Methane efflux in rice paddy field under different irrigation managements

Paddy rice fields may contribute to methane (CH4) emission from soil due to anaerobic conditions after flooding. Alternatives to continuous flooding irrigation in rice have been developed to mitigate CH4 efflux into the atmosphere. This study aims to investigate the effects of irrigation managements in the CH4 efflux during the rice growing season. An experiment was carried out at in Santa Maria, Rio Grande do Sul State, Brazil, during 2007/08 and 2009/10 growing seasons. The treatments were continuous flooding and intermittent irrigation in 2007/08 and continuous flooding, intermittent irrigation and flush irrigation in 2009/10. Intermittent irrigation is effective in mitigating CH4 efflux from rice fields when climatic conditions enable water absence during cultivation, but its efficiency depends on the electrochemical soil conditions during the flooding cycles.

Molecular players shaping the arbuscular mycorrhizal symbiosis of rice

SUMMARY : The arbuscular mycorrhizal (AM) symbiosis is an evolutionarily ancient association between most land plants and Glomeromycotan fungi that is based on the mutual exchange of nutrients between the two partners. Its structural and physiological establishment is a multi-step process involving a tightly regulated signal exchange leading to intracellular colonization of roots by the fungi. Most research on the molecular biology and genetics of symbiosis development has been performed in dicotyledonous model legumes. In these, a plant signaling pathway, the common SYM pathway, has been found to be required for accommodation of both root symbionts rhizobia and AM fungi. Rice, a monocotyledon model and the world's most important staple crop also forms AM symbioses, has been largely ignored for studies of the AM symbiosis. Therefore in this PhD work functional conservation of the common SYM pathway in rice was addressed and demonstrated. Mycorrhiza-specific marker genes were established that are expressed at different stages of AM development and therefore represent readouts for various AM-specific signaling events. These tools were successfully used to obtain evidence for a yet unknown signaling network comprising common SYM-dependent and -independent events. In legumes AM colonization induces common SYM signaling dependent changes in root system architecture. It was demonstrated that also in rice, root system architecture changes in response to AM colonization but these alterations occur independently of common SYM signaling. The rice root system is complex and contains three different root types. It was shown that root type identity influences the quantity of AM colonization, indicating root type specific symbiotic properties. Interestingly, the root types differed in their transcriptional responses to AM colonization and the less colonized root type responded more dramatically than the more strongly colonized root type. Finally, in an independent project a novel mutant, inhospitable (iho), was discovered. It is perturbed at the most early step of AM colonization, namely differentiation of the AM fungal hyphae into a hyphopodium at the root surface. As plant factors required for this early step are not known, identification of the IHO gene will greatly contribute to the advance of mycorrhiza RÉSUMÉ : La symbiose mycorhizienne arbusculaire (AM) est une association évolutionnairement ancienne entre la majorité des plantes terrestres et les champignons du type Glomeromycota, basée sur l'échange mutuel d'éléments nutritifs entre les deux partenaires. Son établissement structural et physiologique est un processus en plusieurs étapes, impliquant des échanges de signaux étroitement contrôlés, aboutissant à la colonisation intracellulaire des racines par le champignon. La plupart des recherches sur la biologie moléculaire et la génétique du développement de la symbiose ont été effectuées sur des légumineuses dicotylédones modèles. Dans ces dernières, une voie de signalisation, la voie SYM, s'est avérée nécessaire pour permettre la mise en place de la symbiose mycorhizienne. Chez les plantes monocotylédones, comme le riz, une des céréales les plus importantes, nourrissant la moitié de la population mondiale, peu de recherches ont été effectuées sur les bases de la cette symbiose. Dans ce travail de thèse, la conservation fonctionnelle de la voie commune SYM chez le riz a été étudiée et démontrée. De plus, des gènes marqueurs spécifiques des différentes étapes du développement de l'AM ont été identifiés, permettant ainsi d'avoir des traceurs de la colonisation. Ces outils ont été utilisés avec succès pour démontrer l'existence d'un nouveau réseau de signalisation, comprenant des éléments SYM dépendant et indépendant. Chez les légumineuses, la colonisation par les AM induit des changements dans l'architecture du système racinaire, via la signalisation SYM dépendantes. Cependant chez le riz, il a été démontré que l'architecture de système racinaire changeait suite à la colonisation de l'AM, mais ceux, de façon SYM indépendante. Le système racinaire du riz est complexe et contient trois types différents de racines. Il a été démontré que le type de racine pouvait influencer l'efficacité de la colonisation par l'AM, indiquant que les racines ont des propriétés symbiotiques spécifiques différentes. De façon surprenante, les divers types de racines répondent de différemment suite à colonisation par l'AM avec des changements de la expression des gènes. Le type de racine le moins colonisé, répondant le plus fortement a la colonisation, et inversement. En parallèle, dans un projet indépendant, un nouveau mutant, inhospitable (iho), a été identifié. Ce mutant est perturbé lors de l'étape la plus précoce de la colonisation par l'AM, à savoir la différentiation des hyphes fongiques de l'AM en hyphopodium, à la surface des racines. Les facteurs d'origine végétale requis pour cette étape étant encore inconnus, l'identification du gène IHO contribuera considérablement a accroître nos connaissance sur les bases de la mise en place de cette symbiose.

