The chemical properties of soil for alfalfa production after biofertiliser application

The objective of this study was to evaluate the use of biofertilisers for the production of alfalfa shoot, root and nodule dry matter, and also, to evaluate the chemical properties of the soil. This study was conducted in the greenhouse of the Support Department, Animal Production and Health, Faculty of Veterinary Medicine/UNESP, Aracatuba - SP, from May to October 2010. The experimental design was completely randomised with six biofertiliser doses (0, 25, 50, 100, 200 and 400 m(3) ha(-1)) and five replicates. The biofertiliser doses were the primary treatments and the cuts (five) were subplots. The cuts were performed, on average, every 27 days at 10 cm above the soil. At the end of the experiment, the roots, nodules and soil from all experimental units were collected for chemical analysis. We observed a linear increase in dry matter production of the shoots relative to the doses studied. The dry matter production of the roots and nodules was not significantly different. The chemical properties of the soil significantly improved for calcium and magnesium as well as the sum of bases and base saturation with biofertiliser application. Biofertilisers can be used for agricultural production and favourably alter the soil characteristics.

Nutrient accumulation and biomass production of alfafa after soil amendment with silicates

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

NUTRITIONAL STATUS AND DRY MASS PRODUCTION OF ALFALFA IN FUNCTION OF POTASSIUM RATES IN TWO SOILS

Potassium fertilization is very important to alfalfa crop in terms of yield, quality and persistence of forage, especially on soils naturally poor K. Thus, to assess the effects of K fertilization in alfalfa production and nutritional status, was carried out an experiment in a greenhouse using samples of a Dystrophic Oxisol medium texture (LV) (0.6 mmol(c) dm(-3) K) and a Dystrophic Ultisol sandy/medium texture (PVA) (2.2 mmol(c) dm(-3) K). A completely randomized design in a factorial arrangement 6 x 2 (six K rates and two soils) was used, with four replications. The K rates used were: 0, 25, 50, 100, 150 and 200 mg kg(-1) K. Potassium fertilization increased K content in soil and shoots. Dry matter production was increased with the K addition. However, in the PVA, this occurred only in the second cut. In LV, potassium fertilization increased N concentration in alfalfa shoots in both cuts. Plants with K concentration around 10 g kg(-1) had typical symptoms of this nutrient deficiency. The K critical levels of K in soil and shoots were 1.8 mmolc dm(-3) and 16.7 g kg(-1), respectively.

Escarificado en siembra directa

Para obtener información respecto de los posibles recorridos que realiza un flujo de agua y su distribución espacial bajo la labor de escarificado en siembra directa y comprobar su persistencia al 1 ½ año de haber sido efectuada, se realizó un ensayo en Zavalla, Santa Fe (60° 53' O y 33° 01' S) en un Argiudol vértico bajo cultivo de Medicago sativa. T0 fue siembra directa sin escarificar, T1 escarificado antes de sembrar y T2 escarificado al año de la siembra de la alfalfa. Se simuló la lluvia irrigando con una solución de 1g.l-1 de azul de metileno. Se fotografiaron perfiles sucesivos en cada tratamiento. Los sectores coloreados de las fotografías fueron delimitados, digitalizados y medidas las áreas y longitudes verticales. El movimiento descendente tuvo lugar mayoritariamente como flujos preferenciales que fueron iniciados por las fisuras, las raíces creciendo dentro y fuera del área removida y las grietas del suelo. En todos los tratamientos los flujos atravesaron el límite A / B1, pero en ningún caso alcanzaron el B2. La configuración de los patrones de distribución permitió comprobar la mayor capacidad de conducción hidráulica y persistencia de la labor de escarificado sobre siembra directa.

Efecto del régimen de riego en el rendimiento de alfalfa para corte en el Valle Central de Catamarca, Argentina

En la provincia de Catamarca existe una gran superficie cultivada con alfalfa bajo riego. La falta de conocimiento de los requerimientos hídricos del cultivo en esta zona es un grave inconveniente. Con el objeto de subsanar este problema, se realizó en el Valle Central un ensayo de riego con un diseño en bloques completamente al azar, con cuatro tratamientos y cuatro repeticiones. Los tratamientos consistieron en la reposición de humedad al suelo a distintos umbrales: 75, 50, 25 y 0% de la capacidad de almacenamiento total de agua del suelo. Se evaluó la producción de materia seca en cada tratamiento durante siete cortes. Los resultados mostraron que al regar con un umbral del 75 o del 50% se obtienen rendimientos promedio similares de materia seca: 25.674,4 kg.ha-1.año-1 y 24.215,4 kg.ha-1.año-1 respectivamente. El trabajo muestra que hay diferencias significativas (p < 0,0001) entre las medias de estos tratamientos con respecto a los otros dos, que tienen rendimientos inferiores: 16.366,7 kg.ha-1.año-1 para el umbral de riego del 25% y de 9.970,9 kg.ha-1.año-1 para un umbral del 0%. Se concluyó que la producción del cultivo disminuye considerablemente con intervalos de riego muy prolongados.

