Plants in constructed wetlands serve as carriers for attached microbial growth. They mainly transfer oxygen and release exudates into the root zone. As a result, an area exists around the roots (rhizosphere) in which bacteria are stimulated by root growth. Our goals were to ascertain whether stimulating the microbial community only has a local effect on the rhizoplane, and to establish the importance of this stimulation for wastewater purification in the root zone. Observations were carried out in a planted and an unplanted laboratory batch reactor incubated with an artificial wastewater with a high concentration of ammonia. Samples were taken at intervals of 10 mm away from the roots. The chemical and physical conditions and enzyme activities in soil sections at various distances from the roots affecting the efficiency of microbial nitrogen removal were characterized. An influence on the nitrification and denitrification process was detected via several parameters up to a range of different root distances: microbial parameters such as the total bacterial number, respiratory activity, protein and DNA amount seem to be affected by roots up to a distance of 50 mm from the roots, whereas the oxygen concentration, DOC and redox potential are unaffected at a distance exceeding 20-30 mm. This indicates that improved nitrogen removal is also possible in the wider root surroundings. Given the average root-to-root distance of 35 mm, the root-influenced area could therefore be expanded to the whole rooted zone in a constructed wetland.

Revegetation of bauxite residue is hampered by a lack of understanding of the limitations imposed on plant growth in highly saline and alkaline growth mediums. In this study, several of these growth limiting factors were investigated. The toxicity of the hydroxyl ion (OH-) was examined using a solution culture system developed to allow studies at high pH without nutritional limitations. Also using this solution culture system, the effect of the high Na and Mg concentrations of bauxite residue on the Ca nutrition of plants was investigated. As the toxicity of Al at high pH is not known, a study was conducted to examine the rhizotoxicity of aluminate (Al(OH)4-) and polycationic Al at high pH. The ability of plant roots to reduce rhizosphere pH in bauxite residue was also considered. A novel gypsum application method was assessed for its efficiency at improving the Ca status of bauxite residue. Manual adjustment, ion exchange resins and automated titration were examined for their suitability for nutrient solution pH control in alkaline conditions. For short-term studies, it was found that a solution without supply of Cu, Fe, Mn and Zn, and aerated with CO2 depleted air, greatly reduced nutrient precipitation at high pH, thus eliminating nutritional differences between treatments. Manual pH adjustment and the use of ion exchange resins as pH buffers were unsuitable methods of pH control. In contrast, pH control by automated titration had little effect on solution composition while maintaining constant pH. The solution culture system was used to examine OH- toxicity in mungbeans (Vigna radiata (L.) Wilczek cv. Emerald), with root length reduced at a bulk solution pH of 8.5 and greater. The effect of Ca activity ratio (CAR) and pH on Ca uptake by mungbeans and Rhodes grass (Chloris gayana cv. Pioneer) in Na dominated solution cultures and in soil was investigated. Changes in pH in the alkaline range were shown to have no effect on the critical CAR of 0.024 (corresponding to 90 % relative root length) for mungbeans grown in solution culture. Results from soil grown mungbeans confirmed those from solution culture, with a critical CAR of 0.025. A critical CAR of 0.034 was also established for soil grown Rhodes grass. However, using dilute nutrient solutions dominated by Mg at pH 9.0, root growth was found to be more limited than had been observed for Na solutions, with growth reduced beneath a critical CAR of 0.050. Using a CAR equation modified with plasma membrane binding constants (to incorporate the differing antagonistic effects of Mg and Na), new critical CAR values were calculated for Na (0.56) and Mg (0.44) dominated solutions. This modified CAR equation permits the calculation of CAR irrespective of the dominant salt present. Solubilities of various gypsum sources and size fractions in seawater were studied to investigate the effectiveness of gypsum addition to the residue sand pipeline, rather than as a direct field application. The dissolution rate constant varied with gypsum source (analytical grade (AR) > phosphogypsum (PG) > mined gypsum (MG)) due to reactivity and surface area differences, generally reaching saturation within 15 s (AR) to 30 min (MG > 2.0). The ability of bauxite residue to remove Ca from solution (due to cation exchange and precipitation) was also examined; the quantity of the total solution Ca adsorbed was found to be small (5 %). These low rates of solution Ca adsorption, comparatively rapid dissolution rates, and long pumping times (20 min), preclude the application of gypsum to the residue sand/seawater slurry as a method for residue amelioration. Dilute, alkaline (pH 9.5) nutrient solutions were used to investigate the effects of aluminate (Al(OH)4-) on mungbean root growth. Although root growth in Al(OH)4- solutions was slightly limited, the symptoms associated with this growth reduction were observed to be similar to those caused by the Al13 polycation at concentrations lower than that which can be detected. Also, when roots displaying these symptoms were transferred to fresh Al(OH)4- solutions, no root tip lesions were observed, and root hair growth on the lateral roots improved. Thus, Al(OH)4- is considered to be non-toxic, with the observed reduction in root growth in solutions containing Al(OH)4- due to the gradual formation of toxic Al13 in the bulk nutrient solution resulting from the acidification of the alkaline nutrient solution by the plant roots. The effect of Mn deficiency in Rhodes grass and of legume inoculation in lucerne (Medicago sativa L. cv. Hunter River), on the rhizosphere pH of plants grown in highly alkaline bauxite residue was investigated. In response to Mn deficiency in residue sand, Rhodes grass was observed to increase acidification of its rhizosphere (being up to 1.22 pH units lower than the bulk soil). Due to its ability to fix atmospheric N2 rather than relying on soil N (NO3-) reserves, inoculated lucerne (1.75 pH unit decrease) was also found to acidify its rhizosphere to a greater extent than non-inoculated lucerne (1.16 pH unit decrease).

