Theobroma cacao L. Transformation

Cocoa is the main cash crop of developing countries in the tropics, second only to coffee. Theobroma cacao L., the source of cocoa, is an essential woody crop and the centerpiece of the multi-billion dollar chocolate industry. Scientists have been working to breed varieties resistant to pests and diseases. However, Theobroma cacao L. is slow, expensive, and requires a lot of land to grow. The availability of cacao genome sequence data provides an opportunity to apply new breeding strategies or biotechnological approaches to improve Theobroma cacao L. genetic improvement. Scientists have proposed a strategy for generating stable transgenic Theobroma cacao L.. However, even this process takes several years to produce a mature tree that can be used to experimentally determine the effects of transgene overexpression or knockdown.

Fig. 1. Workflow diagram for transient transformation of cacao leaf tissue. (Fister A S, et al, 2016)

Solutions

Lifeasible is an industry leader in plant genetic transformation and is proud to offer cutting-edge genetic transformation services for Theobroma cacao L.. Our experts use Agrobacterium tumefaciens to genetically transform cocoa leaf cells. We aim to help our customers produce Theobroma cacao L. with enhanced resistance to pests and diseases.

We offer protocols for Agrobacterium-mediated transient transformation of isolated cocoa leaf tissues. Our transformation process involves co-culturing leaf strips of selected Theobroma cacao L. clones with Agrobacterium superb strain A281-Kan. This strain carries the wild-type Ti plasmid and the add-on plasmid pGPTV-Kan, which conferred kanamycin resistance to the transformed plant cells upon neomycin phosphotransferase II (nptII) gene integration and expression. By careful screening of kanamycin-containing healing tissue culture media, we can identify and isolate transformed cells with enhanced resistance.

We use green fluorescent protein (GFP) expression by observation and quantitative real-time PCR (qRT-PCR) to indicate successful transformation. Transformation efficiency is validated on cocoa leaves at specific developmental stages to ensure consistent and reliable results. Our transient transformation protocols and subsequent functional analyses can be completed within a week, ensuring high transformation success rates for efficient gene expression and characterization in cocoa leaves.

Applications of Our Theobroma cacao L. Transformation

• Pathogen bioassays for disease resistance studies: Our Theobroma cacao L. transformation service can help you overexpress genes associated with defense mechanisms and assess their impact on pathogen resistance.
• Accelerated functional genomics studies: We can help you perform functional genomics studies more efficiently. Leaves expressing transgenes of interest can be used for various downstream analyses, such as metabolic studies and gene expression analysis.
• Accelerating genetic improvement of Theobroma cacao L.: We can help you identify candidate genes for defense more quickly, thereby accelerating the development of new breeding strategies or biotechnological approaches to improve cocoa trees.

Lifeasible's Theobroma cacao L. genetic transformation services provide a cutting-edge approach to designing disease- and insect-resistant cocoa plants. Through continuous innovation and collaboration, we strive to revolutionize Theobroma cacao L. production and promote sustainable growth in the cocoa industry. If you are interested in our solutions, please contact us for technical consultation and quotation.