Importance of root respiration for water deficit tolerance in soybean

O objetivo deste trabalho foi melhor compreender os mecanismos radiculares que tornam a cultivar de soja Embrapa 48 mais tolerante à seca que a cultivar BR16, e testar a hipótese de que diferenças na respiração radicular entre estes genótipos contribuem para esta tolerância diferencial. Foram conduzidos dois experimentos, no primeiro as plantas foram avaliadas em condições de plena irrigação (controle) ou submetidas a déficit hídrico, imposto pela suspensão da irrigação, até alcançarem um potencial hídrico na antemanhã (Ψam) de –1,0 ± 0,1 MPa e -1,5 ± 0,1 MPa, valores que caracterizam uma condição de estresse moderado e severo em soja, respectivamente. Além desses tratamentos, um quarto grupo foi mantido para serem reirrigadas após alcançarem o déficit severo. O segundo experimento foi realizado para avaliar o efeito do ácido abscísico (ABA) e de seu inibidor (fluridona) na tolerância à seca nas cultivares de soja Embrapa 48 e BR 16. O ensaio foi conduzido como descrito acima, com a diferença que o ABA (50μM) ou a fluridona (50 μM) foram aspergidas nas folhas e na região do coleto a partir da imposição do estresse. A cultivar tolerante retardou a desidratação foliar apresentando maior fotossíntese líquida no estresse severo e após a reirrigação, além de maior eficiência do uso da água. A cultivar tolerante também possuiu maior respiração radicular mas não alterou sua arquitetura radicular sob estresse, excluindo um papel do sistema radicular como um mecanismo de escape ao estresse. Maior nível de mecanismos antioxidativos radiculares contribuem para a tolerância à seca, como aumento do teor de ascorbato e de antioxidantes enzimáticos como catalase, APX, SOD e GR. A maior respiração durante o estresse hídrico na cultivar tolerante foi associado com alterações na expressão gênica. Além disso, a cultivar Embrapa 48 obteve maior atividade de algumas enzimas respiratórias analisadas. O estresse hídrico afetou diferentemente vários hormônios radiculares, aumentando ABA e reduzindo AIA e MeJA. O estresse hídrico promoveu aumentos nos teores de ácidos orgânicos e aminoácidos nas raízes da cultivar tolerante, e menores níveis de NAD+ sob estresse severo e reirrigação. A aplicação exógena de ABA reduziu a fotossíntese e aumentou a respiração radicular sob estresse e afetou diferentemente o metabolismo de aminoácidos e açúcares entre vias cultivares com diferenças na resposta de tolerância estudados aqui. A fluridona reduziu a fotossíntese e respiração nos dois genótipos e afetou pouco os níveis de ABA nas raízes. Além disso, de forma geral, a aplicação de fluridona proporcionou aumentos nos teores de aminoácidos e ácidos orgânicos em ambas cultivares. A partir dos diferentes testes realizados para estudar o mecanismo diferencial de tolerância à deficiência hídrica entre as duas cultivares, verificou-se que para as raízes de soja as maiores taxas respiratórias e o eficiente metabolismo antioxidativo foram essenciais para maior tolerância da cultivar Embrapa 48.This study aimed to better understand the root mechanisms in soybean that render cultivar Embrapa 48 drought tolerance, in comparison with drought-sensitive BR16, and to test if root respiration is related to differential drought responses observed in both cultivars. Two experiments were conducted simultaneously. First, plants were evaluated under regular irrigation conditions (control), and under water deficit imposed by withholding irrigation until the pre-dawn leaf water potential reached (Ψam) –1.0 ± 0.1 MPa (moderate stress) or –1.5 ± 0.1 MPa (severe stress). Severe stressed plants from both cultivars were also evaluated after their complete rehydration. Second, effects of abscisic acid (ABA) and fluridone (ABA biosynthesis inhibitor) on drought tolerance were tested. ABA (50μM) or fluridone (50μM) were sprayed daily onto leaves and stems of both cultivars when moderate and severe drought stress imposition started. The drought tolerant cultivar presented a delay in leaf water dehydration, and higher values of net photosynthesis rate and water use efficiency under severe stress, and after rehydration. Drought tolerant cultivar also presented greater root respiration, but no changes in root architecture under stress were observed, excluding the importance of root system in drought avoidance. Higher levels of root antioxidant mechanism improve drought tolerance, as seen by the increase in ascorbate content and activities of antioxidative enzymes catalase, APX, SOD and GR. Under stress, the higher level of respiration observed in the drought tolerant cultivar was associated with changes in gene expression. Moreover, Embrapa 48 had greater activities of some respiratory enzymes evaluated. Water stress influenced differently plant hormone levels in roots, such as increase in ABA and decrease in IAA and MeJA. There was an increase of levels of organic acids and amino acids, and a decrease of NAD+ in roots of the tolerant cultivar, under severe stress and after rehydration. Exogenous application of ABA decreased photosynthesis and increased root respiration under stress, and differently affected amino acid and sugar metabolism in both cultivars. Fluridone decreased photosynthesis and root respiration of both cultivars, and little affected ABA levels in roots. In addition, fluridone led to an increase of amino acids and organic acids in roots of both cultivars. Altogether, our data indicate that higher levels of root respiration and an efficient antioxidant metabolism were essential to provide soybean cultivar Embrapa 48 greater tolerance to dehydration.Conselho Nacional de Desenvolvimento Científico e Tecnológic

