Micropropagation of Iraqi Garlic (Allium sativum L.) Through Tissue Culture Method and Assessment of Genetic Stability Using RAPD Markers

Garlic (Allium sativum L.) is a valuable medicinal and culinary crop in Iraq, facing yield limitations due to vegetative propagation and disease.Tissue culture technique offers a viable solution for rapid propagation.This study aimed to micro-propagate Iraqi garlic and evaluate the genetic stability of regenerated plants using randomly amplified polymorphic DNA (RAPD) molecular markers.Garlic explants were cultured on Murashige and Skoog's (MS) medium with varying hormone combinations for callus induction, shoot regeneration, and rooting.Regenerated microplants were acclimatized in sterile substrate.DNA was extracted from leaf tissues, and genetic stability was assessed using RAPD markers.Optimal sterilization of Iraqi garlic was achieved with 0.8% sodium hypochlorite (NaOCl) for 20 minutes, resulting in 64% clean explants.MS medium supplemented with 5.0 mg/L 6-benzyladenine (BA) resulted in an 80% callus response and an average of 6.6 shoots per explant.Meanwhile, 2.5 mg/L BA produced the longest shoot length (1.80 cm).RAPD analysis using the OPC-03 primer showed 10 consistent bands (1835-1910 bp), confirming the genetic stability of regenerated plants.Maximum induction (70%) and fresh weight (286.3 mg) occurred with 100 M 2,4dichlorophenoxyacetic acid (2,4-D).The use of liquid medium enhanced bud formation, with an average of 24.8 buds per explant, and achieved acclimatization rates of up to 80%.Overall, tissue culture proved to be an effective method for garlic propagation.The confirmed genetic stability of regenerated plants indicates strong potential for agricultural applications and future genetic improvement programs.