix and supplementation of a culture medium with PSK effectively stimulated cell division and further development of Tartary buckwheat protoplast cultures along with the plant regeneration. Together, these results provide the first evidence of developing a protoplast-to-plant system from the MC of Fagopyrum tataricum used as source material. These findings suggest that Tartary buckwheat’s protoplast cultures have potential implications for the species’ somatic hybridization and genetic improvement.

APA, Harvard, Vancouver, ISO, and other styles

34

Li, Nan, Jie Lu, Zirui Wang, et al. "Improving the regeneration rate of deep lethal mutant protoplasts by fusion to promote efficient L-lysine fermentation." BMC Biotechnology 23, no. 1 (2023). http://dx.doi.org/10.1186/s12896-023-00792-8.

Full text

Abstract:

Abstract Background L-lysine is widely used for feed and special diet products. The transformation of fermentation strains plays a decisive role in the development of these industries. Based on the mutation breeding theory and metabolic engineering methods, this study aimed to improve the regeneration rate of high-lethality protoplasts by combining multiple mutagenesis and homologous cell fusion techniques to efficiently concentrate multiple dominant mutations and optimize the L-lysine production strain Escherichia coli QDW. Results In order to obtain the best protoplasts, the optimal enzymolysis time was selected as 4 h. The optimal lysozyme concentration was estimated at 0.8 mg/mL, because the protoplast formation rate and regeneration rate reached 90% and 30%, respectively, and their product reached the maximum. In this study, it was necessary that UV mutagenesis be excessive to obtain an expanded mutation library. For high lethality protoplasts, under the premise of minimal influence on its recovery, the optimal time for UV mutagenesis of protoplasts was 7 min, and the optimal time for thermal inactivation of protoplasts at 85 ℃ was 30 min. After homologous fusion, four fusion strains of E. coli were obtained, and their stability was analyzed by flow cytometry. The L-lysine yield of QDW-UH3 increased by 7.2% compared with that of QDW in a fermentation experiment, which promoted the expression of key enzymes in L-lysine synthesis, indicating that the combination of ultraviolet mutagenic breeding and protoplast fusion technology improved the acid-production level of the fusion strain. Conclusion This method provides a novel approach for the targeted construction of microbial cell factories.

APA, Harvard, Vancouver, ISO, and other styles

35

Qian, Keqing, Zhengxiang Qi, Anran Xu, Xiao Li, Bo Zhang, and Yu Li. "Interspecies hybridization between Auricularia cornea cv. Yu Muer and Auricularia heimuer cv. Bai Muer through protoplast fusion." Frontiers in Microbiology 14 (October 30, 2023). http://dx.doi.org/10.3389/fmicb.2023.1280420.

Full text

Abstract:

Color variations in cultivated edible mushrooms present novel and potentially valuable alternatives to the research and cultivation industries. We collected, identified, and domesticated a white strain of Auricularia cornea and a white strain of Auricularia heimuer from China. However, due to an unstable phenotype and stricter requirements on environment and management technology, the production and utilization of Auricularia heimuer cv. Bai Muer make slow progress. Outcrossing is an essential means to broaden the intraspecific genetic resources to expand the gene pool and compensate for the limitations of related species hybridization. In this study, interspecies hybridization between Auricularia cornea cv. Yu Muer and Auricularia heimuer cv. Bai Muer was conducted using polyethylene glycol (PEG)-induced double-inactivated protoplast fusion. Apart from the functional complementation of double-inactivated protoplasts, the hybrids were characterized by colony morphology, antagonistic test, primordial morphology, and polymerase chain reaction (PCR) fingerprinting. The results suggested that the hybrids and their parents showed significant differences in their colony morphology. Moreover, positive barrage reactions were observed between each parent and hybrid. Inter-simple sequence repeat (ISSR) and start codon targeted (SCoT) profile analysis of fusants and parents depicted that fusants contained polymorphic bands, which indicated the rearrangement and deletion of deoxyribonucleic acid (DNA) in the fusants. Yellowish-white primordia were obtained from two hybrids. Protoplast fusion may reinforce the genetic potential and provide an ideal alternative for breeding albino Auricularia.