Loss of ammonia from nitrogen fertilizers applied to maize and soybean straw

In Brazilian agriculture, urea is the most commonly used nitrogen (N) source, in spite of having the disadvantage of losing considerable amounts of N by ammonia-N volatilization. The objectives of this study were to evaluate: N lossby ammonia volatilization from: [urea coated with copper sulfate and boric acid], [urea coated with zeolite], [urea+ammonium sulfate], [urea coated with copper sulfate and boric acid+ammonium sulfate], [common urea] and [ammonium nitrate]; and the effect of these N source son the maize yield in terms of amount and quality. The treatments were applied to the surface of a soil under no-tillage maize, in two growing seasons. The first season (2009/2010) was after a maize crop (maize straw left on the soil surface) and the second cycle (2012/2011) after a soybean crop. Due to the weather conditions during the experiments, the volatilization of ammonia-N was highest in the first four days after application of the N sources. Of all urea sources, under volatilization-favorable conditions, the loss of ammonia from urea coated with copper sulfate and boric acid was lowest, while under high rainfall, the losses from the different urea sources was similar, i.e., an adequate rainfall was favorablet o reduce volatilization. The ammonia volatilization losses were greatest in the first four days after application. Maize grain yield differed due to N application and in the treatments, but this was only observed with cultivation of maize crop residues in 2009/2010. The combination of ammonium+urea coated with copper sulfate and boric acid optimized grain yield compared to the other urea treatments. The crude protein concentration in maize was not influenced by the technologies of urea coating.

Upland rice under no-tillage preceded by crops for soil cover and nitrogen fertilization

The grain yield of upland rice under no-tillage has been unsatisfactory and one reason could be the nitrate/ammonium balance in the soil. Cover crops and nitrogen fertilization can be used to change the nitrate/ammonium relation in the soil and improve conditions for the development of upland rice in the no-tillage (NT) system. The aim was to study the effect of cover crops and nitrogen sources on grain yield of upland rice under no tillage. The study was carried out on the Fazenda Experimental Lageado, in Botucatu, State of São Paulo, Brazil, in an Oxisol area under no-tillage for six years. The experiment was arranged in a randomized block split-plot design with four replications. The plots consisted of six cover crop species (Brachiaria brizantha, B. decumbens, B. humidicola, B. ruziziensis, Pennisetum americanum, and Crotalaria spectabilis) and the split-plots of seven forms of N fertilizer management. Millet is the best cover crop to precede upland rice under NT. The best form of N application, as nitrate, is in split rates or total rate at topdressing or an ammonium source with or without a nitrification inhibitor, in split doses. When the cover crops C. spectabilis, B. brizantha, B. decumbens, B. humidicola, and B. ruziziensis preceded rice, they induced the highest grain yield when rice was fertilized with N as ammonium sulfate source + nitrification inhibitor in split rates or total dose at topdressing.

Iron oxidation on the surface of adventitious roots and its relation to aerenchyma formation in rice genotypes

Establishment of the water layer in an irrigated rice crop leads to consumption of free oxygen in the soil which enters in a chemical reduction process mediated by anaerobic microorganisms, changing the crop environment. To maintain optimal growth in an environment without O2, rice plants develop pore spaces (aerenchyma) that allow O2 transport from air to the roots. Carrying capacity is determined by the rice genome and it may vary among cultivars. Plants that have higher capacity for formation of aerenchyma should theoretically carry more O2 to the roots. However, part of the O2 that reaches the roots is lost due to permeability of the roots and the O2 gradient created between the soil and roots. The O2 that is lost to the outside medium can react with chemically reduced elements present in the soil; one of them is iron, which reacts with oxygen and forms an iron plaque on the outer root surface. Therefore, evaluation of the iron plaque and of the formation of pore spaces on the root can serve as a parameter to differentiate rice cultivars in regard to the volume of O2 transported via aerenchyma. An experiment was thus carried out in a greenhouse with the aim of comparing aerenchyma and iron plaque formation in 13 rice cultivars grown in flooded soils to their formation under growing conditions similar to a normal field, without free oxygen. The results indicated significant differences in the volume of pore spaces in the roots among cultivars and along the root segment in each cultivar, indicating that under flooded conditions the genetic potential of the plant is crucial in induction of cell death and formation of aerenchyma in response to lack of O2. In addition, the amount of Fe accumulated on the root surface was different among genotypes and along the roots. Thus, we concluded that the rice genotypes exhibit different responses for aerenchyma formation, oxygen release by the roots and iron plaque formation, and that there is a direct relationship between porosity and the amount of iron oxidized on the root surface.