Aislamiento y caracterización de bacterias endosimbióticas procedentes de suelos contaminados por cobre en Chile

El uso intensivo de compuestos de cobre como herbicidas y fungicidas provoca la contaminación de suelos de uso agrícola debido a la acumulación de este metal en las capas más superficiales del suelo. Se sabe que la presencia de cobre y otros metales pesados afecta negativamente a las interacciones simbióticas que se establecen entre bacterias diazotróficas de los géneros Rhizobium, Sinorhizobium y Bradyrhizobium y leguminosas de interés agrícola (Laguerre et al., 2006). El objetivo de este trabajo es estudiar la diversidad de cepas endosimbióticas de leguminosas en suelos agrícolas chilenos que presentan un elevado contenido en cobre como resultado de la contaminación con residuos de extracciones mineras. Además, se pretende caracterizar el nivel de resistencia a cobre en las cepas aisladas con objeto de identificar aquellas altamente eficientes que puedan ser utilizadas como inoculantes microbianos. Para ello, se han prospectado 9 suelos agrícolas de las regiones III, V y VI de Chile con contenidos muy variables de metales. Utilizando estos suelos como inóculos de plantas trampa de leguminosas se ha obtenido una colección de 362 cepas aisladas de nódulos de guisante (Pisum sativum), judía (Phaseolus vulgaris) y alfalfa (Medicago sativa). Los análisis filogenéticos y los ensayos de resistencia a cobre realizados han permitido caracterizar y seleccionar aquellas cepas con mayores niveles de resistencia a este metal. Los resultados demuestran que los suelos altamente contaminados por cobre poseen una menor diversidad de bacterias endosimbióticas; las cepas más resistentes han sido aisladas de los suelos con niveles de contaminación intermedia. Los análisis fenotípicos y moleculares realizados sobre las cepas más resistentes han demostrado la existencia de sistemas de resistencia a cobre inducibles por este metal y potencialmente implicados en su homeostasis.

Identification of lumichrome as a Sinorhizobium enhancer of alfalfa root respiration and shoot growth

Sinorhizobium meliloti bacteria produce a signal molecule that enhances root respiration in alfalfa (Medicago sativa L.) and also triggers a compensatory increase in whole-plant net carbon assimilation. Nuclear magnetic resonance, mass spectrometry, and ultraviolet–visible absorption identify the enhancer as lumichrome, a common breakdown product of riboflavin. Treating alfalfa roots with 3 nM lumichrome increased root respiration 21% (P < 0.05) within 48 h. A closely linked increase in net carbon assimilation by the shoot compensated for the enhanced root respiration. For example, applying 5 nM lumichrome to young alfalfa roots increased plant growth by 8% (P < 0.05) after 12 days. Soaking alfalfa seeds in 5 nM lumichrome before germination increased growth by 18% (P < 0.01) over the same period. In both cases, significant growth enhancement (P < 0.05) was evident only in the shoot. S. meliloti requires exogenous CO2 for growth and may benefit directly from the enhanced root respiration that is triggered by lumichrome. Thus Sinorhizobium–alfalfa associations, which ultimately form symbiotic N2-reducing root nodules, may be favored at an early developmental stage by lumichrome, a previously unrecognized mutualistic signal. The rapid degradation of riboflavin to lumichrome under many physiological conditions and the prevalence of riboflavin release by rhizosphere bacteria suggest that events demonstrated here in the S. meliloti–alfalfa association may be widely important across many plant–microbe interactions.

NADH-Glutamate Synthase in Alfalfa Root Nodules. Genetic Regulation and Cellular Expression1

