title: Use of Chenopodium murale L. transgenic hairy root in vitro culture system as a new tool for allelopathic assays.
abstract: We investigated Chenopodium murale transgenic hairy root in vitro culture system as a new tool for allelopathic assays. Transgenic hairy roots were induced by Agrobacterium rhizogenes A4M70GUS from roots, cotyledons, leaves, and internodes of C. murale seedlings. Roots were found to be the best target explants, providing transformation efficiency of up to 11.1%. Established hairy root clones differed in their morphology and growth potential. Molecular characterization of these clones was carried out by PCR, RT-PCR and histochemical GUS analyses. No differences in rol gene expression were observed. Liquid culture system of characterized hairy root clones was maintained for over 2 years. Six hairy root clones were selected for assaying the allelopathic effect of their growth medium against germination and seedling elongation of wheat and lettuce test plants. The inhibitory potential varied depending on the hairy root clone. Some transgenic clones showed significantly higher inhibition compared to wild-type roots. These results revealed that hairy roots as an independent system synthesize some bioactive substances with allelopathic activity and exude them into the growth medium. Concentrations of caffeic, ferulic and p-coumaric acids (0.07-2.85 μmol/L) identified by HPLC analysis in the growth media were at least 1000 times lower than the inhibitory active concentration (5 mmol/L) of pure grade phenolic acids, suggesting that they have a limited role in the allelopathic phenomena of C. murale. The presented hairy root system appears to be a suitable tool for further investigation of the potential and nature of root-mediated allelopathic interference of C. murale.
authors: Institute for Biological Research "Siniša Stanković", University of Belgrade, Despot Stefan Blvd. 142, 11060 Belgrade, Serbia. mitic.nevena@ibiss.bg.ac.rs
authors: Nevena
authors: N
authors: Mitić
authors: Slavica
authors: S
authors: Dmitrović
authors: Mirka
authors: M
authors: Djordjević
authors: Snežana
authors: S
authors: Zdravković-Korać
authors: Radomirka
authors: R
authors: Nikolić
authors: Martin
authors: M
authors: Raspor
authors: Tatjana
authors: T
authors: Djordjević
authors: Vuk
authors: V
authors: Maksimović
authors: Suzana
authors: S
authors: Zivković
authors: Dijana
authors: D
authors: Krstić-Milošević
authors: Mariana
authors: M
authors: Stanišić
authors: Slavica
authors: S
authors: Ninković
mesh_terms: D060054:Agrobacterium; D027462:Chenopodium / Q000737:chemistry* / Q000166:cytology* / Q000235:genetics; D002999:Clone Cells; D018548:Cotyledon; D003470:Culture Media; D046508:Culture Techniques / Q000379:methods*; D018506:Gene Expression Regulation, Plant; D014644:Genetic Variation; D010675:Pheromones / Q000032:analysis*; D018515:Plant Leaves; D018517:Plant Roots / Q000737:chemistry* / Q000166:cytology* / Q000254:growth & development / Q000382:microbiology; D030821:Plants, Genetically Modified; D036226:Seedlings; D014170:Transformation, Genetic
chemical_list: D003470:Culture Media; D010675:Pheromones