Revegetation of bauxite residue is hampered by a lack of understanding of the limitations imposed on plant growth in highly saline and alkaline growth mediums. In this study, several of these growth limiting factors were investigated. The toxicity of the hydroxyl ion (OH-) was examined using a solution culture system developed to allow studies at high pH without nutritional limitations. Also using this solution culture system, the effect of the high Na and Mg concentrations of bauxite residue on the Ca nutrition of plants was investigated. As the toxicity of Al at high pH is not known, a study was conducted to examine the rhizotoxicity of aluminate (Al(OH)4-) and polycationic Al at high pH. The ability of plant roots to reduce rhizosphere pH in bauxite residue was also considered. A novel gypsum application method was assessed for its efficiency at improving the Ca status of bauxite residue. Manual adjustment, ion exchange resins and automated titration were examined for their suitability for nutrient solution pH control in alkaline conditions. For short-term studies, it was found that a solution without supply of Cu, Fe, Mn and Zn, and aerated with CO2 depleted air, greatly reduced nutrient precipitation at high pH, thus eliminating nutritional differences between treatments. Manual pH adjustment and the use of ion exchange resins as pH buffers were unsuitable methods of pH control. In contrast, pH control by automated titration had little effect on solution composition while maintaining constant pH. The solution culture system was used to examine OH- toxicity in mungbeans (Vigna radiata (L.) Wilczek cv. Emerald), with root length reduced at a bulk solution pH of 8.5 and greater. The effect of Ca activity ratio (CAR) and pH on Ca uptake by mungbeans and Rhodes grass (Chloris gayana cv. Pioneer) in Na dominated solution cultures and in soil was investigated. Changes in pH in the alkaline range were shown to have no effect on the critical CAR of 0.024 (corresponding to 90 % relative root length) for mungbeans grown in solution culture. Results from soil grown mungbeans confirmed those from solution culture, with a critical CAR of 0.025. A critical CAR of 0.034 was also established for soil grown Rhodes grass. However, using dilute nutrient solutions dominated by Mg at pH 9.0, root growth was found to be more limited than had been observed for Na solutions, with growth reduced beneath a critical CAR of 0.050. Using a CAR equation modified with plasma membrane binding constants (to incorporate the differing antagonistic effects of Mg and Na), new critical CAR values were calculated for Na (0.56) and Mg (0.44) dominated solutions. This modified CAR equation permits the calculation of CAR irrespective of the dominant salt present. Solubilities of various gypsum sources and size fractions in seawater were studied to investigate the effectiveness of gypsum addition to the residue sand pipeline, rather than as a direct field application. The dissolution rate constant varied with gypsum source (analytical grade (AR) > phosphogypsum (PG) > mined gypsum (MG)) due to reactivity and surface area differences, generally reaching saturation within 15 s (AR) to 30 min (MG > 2.0). The ability of bauxite residue to remove Ca from solution (due to cation exchange and precipitation) was also examined; the quantity of the total solution Ca adsorbed was found to be small (5 %). These low rates of solution Ca adsorption, comparatively rapid dissolution rates, and long pumping times (20 min), preclude the application of gypsum to the residue sand/seawater slurry as a method for residue amelioration. Dilute, alkaline (pH 9.5) nutrient solutions were used to investigate the effects of aluminate (Al(OH)4-) on mungbean root growth. Although root growth in Al(OH)4- solutions was slightly limited, the symptoms associated with this growth reduction were observed to be similar to those caused by the Al13 polycation at concentrations lower than that which can be detected. Also, when roots displaying these symptoms were transferred to fresh Al(OH)4- solutions, no root tip lesions were observed, and root hair growth on the lateral roots improved. Thus, Al(OH)4- is considered to be non-toxic, with the observed reduction in root growth in solutions containing Al(OH)4- due to the gradual formation of toxic Al13 in the bulk nutrient solution resulting from the acidification of the alkaline nutrient solution by the plant roots. The effect of Mn deficiency in Rhodes grass and of legume inoculation in lucerne (Medicago sativa L. cv. Hunter River), on the rhizosphere pH of plants grown in highly alkaline bauxite residue was investigated. In response to Mn deficiency in residue sand, Rhodes grass was observed to increase acidification of its rhizosphere (being up to 1.22 pH units lower than the bulk soil). Due to its ability to fix atmospheric N2 rather than relying on soil N (NO3-) reserves, inoculated lucerne (1.75 pH unit decrease) was also found to acidify its rhizosphere to a greater extent than non-inoculated lucerne (1.16 pH unit decrease).

