Rice straw incorporated just before soil flooding increases acetic acid formation and decreases available nitrogen

Incorporation of rice straw into the soil just before flooding for water-seeded rice can immobilize mineral nitrogen (N) and lead to the production of acetic acid harmful to the rice seedlings, which negatively affects grain yield. This study aimed to evaluate the formation of organic acids and variation in pH and to quantify the mineral N concentration in the soil as a function of different times of incorporation of rice straw or of ashes from burning the straw before flooding. The experiment was carried out in a greenhouse using an Inceptisol (Typic Haplaquept) soil. The treatments were as follows: control (no straw or ash); incorporation of ashes from previous straw burning; rice straw incorporated to drained soil 60 days before flooding; straw incorporated 30 days before flooding; straw incorporated 15 days before flooding and straw incorporated on the day of flooding. Experimental units were plastic buckets with 6.0 kg of soil. The buckets remained flooded throughout the trial period without rice plants. Soil samples were collected every seven days, beginning one day before flooding until the 13th week of flooding for determination of mineral N- ammonium (NH4+) and nitrate (NO3-). Soil solution pH and concentration of organic acids (acetic, propionic and butyric) were determined. All NO3- there was before flooding was lost in approximately two weeks of flooding, in all treatments. There was sigmoidal behavior for NH4+ formation in all treatments, i.e., ammonium ion concentration began to rise shortly after soil flooding, slightly decreased and then went up again. On the 91st day of flooding, the NH4+ concentrations in soil was 56 mg kg-1 in the control treatment, 72 mg kg-1 for the 60-day treatment, 73 mg kg-1 for the 30-day treatment and 53 mg kg-1 for the ash incorporation treatment. These ammonium concentrations correspond to 84, 108, 110 and 80 kg ha-1 of N-NH4+, respectively. When the straw was incorporated on the day of flooding or 15 days before, the concentration of N-NH4+ in the soil was 28 and 54 mg kg-1, equivalent to an accumulation of 42 and 81 kg ha-1 of N-NH4+, respectively. There was formation of acetic acid in which toxic concentrations were reached (7.2 mmol L-1) on the 15th day of flooding only for the treatment with straw incorporated on the day of flooding. The pH of the soil solution of all the treatments increased after flooding and this increase was faster in the treatments with incorporation of straw, followed by the ash treatment and then the control. After 60 days of flooding, however, the pH values were around 6.5 for all treatments, except for the control, which reached a pH of 6.3. Rice straw should be incorporated into the soil at least 30 days before flooding; otherwise, it may immobilize part of the mineral N and produce acetic acid in concentrations toxic to rice seedlings.

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.

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.

THE RESURRECTION PLANT TRIPOGON SPICATUS (POACEAE) HARBORS A DIVERSITY OF PLANT GROWTH PROMOTING BACTERIA IN NORTHEASTERN BRAZILIAN CAATINGA

Plant species that naturally occur in the Brazilian Caatinga(xeric shrubland) adapt in several ways to these harsh conditions, and that can be exploited to increase crop production. Among the strategic adaptations to confront low water availability, desiccation tolerance stands out. Up to now, the association of those species with beneficial soil microorganisms is not well understood. The aim of this study was to characterize Tripogon spicatusdiazotrophic bacterial isolates from the Caatingabiome and evaluate their ability to promote plant growth in rice. Sixteen bacterial isolates were studied in regard to their taxonomic position by partial sequencing of the 16S rRNA gene, putative diazotrophic capacity, in vitro indole-acetic acid (IAA) production and calcium phosphate solubilization, metabolism of nine different C sources in semi-solid media, tolerance to different concentrations of NaCl to pHs and intrinsic resistance to nine antibiotics. Finally, the ability of the bacterial isolates to promote plant growth was evaluated using rice (Oryza sativa) as a model plant. Among the 16 isolates evaluated, eight of them were classified as Enterobacteriaceae members, related to Enterobacter andPantoeagenera. Six other bacteria were related toBacillus, and the remaining two were related toRhizobiumand Stenotrophomonas.The evaluation of total N incorporation into the semi-solid medium indicated that all the bacteria studied have putative diazotrophic capacity. Two bacteria were able to produce more IAA than that observed for the strain BR 11175Tof Herbaspirillum seropedicae.Bacterial isolates were also able to form a microaerophilic pellicle in a semi-solid medium supplemented with different NaCl concentrations up to 1.27 mol L-1. Intrinsic resistance to antibiotics and the metabolism of different C sources indicated a great variation in physiological profile. Seven isolates were able to promote rice growth, and two bacteria were more efficient than the reference strainAzospirillum brasilense, Ab-V5. The results indicate the potential of T. spicatus as native plant source of plant growth promoting bacteria.

