HAL Id: hal-01268678
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Submitted on 3 Jun 2020
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Bacterial acid phosphatase activity and localization in [i]Phaseolus vulgaris[/i] nodules
Laurie Amenc, Jean-Jacques Drevon
To cite this version:
Laurie Amenc, Jean-Jacques Drevon. Bacterial acid phosphatase activity and localization in [i]Phaseolus vulgaris[/i] nodules. 2. IBEMPA Conference ”Microorganisms for future agriculture”, Sep 2013, Seville, Spain. Universitad de Sevilla, 527 p., 2013. �hal-01268678�
Localizing bacterial acid phosphatase activity is important to understand the role of bacteria for P availability in legume nodules. Using in situ RT-PCR and immuno-localization methodologies, we show increased transcript and protein levels of bacterial acid phosphatase within infected cells of common bean nodules.
Results
Conclusion
To our knowledge, this in situ RT-PCR result is the first description of a rhizobial transcript within infected cells in legume nodules. From the variations of the signal intensity according to P supply, we can supposed that bacteroids are P-deprived under P deficiency, and that their P-induced acid phosphatase may contribute to the adaptation of the legume-rhizobia symbioses to low-P soils.
References: Bargaz, A, et al. (2012). J. Exp. Bot. 63: 4723-4730. ; van Aarle et al., (2010). Mycorrhiza 17, 487–494. ; Hernandez, G. and Drevon, J.J. (1991). J. Plant Physiol. 138: 587-590.
Figure 2: Localization of transcripts (green fluorescent signal) of rhizobial acid phosphatase in nodule transverse sections of the tolerant common bean RIL115 (A-C) and the less efficient common bean RIL147 (B-D) as a function of P treatments: 250 µmol Pi week-1 pl-1 as
sufficient (A-B) versus 75 µmol Pi.week-1 pl-1 (C-D) as deficient.
Materials & methods
• Common bean recombinant inbred lines (RILs) were inoculated with Rhizobium tropici CIAT899, grown in hydro-aeroponic culture (Hernandez & Drevon 1991)
• Standard nodules (3-5 mm diameter) were harvested at flowering stage and immediately fixed
• In situ RT-PCR was performed on sections of the fixed nodules (Bargaz et al. 2012)
• Specific primers for bacterial acid phosphatase genes were designed online at Kazusa DNA Research
Institute (http://genome.microbedb.jp/rhizobase/) with
the known mRNA sequences of the homologous g e n e s o f R h i z o b i u m , M e s o r h i z o b i u m a n d Sinorhizobium spp. Nodule B) RT- PCR, immunological reaction C) ELF 97 A) Fixation, inclusion, sections 50 µM
(vibratom)
Agarose low melting
Bacterial acid phosphatase activity and localisation
in Phaseolus vulgaris nodules
L. Amenc, J.J. Drevon
amenc@supagro.inra.fr
INRA, UMR Eco&Sols, 2 Place Viala, 34060 Montpellier Cedex 2, France.
D B
C A
Bacterial acid phosphatase transcripts were primarily localized in infected cells, with some evident in vascular traces. The fluorescent signal intensity in infected cells was decreased under P deficiency (C-D) and was higher for the tolerant RIL115 (A-C) than for the less efficient RIL147 (B-D). Interestingly, the gene expression varied among infected cells. This suggest that P availability for bacteroids may depend upon the infected cell localization within the infected zone.
Acid phosphatase proteins were localized in inner cortex, distributing zone and infected cells (data not shown).
A D C B 250 µmol Pi week-1 pl-1 RIL 115 RIL 147 75 µmol Pi week-1 pl-1 115 Tolerant 147 Less efficient
Experimental procedure
Figure 1: Production and in situ RT-PCR methodology for nodules.
