| Title: | Use of Chenopodium murale L. transgenic hairy root in vitro culture system as a new tool for allelopathic assays |
| Author(s): | Mitic N; Dmitrovic S; Djordjevic M; Zdravkovic-Korac S; Nikolic R; Raspor M; Djordjevic T; Maksimovic V; Zivkovic S; Krstic-Milosevic D; Stanisic M; Ninkovic S; |
| Address: | "Institute for Biological Research 'Sinisa Stankovic', University of Belgrade, Despot Stefan Blvd. 142, 11060 Belgrade, Serbia. mitic.nevena@ibiss.bg.ac.rs" |
| DOI: | 10.1016/j.jplph.2012.04.009 |
| ISSN/ISBN: | 1618-1328 (Electronic) 0176-1617 (Linking) |
| 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 mumol/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" |
| Keywords: | "Agrobacterium Chenopodium/*chemistry/*cytology/genetics Clone Cells Cotyledon Culture Media Culture Techniques/*methods Gene Expression Regulation, Plant Genetic Variation Pheromones/*analysis Plant Leaves Plant Roots/*chemistry/*cytology/growth & develop;" |
| Notes: | "MedlineMitic, Nevena Dmitrovic, Slavica Djordjevic, Mirka Zdravkovic-Korac, Snezana Nikolic, Radomirka Raspor, Martin Djordjevic, Tatjana Maksimovic, Vuk Zivkovic, Suzana Krstic-Milosevic, Dijana Stanisic, Mariana Ninkovic, Slavica eng Comparative Study Research Support, Non-U.S. Gov't Germany 2012/07/04 J Plant Physiol. 2012 Aug 15; 169(12):1203-11. doi: 10.1016/j.jplph.2012.04.009. Epub 2012 Jun 28" |
