Hypoxic stress tolerance in soybean : the effects of nitrate.

Orientador: Ladaslav SodekTese (doutorado) - Universidade Esstadual de Campinas, Instituto de BiologiaResumo: A soja é a leguminosa de maior importância econômica no Brasil com produção de 72,23 milhões de toneladas na safra de 2010/2011. O cultivo de soja em regiões que sofrem alagamento é uma das principais causas de perda de produtividade no Brasil. Nestas condições, ocorre redução na taxa de oxigênio do solo, e o sistema radicular da planta entra em hipoxia prejudicando o desenvolvimento e a produtividade das plantas. Existem evidências de que a adição de nitrato em condições de alagamento do sistema radicular (hipoxia) promove tolerância ao estresse, entretanto os mecanismos bioquímicos que envolvem essa tolerância ainda não estão totalmente elucidados. Analisando as alterações no metabolismo de nitrogênio em hipoxia, em plantas submetidas a diferentes condições experimentais, observou-se redução do conteúdo de nitrato dentro da planta, aumento no teor de aminoácidos solúveis totais, aumento na composição dos aminoácidos livres: Ala e Gaba, e redução de Asn, em plantas não noduladas e noduladas, além de redução no conteúdo de ureídeos totais em plantas noduladas. A adição de nitrato promoveu maior conteúdo de nitrato nas raízes, seiva do xilema e nódulos, maior teor de aminoácidos solúveis totais, maior conteúdo de proteínas solúveis totais nas raízes e menor nas folhas. A atividade da enzima redutase do nitrato in vivo em condições de hipoxia reduziu nas raízes das plantas não noduladas e nas raízes e folhas das plantas noduladas. A inclusão de K15NO3 no meio demonstrou que o nitrato é, de fato, absorvido em condições de hipoxia, pois as raízes, folhas e nódulos todos incorporaram o 15N. Entretanto, absorção do nitrato sob hipoxia foi bem menor do que sob normoxia. Tanto em normoxia como hipoxia as raízes apresentaram o maior grau de enriquecimento com 15N enquanto que a incorporação nos nódulos foi menor. Houve incorporação de 15N em aminoácidos em todos os tecidos, o que permite concluir que o nitrato não é apenas absorvido pelas raízes sob hipoxia mas também assimilado e transportado para a parte aérea. Os aminoácidos mais próximos ao processo de assimilação de nitrato, Asp, Glu, Ala e Asn, foram os que mais incorporaram o 15N. No entanto, em geral um grau menor de incorporação foi encontrado sob hipoxia. A adição de nitrato, no dia 7, na solução do vaso de cinco cultivares de soja cujo sistema radicular estava alagado promoveu, na cultivar IAC Foscarin-31, maior crescimento da planta quando comparada às plantas com nitrato desde o início do experimento; na cultivar IAC-23, maior massa seca do sistema radicular; nas cultivares IAC-17 e IAC-18 foi evidenciado o mesmo crescimento da planta que as demais com nitrato; e na cultivar IAC-24, houve menor crescimento da planta. A adição também promoveu maior formação de aerênquima no caule, na raiz principal e adventícia, além de estimular a maior formação de raízes adventícias. Os dados sugerem que em condições de hipoxia, embora em quantidades bastante reduzidas e num processo mais lento, o nitrato é absorvido pelas raízes, sendo parte, convertida a aminoácidos e menos de 40% convertidos a nitrito, o qual é eliminado para o meio. A redução do nitrato via redutase do nitrato, passo inicial de sua metabolização, não é um passo limitante, uma vez que há produção de nitrito, produto da atividade da enzima. Com a adição de nitrato na solução do vaso de cinco cultivares de soja foram observadas tendências para suprir a ausência de nitrogênio e manter o desenvolvimento da plantas em condições de alagamento. A cultivar IAC Foscarin-31 e IAC-24 apresentaram crescimento geral da planta, sendo maior o crescimento observado na cultivar IAC Foscarin-31. Na cultivar IAC-23 houve maior crescimento do sistema radicular; na cultivar IAC-17, da parte aérea, assim como na IAC-18Abstract: Soybean is a legume of great economic importance in Brazil with a production of some 72.23 million tons (2010/2011 harvest). One of the main causes of diminished yields of the cultivars produced in Brazil is their cultivation in regions subject to flooding. Under such conditions reduced availability of oxygen in the soil leads to hypoxia of the root system which impairs plant development and yield. There is evidence that the presence of nitrate during flooding can improve tolerance to hypoxic stress. However, the biochemical mechanisms underlying this phenomenon have not been fully elucidated. After analysis of the changes in nitrogen metabolism that occur under hypoxia, in plants subjected to a variety of experimental conditions, it was observed that there was a reduction in plant nitrate content, an increase in total soluble amino acids, an increase in the relative abundance of free Ala, Gaba, and a reduction in Asn, in both nodulated and non-nodulated plants, as well as a reduction of ureides in nodulated plants. The addition of nitrate during flooding resulted in a higher nitrate content of the root, nodule and xylem sap. Total soluble amino acids and soluble proteins also increased in the root under these conditions but decreased in the leaf. Under hypoxia the in vivo nitrate reductase activity declined in the roots of non-nodulated plants and in both the roots and leaves of nodulated plants. The inclusion of K15NO3 in the nutrient solution confirmed the uptake of nitrate under hypoxia, since roots, leaves and nodules became labelled. However, uptake under hypoxia was much lower than that observed under normoxia. Under both normoxia and hypoxia the roots showed the highest degree of 15N enrichment while the nodules showed the weakest. Incorporation of 15N in amino acids of all tissues shows that 15NO3 was not only taken up by the roots under hypoxia but that it was assimilated and transported to the shoot. The amino acids considered closest to N assimilation, Asp, Glu, Ala and Asn, were the most highly labelled. Nevertheless, lower levels of incorporation were generally found under hypoxia. The addition of nitrate to the hydroponic nutrient solution of five soybean cultivars, 7 days after flooding the root system, stimulated greater growth of the cultivar IAC Foscarin-31 as compared to plants with nitrate from the beginning of the experiment. It also stimulated greater root dry mass of the cultivar IAC-23, produced a similar growth increase of cultivars IAC-17 and IAC-18 compared to those with nitrate from the beginning, but resulted in less growth for the cultivar IAC-24. The addition of nitrate also stimulated formation of adventitious roots as well as aerenchyma which formed on both the stem and the main and adventitious roots. The data show that under conditions of hypoxia, nitrate is taken up by the roots although at a much lower rate than under normoxia, and in part is assimilated into amino acids while nearly 40% is reduced to nitrite which is excreted to the surrounding medium. The reduction of nitrate by the enzyme NR, the first step of nitrate metabolism, does not appear to be limiting since large quantities of the reaction product, nitrite, accumulate. With the addition of nitrate to the flooding medium of five soybean cultivars there was a tendency to overcome the absence of nitrogen and maintain plant development under flooded conditions. Of the five cultivars studied, IAC Foscarin-31 and IAC-24 responded positively through overall plant growth, with Foscarin-31 showing the greatest growth. Cultivar IAC-23 responded with greater root growth while IAC-17 and IAC-18 presented greater shoot growthDoutoradoBiologia VegetalDoutor em Biologia Vegeta