EFICIÊNCIA DE MÉTODOS ANALÍTICOS NA PREDIÇÃO DA DISPONIBILIDADE DE FÓSFORO PARA ARROZ IRRIGADO EM SOLOS CATARINENSES

Os solos cultivados com arroz irrigado no Estado de Santa Catarina apresentam características mineralógicas e químicas distintas dos solos utilizados nos estudos de calibração do extrator Mehlich-1, para estimar a disponibilidade de fósforo do solo para as plantas. Desse modo, o objetivo deste trabalho foi avaliar a eficiência dos extratores Mehlich-1, Mehlich-3, Resina em lâminas, Olsen e EDTA na predição da disponibilidade P para plantas de arroz cultivadas em sistema de alagamento em solos representativos de lavouras orizícolas do Estado de Santa Catarina. O experimento foi conduzido em casa de vegetação, com cinco solos oriundos de várias lavouras orizícolas, em 2014. Foram adicionados aos solos quatro concentrações de P, correspondendo a 0, ¼, ½ e 1,0 vez a quantidade de P para atingir 0,2 mg L-1 de P na solução do solo (com base na capacidade de adsorção de cada solo). Foram usadas três repetições por tratamento, dispostas no delineamento completamente casualizado. Após cinco dias da adição dos níveis de P, as unidades experimentais constituídas por vasos de 8 L contendo 3,5 kg de solo seco foram amostradas para determinar o P disponível do solo por cinco métodos químicos. Na sequência, essas foram alagadas e cultivadas com seis plantas de arroz por 46 dias, quando se quantificaram a massa de matéria seca e o P absorvido pela parte aérea das plantas. A eficácia dos métodos Mehlich-1, Mehlich-3, Resina em lâminas, Olsen e EDTA foi avaliada por correlação linear entre a quantidade de P extraída dos solos e a quantidade absorvida pelas plantas. A quantidade de P absorvida pelas plantas de arroz aumentou com o acréscimo das doses de P aplicadas, variando com o tipo de solo. A maior recuperação do P adicionado aos solos foi obtida pelo extrator EDTA, seguido pelo Olsen. O P extraído pelo EDTA e Mehlich-3 evidenciou melhor correlação com quantidade de P absorvida pelo arroz. O agrupamento dos solos segundo características relacionadas à adsorção de P no solo melhora a eficiência de todos os métodos em predizer a disponibilidade de P em solos cultivados com arroz no sistema de inundação.

Resíduo de Curtumes como Fonte de Nitrogênio para Trigo e Arroz em Sucessão

RESUMO O processo de estabilização necessário para o curtimento das peles, na indústria coureira, gera grande quantidade de resíduo sólido colagênico com elevado potencial poluidor. Esse resíduo contém elevado teor de nitrogênio, o que confere ao resíduo potencial para uso agrícola. Para avaliar a viabilidade da aplicação de rejeito da indústria do couro, como fonte nitrogenada para as culturas do trigo e arroz de sequeiro, foi implantado um experimento em casa de vegetação com cinco doses de N provenientes do colágeno (0, 225, 450, 675 e 900 mg kg-1) e três tratamentos adicionais (controle, ureia e ureia reposta no segundo cultivo). A aplicação de até 675 mg kg-1 de N via colágeno propiciou produção de matéria seca de plantas de trigo similar à aplicação de N mineral e maior massa de grãos de trigo em todas as doses estudadas (14 a 41 % superiores à aplicação de N mineral). Como fonte residual, a aplicação do colágeno proporcionou níveis de N no solo suficientes para obter massa de grãos de arroz nas doses mais elevadas (acima de 675 mg kg-1de N via colágeno), sem que os teores de Cr foliares sejam considerados tóxicos. O colágeno pode ser boa alternativa para a implantação da cultura do arroz após a colheita de trigo, sem prejuízos de produtividade e necessidade de reposição de fertilizantes nitrogenados, se aplicado na dose 675 mg kg-1 de N fornecido via colágeno.

