O conhecimento do comportamento vegetal quanto ao uso de nutrientes permite manusear ou modificar o sistema de cultivo para melhorar a eficiência na utilização de nitrogênio. O fornecimento de potássio para as plantas estimula o aproveitamento do nitrogênio, possibilitando que sua absorção, assimilação e, conseqüentemente, a produtividade sejam aumentadas. Com base nestes fatos, o objetivo deste estudo foi avaliar o efeito de combinações de doses de nitrogênio e de potássio na nutrição, no teor de clorofila e na atividade da enzima redutase do nitrato em plantas de trigo (Triticum aestivum L.). O experimento foi conduzido em casa de vegetação do Departamento de Ciência do Solo da ESALQ/USP, localizada no município de Piracicaba-SP, utilizando-se vasos de três litros onde foram acondicionadas amostras da camada 0-20 cm de um Argissolo Vermelho Amarelo Abrúptico. Utilizou-se esquema fatorial 52 fracionado, com combinações de cinco doses de nitrogênio (0; 70; 140; 210 e 280 mg dm -3) e de cinco doses de potássio (0; 50; 100; 150 e 200 mg dm-3), as quais foram distribuídas segundo delineamento experimental de blocos aleatorizados, com quatro repetições. As leituras SPAD das folhas do terço-médio das plantas de trigo foram realizadas 30 dias após a semeadura. Aos setenta e nove dias após a semeadura foi realizada determinação da atividade da enzima redutase do nitrato. Aos 80 dias após a semeadura, as plantas foram colhidas, separada em parte aérea, espiga e raízes, secadas, pesadas e submetidas à determinação da concentração de nutrientes e acúmulo de nitrogênio e potássio. Os resultados foram analisados por meio do programa estatístico SAS e mostraram que o equilíbrio entre o fornecimento de nitrogênio e potássio foi essencial para o crescimento adequado das plantas de trigo. Ficou evidente que a combinação entre as doses de nitrogênio e de potássio interagiu positivamente na produção de fitomassa e na concentração de nitrogênio, fósforo, potássio, cálcio e enxofre na parte aérea das plantas; o nitrogênio foi determinante para a concentração de magnésio no tecido vegetal. A interação entre as doses de nitrogênio e de potássio influenciou positivamente na produção de fitomassa e na concentração de nitrogênio na espiga. A concentração de potássio na espiga dependeu apenas do fornecimento das doses de nitrogênio. A produção de fitomassa das raízes foi incrementada com o fornecimento das doses de nitrogênio e de potássio, sendo suas concentrações de nitrogênio, magnésio e de enxofre influenciadas pelo suprimento de nitrogênio. A concentração de potássio nas raízes diminuiu e a de cálcio aumentou com as doses de nitrogênio, conforme equação de segundo grau. O fornecimento de potássio promoveu decréscimos lineares na concentração de magnésio na parte aérea e nas raízes das plantas de trigo. O nitrogênio aumentou a atividade da redutase do nitrato até as doses intermediárias, diminuindo nas doses elevadas. O teor de clorofila das folhas do terço-médio da planta foi influenciado pela interação entre as doses de nitrogênio e potássio e refletiu diretamente na produtividade de fitomassa da parte aérea e de espigas, assim como no acúmulo de nitrogênio pelas plantas de trigo. / The knowledge about plant nutrition allows managing crop systems in order to improve the nitrogen use efficiency. Supplying potassium to plants stimulates the efficiency use of nitrogen through plant uptake and assimilation, consequently increasing crop yields. Based on these facts, the aim of this study was to evaluate the effect of combined rates of nitrogen and potassium on the nutrition, chlorophyll content, and nitrate reductase activity in wheat (Triticum aestivumaestivum L.) plants. The experiment was conducted in a greenhouse of the Soil Science Department of the ESALQ/USP in Piracicaba State of São Paulo, Brazil. Three-liter pots were filled with 0-20 cm depth samples of Arenic Hapludult. A fractionated 52 factorial was used, with combinations of five nitrogen rates (0; 70; 140; 210 e 280 mg dm-3) and five potassium rates (0; 50; 100; 150 e 200 mg dm-3), which were set in a randomized block design with four replications. Thirty days after seeding SPAD values readings were taken on the leaves. Nitrate reductase activity was determined seventy-nine days after seeding and one day later plants were harvested and separated in shoot, roots, and ears. These plant materials were weighted and submitted to nutrient content and accumulation of nitrogen and potassium. Statistical analyses were performed by means of SAS software. Results presented that the balance between nitrogen and potassium supplies was essential to adequate wheat plant growth. It was clear that the combination of nitrogen and potassium rates had a positive influence on wheat dry mass yield as well as on nitrogen, phosphorus, potassium, calcium, and sulphur contents in plant shoot. Nitrogen was essential to maintain adequate magnesium content in plant tissues. The interaction between nitrogen and potassium rates was important for plant yield and ear nitrogen content. Potassium content in the ears was affected only by nitrogen rates. Root dry matter yield increased with rates of nitrogen and potassium. Nitrogen, magnesium and sulphur contents in the roots were influenced only by nitrogen application. Potassium concentration in the roots reduced and calcium concentration increased with nitrogen rates according to a quadratic equation. Potassium supply caused linear decreases in magnesium content of shoots and roots. Nitrogen increased nitrate reductase activity up to the medium rates of application and reduced for the highest rates. Chlorophyll content in the leaves was influenced by the interaction between nitrogen and potassium rates, reflecting on the dry matter yield of plant shoot and ears, as well as on nitrogen accumulation in the wheat plants.