Utilization Of (15)no3 (-) By Nodulated Soybean Plants Under Conditions Of Root Hypoxia.

Waterlogging of soils is common in nature. The low availability of oxygen under these conditions leads to hypoxia of the root system impairing the development and productivity of the plant. The presence of nitrate under flooding conditions is regarded as being beneficial towards tolerance to this stress. However, it is not known how nodulated soybean plants, cultivated in the absence of nitrate and therefore not metabolically adapted to this compound, would respond to nitrate under root hypoxia in comparison with non-nodulated plants grown on nitrate. A study was conducted with (15)N labelled nitrate supplied on waterlogging for a period of 48 h using both nodulated and non-nodulated plants of different physiological ages. Enrichment of N was found in roots and leaves with incorporation of the isotope in amino acids, although to a much smaller degree under hypoxia than normoxia. This demonstrates that nitrate is taken up under hypoxic conditions and assimilated into amino acids, although to a much lesser extent than for normoxia. The similar response obtained with nodulated and non-nodulated plants indicates the rapid metabolic adaptation of nodulated plants to the presence of nitrate under hypoxia. Enrichment of N in nodules was very much weaker with a distinct enrichment pattern of amino acids (especially asparagine) suggesting that labelling arose from a tissue source external to the nodule rather than through assimilation in the nodule itself.20287-9

Nitrogen fertilization benefits soybean under flooding conditions

ABSTRACT The aim of this study was to compare the growth of five cultivars of soybean produced in the State of São Paulo under conditions of flooding and evaluate the response of the plants to nitrate under such conditions. The study involved the analysis of leaf area, shoot dry mass, volume of the root system and root dry mass, using the soybean cultivars IAC Foscarin-31, IAC-23, IAC-17, IAC-24 and IAC-18, as well as a histological analysis of the stem and main root of the cultivar IAC-23, after being maintained for 28 days under flooding conditions. All cultivars were submitted to the following treatments: flooding of the root system without nitrate; flooding in the presence of nitrate; and flooding with nitrate added seven days after flooding. Our data indicate that the choice of cultivar and the use of nitrate should be taken into account for the planting of soybean in regions subject to flooding. The presence of nitrate during flooding was beneficial for all five cultivars tested. The cultivar IAC-24 presented the highest growth in the presence of nitrate. IAC Foscarin-31 was the most responsive cultivar to the addition of nitrate seven days after flooding, when considering plant growth. The presence of aerenchyma on the stem and main root of the flooded plants was observed for all treatments. However, variation occurred in the arrangement of parenchyma cells, this being more compact in the treatments with nitrate.RESUMO Este trabalho teve como objetivo comparar o desenvolvimento de cinco cultivares de soja produzidas no Estado de São Paulo, em situação de alagamento do sistema radicular, e avaliar a resposta das plantas ao nitrato nessa situação. No trabalho, foram analisadas a área foliar, a massa seca da parte aérea, o volume do sistema radicular e a massa seca do sistema radicular das cultivares de soja IAC Foscarin-31, IAC-23, IAC-17, IAC-24 e IAC-18, além de análise histológica do caule e da raiz principal da cultivar IAC-23, das plantas mantidas durante 28 dias alagadas. Todas as cultivares foram submetidas aos tratamentos: alagamento do vaso sem nitrato; alagamento com nitrato e alagamento com nitrato adicionado após sete dias de experimento. Os resultados indicam que a escolha da cultivar e a adição de nitrato devem ser levadas em consideração para o plantio em regiões com potencial para alagamento. A presença de nitrato foi benéfica para as cinco cultivares avaliadas. Na cultivar IAC-24 notou-se maior crescimento em presença de nitrato. A adição de nitrato sete dias após alagamento promoveu maior crescimento na cultivar IAC Foscarin-31. A presença de aerênquima no caule e na raiz principal de plantas alagadas foi constatada em todos os tratamentos, porém houve variação no arranjo das células parenquimáticas, o qual foi mais compacto nos tratamentos com nitrato.209Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES