Transformation of elite switchgrass (Panicum virgatum L.) genotypes would facilitate the characterization of genes related to cell wall recalcitrance to saccharification. However, transformation of explants from switchgrass plants has remained difficult. Therefore, the objective of this study was to develop a biolistic transformation protocol for elite genotypes. Three switchgrass genotypes (ST1, ST2, and AL2) were previously selected for tissue culture responsiveness. One genotype, SA37, was selected for further use due to its improved formation of callus amenable to transformation. Various medium sets were compared and a previously published medium set provided cultures with >96 % embryogenic callus, and data on transient and stable gene expression of RFP were used to optimize biolistic parameters, and further validate the switchgrass (PvUbi1) promoter. SA37 proved to be the most transformable, whereas eight transgenic calli on average were recovered per bombardment of 20 calli (40 % efficiency) when using a three-day day preculture step, 0.6 M osmotic adjustment medium, 4,482 kPa rupture disks and 0.4 μm gold particles which traveled 9 cm before hitting the target callus tissue. Regenerability was high, especially for ST2, for which it is possible to recover on average over 400 plants per half-gram callus tissue. It is now possible to routinely and efficiently engineer elite switchgrass genotypes using biolistic transformation.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1007/s00299-013-1531-1 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Nano-assisted delivery tools for plant genetic engineering: a review on recent developments.
Environ Sci Pollut Res Int
December 2024
Department of Pharmaceutical Engineering & Technology, Indian Institute of Technology, Banaras Hindu University, Varanasi, 221005, UP, India.
Conventional approaches like Agrobacterium-mediated transformation, viral transduction, biolistic particle bombardment, and polyethylene glycol (PEG)-facilitated delivery methods have been optimized for transporting specific genes to various plant cells. These conventional approaches in genetically modified crops are dependent on several factors like plant types, cell types, and genotype requirements, as well as numerous disadvantages such as time-consuming, untargeted distribution of genes, and high cost of cultivation. Therefore, it is suggested to develop novel techniques for the transportation of genes in crop plants using tailored nanoparticles (NPs) of manipulative and controlled high-performance features synthesized using green and chemical routes.
View Article and Find Full Text PDFHeterologous Gene Expression in Chlamydomonas reinhardtii Chloroplast by Heterologous Promoters and Terminators, Intercistronic Expression Elements and Minichromosome.
Microb Biotechnol
December 2024
State Key Laboratory of Agricultural Microbiology, Hubei Hongshan Laboratory, Huazhong Agricultural University, Wuhan, China.
Chlamydomonas reinhardtii, a model green alga for expressing foreign proteins, faces challenges in multigene expression and enhancing protein expression level in the chloroplast. To address these challenges, we compared heterologous promoters, terminators and intercistronic expression elements (IEEs). We transformed Chlamydomonas chloroplast with a biolistic approach to introduce vectors containing the NanoLuc expression unit regulated by Chlamydomonas or tobacco promoters and terminators.
View Article and Find Full Text PDFUnlocking regeneration potential: harnessing morphogenic regulators and small peptides for enhanced plant engineering.
Plant J
December 2024
Department of Agronomy, Iowa State University, Ames, Iowa, 50011, USA.
Plant genetic transformation is essential for understanding gene functions and developing improved crop varieties. Traditional methods, often genotype-dependent, are limited by plants' recalcitrance to gene delivery and low regeneration capacity. To overcome these limitations, new approaches have emerged that greatly improve efficiency and genotype flexibility.
View Article and Find Full Text PDFA protocol for in vivo RNA labeling and visualization in tobacco pollen tubes.
STAR Protoc
December 2024
Laboratory of Pollen Biology, Institute of Experimental Botany of the Czech Academy of Sciences, Rozvojová 263, 165 00 Prague, Czech Republic. Electronic address:
Here, we present a protocol for labeling and live visualization of RNA-protein complexes in the form of ribonucleoprotein particles (RNPs) in tobacco pollen tubes. We describe steps for constructing RNA-pp7/MS2 tag and biolistic gene-gun-mediated pollen transformation. We then provide detailed procedures for RNA labeling using PP7 aptamer nascent RNA tagging and a fluorescently labeled Pseudomonas aeruginosa PP7 bacteriophage coat protein (PCP) reporter that binds to PP7 RNA stem loops.
View Article and Find Full Text PDFAdvances in Soybean Genetic Improvement.
Plants (Basel)
October 2024
Departamento de Biotecnología y Bioingeniería, Centro de Investigación y de Estudios Avanzados (CINVESTAV), Ciudad de México 07360, Mexico.
Article Synopsis
- - The soybean is a critical global crop known for its high protein and oil content, essential for nutrition and bioenergy, and the review focuses on recent advances in genetic improvement to enhance resistance to environmental stresses and improve yields and nutritional value.
- - The study highlights various breeding techniques, including traditional methods, marker-assisted selection, and modern biotechnological approaches such as genetic engineering, which aim to create transgenic soybeans with desirable traits like herbicide resistance and pest tolerance.
- - Despite advancements, challenges persist in transformation processes and regulatory issues, but exploring wild soybean varieties and genetic diversity through epigenetics and microbiomes could enhance resilience, supporting food security and sustainable agriculture in the face of climate change.
Want AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!