Revegetation of bauxite residue is hampered by a lack of understanding of the limitations imposed on plant growth in highly saline and alkaline growth mediums. In this study, several of these growth limiting factors were investigated. The toxicity of the hydroxyl ion (OH-) was examined using a solution culture system developed to allow studies at high pH without nutritional limitations. Also using this solution culture system, the effect of the high Na and Mg concentrations of bauxite residue on the Ca nutrition of plants was investigated. As the toxicity of Al at high pH is not known, a study was conducted to examine the rhizotoxicity of aluminate (Al(OH)4-) and polycationic Al at high pH. The ability of plant roots to reduce rhizosphere pH in bauxite residue was also considered. A novel gypsum application method was assessed for its efficiency at improving the Ca status of bauxite residue. Manual adjustment, ion exchange resins and automated titration were examined for their suitability for nutrient solution pH control in alkaline conditions. For short-term studies, it was found that a solution without supply of Cu, Fe, Mn and Zn, and aerated with CO2 depleted air, greatly reduced nutrient precipitation at high pH, thus eliminating nutritional differences between treatments. Manual pH adjustment and the use of ion exchange resins as pH buffers were unsuitable methods of pH control. In contrast, pH control by automated titration had little effect on solution composition while maintaining constant pH. The solution culture system was used to examine OH- toxicity in mungbeans (Vigna radiata (L.) Wilczek cv. Emerald), with root length reduced at a bulk solution pH of 8.5 and greater. The effect of Ca activity ratio (CAR) and pH on Ca uptake by mungbeans and Rhodes grass (Chloris gayana cv. Pioneer) in Na dominated solution cultures and in soil was investigated. Changes in pH in the alkaline range were shown to have no effect on the critical CAR of 0.024 (corresponding to 90 % relative root length) for mungbeans grown in solution culture. Results from soil grown mungbeans confirmed those from solution culture, with a critical CAR of 0.025. A critical CAR of 0.034 was also established for soil grown Rhodes grass. However, using dilute nutrient solutions dominated by Mg at pH 9.0, root growth was found to be more limited than had been observed for Na solutions, with growth reduced beneath a critical CAR of 0.050. Using a CAR equation modified with plasma membrane binding constants (to incorporate the differing antagonistic effects of Mg and Na), new critical CAR values were calculated for Na (0.56) and Mg (0.44) dominated solutions. This modified CAR equation permits the calculation of CAR irrespective of the dominant salt present. Solubilities of various gypsum sources and size fractions in seawater were studied to investigate the effectiveness of gypsum addition to the residue sand pipeline, rather than as a direct field application. The dissolution rate constant varied with gypsum source (analytical grade (AR) > phosphogypsum (PG) > mined gypsum (MG)) due to reactivity and surface area differences, generally reaching saturation within 15 s (AR) to 30 min (MG > 2.0). The ability of bauxite residue to remove Ca from solution (due to cation exchange and precipitation) was also examined; the quantity of the total solution Ca adsorbed was found to be small (5 %). These low rates of solution Ca adsorption, comparatively rapid dissolution rates, and long pumping times (20 min), preclude the application of gypsum to the residue sand/seawater slurry as a method for residue amelioration. Dilute, alkaline (pH 9.5) nutrient solutions were used to investigate the effects of aluminate (Al(OH)4-) on mungbean root growth. Although root growth in Al(OH)4- solutions was slightly limited, the symptoms associated with this growth reduction were observed to be similar to those caused by the Al13 polycation at concentrations lower than that which can be detected. Also, when roots displaying these symptoms were transferred to fresh Al(OH)4- solutions, no root tip lesions were observed, and root hair growth on the lateral roots improved. Thus, Al(OH)4- is considered to be non-toxic, with the observed reduction in root growth in solutions containing Al(OH)4- due to the gradual formation of toxic Al13 in the bulk nutrient solution resulting from the acidification of the alkaline nutrient solution by the plant roots. The effect of Mn deficiency in Rhodes grass and of legume inoculation in lucerne (Medicago sativa L. cv. Hunter River), on the rhizosphere pH of plants grown in highly alkaline bauxite residue was investigated. In response to Mn deficiency in residue sand, Rhodes grass was observed to increase acidification of its rhizosphere (being up to 1.22 pH units lower than the bulk soil). Due to its ability to fix atmospheric N2 rather than relying on soil N (NO3-) reserves, inoculated lucerne (1.75 pH unit decrease) was also found to acidify its rhizosphere to a greater extent than non-inoculated lucerne (1.16 pH unit decrease).

