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An anthocyanin marker for direct visualization of plant transformation and its use to study nitrogen-fixing nodule development.

Zhang, Senlei and Kondorosi, Éva and Kereszt, Attila (2019) An anthocyanin marker for direct visualization of plant transformation and its use to study nitrogen-fixing nodule development. JOURNAL OF PLANT RESEARCH, 132 (5). pp. 695-703. ISSN 0918-9440

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Abstract

The development and functioning of the nitrogen fixing symbiosis between legume plants and soil bacteria collectively called rhizobia requires continuous chemical dialogue between the partners using different molecules such as flavonoids, lipo-chitooligosaccharides, polysaccharides and peptides. Agrobacterium rhizogenes mediated hairy root transformation of legumes is widely used to study the function of plant genes involved in the process. The identification of transgenic plant tissues is based on antibiotics/herbicide selection and/or the detection of different reporter genes that usually require special equipment such as fluorescent microscopes or destructive techniques and chemicals to visualize enzymatic activity. Here, we developed and efficiently used in hairy root experiments binary vectors containing the MtLAP1 gene driven by constitutive and tissue-specific promoters that facilitate the production of purple colored anthocyanins in transgenic tissues and thus allowing the identification of transformed roots by naked eye. Anthocyanin producing roots were able to establish effective symbiosis with rhizobia. Moreover, it was shown that species-specific allelic variations and a mutation preventing posttranslational acetyl modification of an essential nodule-specific cysteine-rich peptide, NCR169, do not affect the symbiotic interaction of Medicago truncatula cv. Jemalong with Sinorhizobium medicae strain WSM419. Based on the experiments, it could be concluded that it is preferable to use the vectors with tissue-specific promoters that restrict anthocyanin production to the root vasculature for studying biotic interactions of the roots such as symbiotic nitrogen fixation or mycorrhizal symbiosis.

Item Type: Article
Additional Information: Funding Agency and Grant Number: MTA Biological Research Center (MTA SZBK); Hungarian National Office for Research, Development and Innovation [OTKA 120120/119652, GINOP 2.3.2-15-2016-00014 Evomer, GINOP 2.3.2-15-2016-00015 I-KOM] Funding text: Open access funding provided by MTA Biological Research Center (MTA SZBK). Research in our laboratory was supported by the Hungarian National Office for Research, Development and Innovation through the grant OTKA 120120/119652 (to AK), the GINOP 2.3.2-15-2016-00014 Evomer (to EK) and GINOP 2.3.2-15-2016-00015 I-KOM (to EK).
Uncontrolled Keywords: reporter gene; hairy root; LEGUME SYMBIOSIS; Nodule-specific cysteine-rich (NCR) peptides; Transgenic tissue;
Subjects: Q Science / természettudomány > QK Botany / növénytan
SWORD Depositor: MTMT SWORD
Depositing User: MTMT SWORD
Date Deposited: 24 Nov 2019 14:05
Last Modified: 24 Nov 2019 14:05
URI: http://real.mtak.hu/id/eprint/103698

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