The ability to solubilize insoluble inorganic pho- sphate compounds by Gluconacetobacter diazotrophicus was studied using different cul-ture approaches. Qualitative plate assays using tricalcium phosphate as the sole ...The ability to solubilize insoluble inorganic pho- sphate compounds by Gluconacetobacter diazotrophicus was studied using different cul-ture approaches. Qualitative plate assays using tricalcium phosphate as the sole P-source showed that G. diazotrophicus produced solu-bilization only when aldoses were used as the C-source. Extracellular aldose oxidation via a pyrroloquinoline quinone-linked glucose dehy-drogenase (PQQ-GDH) is the main pathway for glucose metabolism in G. diazotrophicus. In batch cultures with 5 g l-1 of hydroxyapatite as the P-source and glucose as the C-source, more than 98% of insoluble P was solubilized. No solubilization was observed neither using glyc-erol nor culturing a PQQ-GDH mutant of G. di-azotrophicus. Solubilizaton was not affected by adding 100 mmol l-1 of MES buffer. Continuous cultures of G. diazotrophicus showed significant activities of PQQ-GDH either under C or P limi-tation. An intense acidification in the root envi-ronment of tomato and wheat seedlings inocu-lated with a G. diazotrophicus PAL5 was ob-served. Seedlings inoculated with a PQQ-GDH mutant strain of G. diazotrophicus showed no acidification. Our results suggest that G. di-azotrophicus is an excellent candidate to be used as biofertilizer because in addition to the already described plant growth-promoting abili-ties of this organism, it shows a significant mineral phosphate solubilization capacity.展开更多
It has been forecast that the challenge of meeting increased food demand and protecting environmental quality will be won or lost in maize, rice and wheat cropping systems, and that the problem of environmental nitrog...It has been forecast that the challenge of meeting increased food demand and protecting environmental quality will be won or lost in maize, rice and wheat cropping systems, and that the problem of environmental nitrogen enrichment is most likely to be solved by substituting synthetic nitrogen fertilizers by the creation of cereal crops that are able to fix nitrogen symbiotically as legumes do. In legumes, rhizobia present intracellularly in membrane-bound vesicular compartments in the cytoplasm of nodule cells fix nitrogen endosymbiotically. Within these symbiosomes, membrane-bound vesicular compartments, rhizobia are supplied with energy derived from plant photosynthates and in return supply the plant with biologically fixed nitrogen, usually as ammonia. This minimizes or eliminates the need for inputs of synthetic nitrogen fertilizers. Recently we have demonstrated, using novel inoculation conditions with very low numbers of bacteria, that cells of root meristems of maize, rice, wheat and other major non-legume crops, such as oilseed rape and tomato, can be intracellularly colonized by the non-rhizobial, non-nodulating, nitrogen fixing bacterium, Gluconacetobacter diazotrophicus that naturally occurs in sugarcane. G. diazotrophicus expressing nitrogen fixing (nifH) genes is present in symbiosome-like compartments in the cytoplasm of cells of the root meristems of the target cereals and non-legume crop species, somewhat similar to the intracellular symbiosome colonization of legume nodule cells by rhizobia. To obtain an indication of the likelihood of adequate growth and yield, of maize for example, with reduced inputs of synthetic nitrogen fertilizers, we are currently determining the extent to which nitrogen fixation, as assessed using various methods, is correlated with the extent of systemic intracellular colonization by G. diazotrophicus, with minimal or zero inputs.展开更多
文摘The ability to solubilize insoluble inorganic pho- sphate compounds by Gluconacetobacter diazotrophicus was studied using different cul-ture approaches. Qualitative plate assays using tricalcium phosphate as the sole P-source showed that G. diazotrophicus produced solu-bilization only when aldoses were used as the C-source. Extracellular aldose oxidation via a pyrroloquinoline quinone-linked glucose dehy-drogenase (PQQ-GDH) is the main pathway for glucose metabolism in G. diazotrophicus. In batch cultures with 5 g l-1 of hydroxyapatite as the P-source and glucose as the C-source, more than 98% of insoluble P was solubilized. No solubilization was observed neither using glyc-erol nor culturing a PQQ-GDH mutant of G. di-azotrophicus. Solubilizaton was not affected by adding 100 mmol l-1 of MES buffer. Continuous cultures of G. diazotrophicus showed significant activities of PQQ-GDH either under C or P limi-tation. An intense acidification in the root envi-ronment of tomato and wheat seedlings inocu-lated with a G. diazotrophicus PAL5 was ob-served. Seedlings inoculated with a PQQ-GDH mutant strain of G. diazotrophicus showed no acidification. Our results suggest that G. di-azotrophicus is an excellent candidate to be used as biofertilizer because in addition to the already described plant growth-promoting abili-ties of this organism, it shows a significant mineral phosphate solubilization capacity.
文摘It has been forecast that the challenge of meeting increased food demand and protecting environmental quality will be won or lost in maize, rice and wheat cropping systems, and that the problem of environmental nitrogen enrichment is most likely to be solved by substituting synthetic nitrogen fertilizers by the creation of cereal crops that are able to fix nitrogen symbiotically as legumes do. In legumes, rhizobia present intracellularly in membrane-bound vesicular compartments in the cytoplasm of nodule cells fix nitrogen endosymbiotically. Within these symbiosomes, membrane-bound vesicular compartments, rhizobia are supplied with energy derived from plant photosynthates and in return supply the plant with biologically fixed nitrogen, usually as ammonia. This minimizes or eliminates the need for inputs of synthetic nitrogen fertilizers. Recently we have demonstrated, using novel inoculation conditions with very low numbers of bacteria, that cells of root meristems of maize, rice, wheat and other major non-legume crops, such as oilseed rape and tomato, can be intracellularly colonized by the non-rhizobial, non-nodulating, nitrogen fixing bacterium, Gluconacetobacter diazotrophicus that naturally occurs in sugarcane. G. diazotrophicus expressing nitrogen fixing (nifH) genes is present in symbiosome-like compartments in the cytoplasm of cells of the root meristems of the target cereals and non-legume crop species, somewhat similar to the intracellular symbiosome colonization of legume nodule cells by rhizobia. To obtain an indication of the likelihood of adequate growth and yield, of maize for example, with reduced inputs of synthetic nitrogen fertilizers, we are currently determining the extent to which nitrogen fixation, as assessed using various methods, is correlated with the extent of systemic intracellular colonization by G. diazotrophicus, with minimal or zero inputs.