Loss and re-establishment of desiccation tolerance in the germinated seeds of Sesbania virgata (Cav.) (Pers.)

This research aimed to investigate the cellular alterations during the loss and re-establishment of desiccation tolerance (DT) in germinated Sesbania virgata seeds. The loss of DT was characterized in germinated seeds with increasing radicle lengths (1, 2, 3, 4 and 5 mm) when subjected to dehydration in silica gel, followed by rehydration. To re-establish DT, the germinated seeds were incubated for 72h in polyethylene glycol (PEG, -2.04 MPa) with or without ABA (100 μM) before dehydration in silica gel. Cell viability was assessed by seedling survival, and DNA integrity was evaluated by gel electrophoresis. Seeds with 1 mm radicle length survived dehydration to the original moisture content (MC) of the dry seed (approximately 10%). PEG treatment was able to re-establish DT, at least partially, with 2, 3 and 4 mm but not in 5 mm radicle lengths. Germinated seeds treated with PEG+ABA performed better than those treated only with PEG, and DT was re-established even in germinated seeds with a 5 mm radicle length. Among the PEG-treated germinated seeds dehydrated to 10% MC, DNA integrity was maintained only in those with a 1 mm radicle length.Esta pesquisa objetivou investigar as alterações celulares durante a perda e o restabelecimento da tolerância à dessecação (TD) em sementes germinadas de Sesbania virgata. A perda da TD foi caracterizada em sementes germinadas com 1, 2, 3, 4 e 5 mm de comprimento de radícula, submetidas à desidratação em sílica gel seguida de reidratação. Para restabelecer a TD, as sementes germinadas foram incubadas por 72h em PEG (-2,04 MPa) com e sem ABA (100 μM) antes da secagem em sílica gel. A viabilidade celular foi avaliada pela sobrevivência de plântulas e a integridade do DNA foi avaliada por meio de eletroforese em gel. Sementes com 1 mm de radícula sobreviveram à secagem até o teor de água original (aproximadamente 10%). O tratamento com PEG foi eficiente para restabelecer a TD, parcialmente, em sementes com 2, 3 e 4 mm, exceto com 5 mm de radícula. Sementes germinadas tratadas com PEG+ABA apresentaram melhor desempenho em relação às sementes sem ABA, sendo que a TD foi restabelecida em sementes com 5 mm de radícula. Dentre as sementes tratadas com PEG e secas até 10% de teor de água, a integridade do DNA foi mantida em sementes com 1 mm de radícula

Osmopriming on Sesbania virgata (CAV.) PERS (Fabaceae) seeds

This study evaluated the effect of the osmopriming on germination and vigour of Sesbania virgata seeds. Seeds were chemically scarified in concentrated sulphuric acid for 40 minutes and put to germinate either directly or after being submitted to osmopriming, drying and accelerated aging. Osmopriming was carried out with polyethylene glycol solutions (PEG 8000) at the following osmotic potentials-0.2; -0.4; -0.6 and -0.8 MPa for 12, 24 and 48 hours. After osmopriming, seeds were dried in silica gel until the initial moisture content was reached, and then submitted to the accelerated aging (48 h/100% RH). The effects of osmopriming and accelerated aging were evaluated through germination test, first counting germination and germination speed index. The osmopriming, followed or not by accelerated aging, positively influenced germination and vigour of Sesbania virgata seeds

Genetic Variability and Symbiotic Efficiency of Erythrina velutina Willd. Root Nodule Bacteria from the Semi-Arid Region in Northeastern Brazil

