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A study on the creation and growth of hairy root in luffa cylindrical produced by the insemination of various agrobacterium rhizogenes species by tissue calture
1. pasteur institute of IRAN 2. Islamic Azad University،Science And Research Branch 3. pasteur institute of IRAN 4. Islamic Azad University،Science And Research Branch 6. pasteur institute of IRAN
Abstract
Introduction
• luffin, a protease inhibitor, is a plant metabolite that has antibacterial and antifungal properties that is more produced in plant roots.the antibacterial activity of luffin has been experimentedon several bacteria including: staphylococcus aureus, bacillus subtilis and salmonella typhi. thus, applications of luffinhad been investigated in pharmaceutical industry to control the bacterial infections. studies have shown that chemical synthesize of luffin is not convenient and manipulation of plant to produce higher amount of luffin is more promising. biotechnological approaches have been used to enhance manufacturing of the valuable pharmaceutical metabolite in medicinal plants such as generation of hairy roots . application of agrobacterium rhizogenesto produce hairy roots is an alternative method to increase particular pharmaceutical secondary metabolite invitro.
Methods
In order to investigate the ability of bacteria to increase production of luffin we had tested 3 different agrobacterium rhizogenes strains: 15834, 9534 and a4.
the germinated stems from luffa cylindrical (l) m. roem seeds were incubated with 3 above bacterial culture for 48 hours.
the transgenic plant were selected on murashigskoog media containing cefotaxime antibiotic. the transgenes roots were confirmed by tracking the rolb gene situated on tdna of agrobacteria using its specific primers and pcr technique. the transgenic roots were grown and rna were analyzed to detect luffin expression by qrt pcr.
Results
Interestingly, between 3 agrobacteria the most root biomass and luffin gene expression were seen in transgenic roots grew following incubation with 15834 a.rhizogenes strain. moreover, the 9534 and a4a.rhizogenes strains were orderly induced the production of luffin protein in transgenic root samples in comparison to non transgenic plant.
Conclusion
This study showed that by choosing appropriate agrobacterium rhizogenesstrainit is possible to increase the production of valuable pharmaceutical metabolites using hairy root system.
