Agrobacterium rhizogenes mediated transformation of Ficus carica L. for the efficient production of secondary metabolites

Abstract

BACKGROUND Ficus carica L., an ancient source of food and medicines, is rich in valuable nutritional and secondary compounds with antioxidant, antimicrobial, and anticancer effects. The present study is the first attempt to examine hairy root (HR) induction of F. carica (Sabz and Siah) by inoculating the 3‐week‐old shoots and leaves with different strains of Agrobacterium rhizogenes and also to investigate methyl jasmonate (MeJA) elicitation of HRs to produce a fast and high‐yield production method for secondary metabolites. RESULTS The maximum transformation rate (100%) was achieved by inoculating the shoots with Agrobacterium rhizogenes strain A7. Siah HRs elicited with 100 and 200 μmol L−1 MeJA and Sabz HRs with 100 μmol L−1 MeJA showed the highest total phenolic content. The highest flavonoid content was 3.935 mg QE g−1 DW in Siah HRs treated with 200 μmol L−1 MeJA and 2.762 mg QE g−1 DW in Sabz HRs treated with 300 μmol L−1 MeJA. The 2, 2‐diphenyl‐1‐picrylhydrazyl (DPPH) radical scavenging capacity and ferric reducing antioxidant power (FRAP) value of HRs were affected by MeJA treatments. Methyl jasmonate elicitation also significantly enhanced the content of six phenolic acids (gallic acid, caffeic acid, chlorogenic acid, coumaric acid, rosmarinic acid, and cinnamic acid) and three flavonoids (rutin, quercetin, and apigenin). Thymol, a monoterpene phenol, was the main HR compound detected in gas chromatography mass spectrometry (GC–MS) analysis of the essential oils. CONCLUSION Induction of HRs and elicitation of F. carica HRs by MeJA resulted in a significant increase in the production of important phenolic compounds and a significant increase in antioxidant capacity. © 2020 Society of Chemical Industry

MeJA Elicitation of Chicory Hairy Roots Promotes Efficient Increase of 3,5-diCQA Accumulation, a Potent Antioxidant and Antibacterial Molecule

“…Similar results have been obtained in HRCs of Aster scaber [ 35 ] in which, the yields of chlorogenic acid were doubled with MeJA used as an elicitor. Similarly, in HRC of Ficus carica grown in the presence of MeJA, the production of CQA was increased by 4.4-fold [ 39 ]. Other elicitors such as salicylic acid (SA), yeast extract (YE) or cyclodextrin (CD) could be used but MeJA is a more general inducer of the production of specialized metabolites than the others, in particular on the metabolism of phenolic compounds.…”

Hairy Root Cultures as a Source of Phenolic Antioxidants: Simple Phenolics, Phenolic Acids, Phenylethanoids, and Hydroxycinnamates

“…The HRs of cv. Sabz, after the elicitation with MeJa (200 μM), accumulated gallic (0.19 mg/g DW), caffeic (0.28 mg/g DW), chlorogenic (1.63 mg/g DW), and coumaric (0.23 mg/g DW) acids [ 51 ]. The same root culture elicited with the Piriformospora indica fungal cell extract or culture filtrate, at a concentration range of 2–6% ( v / v ), demonstrated an enhanced production of phenolic acids (up to 7.49 mg/g of gallic acid, up to 0.83 mg/g of caffeic acid, up to 1.58 mg/g of 5-CQA, and up to 0.94 mg/g of coumaric acid, as calculated on a DW basis).…”

Piriformospora indica based elicitation for overproduction of phenolic compounds by hairy root cultures of Ficus carica

“…HR lines were excised from induction sites and sub-cultured in fresh solid media (½ MS medium with gradually decrease in cefotaxime concentration) every 2-3 weeks. After HR establishment and optimization of culturing (Amani et al, 2020), the 300 mg fresh weight of the hairy root line with the highest growth rate (superior HR line) were transferred to 30 mL hormone-free ½ MS liquid medium and placed on a rotary shaker (120 rpm) under darkness at 25±2 ºC for further experiments.…”

“…The HPLC standards of phenolic compounds (purity >98%) were purchased from Merck (Darmstadt, Germany). HPLC analysis of nine Manufacturing Company) as described in our previous study (Amani et al, 2020). Analysis of reverse phase liquid chromatographic was done using analytical column of octadecylsilane (Shim-pack VP-ODS 25 cm long × 4.6 mm diameter, particle size 5 μm, Japan).…”

“…2). As a result of previous study (Amani et al, 2020), SW8 HR line was screened as a superior HR line with the highest growth rate and high branching morphology and sub-cultured regularly over 10 months for further experiments. The observation of 780-bp DNA fragment with bacterial plasmid DNA as positive control confirmed the presence of rolB gene in HRs lines.…”