ABSTRACT Legume-rhizobia symbiosis is a cross-kingdom association that results in large amounts of nitrogen incorporated in food webs. For the Brazilian semi-arid region, data on genetic variability and symbiotic efficiency of Papilionoidae rhizobial communities are very scarce. The aim of this study was to evaluate the genetic variability and the symbiotic efficiency of eight rhizobial isolates obtained from “mulungu” (Erythrina velutina Willd.) nodules. For 16S rRNA gene sequencing, the genomic DNA was extracted using a commercial kit, amplified with universal primers, and subjected to sequencing reactions. For the isolate ESA 71, PCR amplifications for nodC and nodA genes were attempted. Rhizobial efficiency was assessed by two greenhouse experiments. The first assay was carried out under gnotobiotic conditions, with sterile sand as a substrate; the second experiment was conducted in a non-sterile soil. For both experiments, the inoculation treatments consisted of a single inoculation of each isolate, in addition to a treatment with Bradyrhizobium elkanii BR 5609 as a reference strain. Furthermore, two non-inoculated control treatments, supplied and not supplied with mineral N, were also evaluated. Bacterial identification indicated that both α and β-rhizobia could be found in “mulungu” root nodules. Three isolates where classified within the Rhizobium genus, four bacteria belonged to Bradyrhizobium and one isolate clustered with Burkholderia. Positive amplification of an intragenic fragment of the nodA gene using a primer set to β-rhizobia could be found for ESA 71 (Burkholderia). All bacterial isolates were effective in colonizing “mulungu” roots. In the first experiment, all inoculated treatments and N fertilization increased the N concentration in “mulungu” shoot tissues. For total N in the shoots, the isolates ESA 70, ESA 72, and ESA 75 stood out. In the non-sterile substrate experiment, the isolates ESA 70, ESA 71, ESA 72, and ESA 75, together with the reference strains, induced increases in the shoot N concentration and total accumulation compared to the absolute control. The results indicate that “mulungu” is able to establish associations with efficient α and β-rhizobia in Brazilian semi-arid soils

Mechanism and control of Genipa americana seed germination

Genipa americana (Rubiaceae) is important for restoration of riparian forest in the Brazilian Cerrado. The objective was to characterize the mechanism and control of germination of G. americana to support uniform seedling production. Morphology and morphometrics of seeds, embryo and endosperm were assessed by light and scanning electron microscopy during germination. Imbibition and germination curves were generated and over the same time interval endosperm digestion and resistance were measured by puncture force analysis and activity assay of endo-beta-mannanase (EBM) in water and in abscisic acid (ABA). The gene encoding for EBM was partially cloned and its expression monitored by quantitative real-time-polymerase chain reaction. Embryos displayed growth prior to radicle protrusion. A two-phase increase in EBM activity coincided with the two stages of weakening of the micropylar endosperm. The second stage also coincided with growth of the embryo prior to radicle protrusion. Enzyme activity was initiated in the micropylar endosperm but spread to the lateral endosperm. ABA completely inhibited germination by inhibiting embryo growth, the second stage of weakening and expression of the EBM gene, but EBM activity was not significantly inhibited. This suggests that a specific isoform of the enzyme is involved in endosperm weakening. EBM may cause a general softening of micropylar endosperm cell walls, allowing the embryo to puncture the endosperm as the driving force of the decrease in puncture force.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

Bioenergia: desenvolvimento, pesquisa e inovação

Com 27 trabalhos produzidos por pesquisadores do Instituto de Pesquisa em Bioenergia (Bioen), da Unesp, este livro oferece uma ampla visão sobre as áreas que compõem o segmento. Seu principal objetivo é contribuir para melhorar a compreensão dos vários aspectos da bioenergia, em especial no Brasil, que figura entre os países com maior nível de desenvolvimento tecnológico no setor. Os artigos abordam uma série abrangente de questões relacionadas à bioenergia, como a construção genética das plantas de cana-de-açúcar visando ao aumento de produtividade, a disseminação de sementes para estimular a propagação de espécies com potencial energético, etapas de produção de bioenergia, usos do combustível e seus efeitos nos diversos tipos de motores. Agrupados por assunto, os textos estão distribuídos em cinco partes: Biomassa para bioenergia; Produção de biocombustíveis; Utilização de bioenergia; Biorrefinaria, alcoolquímica e oleoquímica e Sustentabilidade dos biocombustíveis