Oryza rufipogon Transformation

Oryza sativa L. is one of the most important crops in the world, providing staple food for more than half of the global population. Breeding new rice varieties with high tolerance to environmental changes caused by global warming is an urgent goal. Oryza rufipogon Griff. is a closely related wild species of rice, which retains a large number of high-quality genes and has the advantages of large biomass, self-hybrid dominance, high resistance to diseases and pests, and adaptability to adverse environments, such as drought, inundation, freezing, cold, high temperature, weather extremes, and salinity of the soil. Using Oryza rufipogon Griff. to create new rice germplasm with excellent characteristics is a powerful way to solve the food problem. However, Oryza rufipogon Griff. also has some undesirable traits, such as prostrate growth, plant height, long awns, grain drop, and long tasseling period. Traditional crossbreeding cannot eliminate these undesirable traits quickly, and it is difficult to obtain the good traits of the parents simultaneously. With the development of molecular biology, targeted modification of wild rice-related genes using genome editing technology is a fast and direct way to create new rice germplasm resources.

Fig. 1. Agrobacterium-mediated transformation of wild Oryza. (himizu-Sato S et al., 2020)

Solutions

Lifeasible is an industry leader in plant genetic transformation and is proud to offer cutting-edge genetic transformation services for Oryza rufipogon. We have established several technology platforms to transfer genes into rice, including particle bombardment, polyethylene glycol-mediated, and Agrobacterium-mediated methods. With the modified Agrobacterium-mediated approach, transformation efficiency becomes high, and transgenes with single or low copy number or large DNA fragments with defined ends can be integrated into the host genome.

We offer two solutions for the Oryza rufipogon genetic transformation:

• Agrobacterium-mediated genetic transformation of Oryza rufipogon using immature embryos. This method is convenient in that we can collect and store enough mature seeds to conduct experiments at any time.
• Agrobacterium-mediated genetic transformation of Oryza rufipogon using guillotine tissue induced from the shield tissue of embryos in mature seeds. This approach can be applied to a broader range of cultivated rice.

Our state-of-the-art technology and expertise have enabled us to introduce cold tolerance and disease resistance genes into Oryza rufipogon, leading to the development of new rice varieties more tolerant to environmental changes.

Transferable Varieties

• O. mfipogon Griff.
• O. rufipogon Griff.
• O. longistaminata
• O. glaberrima

Genetic Transformation Workflow of Oryza rufipogon

Through years of meticulous experimentation and research, we have successfully designed genetic transformation programs suitable for a wide range of Oryza rufipogon.

• Identification of Target Genes and Vector Construction

We will first identify the target genes in wild rice. Once identified, a suitable vector (usually Agrobacterium rhizogenes) will be selected to deliver the desired genetic material into plant cells.

• Healing Tissue Induction and Succession Cultures / Plant Regeneration from Immature Embryos

Depending on the project requirements, we will select the use of shield tissue from immature embryos or embryos from mature seeds. We ensure high induction rates.

• Agrobacterium-mediated transformation

After induction of the healing tissue, Agrobacterium carrying the desired genes is introduced. Our protocols are customized to maximize transformation efficiency while ensuring the genetic stability of the transformed cells.

• Screening and Characterization

We employ a rigorous screening process involving antibiotic resistance markers or reporter genes such as GFP (green fluorescent protein).

• Regeneration and Hardening

Transformed healing tissues are directed to plant regeneration. Once seedlings are formed, they undergo a hardening process to prepare them for transplantation and further growth under natural conditions.

• Validation and Analysis

The final step involves molecular analyses such as PCR, Southern blotting, and phenotypic evaluation to verify the success of the transformation and to ensure the stability and expression of the introduced genes.

Why Choose Us

• Transformable varieties of Oryza rufipogon, some still under development.
• We have a mature Oryza rufipogon transformation system with optimized transformation protocols.
• Just provide the gene ID or sequence.
• High positivity rate and best efforts to ensure success.

Lifeasible offers state-of-the-art genetic transformation services for wild Oryza species, providing growers and breeders with the tools to unlock the full potential of this extraordinary crop. If you are interested in our solutions, please contact us for technical consultation and quotation.