Isolation and Characterization of Plant Growth-Promotion Diazotrophic Endophytic Bacteria Associated to Sugarcane (Saccharum officinarum L.) Grown in Paraíba, Brazil

Abstract: Sugarcane is an important Brazilian commodity, being usually cultivated in soils with low natural fertility. This study aimed to isolate diazotrophic endophytes from sugarcane tissues and evaluate the morphological and physiological characteristics of their colonies as well as their plant growth-promoting (PGP) traits in select diazotrophic endophytic bacteria. Fifty-six bacterial isolates were identified in the sugarcane tissues, and these isolates presented distinct morphological and physiological traits. A total of thirty-five bacterial isolates were biochemically evaluated. Overall, Bacillus was the dominant genus. Isolates of Methylobacterium spp. and Brevibacillus agri were present only in leaves, while Herbaspirillum seropedicae occurred only in stems. Except to IPA-CF45A, all isolates were nitrogenase positive. All endophytes exhibit production of indol 3-acetic acid. Over 50% of endophytes solubilize phosphate, release N-acyl homoserine lactones, and present the activity of 1-aminocyclopropane-1-carboxylic acid deaminase, catalase, lipase and protease. The network analysis showed that isolates belonged to Burkholderia, Herbaspirillum, and Methylobacterium interact with Bacillus. Bacterial endophytes exhibited distinct morphological, physiological, and PGP traits that are useful for sustainable agriculture, highlighting the isolates IPA-CC33, IPA-CF65, IPA-CC9 and IPA-CF27. Further studies on the effects of these diazotrophic endophytes and their potential for providing microbial inoculants for improving sugarcane fields will provide valuable information to maintain the sustainability and environment quality.

Distinct bacterial community structure and composition along different cowpea producing ecoregions in Northeastern Brazil.

Soil microbial communities represent the largest biodiversity on Earth, holding an important role in promoting plant growth and productivity. However, the knowledge about how soil factors modulate the bacteria community structure and distribution in tropical regions remain poorly understood, mainly in different cowpea producing ecoregions belonging to Northeastern Brazil. This study addressed the bacterial community along three different ecoregions (Mata, Sertão, and Agreste) through the16S rRNA gene sequencing. The results showed that soil factors, such as Al3+, sand, Na+, cation exchange excel, and total organic C, influenced the bacterial community and could be a predictor of the distinct performance of cowpea production. Also, the bacterial community changed between different ecoregions, and some keystone groups related to plant-growth promotion, such as Bradyrhizobium, Bacillales, Rhizobiales, and Solibacillus, were correlated to cowpea yield, so revealing that the soil microbiome has a primordial role in plant productivity. Here, we provide evidence that bacterial groups related to nutrient cycling can help us to increase cowpea efficiency and we suggest that a better microbiome knowledge can contribute to improving the agricultural performance.

Inoculation of arbuscular mycorrhizal fungi as a strategy to improve annatto (Bixa orellana L. ) growth

This study aimed to assess the occurrence of arbuscular mycorrhizal fungi (AMF) in annatto (Bixa orellana L.) cultivars and their response to AMF inoculation using biometric parameters. The occurrence surveys were conducted in annatto fields in three municipalities from Pernambuco Forest Zone: Lagoa de Itaenga, Gloria de Goitá, and Vitoria de Santo Antão, and in four cultivars (Red Piave, Green Piave, Red Peruvian Paulista, and Green Peruvian Paulista). In a greenhouse, biometric parameters of annatto seedlings of Red Piave, Red Peruvian Paulista, Embrapa-36, and Embrapa-37 cultivars inoculated with AMF isolated from annatto fields. The Red Piave cultivar exhibited greater root colonization than the Green Peruvian Paulista in the Lagoa de Itaenga and Vitoria de Santo Antão municipalities. The cultivar Red Piave showed a more beneficial association with AMF in plants and soil than cultivar Green Peruvian Paulista did, in both Lagoa de Itaenga and Vitoria de Santo Antão. AMF inoculation was effective in promoting the growth of annatto plants, particularly those inoculants with S. heterogama and C. etunicatum.

Seed size influences the promoting activity of rhizobia on plant growth, nodulation and N fixation in lima bean / O tamanho da semente influencia a atividade promotora dos rizóbios sobre o crescimento de plantas, nodulação e fixação de N em feijão-fava

ABSTRACT: This study evaluated the activity of rhizobia isolates inoculated in large (18 mm) and small (11 mm) seeds on lima bean growth, nodulation and N fixation. Selected rhizobia isolates were compared with a reference strain CIAT899 and two controls without inoculation. Large seeds contributed for highest plant growth, nodulation and N fixation than small seeds. The isolates UFPI-59, UFPI-18 and UFPI-38 promoted the highest values of shoot and root dry weight, respectively. The isolates UFPI-32 promoted the highest values of nodule number, while UFPI-59 promoted the highest values of nodule dry weight. The isolates UFPI-38 and UFPI-59 promoted the highest accumulation of N. This study showed that seed size really influences lima bean growth, nodulation and BNF. Considering rhizobia isolates, UFPI-59, UFPI-38, and UFPI-18 contributed for plant growth, promoted better nodulation and effectiveness on biological N fixation.

RESUMO: Este estudo avaliou a atividade de isolados de rizóbios inoculados em sementes pequenas (11 mm) e grandes (18 mm) sobre o crescimento, nodulação e fixação de N de feijão-fava. Isolados selecionados de rizóbios foram comparados com uma estirpe de referência CIAT899 e dois controles sem inoculação. Sementes grandes contribuíram para maior crescimento de plantas, nodulação e fixação de N do que sementes pequenas. Os isolados UFPI-59, UFPI-18 e UFPI-38 promoveram os maiores valores de massa seca de parte aérea e raízes. Os isolados UFPI-32 e UFPI-59 promoveram os maiores valores de número e massa de nódulos, respectivamente. Os isolados UFPI-38 e UFPI-59 promoveram maior acúmulo de N. Este estudo mostrou que o tamanho de sementes influencia o crescimento, nodulação e fixação de N em feijão-fava. Considerando os isolados de rizóbios, UFPI-59, UFPI-38 e UFPI-18 contribuíram para o crescimento de plantas, promoveram melhor nodulação e eficiência na fixação de N.

Changes induced by co-inoculation in nitrogen-carbon metabolism in cowpea under salinity stress

ABSTRACT To mitigate the deleterious effects of abiotic stress, the use of plant growth-promoting bacteria along with diazotrophic bacteria has been increasing. The objectives of this study were to investigate the key enzymes related to nitrogen and carbon metabolism in the biological nitrogen fixation process and to elucidate the activities of these enzymes by the synergistic interaction between Bradyrhizobium and plant growth-promoting bacteria in the absence and presence of salt stress. Cowpea plants were cultivated under axenic conditions, inoculated with Bradyrhizobium and co-inoculated with Bradyrhizobium sp. and Actinomadura sp., Bradyrhizobium sp. and Bacillus sp., Bradyrhizobium sp. and Paenibacillus graminis, and Bradyrhizobium sp. and Streptomycessp.; the plants were also maintained in the absence (control) and presence of salt stress (50 mmolL-1 NaCl). Salinity reduced the amino acids, free ammonia, ureides, proteins and total nitrogen content in nodules and increased the levels of sucrose and soluble sugars. The co-inoculations responded differently to the activity of glutamine synthetase enzymes under salt stress, as well as glutamate synthase, glutamate dehydrogenase aminating, and acid invertase in the control and salt stress. Considering the development conditions of this experiment, co-inoculation with Bradyrhizobium sp. and Bacillus sp. in cowpea provided better symbiotic performance, mitigating the deleterious effects of salt stress.

Protist species richness and soil microbiome complexity increase towards climax vegetation in the Brazilian Cerrado.

Biodiversity underlies ecosystem functioning. While aboveground biodiversity is often well studied, the belowground microbiome, in particular protists, remains largely unknown. Indeed, holistic insights into soil microbiome structures in natural soils, especially in hyperdiverse biomes such as the Brazilian Cerrado, remain unexplored. Here, we study the soil microbiome across four major vegetation zones of the Cerrado, ranging from grass-dominated to tree-dominated vegetation with a focus on protists. We show that protist taxon richness increases towards the tree-dominated climax vegetation. Early successional habitats consisting of primary grass vegetation host most potential plant pathogens and least animal parasites. Using network analyses combining protist with prokaryotic and fungal sequences, we show that microbiome complexity increases towards climax vegetation. Together, this suggests that protists are key microbiome components and that vegetation succession towards climax vegetation is stimulated by higher loads of animal and plant pathogens. At the same time, an increase in microbiome complexity towards climax vegetation might enhance system stability.

Archaea diversity in vegetation gradients from the Brazilian Cerrado

Abstract We used 16S rRNA sequencing to assess the archaeal communities across a gradient of Cerrado. The archaeal communities differed across the gradient. Crenarcheota was the most abundant phyla, with Nitrosphaerales and NRPJ as the predominant classes. Euryachaeota was also found across the Cerrado gradient, including the classes Metanocellales and Methanomassiliicoccaceae.

Antioxidant response of cowpea co-inoculated with plant growth-promoting bacteria under salt stress

Abstract Soil salinity is an important abiotic stress worldwide, and salt-induced oxidative stress can have detrimental effects on the biological nitrogen fixation. We hypothesized that co-inoculation of cowpea plants with Bradyrhizobium and plant growth-promoting bacteria would minimize the deleterious effects of salt stress via the induction of enzymatic and non-enzymatic antioxidative protection. To test our hypothesis, cowpea seeds were inoculated with Bradyrhizobium or co-inoculated with Bradyrhizobium and plant growth-promoting bacteria and then submitted to salt stress. Afterward, the cowpea nodules were collected, and the levels of hydrogen peroxide; lipid peroxidation; total, reduced and oxidized forms of ascorbate and glutathione; and superoxide dismutase, catalase and phenol peroxidase activities were evaluated. The sodium and potassium ion concentrations were measured in shoot samples. Cowpea plants did not present significant differences in sodium and potassium levels when grown under non-saline conditions, but sodium content was strongly increased under salt stress conditions. Under non-saline and salt stress conditions, plants co-inoculated with Bradyrhizobium and Actinomadura or co-inoculated with Bradyrhizobium and Paenibacillus graminis showed lower hydrogen peroxide content in their nodules, whereas lipid peroxidation was increased by 31% in plants that were subjected to salt stress. Furthermore, cowpea nodules co-inoculated with Bradyrhizobium and plant growth-promoting bacteria and exposed to salt stress displayed significant alterations in the total, reduced and oxidized forms of ascorbate and glutathione. Inoculation with Bradyrhizobium and plant growth-promoting bacteria induced increased superoxide dismutase, catalase and phenol peroxidase activities in the nodules of cowpea plants exposed to salt stress. The catalase activity in plants co-inoculated with Bradyrhizobium and Streptomyces was 55% greater than in plants inoculated with Bradyrhizobium alone, and this value was remarkably greater than that in the other treatments. These results reinforce the beneficial effects of plant growth-promoting bacteria on the antioxidant system that detoxifies reactive oxygen species. We concluded that the combination of Bradyrhizobium and plant growth-promoting bacteria induces positive responses for coping with salt-induced oxidative stress in cowpea nodules, mainly in plants co-inoculated with Bradyrhizobium and P. graminis or co-inoculated with Bradyrhizobium and Bacillus.

Changes induced by co-inoculation in nitrogen-carbon metabolism in cowpea under salinity stress.

