GENETIC TRANSFORMATION AND GENOME EDITING: BIOTECHNOLOGICAL TOOLS TO MITIGATE THE EFFECT OF BIOTIC AND ABIOTIC STRESSES IN SUGARCANE CULTIVATION.

ENRIQUE, RAMÓN; BUDEGUER, FLORENCIA; OSTENGO, SANTIAGO; NOGUERA, ALDO

CABA

Conferencia; 27th ANNUAL ICABR CONFERENCE; 2023

ICARB

Sugarcanecultivation is faced with various biotic and abiotic stresses that negativelyimpact its productivity. Traditional breeding methods are laborious and slowdue to the genetic complexity and low fertility of sugarcane. Geneticengineering, specifically genetic transformation (GT) and genome editing (GE),offer biotechnological tools to overcome these challenges and develop improvedsugarcane varieties. In this study, we aimed to introduce the HaHB11 gene from sunflower into an elitesugarcane variety (TUC 03-12) through GT and edit the als alleles of the same variety using CRISPR-Cas technology toconfer herbicide resistance. The GT experiments involved bombarding embryogeniccalli with a genetic construct containing the HaHB11 gene. Out of 1735 transformed calli, 117 transgenic linesresistant to geneticin were obtained, exhibiting increased Emergence VelocityIndex (EVI), stem length, and root development compared to non-transformedplants. These phenotypic changes observed under normal growth conditionssuggest that these transgenic lines could be more tolerant to face abioticstresses. For GE experiments, calli were bombarded with ribonucleoproteins(RNPs) to edit the als alleles. Tenpotentially edited lines resistant to the herbicide Bispyribac sodium were obtained.Preliminary analysis showed partial recovery of Acetolactate Synthase (ALS)enzyme activity in some edited lines. These results demonstrate the successfulgenetic transformation of sugarcane using the HaHB11 gene and the generation of herbicide-resistant sugarcanelines through genome editing of the alsalleles. Further studies are needed to verify the functionality of theintroduced/edited genes and evaluate their effectiveness under stressconditions in the field. These biotechnological approaches have the potentialto contribute to the development of stress-tolerant sugarcane varieties, thusenhancing its productivity and sustainability in the face of challengingenvironmental conditions.