Agrobacterium-mediated transformation is a suitable method to transform different cultivars using different systems of A. tumefaciens strains and binary vectors as well as selection cassettes. We describe here a detailed protocol for two cultivars, one naked and one husked, using the AGL1 strain and the pGreen vector containing the nptII selection cassette ( http://www.pgreen.ac.uk/ ), suitable for oat as well as other cereals. The pGreen vector system was recently developed for pBract ( http://www.bract.org/ ) and its transformation ability for cereals was proved. Assuming our experience and the latest knowledge on Agrobacterium-mediated transformation of cereals, we suggest using in the protocol one of the newly developed pBract or pCAMBIA ( http://www.cambia.org/daisy/cambia/ ) vector systems which carry different selection cassettes. The commonly used selection genes nptII, bar, and hpt were proved to be applicable for oat transformation and might be used as needed.
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http://dx.doi.org/10.1007/978-1-4939-1695-5_11 | DOI Listing |
BMC Plant Biol
January 2025
National Key Laboratory for Germplasm Innovation & Utilization of Horticultural Crops, Zhengzhou Fruit Research Institute, Chinese Academy of Agricultural Sciences, Zhengzhou, 450009, China.
Background: Red Actinidia arguta has recently become highly popular because of its red appearance resulting from anthocyanin accumulation, and has gradually become an important breeding direction. However, regulators involved in anthocyanin biosynthesis have not been fully characterized in A. arguta.
View Article and Find Full Text PDFFunct Integr Genomics
January 2025
Department of Botany, Maharshi Dayanand University, Rohtak, 124001, India.
Despite years of progress in biotechnology, altering the genetic makeup of many plant species, especially their plastids, remains challenging. The existence of a cell wall poses a significant obstacle to the effectual transportation of biomolecules. Developing efficient methods to introduce genes into plant cells and organelles without causing harm is an ongoing area of research.
View Article and Find Full Text PDFPhysiol Plant
January 2025
Institute of Biochemistry and Biology, University of Potsdam, Potsdam-Golm, Germany.
Genetic transformation is a powerful tool in plant biotechnology. However, its application is limited to species that are well-studied and easy to transform. There is a critical need to establish transformation protocols for non-model species.
View Article and Find Full Text PDFPlant Cell Physiol
January 2025
Plant Genetic Engineering Laboratory, Department of Biotechnology, Bharathiar University, Coimbatore, 641046, India.
Carnosol (CO) and carnosic acid (CA) are pharmaceutically important diterpenes predominantly produced in members of Lamiaceae, Salvia officinalis (garden sage), Salvia fruticosa and Rosmarinus officinalis. Nevertheless, availability of these compounds in plant system is very low. In an effort to improve the in planta content of these diterpenes in garden sage, SmERF6 (Salvia miltiorrhiza Ethylene Responsive Factor 6) transcription factor was expressed heterologously.
View Article and Find Full Text PDFBio Protoc
January 2025
Department of Biochemistry, Microbiology and Biotechnology, Kenyatta University, Nairobi, Kenya.
Agrobacterium-mediated gene transformation method is a vital molecular biology technique employed to develop transgenic plants. Plants are genetically engineered to develop disease-free varieties, knock out unsettling traits for crop improvement, or incorporate an antigenic protein to make the plant a green factory for edible vaccines. The method's robustness was validated through successful transformations, demonstrating its effectiveness as a standard approach for researchers working in plant biotechnology.
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