Salt and temperature tolerance of Sinorhizobium strains isolated from dry environments in Alentejo
P. Fareleira1, N. Matos1, E. Ferreira2 & J. F. Marques
RESUMO
O trabalho teve como principal objectivo a obtenção de estirpes de rizóbio adequadas à nodulação de luzernas anuais e adaptadas a sobreviver nas condições ambientais susceptíveis de ocorrer em solos degradados. Dado que as populações de rizóbio apresentam variabilidade considerável no que respeita à tolerância a factores ambientais, os estudos focaram-se na pesquisa e na selecção de estirpes resistentes a condições adversas. Efectuaram-se colheitas de solos em diversas zonas no sul do País, seleccionando-se locais afectados por secura, temperaturas elevadas e, pontualmente, salinidade. A partir destes solos, isolaram-se estirpes de rizóbio, usando como planta hospedeira a luzerna anual Medicago polymorpha.
Estudaram-se os efeitos de condições de stresse ambiental, como a salinidade e as altas temperaturas, no crescimento das estirpes isoladas. Dos 41 isolamentos analisados, 11 apresentaram crescimento em meio contendo 1,4 M de cloreto de sódio e suplementado com 10% de extracto de terra, e 22 cresceram quando incubadas a 45 ºC em meio sem aditivos. Três estirpes mostraram ter capacidade para crescer sob os efeitos conjuntos da salinidade e da alta temperatura.
A análise de extractos etanólicos de estirpes tolerantes à salinidade revelou, na maior parte dos casos, a acumulação, induzida pelo sal, dos solutos compatíveis de Sinorhizobium: o dipéptido N - acetilglutaminilglutamina amida, vários tipos de betaínas, trealose, glutamato e prolina.
A observação, por NMR de 31P in vivo, de uma estirpe tolerante ao sal, proveniente de um solo xistoso de baixo teor em fósforo, mostrou a presença de níveis elevados de reservas intracelulares de fosfato inorgânico (polifosfato), sugerindo um bom potencial para utilização em solos onde os riscos de salinização se conjuguem com deficiências em fósforo assimilável.
]]> ABSTRACT
The main objective of this work was to obtain rhizobial strains able to nodulate annual medics and highly adapted to the environmental conditions present in degraded soils. Since populations of rhizobia vary in their tolerance to environmental factors, screening for resistant strains was pursued. Soil samples were collected in different regions in Southern Portugal, severely affected by drought, high temperatures, and, in some places, salinity; rhizobial strains were isolated using Medicago polymorpha as trap-host.
The effects of environmental stressful conditions, such as salinity or high temperatures, in the growth of the isolated strains were studied. Among the 41 isolates that were ana-lysed here, 11 showed good growth capacity in the presence of 1.4 M sodium chloride and 10% soil extract, and 22 could grow upon incubation at 45 ºC. Three strains were able to grow under the simultaneous effects of salinity and high temperature.
NMR analysis of ethanolic cell-free extracts of salt tolerant strains showed that, in most cases, cell growth in medium containing high salt concentrations resulted in the accumulation of the compatible solutes described for Sinorhizobium: the dipeptide N-acetylglutaminylglutamine amide, betaines, trehalose, glutamate, and proline.
Analysis by in vivo 31P-NMR of a salt tolerant strain originated from a schistous soil with low-phosphorus content, revealed the presence of high levels of intracellular inorganic phosphate reserves (polyphosphates). This suggests a high potential for the utilization of the strain in soils affected by both salinity and phosphorus deficiency.
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1 Estação Agronómica Nacional, Departamento de Ciência do Solo, Quinta do Marquês, Av. da República, 2784-505 Oeiras, e-mail: paula.fareleira@oniduo.pt;
2 Estação Florestal Nacional, Departamento de Ecologia, Recursos Naturais e Ambiente, Quinta do Marquês, Av. da República, 2784-505 Oeiras
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