To mitigate the deleterious effects of abiotic stress, the use of plant growth-promoting bacteria along with diazotrophic bacteria has been increasing. The objectives of this study were to investigate the key enzymes related to nitrogen and carbon metabolism in the biological nitrogen fixation process and to elucidate the activities of these enzymes by the synergistic interaction between Bradyrhizobium and plant growth-promoting bacteria in the absence and presence of salt stress. Cowpea plants were cultivated under axenic conditions, inoculated with Bradyrhizobium and co-inoculated with Bradyrhizobium sp. and Actinomadura sp., Bradyrhizobium sp. and Bacillus sp., Bradyrhizobium sp. and Paenibacillus graminis, and Bradyrhizobium sp. and Streptomycessp.; the plants were also maintained in the absence (control) and presence of salt stress (50mmolL-1 NaCl). Salinity reduced the amino acids, free ammonia, ureides, proteins and total nitrogen content in nodules and increased the levels of sucrose and soluble sugars. The co-inoculations responded differently to the activity of glutamine synthetase enzymes under salt stress, as well as glutamate synthase, glutamate dehydrogenase aminating, and acid invertase in the control and salt stress. Considering the development conditions of this experiment, co-inoculation with Bradyrhizobium sp. and Bacillus sp. in cowpea provided better symbiotic performance, mitigating the deleterious effects of salt stress.

Antioxidant response of cowpea co-inoculated with plant growth-promoting bacteria under salt stress.

Soil salinity is an important abiotic stress worldwide, and salt-induced oxidative stress can have detrimental effects on the biological nitrogen fixation. We hypothesized that co-inoculation of cowpea plants with Bradyrhizobium and plant growth-promoting bacteria would minimize the deleterious effects of salt stress via the induction of enzymatic and non-enzymatic antioxidative protection. To test our hypothesis, cowpea seeds were inoculated with Bradyrhizobium or co-inoculated with Bradyrhizobium and plant growth-promoting bacteria and then submitted to salt stress. Afterward, the cowpea nodules were collected, and the levels of hydrogen peroxide; lipid peroxidation; total, reduced and oxidized forms of ascorbate and glutathione; and superoxide dismutase, catalase and phenol peroxidase activities were evaluated. The sodium and potassium ion concentrations were measured in shoot samples. Cowpea plants did not present significant differences in sodium and potassium levels when grown under non-saline conditions, but sodium content was strongly increased under salt stress conditions. Under non-saline and salt stress conditions, plants co-inoculated with Bradyrhizobium and Actinomadura or co-inoculated with Bradyrhizobium and Paenibacillus graminis showed lower hydrogen peroxide content in their nodules, whereas lipid peroxidation was increased by 31% in plants that were subjected to salt stress. Furthermore, cowpea nodules co-inoculated with Bradyrhizobium and plant growth-promoting bacteria and exposed to salt stress displayed significant alterations in the total, reduced and oxidized forms of ascorbate and glutathione. Inoculation with Bradyrhizobium and plant growth-promoting bacteria induced increased superoxide dismutase, catalase and phenol peroxidase activities in the nodules of cowpea plants exposed to salt stress. The catalase activity in plants co-inoculated with Bradyrhizobium and Streptomyces was 55% greater than in plants inoculated with Bradyrhizobium alone, and this value was remarkably greater than that in the other treatments. These results reinforce the beneficial effects of plant growth-promoting bacteria on the antioxidant system that detoxifies reactive oxygen species. We concluded that the combination of Bradyrhizobium and plant growth-promoting bacteria induces positive responses for coping with salt-induced oxidative stress in cowpea nodules, mainly in plants co-inoculated with Bradyrhizobium and P. graminis or co-inoculated with Bradyrhizobium and Bacillus.

Archaea diversity in vegetation gradients from the Brazilian Cerrado.

We used 16S rRNA sequencing to assess the archaeal communities across a gradient of Cerrado. The archaeal communities differed across the gradient. Crenarcheota was the most abundant phyla, with Nitrosphaerales and NRPJ as the predominant classes. Euryachaeota was also found across the Cerrado gradient, including the classes Metanocellales and Methanomassiliicoccaceae.

Distinct bacterial communities across a gradient of vegetation from a preserved Brazilian Cerrado.

