In rhizobial symbiosis with legume plant hosts, the symbiotic tissue in the root nodules of indeterminate type is localized to the basal part of the nodule where the symbiotic zones contain infected cells (IC) interspersed with uninfected cells (UC) that are devoid of rhizobia. Although IC are easily distinguished in nodule sections using standard histochemical techniques, their observation in intact nodules is hampered by nodule tissue characteristics. Tagging of Rhizobium leguminosarum bv. viciae strain 128C30 with a constitutively expressed gene for green fluorescent protein (nonshifted mutant form cycle3) in combination with the advantages of the tiny nodules formed by Vicia tetrasperma (L.) SCHREB . allowed for vital observation of symbiotic tissue using fluorescence microscopy. Separation of a red-shifted background channel and digital image stacking along z-axis enabled us to construct a nodule image in a classical fluorescence microscopy of nodules exceeding 1 mm in diameter. In parallel, visualization of nodule bacteria inside the symbiotic tissue by confocal microscopy at the excitation wavelength 488 nm clearly distinguished IC/UC pattern in the nodule virtual sections and revealed red-shifted fluorescence of nonrhizobial origin. This signal was located on the periphery of IC and increased with their degradation, thus suggesting accumulation of secondary metabolites, presumably flavonoids. The simultaneous detection of bacteria and secondary metabolites can be used for monitoring changes to intact nodule physiology in the model legumes. The advantage of V. tetrasperma as a suggested laboratory model for pea cross-inoculation group has been demonstrated.
- MeSH
- fluorescenční mikroskopie MeSH
- kořenové hlízky rostlin fyziologie chemie mikrobiologie MeSH
- půdní mikrobiologie MeSH
- Rhizobium leguminosarum cytologie fyziologie genetika chemie MeSH
- symbióza MeSH
- vikev cytologie fyziologie chemie mikrobiologie MeSH
- zelené fluorescenční proteiny analýza genetika metabolismus MeSH
- MeSH
- antibióza MeSH
- Fabaceae fyziologie MeSH
- isoflavony analogy a deriváty MeSH
- Rhizobium leguminosarum genetika MeSH
- symbióza MeSH
- Publikační typ
- srovnávací studie MeSH
No significant differences in the acetylene-reducing activity and evolution of H2 and CO2 from nodulated roots of Pisum sativum inoculated with soil Rhizobium populations from two soils with different acidities (Ruzynĕ soil 7.6; Lukavec soil 4.9) were observed. Rhizobium population from Lukavec soil formed nodules, exhibiting a higher H2 evolution. Co-inoculation with the Hup+ strain 128C30 (7 x 10(7) cells per seedling) eliminated, to some extent, the effect of soil populations on physiological activity.
- MeSH
- acetylen metabolismus MeSH
- Fabaceae metabolismus mikrobiologie MeSH
- fixace dusíku * MeSH
- léčivé rostliny * MeSH
- nitrogenasa metabolismus MeSH
- oxid uhličitý metabolismus MeSH
- půdní mikrobiologie * MeSH
- Rhizobium leguminosarum metabolismus MeSH
- Rhizobium růst a vývoj metabolismus MeSH
- symbióza MeSH
- vodík metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
