| مستخلص: |
Highlights • In this study a reliable gene transfer system via modifying the co-cultivation medium using different strains of Agrobacterium rhizogenes and explants were established in C. intybus for the first time. • Different co-cultivation media including MS basal medium and MS medium lacking KNO 3 , NH 4 NO 3 , KH 2 PO 4 , CaCl 2 and MgSO 4 respectively was used. • Observed an increase in hairy roots induction percentage, biomass accumulation and secondary metabolites content by all of strains. • Maximum hairy roots induction occurred by A 4 strain when a MS medium without KNO 3 was used. • The high frequency of hairy roots induction with ATCC11325 and ATCC15834 strains was achieved when KH 2 PO 4 was removed of MS medium. Abstract Transformed hairy roots technology provides new opportunities for mass production of pharmaceutical metabolites. Hairy roots culture has been considered as an alternative method for producing medicinal biomolecule s in plants. Chicory (Cichorium intybus L.) is a medicinal herb from Asteraceae family. It contains many important metabolites including chicoric acid, inulin, scoline, coumarin, and flavonoids. In this study, for the first time, a reliable gene transfer system via modifying co-cultured medium using different strains of Agrobacterium rhizogenes and explants were established for C. intybus. Different co-culture media were used for this purpose, including MS basal medium and MS medium without either of KNO 3 , NH 4 NO 3 , KH 2 PO 4 , CaCl 2 , and MgSO 4. The results showed increases in hairy roots induction percentage, biomass production, and phenolic content of the hairy roots. Maximum hairy root induction percentage obtained by using A 4 strain when a KNO 3 -free MS medium was used. High frequency of hairy root induction percentage with ATCC11325 and ATCC15834 strains was achieved when MS medium had KH 2 PO 4 removed. In all of the experiments, very low hairy root induction was observed when NH 4 NO 3 was removed from co-culture MS medium. Molecular confirmation of transgenic hairy roots was done with PCR using gene-specific primers for rol B gene. Total phenols, flavonoids, anthocyanins and chicoric acid contents of hairy root clones and the control (untransformed) roots were also determined. The results indicated significant increases in total metabolites content of hairy root clones induced by A 4 , ATCC11325 and ATCC15834 strains with some macronatrient–free co-culture MS media and also directly-regenerated shoots from hairy roots were spontaneous without using any plant growth regulators. [ABSTRACT FROM AUTHOR] |
