Use of Chenopodium murale L. transgenic hairy root in vitro culture system as a new tool for allelopathic assays

• Published in: Journal of plant physiology Vol. 169; no. 12; pp. 1203 - 1211
• Main Authors: Mitić, Nevena, Dmitrović, Slavica, Djordjević, Mirka, Zdravković-Korać, Snežana, Nikolić, Radomirka, Raspor, Martin, Djordjević, Tatjana, Maksimović, Vuk, Živković, Suzana, Krstić-Milošević, Dijana, Stanišić, Mariana, Ninković, Slavica
• Format: Journal Article
• Language: English
• Published: Munich Elsevier GmbH 15-08-2012; Elsevier
• Subjects: Agrobacterium; Agrobacterium rhizogenes; Allelopathy; Assaying; Biological and medical sciences; Chenopodium - chemistry; Chenopodium - cytology; Chenopodium - genetics; Chenopodium murale; Clone Cells; Cotyledon; Culture; Culture Media; Culture Techniques - methods; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation, Plant; Genetic transformation; Genetic Variation; Hairy roots; In vitro testing; Phenolic acids; Pheromones - analysis; Physiology; Phytopathology. Animal pests. Plant and forest protection; Plant Leaves; Plant physiology and development; Plant Roots - chemistry; Plant Roots - cytology; Plant Roots - growth & development; Plant Roots - microbiology; Plants (organisms); Plants, Genetically Modified; Rol genes; Roots; Seedlings; Transformation, Genetic; Transgenic; Triticum aestivum; Chenopodium murale; Genetic transformation; Allelopathy; Rol genes; Agrobacterium rhizogenes; Hairy roots; Phenolic acids; Hairy root; Weed; Plant pathogen; Phenolic acid; In vitro; System; Chenopodiaceae; Gene; Plant physiology; Dicotyledones; Angiospermae; Bacteria; Spermatophyta; Rhizobiaceae; Genetically modified organism
• ISSN: 0176-1617; 1618-1328
• DOI: 10.1016/j.jplph.2012.04.009

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 (0.07–2.85μmol/L) identified by HPLC analysis in the growth media were at least 1000 times lower than the inhibitory active concentration (5mmol/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.