APA, Harvard, Vancouver, ISO, and other styles

36

Zhu, Lei. "Targeted Gene Knockouts by Protoplast Transformation in the Moss Physcomitrella patens." Frontiers in Genome Editing 3 (December 17, 2021). http://dx.doi.org/10.3389/fgeed.2021.719087.

Full text

Abstract:

Targeted gene knockout is particularly useful for analyzing gene functions in plant growth, signaling, and development. By transforming knockout cassettes consisting of homologous sequences of the target gene into protoplasts, the classical gene targeting method aims to obtain targeted gene replacement, allowing for the characterization of gene functions in vivo. The moss Physcomitrella patens is a known model organism for a high frequency of homologous recombination and thus harbors a remarkable rate of gene targeting. Other moss features, including easy to culture, dominant haploidy phase, and sequenced genome, make gene targeting prevalent in Physcomitrella patens. However, even gene targeting was powerful to generate knockouts, researchers using this method still experienced technical challenges. For example, obtaining a good number of targeted knockouts after protoplast transformation and regeneration disturbed the users. Off-target mutations such as illegitimate random integration mediated by nonhomologous end joining and targeted insertion wherein one junction on-target but the other end off-target is commonly present in the knockouts. Protoplast fusion during transformation and regeneration was also a problem. This review will discuss the advantages and technical challenges of gene targeting. Recently, CRISPR-Cas9 is a revolutionary technology and becoming a hot topic in plant gene editing. In the second part of this review, CRISPR-Cas9 technology will be focused on and compared to gene targeting regarding the practical use in Physcomitrella patens. This review presents an updated perspective of the gene targeting and CRISPR-Cas9 techniques to plant biologists who may consider studying gene functions in the model organism Physcomitrella patens.

APA, Harvard, Vancouver, ISO, and other styles

37

Hospet, Ravichandra, Devarajan Thangadurai, Natália Cruz-Martins, et al. "Genome shuffling for phenotypic improvement of industrial strains through recursive protoplast fusion technology." Critical Reviews in Food Science and Nutrition, October 1, 2021, 1–10. http://dx.doi.org/10.1080/10408398.2021.1983763.

Full text

APA, Harvard, Vancouver, ISO, and other styles

38

"Breeding of Producing L-Valine Strain Brevibacterium flavum NJ112 by Protoplast Fusion Technology." Asian Journal of Chemistry 25, no. 3 (2013). http://dx.doi.org/10.14233/ajchem.2013.13878.

Full text

APA, Harvard, Vancouver, ISO, and other styles

39

Nie, Hushuai, Yali Wang, Chengcheng Wei, et al. "Embryogenic Calli Induction and Salt Stress Response Revealed by RNA-Seq in Diploid Wild Species Gossypium sturtianum and Gossypium raimondii." Frontiers in Plant Science 12 (August 25, 2021). http://dx.doi.org/10.3389/fpls.2021.715041.

Full text

Abstract:

Wild cotton species can contribute to a valuable gene pool for genetic improvement, such as genes related to salt tolerance. However, reproductive isolation of different species poses an obstacle to produce hybrids through conventional breeding. Protoplast fusion technology for somatic cell hybridization provides an opportunity for genetic manipulation and targeting of agronomic traits. Transcriptome sequencing analysis of callus under salt stress is conducive to study salt tolerance genes. In this study, calli were induced to provide materials for extracting protoplasts and also for screening salt tolerance genes. Calli were successfully induced from leaves of Gossypium sturtianum (C1 genome) and hypocotyls of G. raimondii (D5 genome), and embryogenic calli of G. sturtianum and G. raimondii were induced on a differentiation medium with different concentrations of 2, 4-D, KT, and IBA, respectively. In addition, embryogenic calli were also induced successfully from G. raimondii through suspension cultivation. Transcriptome sequencing analysis was performed on the calli of G. raimondii and G. sturtianum, which were treated with 200 mM NaCl at 0, 6, 12, 24, and 48 h, and a total of 12,524 genes were detected with different expression patterns under salt stress. Functional analysis showed that 3,482 genes, which were differentially expressed in calli of G. raimondii and G. sturtianum, were associated with biological processes of nucleic acid binding, plant hormone (such as ABA) biosynthesis, and signal transduction. We demonstrated that DEGs or TFs which related to ABA metabolism were involved in the response to salt stress, including xanthoxin dehydrogenase genes (ABA2), sucrose non-fermenting 1-related protein kinases (SnRK2), NAM, ATAT1/2, and CUC2 transcription factors (NAC), and WRKY class of zinc-finger proteins (WRKY). This research has successfully induced calli from two diploid cotton species and revealed new genes responding to salt stress in callus tissue, which will lay the foundation for protoplast fusion for further understanding of salt stress responses in cotton.

