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Typescript (photocopy). / Digitized by Kansas Correctional Industries

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There is accumulating evidence that arbuscular mycorrhizal fungi (AMF) produce a glycoprotein called glomalin, which has the potential to increase soil carbon (C) and nitrogen (N) storage, thereby reducing soil emissions of carbon dioxide (CO2) and nitrous oxide (N2O) into the atmosphere. However, other soil microorganisms such as plant growth-promoting rhizobacteria (PGPR) that interact with AMF could indirectly influence glomalin production. The objectives of this study were to determine the effects of AMF and PGPR interactions on glomalin production and identify possible combinations of these organisms that could enhance C and N storage in the rhizosphere. The effects of AMF and PGPR interactions on pea (Pisum sativum L.) growth and correlations between glomalin production and plant growth also were assessed.<p> A series of growth chamber and laboratory experiments were conducted to examine the effect of fungal and host plant species on glomalin production by comparing the amounts of glomalin produced by Glomus clarum, G. intraradices, and G. mosseae in association with corn (Zea mays L.), in addition to examining differences in the ability of corn, pea, and wheat (Triticum aestivum L.) to support glomalin production by G. intraradices. There were no significant differences in glomalin production [measured in the rhizosphere as Bradford-reactive soil protein (BRSP)] by the three AMF species, whereas host plant significantly affected glomalin production. Specifically, higher BRSP concentrations were found in the rhizosphere of corn as compared to pea and wheat.<p> Additionally, the effect of long-term storage on the growth promoting traits of the PGPR strains selected; namely, Pseudomonas cepacia R55 and R85, P. aeruginosa R75, P. putida R105, and P. fluorescence R111 were investigated. These bacterial strains previously had been identified as PGPR, but had since undergone approximately twenty years of storage at -80¢ªC; thus, it was necessary to confirm that these strains had retained their plant growth promoting characteristics. Apparently, long-term storage had no significant adverse effect on the PGPR strains as all strains increased the total biomass of wheat significantly and demonstrated antagonism against fungal pathogens.<p> The possibility that spore-associated bacteria (SAB) could influence AMF associations, thereby affecting glomalin production, and subsequent crop yield potential was assessed. This was achieved by first isolating bacteria from disinfested spores of the AMF species and determining their potential as PGPR for wheat. According to fatty acid methyl ester (FAME) profiles, four genera of bacteria were isolated from AMF spores namely; Arthrobacter, Bacillus, Micrococcus, and Paenibacillus, of which Bacillus species were the most common SAB. None of these isolates, however, showed growth promoting abilities on wheat.<p> Based on the preliminary findings, the combined effects of the three AMF species and the five PGPR strains were examined on plant growth and glomalin production under gnotobiotic conditions using pea as the host plant. Interactions between G. intraradices and R75, R85, or R105 resulted in increased BRSP concentration in the mycorrhizosphere of pea. Additionally, significant interactions were observed between the AMF species and PGPR strains on BRSP concentration in pea rhizosphere under non-sterile conditions. As observed under sterile conditions, the co-inoculation of pea with G. intraradices and R75 or R85 increased BRSP concentrations in the rhizosphere of pea grown in non-sterile soil, although interaction effects were not significantly different from the control or when G. intraradices was applied alone. Significant AMF and PGPR interactions were observed to affect AMF colonization; however, the combination of these organisms did not significantly affect pea growth, nutrient uptake, and C and N storage in the plant rhizosphere. No correlations were detected between glomalin-related soil protein (GRSP), pea growth, nutrient concentrations in the plant tissue, and soil organic C and N content. This study demonstrated that although the potential exists to manipulate certain AMF and PGPR to enhance glomalin production, co-inoculation of AMF and PGPR did not enhance plant growth or C and N storage beyond that achieved by inoculation of either organism.