<p>This investigation addresses how photosynthetic efficiency is affected when pea (Pisum sativum) plants are restricted to a sole nitrogen source (i.e. ammonium or nitrate). The pea plants were watered with different nutrient solutions without NO3- or NH4+ for different time-periods in order to assay for nitrogen content. The soluble ammonium and nitrate content was measured throughout the entire growth period. No major differences were observed in nitrogen content during the starvation period up to 25 days. For technical reasons, cultivation of plants could not be extended beyond this time. The chloroplasts and thylakoids were isolated after 25 days and assayed for chlorophyll contents and photosynthetic activity.</p><p>The outcome of these tests indicates a small but unambiguous decrease in the photosynthesis activity for all treatments, relative the control.</p>

This investigation addresses how photosynthetic efficiency is affected when pea (Pisum sativum) plants are restricted to a sole nitrogen source (i.e. ammonium or nitrate). The pea plants were watered with different nutrient solutions without NO3- or NH4+ for different time-periods in order to assay for nitrogen content. The soluble ammonium and nitrate content was measured throughout the entire growth period. No major differences were observed in nitrogen content during the starvation period up to 25 days. For technical reasons, cultivation of plants could not be extended beyond this time. The chloroplasts and thylakoids were isolated after 25 days and assayed for chlorophyll contents and photosynthetic activity. The outcome of these tests indicates a small but unambiguous decrease in the photosynthesis activity for all treatments, relative the control.

Thesis (Ph. D.)--University of Missouri-Columbia, 2001. / Typescript. Vita. Includes bibliographical references. Also available on the Internet.

Thesis (Ph. D.)--University of Missouri-Columbia, 2001. / Typescript. Vita. Includes bibliographical references. Also available on the Internet.