Revegetation of bauxite residue is hampered by a lack of understanding of the limitations imposed on plant growth in highly saline and alkaline growth mediums. In this study, several of these growth limiting factors were investigated. The toxicity of the hydroxyl ion (OH-) was examined using a solution culture system developed to allow studies at high pH without nutritional limitations. Also using this solution culture system, the effect of the high Na and Mg concentrations of bauxite residue on the Ca nutrition of plants was investigated. As the toxicity of Al at high pH is not known, a study was conducted to examine the rhizotoxicity of aluminate (Al(OH)4-) and polycationic Al at high pH. The ability of plant roots to reduce rhizosphere pH in bauxite residue was also considered. A novel gypsum application method was assessed for its efficiency at improving the Ca status of bauxite residue. Manual adjustment, ion exchange resins and automated titration were examined for their suitability for nutrient solution pH control in alkaline conditions. For short-term studies, it was found that a solution without supply of Cu, Fe, Mn and Zn, and aerated with CO2 depleted air, greatly reduced nutrient precipitation at high pH, thus eliminating nutritional differences between treatments. Manual pH adjustment and the use of ion exchange resins as pH buffers were unsuitable methods of pH control. In contrast, pH control by automated titration had little effect on solution composition while maintaining constant pH. The solution culture system was used to examine OH- toxicity in mungbeans (Vigna radiata (L.) Wilczek cv. Emerald), with root length reduced at a bulk solution pH of 8.5 and greater. The effect of Ca activity ratio (CAR) and pH on Ca uptake by mungbeans and Rhodes grass (Chloris gayana cv. Pioneer) in Na dominated solution cultures and in soil was investigated. Changes in pH in the alkaline range were shown to have no effect on the critical CAR of 0.024 (corresponding to 90 % relative root length) for mungbeans grown in solution culture. Results from soil grown mungbeans confirmed those from solution culture, with a critical CAR of 0.025. A critical CAR of 0.034 was also established for soil grown Rhodes grass. However, using dilute nutrient solutions dominated by Mg at pH 9.0, root growth was found to be more limited than had been observed for Na solutions, with growth reduced beneath a critical CAR of 0.050. Using a CAR equation modified with plasma membrane binding constants (to incorporate the differing antagonistic effects of Mg and Na), new critical CAR values were calculated for Na (0.56) and Mg (0.44) dominated solutions. This modified CAR equation permits the calculation of CAR irrespective of the dominant salt present. Solubilities of various gypsum sources and size fractions in seawater were studied to investigate the effectiveness of gypsum addition to the residue sand pipeline, rather than as a direct field application. The dissolution rate constant varied with gypsum source (analytical grade (AR) > phosphogypsum (PG) > mined gypsum (MG)) due to reactivity and surface area differences, generally reaching saturation within 15 s (AR) to 30 min (MG > 2.0). The ability of bauxite residue to remove Ca from solution (due to cation exchange and precipitation) was also examined; the quantity of the total solution Ca adsorbed was found to be small (5 %). These low rates of solution Ca adsorption, comparatively rapid dissolution rates, and long pumping times (20 min), preclude the application of gypsum to the residue sand/seawater slurry as a method for residue amelioration. Dilute, alkaline (pH 9.5) nutrient solutions were used to investigate the effects of aluminate (Al(OH)4-) on mungbean root growth. Although root growth in Al(OH)4- solutions was slightly limited, the symptoms associated with this growth reduction were observed to be similar to those caused by the Al13 polycation at concentrations lower than that which can be detected. Also, when roots displaying these symptoms were transferred to fresh Al(OH)4- solutions, no root tip lesions were observed, and root hair growth on the lateral roots improved. Thus, Al(OH)4- is considered to be non-toxic, with the observed reduction in root growth in solutions containing Al(OH)4- due to the gradual formation of toxic Al13 in the bulk nutrient solution resulting from the acidification of the alkaline nutrient solution by the plant roots. The effect of Mn deficiency in Rhodes grass and of legume inoculation in lucerne (Medicago sativa L. cv. Hunter River), on the rhizosphere pH of plants grown in highly alkaline bauxite residue was investigated. In response to Mn deficiency in residue sand, Rhodes grass was observed to increase acidification of its rhizosphere (being up to 1.22 pH units lower than the bulk soil). Due to its ability to fix atmospheric N2 rather than relying on soil N (NO3-) reserves, inoculated lucerne (1.75 pH unit decrease) was also found to acidify its rhizosphere to a greater extent than non-inoculated lucerne (1.16 pH unit decrease).

