Aug 2026· Plant Cell Tissue and Organ Culture· Vol 166· 0 citations· 62 references
TL;DR
A novel embryonic axis-based regeneration system enables rapid shoot recovery, rooting, and biolistic gene delivery in cacao, substantially shortening the timeline for plant transformation.
Abstract
Theobroma cacao L. genetic transformation still relies predominantly on somatic embryogenesis-based systems, which are often genotype-dependent, labor-intensive, and associated with prolonged culture periods. In this study, we developed an alternative regeneration platform based on mature seed-derived embryonic axes and evaluated its suitability for biolistic DNA delivery. Embryonic axes from the cacao cultivars ‘TSH1188’ and ‘Parazinho’ were cultured on media supplemented with different cytokinins for multiple shoot induction. Among the plant growth regulators tested, 6-benzylaminopurine (BAP) was the most effective, promoting direct organogenesis from the apical dome and producing the highest shoot regeneration frequency and number of shoots per explant at 0.75 mg L⁻¹ for ‘TSH1188’ and 1.0 mg L⁻¹ for ‘Parazinho’. Sensitivity assays using antibiotics and herbicides identified genotype-dependent responses to kanamycin and hygromycin, whereas ammonium glufosinate, imazapyr, and glyphosate caused severe phytotoxic effects and completely inhibited shoot regeneration at relatively low concentrations. Hygromycin at 2.5 mg L⁻¹ (‘Parazinho’) and 5.0 mg L⁻¹ (‘TSH1188’) provided effective inhibition of shoot regeneration while maintaining the recovery of regenerating tissues, supporting its use in future selection protocols. For rooting, indole-3-acetic acid (IAA) at 0.5 mg L⁻¹ promoted the highest rooting efficiency, generating elongated roots and vigorous plantlets without intermediate callus formation. Preliminary biolistic assays using the pCAMBIA2301 vector resulted in localized transient GUS expression in the apical dome, demonstrating that this meristematic region is competent for exogenous DNA delivery. Collectively, these findings identify mature seed-derived embryonic axes as a promising explant for future stable genetic transformation and genome editing in cacao. A novel embryonic axis-based regeneration system enables rapid shoot recovery, rooting, and biolistic gene delivery in cacao, substantially shortening the timeline for plant transformation.
It is concluded that bridging the gap between foundational CRISPR research and its real-world applications is imperative and future efforts should focus on democratizing tools via open-source platforms, advancing delivery systems, and fostering sustainable innovation through synthetic biology integration to fully realize the transformative potential of genome editing in organisms beyond model organisms.
S. Sarsaiya, Archana Jain, Jishuang Chen et al.· Biotechnology Advances· 2 citations
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Zofia Pietrasik, Mikołaj Kapała, Joanna Pietrasik et al.· Cancers· 0 citations
Genetic engineering (GE) and gene editing may endow traits to trees such as increased biomass and the production of novel biomaterials. Long-lived organisms such as trees might be subject to biotechnology-related risks that could be different than those of annual row crops. Those risks could be relevant to production in engineered plantations and beyond plantations to natural forests. Therefore, appropriate risk regulation is important to assure biosafety of commercialized engineered trees. In addition to gene flow via sexual reproduction, vegetative reproduction might play an additional role in environmental "exposure" risk relative to transgene dispersal in GE tree plantations. While vegetative reproduction is beneficial for preserving desired genetic traits during tree propagation, it may lead to proximal clonal spread in the field. Although the environmental risks associated with vegetative reproduction of GE trees are recognized in commercial forestry, there are few field-based environmental risk assessment (ERA) studies on dispersal risks of self-propagated GE trees. GE or gene editing of target genes involved in the vegetative propagation processes may be useful to mitigate environmental risks of clonal spread through vegetative reproduction. This review provides updates for recent field test results of GE and gene edited trees. Gene candidates related to vegetative reproduction including adventitious shooting (AS) and adventitious rooting (AR) are discussed herein as a means to mitigate unintended clonal spread from GE tree plantations.
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MIT News · Artificial Intelligence· news.mit.eduAug 17, 2026