The Cerrado biome in the Sete Cidades National Park, an Ecological Reserve in Northeastern Brazil, has conserved its native biodiversity and presents a variety of plants found in other savannas in Brazil. Despite this finding the soil microbial diversity and community structure are poorly understood. Therefore, we described soil bacterial diversity and distribution along a savanna vegetation gradient taking into account the prevailing environmental factors. The bacterial composition was retrieved by sequencing a fragment of the 16S ribosomal RNA gene. The bacterial operational taxonomic units (OTUs) were assigned to 37 different phyla, 96 classes, and 83 genera. At the phylum level, a core comprised by Proteobacteria, Acidobacteria, Actinobacteria, Firmicutes, Verrucomicrobia and Planctomycetes, was detected in all areas of Cerrado. 'Cerrado stricto sensu' and 'Cerradao' share more similarities between edaphic properties and vegetation and also present more similar bacterial communities, while 'Floresta decidual' and 'Campo graminoide' show the largest environmental differences and also more distinct bacterial communities. Proteobacteria (26%), Acidobacteria (21%) and Actinobacteria (21%) were the most abundant phyla within the four areas. All the samples present similar bacteria richness (alpha diversity) and the observed differences among them (beta diversity) were more related to the abundance of specific taxon OTUs compared to their presence or absence. Total organic C, N and P are the main abiotic factors structuring the bacterial communities. In summary, our findings show the bacterial community structure was clearly different across the Cerrado gradient, but that these environments share a bacterial phylum-core comprising Proteobacteria, Acidobacteria, Actinobacteria, Verrucomicrobia and Planctomycetes with other Brazilian savannas.

Resposta da co-inoculação de bactérias promotoras de crescimento em plantas e Bradyrhizobium sp. em caupi / Co-inoculation response of growth promoting bacterias in plants and Bradyrhizobium sp. in cowpea

Para a região árida e semi-árida do Nordeste brasileiro, o cultivo do caupi (Vigna unguiculata [L.] Walp.) vem se destacando por ser uma cultura adaptável as condições da região. Devido à capacidade do caupi de, em simbiose com o rizóbio, realizar a FBN, a co-inoculação rizóbio e bactérias promotoras de crescimento em plantas (BPCP's) poderá optimizar a fixação do N2 atmosférico dependendo da combinação e compatibilidade das estirpes envolvidas. O objetivo do trabalho foi avaliar a resposta da co-inoculação de BPCP's e Bradyrhizobium sp. como alternativa para optimizar a performance simbiótica e o desenvolvimento do caupi em condições de casa-de-vegetação. O experimento foi instalado em casa de vegetação, utilizando como substrato areia lavada e autoclavada em vasos de Leonard. Os tratamentos foram: 22 estirpes de BPCP's comparadas com a estirpe padrão (BR 3267), no caupi cv. "IPA 206". A colheita foi efetuada aos 35 dias após o plantio (DAP) e foram avaliados as seguintes variáveis: altura de planta (AP) aos 15, 25 e 35 dias, comprimento da raiz (CR), matéria seca da parte aérea (MSPA), matéria seca da raiz (MSR), matéria seca do nódulo (MSN), nodulação específica (NE) e nitrogênio acumulado na parte aérea (Nac). Os resultados mostram que a cultura do caupi respondeu significativamente a inoculação de estirpes de Bacillus, Brevibacillus e Paenibacillus co-inoculados com a estirpe de Bradyrhizobium sp. (BR 3267). A interação dos micro-organismos estudados não influenciou as variáveis, nodulação específica e matéria seca dos nódulos no caupi cv. IPA 206. A estirpe de Paenibacillus graminis (MC 04.21) co-inoculada com Bradyrhizobium sp. (BR 3267) foi superior em relação às demais estirpes avaliadas na produção de nitrogênio acumulado na matéria seca da parte aérea, proporcionando uma melhor performance simbiótica.