Revegetation of bauxite residue is hampered by a lack of understanding of the limitations imposed on plant growth in highly saline and alkaline growth mediums. In this study, several of these growth limiting factors were investigated. The toxicity of the hydroxyl ion (OH-) was examined using a solution culture system developed to allow studies at high pH without nutritional limitations. Also using this solution culture system, the effect of the high Na and Mg concentrations of bauxite residue on the Ca nutrition of plants was investigated. As the toxicity of Al at high pH is not known, a study was conducted to examine the rhizotoxicity of aluminate (Al(OH)4-) and polycationic Al at high pH. The ability of plant roots to reduce rhizosphere pH in bauxite residue was also considered. A novel gypsum application method was assessed for its efficiency at improving the Ca status of bauxite residue. Manual adjustment, ion exchange resins and automated titration were examined for their suitability for nutrient solution pH control in alkaline conditions. For short-term studies, it was found that a solution without supply of Cu, Fe, Mn and Zn, and aerated with CO2 depleted air, greatly reduced nutrient precipitation at high pH, thus eliminating nutritional differences between treatments. Manual pH adjustment and the use of ion exchange resins as pH buffers were unsuitable methods of pH control. In contrast, pH control by automated titration had little effect on solution composition while maintaining constant pH. The solution culture system was used to examine OH- toxicity in mungbeans (Vigna radiata (L.) Wilczek cv. Emerald), with root length reduced at a bulk solution pH of 8.5 and greater. The effect of Ca activity ratio (CAR) and pH on Ca uptake by mungbeans and Rhodes grass (Chloris gayana cv. Pioneer) in Na dominated solution cultures and in soil was investigated. Changes in pH in the alkaline range were shown to have no effect on the critical CAR of 0.024 (corresponding to 90 % relative root length) for mungbeans grown in solution culture. Results from soil grown mungbeans confirmed those from solution culture, with a critical CAR of 0.025. A critical CAR of 0.034 was also established for soil grown Rhodes grass. However, using dilute nutrient solutions dominated by Mg at pH 9.0, root growth was found to be more limited than had been observed for Na solutions, with growth reduced beneath a critical CAR of 0.050. Using a CAR equation modified with plasma membrane binding constants (to incorporate the differing antagonistic effects of Mg and Na), new critical CAR values were calculated for Na (0.56) and Mg (0.44) dominated solutions. This modified CAR equation permits the calculation of CAR irrespective of the dominant salt present. Solubilities of various gypsum sources and size fractions in seawater were studied to investigate the effectiveness of gypsum addition to the residue sand pipeline, rather than as a direct field application. The dissolution rate constant varied with gypsum source (analytical grade (AR) > phosphogypsum (PG) > mined gypsum (MG)) due to reactivity and surface area differences, generally reaching saturation within 15 s (AR) to 30 min (MG > 2.0). The ability of bauxite residue to remove Ca from solution (due to cation exchange and precipitation) was also examined; the quantity of the total solution Ca adsorbed was found to be small (5 %). These low rates of solution Ca adsorption, comparatively rapid dissolution rates, and long pumping times (20 min), preclude the application of gypsum to the residue sand/seawater slurry as a method for residue amelioration. Dilute, alkaline (pH 9.5) nutrient solutions were used to investigate the effects of aluminate (Al(OH)4-) on mungbean root growth. Although root growth in Al(OH)4- solutions was slightly limited, the symptoms associated with this growth reduction were observed to be similar to those caused by the Al13 polycation at concentrations lower than that which can be detected. Also, when roots displaying these symptoms were transferred to fresh Al(OH)4- solutions, no root tip lesions were observed, and root hair growth on the lateral roots improved. Thus, Al(OH)4- is considered to be non-toxic, with the observed reduction in root growth in solutions containing Al(OH)4- due to the gradual formation of toxic Al13 in the bulk nutrient solution resulting from the acidification of the alkaline nutrient solution by the plant roots. The effect of Mn deficiency in Rhodes grass and of legume inoculation in lucerne (Medicago sativa L. cv. Hunter River), on the rhizosphere pH of plants grown in highly alkaline bauxite residue was investigated. In response to Mn deficiency in residue sand, Rhodes grass was observed to increase acidification of its rhizosphere (being up to 1.22 pH units lower than the bulk soil). Due to its ability to fix atmospheric N2 rather than relying on soil N (NO3-) reserves, inoculated lucerne (1.75 pH unit decrease) was also found to acidify its rhizosphere to a greater extent than non-inoculated lucerne (1.16 pH unit decrease).