APA, Harvard, Vancouver, ISO, and other styles

40

El-Gendy, M. M. A. A., A. A. M. El-Bondkly, S. I. A. Mohamed, and A. M. A. El-Bondkly. "Enhancement of heavy metals removal from real electroplating effluent by protoplast fusion technology between different indigenous fungi." International Journal of Environmental Science and Technology, May 27, 2025. https://doi.org/10.1007/s13762-025-06548-5.

Full text

Abstract:

Abstract Among a collection of 64 natural isolates derived from the mycobiome of electroplating effluents screened for heavy metals biosorption, two fungal isolates AHM 21 and AHM 53 identified and designated as Fusarium sp. AHM 21 and Alternaria sp. AHM 53 proved to be potent fungal biosorbents for efficient removal of heavy metals. They were used as parental strains in intergeneric protoplast fusion technique. One of fifteen fusants; recombinant strain RAM 5 showed improved biosorption activity against a wide range of heavy metals compared to its parental strains. The recombinant strain RAM 5 achieved 100% removal of Fe3+, Co2+, Ni2+, Pb2+, and Cd2+ (pH 5.0–7.0 at 25–30 °C); Cr6+, and Zn2+ (5.0 -7.0 at 30 °C); Se4+, Ba2+, V+, and Cu2+ (pH 7.0 at 25–30 °C) and As3+ (pH 5.0 at 25–30 °C) from electroplating wastewater. It also reduced the amounts of Hg2+, total dissolved solids, Cl−, SO4 2−, Na+, and PO4 by 87.90, 77.12, 60.21, 61.05, 62.44, and 71.00%, respectively at pH 7.0 and 30 °C and removed Be2+, Mn2+, K+, chemical oxygen demand, biochemical oxygen demand, total phosphorous, and NO3 − by 96.14, 99.33, 71.00, 82.14, 67.71, 94.50, and 58.43% (pH 5.0 at 25 °C). Interestingly, fusant RAM 5 showed no antiproliferative activity against hepatocellular carcinoma, human breast cancer and normal human skin fibroblast cell lines. The current work provides cheaper, nontoxic, and more environmentally friendly biological treatment based on fungi for metals removal.

APA, Harvard, Vancouver, ISO, and other styles

41

"Plant cell biotechnology and agriculture: impacts and perspectives." Philosophical Transactions of the Royal Society of London. B, Biological Sciences 310, no. 1144 (1985): 215–20. http://dx.doi.org/10.1098/rstb.1985.0109.

Full text

Abstract:

Methods of cell and tissue culture, together with the regeneration of whole plants, are now routine for many plant species and are rapidly becoming key tools in the further development of agriculture and horticulture. At the base of the technology lies the ability to produce large numbers of identical (cloned) plants far more rapidly than with traditional practice. A number of commercially important species, including strawberries, tomatoes, oil palms, Douglas firs and orchids are already being produced in large numbers through tissue culture. A further dimension is provided by techniques of protoplast-fusion-somatichybridization and genetic manipulation. Harnessed to methods of automated mass propagation it should be possible to screen for, and develop much more swiftly, new commercially important varieties than was possible hitherto with conventional plant breeding approaches. In spite of good progress in many aspects of plant tissue culture there is, however, much to be learned about the nature of the system. Techniques of plant tissue culture are by no means yet generally applicable.