Soil salinity is a widespread problem that limits crop yield throughout the world. The accumulation of soluble salts in the soil can inhibit plant growth by increasing the osmotic potential of interstitial water, inducing ion toxicity and nutrient imbalances in plants. Over the last decade, considerable effort has been put into developing economical and effective methods to reclaim these damaged soils. Phytoremediation is a technique that uses plants to extract, contain, immobilize and degrade contaminants in soil. The most common process for salt bioremediation is phytoextraction which uses plants to accumulate salt in the shoots, which is then removed by harvesting the foliage. As developing significant plant biomass in saline soils is an issue, a group of free-living rhizobacteria, called plant growth promoting rhizobacteria (PGPR), can be applied to plant seeds to aid plant growth by alleviating salt stress. The principle objective of this research was to test the efficacy of PGPR in improving the growth of plants on salt-impacted soils through greenhouse and field studies. In this research, previously isolated PGPR strains of Pseudomonas putida. UW3, Pseudomonas putida UW4, and Pseudomonas corrugata CMH3 were applied to barley (Hordeum valgare C.V. AC ranger), oats (Avena sativa C.V. CDC baler), tall wheatgrass (Agropyron elongatum), and tall fescue (festuca arundinacea C.V. Inferno). PGPR effects on plant growth, membrane stability, and photosynthetic activity under salt stress were examined. Greenhouse studies showed that plants treated with PGPR resulted in an increase in plant biomass by up to 500% in salt-impacted soils. Electrolyte leakage assay showed that plants treated with PGPR resulted in 50% less electrolyte leakage from membranes. Several chlorophyll a fluorescence parameters, Fv/Fm, effective quantum yield, Fs, qP, and qN obtained from pulse amplitude modulation (PAM) fluorometry showed that PGPR-treated plants resulted in improvement in photosynthesis under salt stress. Field studies showed that PGPR promoted shoot dry biomass production by 27% to 230%. The NaCl accumulation in plant shoots increased by 7% to 98% with PGPR treatment. The averaged soil salinity level at the CMS and CMN site decreased by 20% and 60%, respectively, during the 2008 field season. However, there was no evidence of a decrease in soil salinity at the AL site. Based on the improvements of plant biomass production and NaCl uptake by PGPR observed in the 2008 field studies, the phytoremediation efficiency on salt-impacted sites is expected to increase by 30-60% with PGPR treatments. Based on the average data of 2007 and 2008 field season, the time required to remove 25% of NaCl of the top 50 cm soil at the CMS, CMN and AL site is estimated to be six, twelve, and sixteen years, respectively, with PGPR treatments. The remediation efficiency is expected to accelerate during the remediation process as the soil properties and soil salinity levels improve over time.

Soil salinity is a widespread problem that limits crop yield throughout the world. The accumulation of soluble salts in the soil can inhibit plant growth by increasing the osmotic potential of interstitial water, inducing ion toxicity and nutrient imbalances in plants. Over the last decade, considerable effort has been put into developing economical and effective methods to reclaim these damaged soils. Phytoremediation is a technique that uses plants to extract, contain, immobilize and degrade contaminants in soil. The most common process for salt bioremediation is phytoextraction which uses plants to accumulate salt in the shoots, which is then removed by harvesting the foliage. As developing significant plant biomass in saline soils is an issue, a group of free-living rhizobacteria, called plant growth promoting rhizobacteria (PGPR), can be applied to plant seeds to aid plant growth by alleviating salt stress. The principle objective of this research was to test the efficacy of PGPR in improving the growth of plants on salt-impacted soils through greenhouse and field studies. In this research, previously isolated PGPR strains of Pseudomonas putida. UW3, Pseudomonas putida UW4, and Pseudomonas corrugata CMH3 were applied to barley (Hordeum valgare C.V. AC ranger), oats (Avena sativa C.V. CDC baler), tall wheatgrass (Agropyron elongatum), and tall fescue (festuca arundinacea C.V. Inferno). PGPR effects on plant growth, membrane stability, and photosynthetic activity under salt stress were examined. Greenhouse studies showed that plants treated with PGPR resulted in an increase in plant biomass by up to 500% in salt-impacted soils. Electrolyte leakage assay showed that plants treated with PGPR resulted in 50% less electrolyte leakage from membranes. Several chlorophyll a fluorescence parameters, Fv/Fm, effective quantum yield, Fs, qP, and qN obtained from pulse amplitude modulation (PAM) fluorometry showed that PGPR-treated plants resulted in improvement in photosynthesis under salt stress. Field studies showed that PGPR promoted shoot dry biomass production by 27% to 230%. The NaCl accumulation in plant shoots increased by 7% to 98% with PGPR treatment. The averaged soil salinity level at the CMS and CMN site decreased by 20% and 60%, respectively, during the 2008 field season. However, there was no evidence of a decrease in soil salinity at the AL site. Based on the improvements of plant biomass production and NaCl uptake by PGPR observed in the 2008 field studies, the phytoremediation efficiency on salt-impacted sites is expected to increase by 30-60% with PGPR treatments. Based on the average data of 2007 and 2008 field season, the time required to remove 25% of NaCl of the top 50 cm soil at the CMS, CMN and AL site is estimated to be six, twelve, and sixteen years, respectively, with PGPR treatments. The remediation efficiency is expected to accelerate during the remediation process as the soil properties and soil salinity levels improve over time.