Revegetation of bauxite residue is hampered by a lack of understanding of the limitations imposed on plant growth in highly saline and alkaline growth mediums. In this study, several of these growth limiting factors were investigated. The toxicity of the hydroxyl ion (OH-) was examined using a solution culture system developed to allow studies at high pH without nutritional limitations. Also using this solution culture system, the effect of the high Na and Mg concentrations of bauxite residue on the Ca nutrition of plants was investigated. As the toxicity of Al at high pH is not known, a study was conducted to examine the rhizotoxicity of aluminate (Al(OH)4-) and polycationic Al at high pH. The ability of plant roots to reduce rhizosphere pH in bauxite residue was also considered. A novel gypsum application method was assessed for its efficiency at improving the Ca status of bauxite residue. Manual adjustment, ion exchange resins and automated titration were examined for their suitability for nutrient solution pH control in alkaline conditions. For short-term studies, it was found that a solution without supply of Cu, Fe, Mn and Zn, and aerated with CO2 depleted air, greatly reduced nutrient precipitation at high pH, thus eliminating nutritional differences between treatments. Manual pH adjustment and the use of ion exchange resins as pH buffers were unsuitable methods of pH control. In contrast, pH control by automated titration had little effect on solution composition while maintaining constant pH. The solution culture system was used to examine OH- toxicity in mungbeans (Vigna radiata (L.) Wilczek cv. Emerald), with root length reduced at a bulk solution pH of 8.5 and greater. The effect of Ca activity ratio (CAR) and pH on Ca uptake by mungbeans and Rhodes grass (Chloris gayana cv. Pioneer) in Na dominated solution cultures and in soil was investigated. Changes in pH in the alkaline range were shown to have no effect on the critical CAR of 0.024 (corresponding to 90 % relative root length) for mungbeans grown in solution culture. Results from soil grown mungbeans confirmed those from solution culture, with a critical CAR of 0.025. A critical CAR of 0.034 was also established for soil grown Rhodes grass. However, using dilute nutrient solutions dominated by Mg at pH 9.0, root growth was found to be more limited than had been observed for Na solutions, with growth reduced beneath a critical CAR of 0.050. Using a CAR equation modified with plasma membrane binding constants (to incorporate the differing antagonistic effects of Mg and Na), new critical CAR values were calculated for Na (0.56) and Mg (0.44) dominated solutions. This modified CAR equation permits the calculation of CAR irrespective of the dominant salt present. Solubilities of various gypsum sources and size fractions in seawater were studied to investigate the effectiveness of gypsum addition to the residue sand pipeline, rather than as a direct field application. The dissolution rate constant varied with gypsum source (analytical grade (AR) > phosphogypsum (PG) > mined gypsum (MG)) due to reactivity and surface area differences, generally reaching saturation within 15 s (AR) to 30 min (MG > 2.0). The ability of bauxite residue to remove Ca from solution (due to cation exchange and precipitation) was also examined; the quantity of the total solution Ca adsorbed was found to be small (5 %). These low rates of solution Ca adsorption, comparatively rapid dissolution rates, and long pumping times (20 min), preclude the application of gypsum to the residue sand/seawater slurry as a method for residue amelioration. Dilute, alkaline (pH 9.5) nutrient solutions were used to investigate the effects of aluminate (Al(OH)4-) on mungbean root growth. Although root growth in Al(OH)4- solutions was slightly limited, the symptoms associated with this growth reduction were observed to be similar to those caused by the Al13 polycation at concentrations lower than that which can be detected. Also, when roots displaying these symptoms were transferred to fresh Al(OH)4- solutions, no root tip lesions were observed, and root hair growth on the lateral roots improved. Thus, Al(OH)4- is considered to be non-toxic, with the observed reduction in root growth in solutions containing Al(OH)4- due to the gradual formation of toxic Al13 in the bulk nutrient solution resulting from the acidification of the alkaline nutrient solution by the plant roots. The effect of Mn deficiency in Rhodes grass and of legume inoculation in lucerne (Medicago sativa L. cv. Hunter River), on the rhizosphere pH of plants grown in highly alkaline bauxite residue was investigated. In response to Mn deficiency in residue sand, Rhodes grass was observed to increase acidification of its rhizosphere (being up to 1.22 pH units lower than the bulk soil). Due to its ability to fix atmospheric N2 rather than relying on soil N (NO3-) reserves, inoculated lucerne (1.75 pH unit decrease) was also found to acidify its rhizosphere to a greater extent than non-inoculated lucerne (1.16 pH unit decrease).