Rice husk ash as corrective of soil acidity

Rice husk ash (RHA) is a by-product from the burning of rice husk that can have favorable effects on the soil in terms of acidity correction. The objectives of this study were to determine the effective calcium carbonate equivalent (ECC) of RHA under field conditions, and establish technical criteria as a basis for estimating the overall ECC of RHA. The 12 treatments of the experiment consisted of 10 RHA dosages (0, 10, 20, 30, 40, 60, 80, 100, 120, and 140 Mg ha-1) and two references, one of which was an absolute control (AC) and the other a plot limed and fertilized according to official recommendations (recommended fertilization - RF). The soil was sampled twice (15 and 210 days after incorporating RHA), in the layers 0.00-0.10 and 0.10-0.20 m, to determine the pH(H2O) and base saturation (V%). The ECC and neutralizing value (NV) of RHA were also determined. The results showed that RHA neutralizes soil acidity, in a faster reaction than conventional limestone, despite a low ECC (around 3 %).

Tissue-adapted invasion strategies of the rice blast fungus Magnaporthe oryzae.

Magnaporthe oryzae causes rice blast, the most serious foliar fungal disease of cultivated rice (Oryza sativa). During hemibiotrophic leaf infection, the pathogen simultaneously combines biotrophic and necrotrophic growth. Here, we provide cytological and molecular evidence that, in contrast to leaf tissue infection, the fungus adopts a uniquely biotrophic infection strategy in roots for a prolonged period and spreads without causing a loss of host cell viability. Consistent with a biotrophic lifestyle, intracellularly growing hyphae of M. oryzae are surrounded by a plant-derived membrane. Global, temporal gene expression analysis used to monitor rice responses to progressive root infection revealed a rapid but transient induction of basal defense-related gene transcripts, indicating perception of the pathogen by the rice root. Early defense gene induction was followed by suppression at the onset of intracellular fungal growth, consistent with the biotrophic nature of root invasion. By contrast, during foliar infection, the vast majority of these transcripts continued to accumulate or increased in abundance. Furthermore, induction of necrotrophy-associated genes during early tissue penetration, previously observed in infected leaves, was not seen in roots. Collectively, our results not only report a global characterization of transcriptional root responses to a biotrophic fungal pathogen but also provide initial evidence for tissue-adapted fungal infection strategies.

Carbon and Nitrogen Mineralization in Soil Combining Sewage Sludge and Straw

ABSTRACT The combined incorporation of sewage sludge (SS) and oat straw (OS) to the soil can increase straw carbon mineralization and microbial nitrogen immobilization. This hypothesis was tested in two laboratory experiments, in which SS was incorporated in the soil with and without OS. One treatment in which only straw was incorporated and a control with only soil were also evaluated. The release of CO2 and mineral N in the soil after organic material incorporation was evaluated for 110 days. The cumulative C mineralization reached 30.1 % for SS and 54.7 % for OS. When these organic materials were incorporated together in the soil, straw C mineralization was not altered. About 60 % of organic N in the SS was mineralized after 110 days. This N mineralization index was twice as high as that defined by Resolution 375/2006 of the National Environmental Council. The combined incorporation of SS and OS in the soil caused an immobilization of microbial N of 5.9 kg Mg-1 of OS (mean 3.5 kg Mg-1). The results of this study indicated that SS did not increase straw C mineralization, but the SS rate should be adjusted to compensate for the microbial N immobilization caused by straw.