We present the first characterization of K+ optimization of N uptake and metabolism in an NH4+-tolerant species, tropical lowland rice (cv. IR-72). 13N radiotracing showed that increased K+ supply reduces futile NH4+ cycling at the plasma membrane, diminishing the excessive rates of both unidirectional influx and efflux. Pharmacological testing showed that low-affinity NH4+ influx may be mediated by both K+ and non-selective cation channels. Suppression of NH4+ influx by K+ occurred within minutes of increasing K+ supply. Increased K+ reduced free [NH4+] in roots and shoots by 50-75%. Plant biomass was maximized on 10 mM NH4+ and 5 mM K+, with growth 160% higher than 10 mM NO3--grown plants, and 220% higher than plants grown at 10 mM NH4+ and 0.1 mM K+. Unlike in NH4+-sensitive barley, growth optimization was not attributed to a reduced energy cost of futile NH4+ cycling at the plasma membrane. Activities of the key enzymes glutamine synthetase (GS) and phosphoenolpyruvate carboxylase (PEPC) were strongly stimulated by elevated K+, mirroring plant growth and protein content. Improved plant performance through optimization of K+ and NH4+ is likely to be of substantial agronomic significance in the world’s foremost crop species.

We present the first characterization of K+ optimization of N uptake and metabolism in an NH4+-tolerant species, tropical lowland rice (cv. IR-72). 13N radiotracing showed that increased K+ supply reduces futile NH4+ cycling at the plasma membrane, diminishing the excessive rates of both unidirectional influx and efflux. Pharmacological testing showed that low-affinity NH4+ influx may be mediated by both K+ and non-selective cation channels. Suppression of NH4+ influx by K+ occurred within minutes of increasing K+ supply. Increased K+ reduced free [NH4+] in roots and shoots by 50-75%. Plant biomass was maximized on 10 mM NH4+ and 5 mM K+, with growth 160% higher than 10 mM NO3--grown plants, and 220% higher than plants grown at 10 mM NH4+ and 0.1 mM K+. Unlike in NH4+-sensitive barley, growth optimization was not attributed to a reduced energy cost of futile NH4+ cycling at the plasma membrane. Activities of the key enzymes glutamine synthetase (GS) and phosphoenolpyruvate carboxylase (PEPC) were strongly stimulated by elevated K+, mirroring plant growth and protein content. Improved plant performance through optimization of K+ and NH4+ is likely to be of substantial agronomic significance in the world’s foremost crop species.

AM plants acquire Pi via two pathways; the direct uptake pathway via plant roots and the AM pathway via external fungal hyphae and colonised cortical cells. It has been assumed that these two pathways are additive and therefore in non-responsive plants the AM pathway is often considered to be non-functional. However, data from ³²P uptake studies indicates that the AM pathway is functional in many non-responsive symbioses and in some instances supplies the majority of plant P. In recent years the high-affinity Pi transporters involved in both direct and AM Pi uptake pathways have been identified. They are expressed at the root epidermis and the symbiotic interface of colonised cortical cells and respond to the P and AM status of the plant. The overall objective of the work described in this thesis was to characterise Pi uptake via the AM pathway in barley, a non-responsive AM host, using an approach which integrated physiological measurements of plant responsiveness and AM contribution with investigations of gene expression and functional characterisation of the plant Pi transporters. A preliminary survey of field-grown barley demonstrated the persistence of AM colonisation under commercial cropping regimes in southern Australia and highlighted the relevance of AM studies to commercial agriculture. Under glasshouse conditions AM colonisation of barley induced depressions in growth and P uptake compared to NM controls. Growth depressions were unrelated to percent colonisation by two AM fungal species and could not readily be explained by fungal C demand; the strong correlation between growth and P content suggested that P was the limiting factor in these experiments. However, a compartmented pot system incorporating ³²P-labelling demonstrated that the AM pathway is functional in colonised barley and, in the interaction with G. intraradices, contributed 48% of total P. This suggested that P flux via the direct uptake pathway is decreased in AM barley. The expression of three Pi transporters, HvPT1, HvPT2 and HvPT8 was investigated in colonised roots. HvPT1 and HvPT2 have previously been localised to the root epidermis and root hairs and are involved in Pi uptake via the direct pathway whilst HvPT8 is an AM-inducible Pi transporter which was localised by in-situ hybridisation to colonised cortical cells. Using promoter::GFP gene fusions the localisation of HvPT8 to arbuscule-containing cortical cells was confirmed in living roots from transgenic barley. Quantitative real-time PCR analysis of the expression of these three Pi transporters indicated that HvPT1 and HvPT2 were expressed constantly, under all conditions regardless of AM colonisation status and indicated that decreased P flux via the direct pathway is not related to expression of these transporters. HvPT8 was induced in AM colonised roots. However, the level of expression was not related to flux via the AM pathway or arbuscular colonisation. The HvPT8 transporter was further characterised by constitutive over-expression in transgenic barley. ³²P uptake assays in excised roots demonstrated increased Pi uptake from low P solution compared to wild-type roots and confirmed that HvPT8 is a functional Pi transporter with high-affinity transport properties. This is the first report of characterisation of an AM-inducible Pi transporter in planta. When these transgenic plants were grown in solution culture there was no increase in growth or P uptake relative to wild-type or transgenic controls and growth in soil and AM colonisation were also unaffected in these transgenic lines. The data presented in this thesis highlights the importance of combined physiological and molecular approaches to characterising plant AM interactions. The persistence of AM colonisation in barley in the field indicates the importance of improving our understanding of symbiotic function in non-responsive plants. Future efforts should be directed towards understanding the signals which regulate P flux via both the direct and AM pathways with the ultimate aim of enhancing AM responsiveness of non-responsive species. Making the direct and AM pathways additive in nonresponsive species should be a key aim of future research. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1313311 / Thesis (Ph.D.) -- University of Adelaide, School of Earth and Environmental Sciences, 2008