NADH-dependent glutamate synthase (NADH-GOGAT; EC 1.4.1.14) is a key enzyme in primary nitrogen assimilation in alfalfa (Medicago sativa L.) root nodules. Here we report that in alfalfa, a single gene, probably with multiple alleles, encodes for NADH-GOGAT. In situ hybridizations were performed to assess the location of NADH-GOGAT transcript in alfalfa root nodules. In wild-type cv Saranac nodules the NADH-GOGAT gene is predominantly expressed in infected cells. Nodules devoid of bacteroids (empty) induced by Sinorhizobium meliloti 7154 had no NADH-GOGAT transcript detectable by in situ hybridization, suggesting that the presence of the bacteroid may be important for NADH-GOGAT expression. The pattern of expression of NADH-GOGAT shifted during root nodule development. Until d 9 after planting, all infected cells appeared to express NADH-GOGAT. By d 19, a gradient of expression from high in the early symbiotic zone to low in the late symbiotic zone was observed. In 33-d-old nodules expression was seen in only a few cell layers in the early symbiotic zone. This pattern of expression was also observed for the nifH transcript but not for leghemoglobin. The promoter of NADH-GOGAT was evaluated in transgenic alfalfa plants carrying chimeric β-glucuronidase promoter fusions. The results suggest that there are at least four regulatory elements. The region responsible for expression in the infected cell zone contains an 88-bp direct repeat.

NADH-Glutamate Synthase in Alfalfa Root Nodules. Immunocytochemical Localization1

In root nodules of alfalfa (Medicago sativa L.), N2 is reduced to NH4+ in the bacteroid by the nitrogenase enzyme and then released into the plant cytosol. The NH4+ is then assimilated by the combined action of glutamine synthetase (EC 6.3.1.2) and NADH-dependent Glu synthase (NADH-GOGAT; EC 1.4.1.14) into glutamine and Glu. The alfalfa nodule NADH-GOGAT protein has a 101-amino acid presequence, but the subcellular location of the protein is unknown. Using immunocytochemical localization, we determined first that the NADH-GOGAT protein is found throughout the infected cell region of both 19- and 33-d-old nodules. Second, in alfalfa root nodules NADH-GOGAT is localized predominantly to the amyloplast of infected cells. This finding, together with earlier localization and fractionation studies, indicates that in alfalfa the infected cells are the main location for the initial assimilation of fixed N2.

Winter Survival of Transgenic Alfalfa Overexpressing Superoxide Dismutase1

To test the hypothesis that enhanced tolerance of oxidative stress would improve winter survival, two clones of alfalfa (Medicago sativa) were transformed with a Mn-superoxide dismutase (Mn-SOD) targeted to the mitochondria or to the chloroplast. Although Mn-SOD activity increased in most primary transgenic plants, both cytosolic and chloroplastic forms of Cu/Zn-SOD had lower activity in the chloroplast SOD transgenic plants than in the nontransgenic plants. In a field trial at Elora, Ontario, Canada, the survival and yield of 33 primary transgenic and control plants were compared. After one winter most transgenic plants had higher survival rates than control plants, with some at 100%. Similarly, some independent transgenic plants had twice the herbage yield of the control plants. Prescreening the transgenic plants for SOD activity, vigor, or freezing tolerance in the greenhouse was not effective in identifying individual transgenic plants with improved field performance. Freezing injury to leaf blades and fibrous roots, measured by electrolyte leakage from greenhouse-grown acclimated plants, indicated that the most tolerant were only 1°C more freezing-tolerant than alfalfa clone N4. There were no differences among transgenic and control plants for tetrazolium staining of field-grown plants at any freezing temperature. Therefore, although many of the transgenic plants had higher winter survival rates and herbage yield, there was no apparent difference in primary freezing injury, and therefore, the trait is not associated with a change in the primary site of freezing injury.

Glycolytic Flux Is Adjusted to Nitrogenase Activity in Nodules of Detopped and Argon-Treated Alfalfa Plants1

To investigate the short-term (30–240 min) interactions among nitrogenase activity, NH4+ assimilation, and plant glycolysis, we measured the concentrations of selected C and N metabolites in alfalfa (Medicago sativa L.) root nodules after detopping and during continuous exposure of the nodulated roots to Ar:O2 (80:20, v/v). Both treatments caused an increase in the ratios of glucose-6-phosphate to fructose-1,6-bisphosphate, fructose-6-phosphate to fructose-1,6-bisphosphate, phosphoenolpyruvate (PEP) to pyruvate, and PEP to malate. This suggested that glycolytic flux was inhibited at the steps catalyzed by phosphofructokinase, pyruvate kinase, and PEP carboxylase. In the Ar:O2-treated plants the apparent inhibition of glycolytic flux was reversible, whereas in the detopped plants it was not. In both groups of plants the apparent inhibition of glycolytic flux was delayed relative to the decline in nitrogenase activity. The decline in nitrogenase activity was followed by a dramatic increase in the nodular glutamate to glutamine ratio. In the detopped plants this was coincident with the apparent inhibition of glycolytic flux, whereas in the Ar:O2-treated plants it preceded the apparent inhibition of glycolytic flux. We propose that the increase in the nodular glutamate to glutamine ratio, which occurs as a result of the decline in nitrogenase activity, may act as a signal to decrease plant glycolytic flux in legume root nodules.