OPTIMIZATION OF PRICKLY PEAR CACTI (Opuntia spp.) MICROPROPAGATION USING AN EXPERIMENTAL DESIGN METHOD

Achraf Mabrouk; Younes Abbas; Younes El Goumi; Ezzouhra El Maaiden; Youssef El Kharrassi; Salwa EL Antry-Tazi; El Houssine EL Mzouri; Malika Fakiri

doi:10.15414/jmbfs.1577

Authors

Achraf Mabrouk Phd student, Laboratory of Agri-food and Health, Faculty of Sciences and Techniques, Hassan 1st University https://orcid.org/
Younes Abbas PhD, University Sultan Moulay Slimane, Polidisciplinary faculty.
Younes El Goumi Doctor, Laboratory of Phyto-Biotechnology, National Agency for Medicinal and Aromatic Plants https://orcid.org/
Ezzouhra El Maaiden PhD student, Laboratory of Agri-food and Health, Faculty of Sciences and Techniques, Hassan 1st University
Youssef El Kharrassi Dr in the Laboratory of Biochemistry and Neurosciences, Department of Biology, University Hassan Ist
Salwa EL Antry-Tazi Researcher at the Forestry Research Center, High Commission for Water and Forests and the fight against desertification
El Houssine EL Mzouri Dr at the National Institute of Agronomic Research, Regional Research Centre of Settat
Malika Fakiri PhD at the Laboratory of Agri-food and Health, Faculty of Sciences and Techniques, Hassan 1st University

DOI:

https://doi.org/10.15414/jmbfs.1577

Keywords:

Disinfection, Opuntia Spp, Micropropagation, Contamination, necrosis, ANA/BAP, Optimal design

Abstract

In Morocco, despite the adaptation of the prickly pear cactus (Opuntia spp) to edaphoclimatic conditions, differing from those that prevail in the countries from which it originates, this introduced species is regressing due to biotic and abiotic stresses that impact its survival. Hence, finding innovative technologies to preserve this species are essential. However, the success of in vitro micropropagation technologies is hinged upon the initial disinfection step. In this work we have tested several solutions and exposure time mixtures for disinfection across three different explants, as well as the adoption of hormonal combinations for the induction/regeneration step that allow for efficient micropropagation of cacti. We aimed to produce healthy regenerable vitroplants capable of effectively preserving themselves under aseptic conditions. We describe a specific protocol based on the type of areoles: (1) cladode without areolas (CWA); (2) areolas with shredded glochids (ASG); and (3) areolas with intact glochids (AIG). For each of these areoles, the disinfectant concentration and contact time were optimized and various hormonal combinations were tested for induction/regeneration. The optimal design meeting I-optimality criterion was used to predict the optimal combination by maximizing desirability. The disinfection combination that used 4.23% calcium hypochlorite (w:v), 0.4% Tween 20 (v:v), and 10 minutes of exposure time reduced the contamination frequency to 0.002% with a necrosis rate of less than 0.33% in explants with AIG. This type was the only one that gave responses in the induction/regeneration phase, which allowed us to predict the best rate of caulogenesis (71.74%) under a hormonal combination of 5.3 mg/L of BenzylAminopurine (BAP) and 0.59 mg/L of 1-Naphthaleneacetic acid (ANA). Furthermore, the highest callogenesis rate of 94.45% was obtained by a hormonal combination of 6 mg/L of BAP and 0.59 mg/L of ANA.Â The explantâ€™s rhizognesis was carried out with a concentration of 0.5 mg / l ANA, before acclimatization on a substrate containing soil and sand in a ratio of 3:1. In conclusion, these protocols provide easy and cost-effective means to multiply cacti in vitro to preserve them in areas where their culture has been destroyed by cochineal.