کنگره بین المللی زیست فناوری گیاهان دارویی و قارچ های کوهی

صفحه اصلی / چهارمین کنگره بین المللی و پنجمین همایش ملی زیست فناوری گیاهان دارویی و قارچ های کوهی

Hairy root Induction of Foeniculum vulgare M.

نویسندگان :

Shirin Alijani¹

1- دانشگاه تهران

کلمات کلیدی :

Agrobacterium rhizogenes،Foeniculum vulgare،Hairy roots،Inplanta،Phenological course

چکیده :

Abstract This study was conducted with the aim of investigating the induction of hairy roots in fennel plant (Foeniculum vulgare Miller.). In this research, the effects of two strains of Agrobacterium rhizogenes A4 and ATCC15834 on three genotypes of Hajiabad, Meshkin Shahr and Fasa respectively with late, mid and early phenological courses and using cotyledon, hypocotyl, root, leaf and stem explants was evaluated. This research was done in two environmental conditions. Induction of hairy roots was done in laboratory environment using MS culture medium containing cefotaxime antibiotic and in greenhouse in culture trays containing cocopeat and perlite (Inplanta). The inoculation time of the explants was 30 minutes, with a concentration of 0.8 in OD600, and the co-cultivation time was recorded as 48 hours. Differences were realized in the frequency of hairy root induction of three types of F.vulgare genotypes and their explants. The highest ferequency of induction in Haji Abad genotype with A4 strain in hypocotyl explant was 63%, the highest ferequency in Meshkin Shahr's genotype with A4 in cotyledon explant was 53% and the highest in Fasa's genotype with ATCC15834 in cotyledon explant was recorded 70%. This percentage in Inplanta method (the direct injection in the tissue), showed the highest percentage of transgenics with 27% belonged to Fasa variety using ATCC15834 strain. Treatment of ascorbic acid and darkness, simultaneously, for the first 7 days after induction reduced the amount of browning of explants. In comparing the average of 5 explants of cotyledon, hypocotyl, leaf, stem and root, cotyledon showed the highest rate of transformation with 70%. Variance analysis of hairy root number transformation percentage between three genotypes, in cotyledon, hypocotyl and leaf explants and two bacterial strains showed a significant difference. Transgenic status in hairy roots was confirmed by PCR and by examining rolC and rolB genes. Thus, hairyroot induction can be applied as an alternative method for the production of secondary metabolites and also as a novel method in metabolic engineering