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Hydro-Physical Properties of a Typic Hapludult under the Effect of Rice Husk Ash

ABSTRACT The combustion of rice husk generates a partially burnt mixture called rice husk ash (RHA) that can be used as a source of nutrients to crops and as a conditioner of soil physical properties. The objective of this study was to evaluate the effect of RHA levels on the hydro-physical properties of a Typic Hapludult. The experimental design was composed of random blocks with four replications, which comprised plots of 24 m2 and treatments with increasing RHA rates: 0, 40, 80 and 120 Mg ha-1. Undisturbed soil samples were collected in the soil layers of 0.00-0.10 and 0.10-0.20 m after nine months of RHA application, using steel cylinders (0.03 m of height and 0.047 m of diameter). These samples were used to determine soil bulk density (Bd), total soil porosity (TP), soil macroporosity (Ma), soil microporosity (Mi) and the available water capacity (AWC). Disturbed soil samples were collected to determine the stability of soil aggregates in water, mean weight diameter of water stable aggregates (MWD), and soil particle size distribution. The results show that, as the RHA rate increased in the soil, Bd values decreased and TP, Ma and MWD values increased. No effect of RHA was found on Mi and AWC values. The effects of RHA on the S parameter (Dexter, 2004), precompression stress and compression index (Dias Junior and Pierce, 1995) values are consistent those shown for density and total porosity. Rice husk ash was shown to be an efficient residue to improve soil physical properties, mainly at rates between 40 and 80 Mg ha-1. Rice husk ash reduces bulk density and increases total porosity, macroporosity and soil aggregation, but does not affect microporosity, field capacity, permanent wilting point, and available water capacity of the soil. The effect of rice husk ash on the S parameter, precompression stress and index compressibility coefficient values are consistent with those observed for the bulk density and total porosity.

Analysis of bioavailable arsenic in rice with whole cell living bioreporter bacteria.

A multiwell plate bioassay was developed using genetically modified bacteria (bioreporter cells) to detect inorganic arsenic extracted from rice. The bacterial cells expressed luciferase upon exposure to arsenite, the activity of which was detected by measurement of cellular bioluminescence. The bioreporter cells detected arsenic in all rice varieties tested, with averages of 0.02-0.15 microg of arsenite equivalent per gram of dry weight and a method detection limit of 6 ng of arsenite per gram of dry rice. This amounted to between approximately 20 and 90% of the total As content reported by chemical methods for the same sample and suggested that a major proportion of arsenic in rice is in the inorganic form. Calibrations of the bioassay with pure inorganic and organic arsenic forms showed that the bacterial cells react to arsenite with highest affinity, followed by arsenate (with 25% response relative to an equivalent arsenite concentration) and trimethylarsine oxide (at 10% relative response). A method for biocompatible arsenic extraction was elaborated, which most optimally consisted of (i) grinding rice to powder, (ii) mixing with an aqueous solution containing pancreatic enzymes, (iii) mechanical shearing, (iv) extraction in mild acid conditions and moderate heat, and (v) centrifugation and pH neutralization. Detection of mainly inorganic arsenic by the bacterial cells may have important advantages for toxicity assessment of rice consumption and would form a good complement to total chemical arsenic determination.

The half-size ABC transporters STR1 and STR2 are indispensable for mycorrhizal arbuscule formation in rice.

The central structure of the symbiotic association between plants and arbuscular mycorrhizal (AM) fungi is the fungal arbuscule that delivers minerals to the plant. Our earlier transcriptome analyses identified two half-size ABCG transporters that displayed enhanced mRNA levels in mycorrhizal roots. We now show specific transcript accumulation in arbusculated cells of both genes during symbiosis. Presently, arbuscule-relevant factors from monocotyledons have not been reported. Mutation of either of the Oryza sativa (rice) ABCG transporters blocked arbuscule growth of different AM fungi at a small and stunted stage, recapitulating the phenotype of Medicago truncatula stunted arbuscule 1 and 2 (str1 and str2) mutants that are deficient in homologous ABCG genes. This phenotypic resemblance and phylogenetic analysis suggest functional conservation of STR1 and STR2 across the angiosperms. Malnutrition of the fungus underlying limited arbuscular growth was excluded by the absence of complementation of the str1 phenotype by wild-type nurse plants. Furthermore, plant AM signaling was found to be intact, as arbuscule-induced marker transcript accumulation was not affected in str1 mutants. Strigolactones have previously been hypothesized to operate as intracellular hyphal branching signals and possible substrates of STR1 and STR2. However, full arbuscule development in the strigolactone biosynthesis mutants d10 and d17 suggested strigolactones to be unlikely substrates of STR1/STR2. Interestingly, rice STR1 is associated with a cis-natural antisense transcript (antiSTR1). Analogous to STR1 and STR2, at the root cortex level, the antiSTR1 transcript is specifically detected in arbusculated cells, suggesting unexpected modes of STR1 regulation in rice.