AM plants acquire Pi via two pathways; the direct uptake pathway via plant roots and the AM pathway via external fungal hyphae and colonised cortical cells. It has been assumed that these two pathways are additive and therefore in non-responsive plants the AM pathway is often considered to be non-functional. However, data from ³²P uptake studies indicates that the AM pathway is functional in many non-responsive symbioses and in some instances supplies the majority of plant P. In recent years the high-affinity Pi transporters involved in both direct and AM Pi uptake pathways have been identified. They are expressed at the root epidermis and the symbiotic interface of colonised cortical cells and respond to the P and AM status of the plant. The overall objective of the work described in this thesis was to characterise Pi uptake via the AM pathway in barley, a non-responsive AM host, using an approach which integrated physiological measurements of plant responsiveness and AM contribution with investigations of gene expression and functional characterisation of the plant Pi transporters. A preliminary survey of field-grown barley demonstrated the persistence of AM colonisation under commercial cropping regimes in southern Australia and highlighted the relevance of AM studies to commercial agriculture. Under glasshouse conditions AM colonisation of barley induced depressions in growth and P uptake compared to NM controls. Growth depressions were unrelated to percent colonisation by two AM fungal species and could not readily be explained by fungal C demand; the strong correlation between growth and P content suggested that P was the limiting factor in these experiments. However, a compartmented pot system incorporating ³²P-labelling demonstrated that the AM pathway is functional in colonised barley and, in the interaction with G. intraradices, contributed 48% of total P. This suggested that P flux via the direct uptake pathway is decreased in AM barley. The expression of three Pi transporters, HvPT1, HvPT2 and HvPT8 was investigated in colonised roots. HvPT1 and HvPT2 have previously been localised to the root epidermis and root hairs and are involved in Pi uptake via the direct pathway whilst HvPT8 is an AM-inducible Pi transporter which was localised by in-situ hybridisation to colonised cortical cells. Using promoter::GFP gene fusions the localisation of HvPT8 to arbuscule-containing cortical cells was confirmed in living roots from transgenic barley. Quantitative real-time PCR analysis of the expression of these three Pi transporters indicated that HvPT1 and HvPT2 were expressed constantly, under all conditions regardless of AM colonisation status and indicated that decreased P flux via the direct pathway is not related to expression of these transporters. HvPT8 was induced in AM colonised roots. However, the level of expression was not related to flux via the AM pathway or arbuscular colonisation. The HvPT8 transporter was further characterised by constitutive over-expression in transgenic barley. ³²P uptake assays in excised roots demonstrated increased Pi uptake from low P solution compared to wild-type roots and confirmed that HvPT8 is a functional Pi transporter with high-affinity transport properties. This is the first report of characterisation of an AM-inducible Pi transporter in planta. When these transgenic plants were grown in solution culture there was no increase in growth or P uptake relative to wild-type or transgenic controls and growth in soil and AM colonisation were also unaffected in these transgenic lines. The data presented in this thesis highlights the importance of combined physiological and molecular approaches to characterising plant AM interactions. The persistence of AM colonisation in barley in the field indicates the importance of improving our understanding of symbiotic function in non-responsive plants. Future efforts should be directed towards understanding the signals which regulate P flux via both the direct and AM pathways with the ultimate aim of enhancing AM responsiveness of non-responsive species. Making the direct and AM pathways additive in nonresponsive species should be a key aim of future research. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1313311 / Thesis (Ph.D.) -- University of Adelaide, School of Earth and Environmental Sciences, 2008