Revegetation of bauxite residue is hampered by a lack of understanding of the limitations imposed on plant growth in highly saline and alkaline growth mediums. In this study, several of these growth limiting factors were investigated. The toxicity of the hydroxyl ion (OH-) was examined using a solution culture system developed to allow studies at high pH without nutritional limitations. Also using this solution culture system, the effect of the high Na and Mg concentrations of bauxite residue on the Ca nutrition of plants was investigated. As the toxicity of Al at high pH is not known, a study was conducted to examine the rhizotoxicity of aluminate (Al(OH)4-) and polycationic Al at high pH. The ability of plant roots to reduce rhizosphere pH in bauxite residue was also considered. A novel gypsum application method was assessed for its efficiency at improving the Ca status of bauxite residue. Manual adjustment, ion exchange resins and automated titration were examined for their suitability for nutrient solution pH control in alkaline conditions. For short-term studies, it was found that a solution without supply of Cu, Fe, Mn and Zn, and aerated with CO2 depleted air, greatly reduced nutrient precipitation at high pH, thus eliminating nutritional differences between treatments. Manual pH adjustment and the use of ion exchange resins as pH buffers were unsuitable methods of pH control. In contrast, pH control by automated titration had little effect on solution composition while maintaining constant pH. The solution culture system was used to examine OH- toxicity in mungbeans (Vigna radiata (L.) Wilczek cv. Emerald), with root length reduced at a bulk solution pH of 8.5 and greater. The effect of Ca activity ratio (CAR) and pH on Ca uptake by mungbeans and Rhodes grass (Chloris gayana cv. Pioneer) in Na dominated solution cultures and in soil was investigated. Changes in pH in the alkaline range were shown to have no effect on the critical CAR of 0.024 (corresponding to 90 % relative root length) for mungbeans grown in solution culture. Results from soil grown mungbeans confirmed those from solution culture, with a critical CAR of 0.025. A critical CAR of 0.034 was also established for soil grown Rhodes grass. However, using dilute nutrient solutions dominated by Mg at pH 9.0, root growth was found to be more limited than had been observed for Na solutions, with growth reduced beneath a critical CAR of 0.050. Using a CAR equation modified with plasma membrane binding constants (to incorporate the differing antagonistic effects of Mg and Na), new critical CAR values were calculated for Na (0.56) and Mg (0.44) dominated solutions. This modified CAR equation permits the calculation of CAR irrespective of the dominant salt present. Solubilities of various gypsum sources and size fractions in seawater were studied to investigate the effectiveness of gypsum addition to the residue sand pipeline, rather than as a direct field application. The dissolution rate constant varied with gypsum source (analytical grade (AR) > phosphogypsum (PG) > mined gypsum (MG)) due to reactivity and surface area differences, generally reaching saturation within 15 s (AR) to 30 min (MG > 2.0). The ability of bauxite residue to remove Ca from solution (due to cation exchange and precipitation) was also examined; the quantity of the total solution Ca adsorbed was found to be small (5 %). These low rates of solution Ca adsorption, comparatively rapid dissolution rates, and long pumping times (20 min), preclude the application of gypsum to the residue sand/seawater slurry as a method for residue amelioration. Dilute, alkaline (pH 9.5) nutrient solutions were used to investigate the effects of aluminate (Al(OH)4-) on mungbean root growth. Although root growth in Al(OH)4- solutions was slightly limited, the symptoms associated with this growth reduction were observed to be similar to those caused by the Al13 polycation at concentrations lower than that which can be detected. Also, when roots displaying these symptoms were transferred to fresh Al(OH)4- solutions, no root tip lesions were observed, and root hair growth on the lateral roots improved. Thus, Al(OH)4- is considered to be non-toxic, with the observed reduction in root growth in solutions containing Al(OH)4- due to the gradual formation of toxic Al13 in the bulk nutrient solution resulting from the acidification of the alkaline nutrient solution by the plant roots. The effect of Mn deficiency in Rhodes grass and of legume inoculation in lucerne (Medicago sativa L. cv. Hunter River), on the rhizosphere pH of plants grown in highly alkaline bauxite residue was investigated. In response to Mn deficiency in residue sand, Rhodes grass was observed to increase acidification of its rhizosphere (being up to 1.22 pH units lower than the bulk soil). Due to its ability to fix atmospheric N2 rather than relying on soil N (NO3-) reserves, inoculated lucerne (1.75 pH unit decrease) was also found to acidify its rhizosphere to a greater extent than non-inoculated lucerne (1.16 pH unit decrease).