Nonspecific PCR amplification by high-fidelity polymerases: implications for next-generation sequencing of AFLP markers.

High-fidelity 'proofreading' polymerases are often used in library construction for next-generation sequencing projects, in an effort to minimize errors in the resulting sequence data. The increased template fidelity of these polymerases can come at the cost of reduced template specificity, and library preparation methods based on the AFLP technique may be particularly susceptible. Here, we compare AFLP profiles generated with standard Taq and two versions of a high-fidelity polymerase. We find that Taq produces fewer and brighter peaks than high-fidelity polymerase, suggesting that Taq performs better at selectively amplifying templates that exactly match the primer sequences. Because the higher accuracy of proofreading polymerases remains important for sequencing applications, we suggest that it may be more effective to use alternative library preparation methods.

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.

Seed-specific silencing of OsMRP5 reduces seed phytic acid and weight in rice.

Phytic acid (PA) is poorly digested by humans and monogastric animals and negatively affects human/animal nutrition and the environment. Rice mutants with reduced PA content have been developed but are often associated with reduced seed weight and viability, lacking breeding value. In the present study, a new approach was explored to reduce seed PA while attaining competitive yield. The OsMRP5 gene, of which mutations are known to reduce seed PA as well as seed yield and viability, was down-regulated specifically in rice seeds by using an artificial microRNA driven by the rice seed specific promoter Ole18. Seed PA contents were reduced by 35.8-71.9% in brown rice grains of transgenic plants compared to their respective null plants (non-transgenic plants derived from the same event). No consistent significant differences of plant height or number of tillers per plant were observed, but significantly lower seed weights (up to 17.8% reduction) were detected in all transgenic lines compared to null plants, accompanied by reductions of seed germination and seedling emergence. It was observed that the silencing of the OsMRP5 gene increased the inorganic P (Pi) levels (up to 7.5 times) in amounts more than the reduction of PA-P in brown rice. This indicates a reduction in P content in other cellular compounds, such as lipids and nucleic acids, which may affect overall seed development. Put together, the present study demonstrated that seed specific silencing of OsMRP5 could significantly reduce the PA content and increase Pi levels in seeds; however, it also significantly lowers seed weight in rice. Discussions were made regarding future directions towards producing agronomically competitive and nutritionally valuable low PA rice.

Rapid and selective surveillance of Arabidopsis thaliana genome annotations with Centrifuge.

Centrifuge is a user-friendly system to simultaneously access Arabidopsis gene annotations and intra- and inter-organism sequence comparison data. The tool allows rapid retrieval of user-selected data for each annotated Arabidopsis gene providing, in any combination, data on the following features: predicted protein properties such as mass, pI, cellular location and transmembrane domains; SWISS-PROT annotations; Interpro domains; Gene Ontology records; verified transcription; BLAST matches to the proteomes of A.thaliana, Oryza sativa (rice), Caenorhabditis elegans, Drosophila melanogaster and Homo sapiens. The tool lends itself particularly well to the rapid analysis of contigs or of tens or hundreds of genes identified by high-throughput gene expression experiments. In these cases, a summary table of principal predicted protein features for all genes is given followed by more detailed reports for each individual gene. Centrifuge can also be used for single gene analysis or in a word search mode. AVAILABILITY: http://centrifuge.unil.ch/ CONTACT: edward.farmer@unil.ch.

Spatio-temporal transcript profiling of rice roots and shoots in response to phosphate starvation and recovery.