The soils of Australia have extensive macro and micronutrient disorders varying greatly in their capacities to provide the chemical nutrients essential for plant growth. Assessment of nutrient availability in soils is important in order to maximise fertilizer efficiency and crop yields and to minimise environmental pollution associated with over fertilisation. Nutrient availability has proven difficult to assess due to the complexity of trace element soil chemistry and plant uptake mechanisms. The relatively new method, Diffusive Gradients in Thin Films ( DGT ), provides the potential to become an alternative soil test that could accurately predict nutrient availability. To date, DGT technology has only been designed for separate assessment of anionic and cationic species in waters or soils typically at concentrations characteristic of highly contaminated systems. In this study a new mixed binding gel ( MBL ) was developed capable of simultaneous assessment of cations and anions in a single assay at concentrations more representative of uncontaminated agricultural soils, sediments and waters. The MBL has the potential to eliminate measurement errors associated with very fine spatial scale changes in element concentrations in these environments. The MBL consisted of ferrihydrite and Chelex - 100 cation exchange resin combined together in a binding gel. Results from the MBL were comparable to experiments performed using individual Chelex gels and ferrihydrite gels that have been shown to work successfully for DGT methodology. To facilitate combined analysis of P and cations by ICP - MS, HCl ( 1 M ) was used for gel elution to minimise interferences from [superscript 14] N [superscript 16] OH or [superscript 15] N [superscript 16] O on [superscript 31] P. All elements tested ( Cd, Cu, Mn, Mo, P and Zn ) were bound successfully to the MBL. DGT measurements obtained using the MBL on agricultural soils correlated well ( r ? = 0.95 ) with measurements obtained using pure Chelex and ferrihydrite binding layers. This suggests that the MBL could be used for simultaneous measurement of cationic and anionic element availability in soils. Performance of the Diffusive Gradient in Thin Films ( DGT ) technique was compared with three other common testing methods ( Colwell, Olsen, Resin ) for available soil P in terms of the ability of each to predict wheat, canola, lupin and barley responsiveness to applied P on 21 Australian agricultural soils. DGT accurately predicted plant responsiveness in > 90 % of the soils used. In contrast the other soil testing methods failed to correctly predict plant response to P on numerous occasions. These observations reveal that the DGT technique with the newly developed MBL can predict plant available P on these soils with greater accuracy than other traditional soil P testing methods and could become a useful tool for predicting P fertilizer requirements. The DGT method using the MBL was also used to test Zn deficiency thresholds for canola and wheat in a manufactured soil ( acid washed sand ). DGT successfully determined the threshold for Zn deficiency in this soil, overcoming detection limit issues usually accompanying such low levels of Zn. This method also provides that potential to assess other micronutrients ( Mn, Cu ) and with further modification potentially assess K. Before DGT can become established as an alternative soil testing method, validation of the performance is required under field conditions. This study has shown that it out performs current common soil testing methods in glasshouse conditions but questions still remain if this will be reflected out in the field. / Thesis (Ph.D.)-- University of Adelaide, School of Earth and Environmental Sciences, 2007.