Functional expression of Sinorhizobium meliloti BetS, a high-affinity betaine transporter, in Bradyrhizobium japonicum USDA110.
Among the Rhizobiaceae, Bradyrhizobium japonicum strain USDA110 appears to be extremely salt sensitive, and the presence of glycine betaine cannot restore its growth in medium with an increased osmolarity (E. Boncompagni, M. Osteras, M. C. Poggi, and D. Le Rudulier, Appl. Environ. Microbiol. 65:2072-2077, 1999). In order to improve the salt tolerance of B. japonicum, cells were transformed with the betS gene of Sinorhizobium meliloti. This gene encodes a major glycine betaine/proline betaine transporter from the betaine choline carnitine transporter family and is required for early osmotic adjustment. Whereas betaine transport was absent in the USDA110 strain, such transformation induced glycine betaine and proline betaine uptake in an osmotically dependent manner. Salt-treated transformed cells accumulated large amounts of glycine betaine, which was not catabolized. However, the accumulation was reversed through rapid efflux during osmotic downshock. An increased tolerance of transformant cells to a moderate NaCl concentration (80 mM) was also observed in the presence of glycine betaine or proline betaine, whereas the growth of the wild-type strain was totally abolished at 80 mM NaCl. Surprisingly, the deleterious effect due to a higher salt concentration (100 mM) could not be overcome by glycine betaine, despite a significant accumulation of this compound. Cell viability was not significantly affected in the presence of 100 mM NaCl, whereas 75% cell death occurred at 150 mM NaCl. The absence of a potential gene encoding Na(+)/H(+) antiporters in B. japonicum could explain its very high Na(+) sensitivity.
Unité Interactions Plantes-Microorganismes et Santé Végétale, CNRS-INRA-Université de Nice Sophia Antipolis, UMR 6192, Faculté des Sciences, Parc Valrose, 06108 Nice Cédex, France.
Alexandre
A
Boscari
Karine
K
Mandon
Marie-Christine
MC
Poggi
Daniel
D
Le Rudulier
D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D001622:Betaine / Q000378:metabolism*; D001693:Biological Transport, Active; D020369:Bradyrhizobium / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism*; D050485:GABA Plasma Membrane Transport Proteins; D015870:Gene Expression; D005798:Genes, Bacterial; D009997:Osmotic Pressure; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D016962:Sinorhizobium meliloti / Q000235:genetics* / Q000378:metabolism*; D012965:Sodium Chloride / Q000494:pharmacology; D014170:Transformation, Genetic
D001426:Bacterial Proteins; D002352:Carrier Proteins; D050485:GABA Plasma Membrane Transport Proteins; D011994:Recombinant Proteins; C072207:betaine plasma membrane transport proteins; D001622:Betaine; D012965:Sodium Chloride