There is accumulating evidence that arbuscular mycorrhizal fungi (AMF) produce a glycoprotein called glomalin, which has the potential to increase soil carbon (C) and nitrogen (N) storage, thereby reducing soil emissions of carbon dioxide (CO2) and nitrous oxide (N2O) into the atmosphere. However, other soil microorganisms such as plant growth-promoting rhizobacteria (PGPR) that interact with AMF could indirectly influence glomalin production. The objectives of this study were to determine the effects of AMF and PGPR interactions on glomalin production and identify possible combinations of these organisms that could enhance C and N storage in the rhizosphere. The effects of AMF and PGPR interactions on pea (Pisum sativum L.) growth and correlations between glomalin production and plant growth also were assessed.<p> A series of growth chamber and laboratory experiments were conducted to examine the effect of fungal and host plant species on glomalin production by comparing the amounts of glomalin produced by Glomus clarum, G. intraradices, and G. mosseae in association with corn (Zea mays L.), in addition to examining differences in the ability of corn, pea, and wheat (Triticum aestivum L.) to support glomalin production by G. intraradices. There were no significant differences in glomalin production [measured in the rhizosphere as Bradford-reactive soil protein (BRSP)] by the three AMF species, whereas host plant significantly affected glomalin production. Specifically, higher BRSP concentrations were found in the rhizosphere of corn as compared to pea and wheat.<p> Additionally, the effect of long-term storage on the growth promoting traits of the PGPR strains selected; namely, Pseudomonas cepacia R55 and R85, P. aeruginosa R75, P. putida R105, and P. fluorescence R111 were investigated. These bacterial strains previously had been identified as PGPR, but had since undergone approximately twenty years of storage at -80¢ªC; thus, it was necessary to confirm that these strains had retained their plant growth promoting characteristics. Apparently, long-term storage had no significant adverse effect on the PGPR strains as all strains increased the total biomass of wheat significantly and demonstrated antagonism against fungal pathogens.<p> The possibility that spore-associated bacteria (SAB) could influence AMF associations, thereby affecting glomalin production, and subsequent crop yield potential was assessed. This was achieved by first isolating bacteria from disinfested spores of the AMF species and determining their potential as PGPR for wheat. According to fatty acid methyl ester (FAME) profiles, four genera of bacteria were isolated from AMF spores namely; Arthrobacter, Bacillus, Micrococcus, and Paenibacillus, of which Bacillus species were the most common SAB. None of these isolates, however, showed growth promoting abilities on wheat.<p> Based on the preliminary findings, the combined effects of the three AMF species and the five PGPR strains were examined on plant growth and glomalin production under gnotobiotic conditions using pea as the host plant. Interactions between G. intraradices and R75, R85, or R105 resulted in increased BRSP concentration in the mycorrhizosphere of pea. Additionally, significant interactions were observed between the AMF species and PGPR strains on BRSP concentration in pea rhizosphere under non-sterile conditions. As observed under sterile conditions, the co-inoculation of pea with G. intraradices and R75 or R85 increased BRSP concentrations in the rhizosphere of pea grown in non-sterile soil, although interaction effects were not significantly different from the control or when G. intraradices was applied alone. Significant AMF and PGPR interactions were observed to affect AMF colonization; however, the combination of these organisms did not significantly affect pea growth, nutrient uptake, and C and N storage in the plant rhizosphere. No correlations were detected between glomalin-related soil protein (GRSP), pea growth, nutrient concentrations in the plant tissue, and soil organic C and N content. This study demonstrated that although the potential exists to manipulate certain AMF and PGPR to enhance glomalin production, co-inoculation of AMF and PGPR did not enhance plant growth or C and N storage beyond that achieved by inoculation of either organism.

Soybean (Glycine max (L.) Merr.) is a subtropical legume that requires root zone temperatures (RZTs) in the 25 to 30$ sp circ$C range for optimal symbiotic activity. The inability of soybean to adapt to cool soil conditions limits its development and yield in short season areas. In particular, nodulation and N$ sb2$ fixation by this subtropical crop species is sensitive to cool (RZT). The objectives of this thesis were to determine whether or not PGPR could be used to help overcome the low RZT inhibition of soybean nodulation, to improve soybean nitrogen fixation and yield under field conditions and to determine the methods by which such increases occurred. The work reported in this thesis has demonstrated that PGPR can increase early season nodulation and total seasonal nitrogen fixation and yield of soybean growing in an area with cool spring soils. The ability of PGPR to stimulate soybean nodulation and growth was shown to be related to their ability to colonize soybean roots, and this was shown to be related to RZT. All steps in early nodulation were stimulated by the presence of PGPR. The beneficial effects of PGPR are exerted through a diffusible molecule excreted into the growth medium. The addition of genistein, a plant-to-bacteria signal molecule already shown to stimulate soybean N$ sb2$ fixation at low RZT, plus PGPR causes increases in soybean nodulation, N$ sb2$ fixation, and growth that were greater than those caused by the addition of PGPR alone, but only at 25 and 17.5$ sp circ$C, and not at 15$ sp circ$C RZT.