Using rice (Oryza sativa) as a model crop species, we performed an in-depth temporal transcriptome analysis, covering the early and late stages of Pi deprivation as well as Pi recovery in roots and shoots, using next-generation sequencing. Analyses of 126 paired-end RNA sequencing libraries, spanning nine time points, provided a comprehensive overview of the dynamic responses of rice to Pi stress. Differentially expressed genes were grouped into eight sets based on their responses to Pi starvation and recovery, enabling the complex signaling pathways involved in Pi homeostasis to be untangled. A reference annotation-based transcript assembly was also generated, identifying 438 unannotated loci that were differentially expressed under Pi starvation. Several genes also showed induction of unannotated splice isoforms under Pi starvation. Among these, PHOSPHATE2 (PHO2), a key regulator of Pi homeostasis, displayed a Pi starvation-induced isoform, which was associated with increased translation activity. In addition, microRNA (miRNA) expression profiles after long-term Pi starvation in roots and shoots were assessed, identifying 20 miRNA families that were not previously associated with Pi starvation, such as miR6250. In this article, we present a comprehensive spatio-temporal transcriptome analysis of plant responses to Pi stress, revealing a large number of potential key regulators of Pi homeostasis in plants.

Progresso genético obtido pelo melhoramento do arroz de sequeiro em 21 anos de pesquisa em Minas Gerais

Este trabalho objetivou estimar o ganho genético obtido pelo programa de melhoramento de arroz de sequeiro (Oryza sativa L.) desenvolvido em Minas Gerais cooperativamente pela Epamig/Embrapa-Centro Nacional de Pesquisa de Arroz e Feijão (CNPAF)/UFLA/UFV, no período de 1974/75 a 1994/95. Para tanto, utilizaram-se os dados dos ensaios comparativos avançados de cultivares e linhagens de arroz de sequeiro conduzidos no referido período. Em virtude da distribuição irregular de chuvas e da resposta diferenciada dos materiais de ciclos diferentes às condições climáticas, optou-se por dividi-los em dois grupos; um contendo os genótipos precoces e o outro os de ciclo médio. Os resultados alcançados mostraram que ocorreu um ganho genético médio anual de 1,26% e de 3,37% em relação ao grupo precoce e ao de ciclo médio, respectivamente. O grupo precoce superou estatisticamente (P<=0,01) em produtividade de grãos o grupo de ciclo médio, indicando que em Minas Gerais deve-se dar preferência ao plantio de cultivares de ciclo curto.

Manejo de água e de fertilizante potássico na cultura de arroz irrigado

O manejo da água de irrigação e as doses e épocas de aplicação de fertilizantes tornam-se aspectos de extrema importância no êxito do aproveitamento das várzeas para o cultivo de arroz (Oryza sativa L.) irrigado ou este seguido de outras espécies. Com o objetivo de comparar distintas formas de manejo de água e de fertilizante potássico no comportamento do arroz irrigado, foram conduzidos experimentos por três anos consecutivos, em um Inceptissolo. Foram estudados os efeitos de manejo de água (MA1 - inundação contínua e MA2 - inundação intermitente seguida de contínua) e o modo de aplicação de fertilizante potássico (K1 - na semeadura; K2 - parcelada e K3 - meia dose parcelada). O manejo de água apresentou efeito mais expressivo sobre o comportamento do arroz que o do fertilizante potássico. A inundação contínua durante todo o ciclo da cultura proporcionou maiores rendimentos de grãos, expressando maiores valores dos parâmetros produtivos, e melhorou a qualidade industrial dos grãos. Com esta irrigação, o parcelamento da adubação potássica aumentou o aproveitamento do fertilizante. Os manejos do fertilizante potássico afetaram diferentemente o comportamento da cultura do arroz nas distintas formas de manejo de água.

Herança da tolerância à toxidez de alumínio em arroz com base em análise dialélica

Objetivando estudar a herança da tolerância à toxidez de alumínio (Al) em arroz (Oryza sativa L.) avaliaram-se três genótipos tolerantes (Guarani, Guaporé e IAC 25) e três sensíveis (CNA 5600, CNA 5615 e IAC 899) à toxidez de Al e seus respectivos cruzamentos F1 em um sistema de cruzamento dialélico, desconsiderando os recíprocos, na concentração de 20 mg/L de Al, em solução nutritiva. Os resultados encontrados evidenciaram que a herança da tolerância à toxidez de Al em arroz é oligogênica, com os alelos dominantes determinando essa tolerância. Os progenitores Guarani, Guaporé e IAC 25 apresentaram maior concentração de alelos dominantes, e CNA 5600, CNA 5615 e IAC 899 maior número de alelos recessivos.