ArticlePlant methods2016
Protocol: transient expression system for functional genomics in the tropical tree Theobroma cacao L.
Article in Plant methods, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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Who cites it
24 citing papers in PubMed, 51 citations in OpenAlex.
- Reduced Susceptibility to Phytophthora in Non-Transgenic Cacao Progeny Through CRISPR-Cas9 Mediated TcNPR3 Mutagenesis.Plant biotechnology journal · 2026Article
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- Improvement and application of vacuum-infiltration system in tomato.Horticulture research · 2024Article
- The cacao gene atlas: a transcriptome developmental atlas reveals highly tissue-specific and dynamically-regulated gene networks in Theobroma cacao L.BMC plant biology · 2024Article
- Critical parameters for robustPlant direct · 2024Article
- A combination of conserved and diverged responses underlies Theobroma cacao's defense response to Phytophthora palmivora.BMC biology · 2024Article
- Interactions of Different Species ofMicroorganisms · 2023Review
- CRISPR/Cas9-Mediated Mutation inGenes · 2023Article
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- Genetic engineering of novel yellow color african violet (Saintpaulia ionantha) produced by accumulation of Aureusidin 6-O-glucoside.Biological procedures online · 2022Article
- The effects of the phytochemistry of cocoa on the food chemistry of chocolate(s) and how disease resistance in cocoa can be improved using CRISPR/Cas9 technology.Food chemistry. Molecular sciences · 2021Review
- A high-efficiency Agrobacterium-mediated transient expression system in the leaves of Artemisia annua L.Plant methods · 2021Article
- Robust, Long-Term, and Exceptionally Sensitive Microneedle-Based Bioimpedance Sensor for Precision Farming.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2021Article
- A simple and efficient agroinfiltration method for transient gene expression in Citrus.Plant cell reports · 2021Article
- Red Clover HDT, a BAHD Hydroxycinnamoyl-Coenzyme A:L-3,4-Dihydroxyphenylalanine (L-DOPA) Hydroxycinnamoyl Transferase That Synthesizes Clovamide and OtherFrontiers in plant science · 2021Article
- Clovamide, a Hydroxycinnamic Acid Amide, Is a Resistance Factor AgainstFrontiers in plant science · 2020Article
- Article
- Development of Agrobacterium-mediated transient expression system in Caragana intermedia and characterization of CiDREB1C in stress response.BMC plant biology · 2019Article
- Resistant and susceptible cacao genotypes exhibit defense gene polymorphism and unique early responses to Phytophthora megakarya inoculation.Plant molecular biology · 2019Article
- Article
Corrections and comments
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Authors and funding
6 authors at 4 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundTheobroma cacao L., the source of cocoa, is a crop of significant economic value around the world. To facilitate the study of gene function in cacao we have developed a rapid Agrobacterium-mediated transient genetic transformation protocol. Here we present a detailed methodology for our transformation assay, as well as an assay for inoculation of cacao leaves with pathogens.
resultsAgrobacterium tumefaciens cultures are induced then vacuum-infiltrated into cacao leaves. Transformation success can be gauged 48 h after infiltration by observation of green fluorescent protein and by qRT-PCR. We clarify the characteristics of cacao leaf stages and demonstrate that our strategy efficiently transforms leaves of developmental stage C. The transformation protocol has high efficacy in stage C leaves of four of eight tested genotypes. We also present the functional analysis of cacao chitinase overexpression using the transient transformation system, which resulted in decreased pathogen biomass and lesion size after infection with Phytophthora tropicalis.
conclusionsLeaves expressing transgenes of interest can be used in subsequent functional genetic assays such as pathogen bioassay, metabolic analysis, gene expression analysis etc. This transformation protocol can be carried out in 1 day, and the transgenes expressing leaf tissue can be maintained in petri dishes for 5-7 days, allowing sufficient time for performance of additional downstream gene functional analysis. Application of these methods greatly increases the rapidity with which candidate genes with roles in defense can be tested.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.