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1.
MethodsX ; 2: 182-91, 2015.
Article in English | MEDLINE | ID: mdl-26150987

ABSTRACT

An optimized method, based on the coupling of two commercial kits, is described for the extraction of soil nucleic acids, with simultaneous extraction and purification of DNA and RNA following a cascade scheme and avoiding the use of harmful solvents. The protocol canmonitor the variations in the recovery yield of DNA and RNA from soils of various types.The quantitative version of the protocol was obtained by testing the starting soil quantity, the grinding parameters and the final elution volumes, in order to avoid saturation of both kits. •A first soil-crushing step in liquid nitrogen could be added for the assessment of fungal parameters.•The protocol was efficienton different tropical soils, including Andosol, while their high contents of clays, including poorly crystalline clays, and Fe and Al oxides usually make the nucleic acid extraction more difficult.•The RNA recovery yield from the previous tropical soils appeared to correlate better to soil respiration than DNA, which is positively influenced by soil clay content.

2.
J Exp Bot ; 63(13): 4723-30, 2012 Aug.
Article in English | MEDLINE | ID: mdl-22771853

ABSTRACT

Although previous studies on N2-fixing legumes have demonstrated the contribution of acid phosphatases to their phosphorus (P) use efficiency under P-deficient growth conditions, localization of these enzymes in bean nodules has not been demonstrated. In this study, phosphoenol pyruvate phosphatase (PEPase) gene transcripts were localized within the nodule tissues of two recombinant inbred lines, RIL115 (P-deficiency tolerant) and RIL147 (P-deficiency sensitive), of Phaseolus vulgaris. Nodules were induced by Rhizobium tropici CIAT899 under hydroaeroponic conditions with a sufficient versus a deficient P supply. The results indicated that PEPase transcripts were particularly abundant in the nodule infected zone and cortex of both RILs. Analysis of fluorescence intensity indicated that nodule PEPase was induced under conditions of P deficiency to a significantly higher extent in RIL147 than in RIL115, and more in the inner cortex (91%) than in the outer cortex (71%) or the infected zone (79%). In addition, a significant increase (39%) in PEPase enzyme activity in the P-deficient RIL147 correlated with an increase (58%) in the efficiency of use in rhizobial symbiosis. It was concluded that nodule PEPase is upregulated under conditions of P deficiency in the P-deficiency-sensitive RIL147, and that this gene may contribute to adaptation of rhizobial symbiosis to low-P environments.


Subject(s)
Acid Phosphatase/genetics , Phaseolus/enzymology , Phosphorus/deficiency , Rhizobium tropici/physiology , Acid Phosphatase/metabolism , Adaptation, Physiological , Inbreeding , Nitrogen/metabolism , Nitrogen Fixation/genetics , Phaseolus/cytology , Phaseolus/genetics , Phaseolus/growth & development , Phosphorus/analysis , Plant Proteins/genetics , Plant Proteins/metabolism , Plant Root Nodulation , Plant Roots/cytology , Plant Roots/enzymology , Plant Roots/genetics , Plant Roots/growth & development , Plant Shoots/cytology , Plant Shoots/enzymology , Plant Shoots/genetics , Plant Shoots/growth & development , RNA, Messenger/genetics , RNA, Plant/genetics , Root Nodules, Plant/cytology , Root Nodules, Plant/enzymology , Root Nodules, Plant/genetics , Root Nodules, Plant/growth & development , Seedlings/cytology , Seedlings/enzymology , Seedlings/genetics , Seedlings/growth & development , Symbiosis
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