Diss. (sammanfattning) Alnarp : Sveriges lantbruksuniv. / Härtill 5 uppsatser.

O uso de bactérias promotoras de crescimento vegetal ou agentes de biocontrole como inoculantes agrícolas é uma alternativa importante e ecologicamente correta, com grandes benefícios na agricultura para substituir, ou ao menos suplementar, a excessiva utilização de fertilizantes e pesticidas. Neste trabalho avaliamos a capacidade de biocontrole e de competência rizosférica de três bactérias com características de promoção de crescimento vegetal (Plant growth promoting - PGP): Bacillus mycoides B38V, Paenibacillus riograndensis SBR5 e Burkholderia sp. 89. As três bactérias avaliadas apresentaram grande versatilidade na utilização de substratos, o que poderia lhes garantir uma vantagem competitiva no ambiente rizosférico. Porém, inconsistências foram observadas nos ensaios em câmara de crescimento, ou seja, as características de PGP e de biocontrole observadas in vitro não se refletiram em benefícios para a planta. A linhagem 89 destacou-se pela produção de um metabólito estável com ampla atividade contra fungos fitopatogênicos. Através de abordagens genômicas e de análises multilocus, descrevemos Burkholderia sp. 89 como uma nova espécie membro do complexo Burkholderia cepacia, denominada de B. catarinensis 89T. O sequenciamento de seu genoma, seguido de uma análise pela ferramenta AntiSMASH, revelou a presença de um agrupamento gênico de peptídeo sintetases não ribossomais (NRPS) relacionadas com a biossíntese do sideróforo ornibactina e um agrupamento híbrido NRPS-policetídeo sintetase responsável pela biossíntese do glicolipopeptideo cíclico com atividade antifúngica burkholdina. Como estratégia de purificação de metabólitos secundários foi utilizada a metodologia da mineração de genoma combinada com fracionamento guiado por bioensaios seguida de análises em espectrômetro de massas. Desta forma, purificamos com sucesso duas variantes de ornibactina, D e F (761 e 789 Da, respectivamente), e detectamos a variante ornibactina B (m/z= 733) e as moléculas sinalizadoras homoserina lactonas C6-HSL, 3OH-C8-HSL e C8-HSL. Análises de espectrometria de massas demonstraram a presença de um grupo de metabólitos com massas de 1240, 1254, 1268, 1216, 1244 e 1272 Da, que, provavelmente, são novas variantes do antifúngico burkoldina. Sendo assim, B. catarinensis 89T possui potencial biotecnológico com possíveis aplicações farmacêuticas e agronômicas para o biocontrole de fungos fitopatogênicos. / The use of plant growth promotion bacteria or biocontrol agents as agricultural inoculants is an important eco-friendly alternative to substitute, or at least supplement, the excessive use of fertilizers and pesticides. In this work, we evaluated the biocontrol potential and rhizosphere competence of three bacteria that had shown plant growth promotion (PGP) abilities: Bacillus mycoides B38V, Paenibacillus riograndensis SBR5 and Burkholderia sp. 89. All three bacteria presented great versatility in their substrate utilization, which could enable them to survive in a competitive rhizosphere environment. However, inconsistencies were observed in the greenhouse experiments, whereas their interesting abilities observed in vitro did not result in benefits to the plants. Strain 89 produces a stable metabolite with a wide range of antifungal activity. Genomic comparisons and multilocus sequence analysis revealed Burkholderia sp. 89 as a new species of the Burkholderia cepacia complex and we described it as B. catarinensis 89T. We sequenced its genome and analyzed it with the AntiSMASH tool. This in silico prediction revealed the presence of a nonribosomal peptide synthetase (NRPS) cluster, which is related to the production of the siderophore ornibactin. Moreover, a hybrid NRPS- polyketide synthetase cluster for the production of the antifungal cyclic glicolipopeptide burkholdin was also found. A genome mining combined with a bioassay-guided fractionation with further mass spectrometry analysis was applied for the purification of these compounds. This approach enabled us to purify and characterize two variants of the siderophore ornibactin, D and F (761 and 789 Da, respectively). Also, we could detect the variant ornibactin B (m/z= 733) and the quorum sensing molecules homoserine lactones C6-HSL, 3OH-C8-HSL and C8-HSL in the supernatant of B. catarinensis 89T. Mass spectrometry analysis showed the presence of a group of metabolites with the masses 1240, 1254, 1268, 1216, 1244 and 1272 Da, which are probably new variants of the antifungal metabolite burkoldin. Therefore, B. catarinensis 89T has a great biotechnological potential for the production of metabolites with pharmaceutical and agricultural applications for the biocontrol of phytopathogenic fungi.