APA, Harvard, Vancouver, ISO, and other styles

42

Bijalwan, Priyanka, and Shilpa . "Plant Tissue Culture- A New Tool for Vegetable Improvement (Indian scenario): A Review." Agricultural Reviews, Of (February 24, 2021). http://dx.doi.org/10.18805/ag.r-1979.

Full text

Abstract:

In vitro culture of plant cells/tissues is now routine using a range of explant types from many of the important vegetable and fruit crops. Successful technologies include isolation, culture of tissues, cells, protoplasts, organs, embryos, ovules, anthers and microspores and regeneration from them of complete plantlets. The development of plant tissue culture technology represents one of the most exciting advances in plant sciences. For example, the prospect of being able to introduce, develop, produce, transfer and conserve the existing gene pool of plant sciences by using tissue culture methods opens up new opportunities for researches and entrepreneurs. The term plant tissue culture should denote in vitro cultivation of plant cells or tissues in an unorganized mass, i.e., callus culture. Plant tissue culture techniques, in combination with recombinant DNA technology, are the essential requirements for the development of transgenic plants. However, culture techniques like anther/pollen/ovule culture, meristem culture can themselves be utilized for crop improvement or may serve as an aid to conventional breeding. In recent, isolated microspore culture has developed as a breeding tool and an experimental system for various genetic manipulations. The inherent potentiality of a plant cell to give rise to a whole plant, a capacity which is often retained even after a cell has undergone final differentiation in the plant body, is described as ‘cellular totipotency’. On the other hand, production of virus-free plants via meristem culture can reduce losses caused by phyto-pathogens. Embryo culture has many potential uses ranging from overcoming seed dormancy to facilitation of inter-specific hybridization. Protoplast fusion technique can be used for the transfer of cytoplasmic male sterility from one species to another in a short period of time. In cabbage, male sterile cybrids are being utilized by seed companies to produce hybrid seeds on commercial scale and at competitive rates. Plant tissue culture and cell culture are providing useful methods for germplasm storage either by low temperature storage of organized tissue, or cryopreservation of cell or embryo culture.

APA, Harvard, Vancouver, ISO, and other styles

43

Shruti, AYUSHI TRIPATHI, and SUSMITA SHUKLA. "METHODS OF GENETIC TRANSFORMATION: MAJOR EMPHASIS TO CROP PLANTS." Journal of microbiology, biotechnology and food sciences, December 4, 2023, e10276. http://dx.doi.org/10.55251/jmbfs.10276.

Full text

Abstract:

Advancements in gene transfer technology have indeed opened up exciting possibilities for more effectively manipulating the genetic makeup of live organisms, ranging from microorganisms to plants and animals. Direct and indirect transformations are the two basic types of gene transfer techniques. Indirect method comprises Agrobacterium mediated method as it involves intermediate host between gene of interest and target and this method is most opted one out of all present. Direct gene transformation methods, on the other hand, do not involve the use of an intermediate host organism. Instead, they rely on physical means to transfer genes between cells. Biolistic transformation uses high-velocity particles to deliver DNA into target cells, while microinjection and macroinjection involve the direct injection of DNA into cells. Protoplast fusion combines the genetic material of two different cells by fusing their protoplasts. Natural methods for gene transfer encompass mechanisms that occur naturally in various organism, includes transposition, conjugation, phage and retroviral transductions and bacterial transformation. Chemical techniques utilize chemical agents to facilitate gene transfer, such as calcium phosphate-mediated transformation, polyethylene glycol (PEG)-mediated transformation, DEAE (Diethylethanolamine)-Dextran-mediated transformation. Genes can be also being transferred using electrical techniques such as electroporation and electrofusion. Crop improvement and trait improvement are now being hastened by the fast-rising number of sequenced plant genomes, information from functional genomics data to understand gene function, innovative gene cloning, and tissue culture techniques. Despite being indispensable, its progress is still hindered by the fact that many plant species and agricultural genotypes exhibit low transformability or are resistant to established tissue culture and regeneration conditions. Here, we review the techniques employed in plant transformation and provide a concise overview of their evolution in agricultural crops, from their first inception to present time.

APA, Harvard, Vancouver, ISO, and other styles

44

Zhang, Meng, Rongkang Tang, Fang-Xia Li, et al. "Optimization of ultrasound-mediated DNA transfer for bacteria and preservation of frozen competent cells." Microbiology Spectrum, November 12, 2024. http://dx.doi.org/10.1128/spectrum.00978-24.

Full text

Abstract:

ABSTRACT The transformation of DNA into cells is the basis of molecular biology. Commonly employed techniques include heat shock transformation, electro-transformation, conjugation, transduction, and protoplast fusion. Recently, ultrasonic transformation technology has been developed to transfer DNA into competent cells. The transformation conditions, such as temperature and ultrasonic power, were preliminarily studied. However, this technique has not been widely applied because competent cells must be prepared de novo . In this study, various factors, such as ultrasonic frequency and power, were optimized for the ultrasonic transformation of Escherichia coli . The study found that the optimal conditions for ultrasonic transformation with a defined ultrasonic transformation vial were a frequency of 28 kHz and a power of 80 W. Meanwhile, this research demonstrated that combining the 42°C heat shock conditions with ultrasonic transformation is the most efficient method compared to using only heat shock. Furthermore, the cryoprotective agent ratio for ultrasonic competent cells was investigated and optimized. These findings provide new insights into enhancing transformation efficiency and lay a foundation for the broader application of ultrasonic transformation. IMPORTANCE Plasmid transformation is widely applicable in gene expression and modification. As an efficient, non-invasive, and gentle method of transformation, ultrasonic transformation provides a novel approach for strain modification. This research presents new strategies for enhancing transformation efficiency and lays the groundwork for expanding the utilization of ultrasonic transformation.

APA, Harvard, Vancouver, ISO, and other styles

45

"Antibiotics: opportunities for genetic manipulation." Philosophical Transactions of the Royal Society of London. B, Biological Sciences 324, no. 1224 (1989): 549–62. http://dx.doi.org/10.1098/rstb.1989.0067.

Full text

Abstract:

New antibiotics can still be discovered by the development of novel screening procedures. Notable successes over the last few years include the monobactams, β- lactamase inhibitors (clavulanic acid) and new glycopeptides in the antibacterial held; antiparasitic agents such as avermectins; and herbicidal antibiotics like bialaphos. In the future we can expect the engineering of genes from ‘difficult’ pathogens, including mycobacteria and fungi, and cancer cells, to provide increasingly useful in vitro targets for the screening of antibiotics that can kill pathogens and tumours. There will also be a greater awareness of the need to reveal the full potential for antibiotic production on the part of microorganisms by the physiologial and/or genetic awakening of ‘silent’ genes. Nevertheless, the supply of natural antibiotics for direct use or chemical modification is not infinite and there will be increasing scope for widening the range of available antibiotics by genetic engineering. ‘Hybrid’ antibiotics have been shown to be generated by the transfer of genes on suitable vectors between strains producing chemically related compounds. More exciting is the possibility of generating novelty by the genetic engineering of the synthases that determine the basic structure of antibiotics belonging to such classes as the β-lactams and polyketides. Research in this area will certainly yield knowledge of considerable scientific interest and probably also of potential applicability. In the improvement of antibiotic titre in actinomycetes, protoplast fusion between divergent selection lines has taken a place alongside random mutation and screening. In some cases the cloning of genes controlling metabolic ‘bottlenecks’ in fungi and actinomycetes will give an immediate benefit in the conversion of accumulated biosynthetic intermediates to the desired end product. However, the main impact of genetic engineering in titre improvement will probably come only after a further use of this technology to understand and manipulate the regulation of antibiotic biosynthesis as a facet of the general challenge of understanding differential gene expression. Streptomyces offers a particularly fertile field for such research, following the isolation of DNA segments that carry groups of closely linked operons for the biosynthesis of and resistance to particular antibiotics, and of genes with pleiotropic effects on morphological differentiation and secondary metabolite formation.

APA, Harvard, Vancouver, ISO, and other styles

Journal articles on the topic 'Protoplast fusion technology'

Create a spot-on reference in APA, MLA, Chicago, Harvard, and other styles