In the arid and semi-arid northeastern Brazil, the cultivation of cowpea has stood out for being an adaptable culture conditions in the region. Because of the ability of cowpea, in symbiosis with rhizobia carry out the FBN, the co-inoculation Rhizobium and growth promoting bacteria in plants can optimize the fixation of atmospheric N2 depending on the combination and compatibility of the strains involved. The objective of this study was to evaluate the response of the inoculation BPCP's and Bradyrhizobium sp. as an alternative to optimize performance and development of symbiotic cowpea under conditions of green-house. The experiment was conducted in a greenhouse, using as substrate washed and sterilized sand in Leonard jars. The treatments were: 22 strains BPCP's compared with the strain (BR 3267), on cowpea cv. IPA 206. At harvest 35 days after planting (DAP) and the following variables were evaluated: plant height (AP) 15, 25 and 35 days, root length (CR), shoot dry matter (MSPA), root dry matter (MSR), nodule dry matter (MSN), specific nodulation (NE) and nitrogen accumulated in shoots (Nac). The results show that the culture of cowpea responded significantly to inoculation of strains of Bacillus, Brevibacillus and Paenibacillus co-inoculated with the strain of Bradyrhizobium sp. (BR 3267). The interaction of microorganisms studied non-influenced the variables specific nodulation and dry matter of nodules in cowpea cv. IPA 206. The strain of Paenibacillus graminis (MC 04/21) coinoculated with Bradyrhizobium sp. (BR 3267) was higher compared to other strains tested in the production of nitrogen accumulated in shoot dry matter, providing a better performance symbiotic.

Triple inoculation with Bradyrhizobium, Glomus and Paenibacillus on cowpea (Vigna unguiculata) [L. ] walp. )development. (au)

The use of microorganisms to improve the availability of nutrients to plants is of great importance to agriculture. This study aimed to evaluate the effect of triple inoculation of cowpea with arbuscular mycorrhizal fungi (AMF), plant growth-promoting bacteria (PGPB) and rhizobia to maximize biological nitrogen fixation (BNF) and promote plant growth. The experiment was conducted in a greenhouse using cowpea plants (Vigna unguiculata L. Walp cv. IPA 206). The treatments included inoculation with strains of Bradyrhizobium sp. (BR 3267 and EI - 6) individually and as a mixture, an absolute control (AC) and mineral nitrogen control (NC), all combined with the presence or absence of native AMF (Glomus etunicatum) and PGPB (Paenibacillus brasilensis - 24) in a 5x2x2 factorial design. All treatments were replicated three times. Contrasts were performed to study the treatment of variables. Inoculation with Bradyrhizobium sp. (BR 3267 and EI - 6) and G. etunicatum favored nitrogen acquisition and phosphorus availability for the cowpea plants. Inoculation with P. brasilensis - 24 increased colonization by Bradyrhizobium sp. and G. etunicatum and promoted cowpea growth, while the nitrogen from symbiosis was sufficient to supply the plants nutritional needs.

Triple inoculation with Bradyrhizobium, Glomus and Paenibacillus on cowpea (Vigna unguiculata [L.] walp.) development.

The use of microorganisms to improve the availability of nutrients to plants is of great importance to agriculture. This study aimed to evaluate the effect of triple inoculation of cowpea with arbuscular mycorrhizal fungi (AMF), plant growth-promoting bacteria (PGPB) and rhizobia to maximize biological nitrogen fixation (BNF) and promote plant growth. The experiment was conducted in a greenhouse using cowpea plants (Vigna unguiculata L. Walp cv. IPA 206). The treatments included inoculation with strains of Bradyrhizobium sp. (BR 3267 and EI - 6) individually and as a mixture, an absolute control (AC) and mineral nitrogen control (NC), all combined with the presence or absence of native AMF (Glomus etunicatum) and PGPB (Paenibacillus brasilensis - 24) in a 5x2x2 factorial design. All treatments were replicated three times. Contrasts were performed to study the treatment of variables. Inoculation with Bradyrhizobium sp. (BR 3267 and EI - 6) and G. etunicatum favored nitrogen acquisition and phosphorus availability for the cowpea plants. Inoculation with P. brasilensis - 24 increased colonization by Bradyrhizobium sp. and G. etunicatum and promoted cowpea growth, while the nitrogen from symbiosis was sufficient to supply the plants nutritional needs.

Nitrate levels and stages of growth in hypernodulating mutants pf Lupinus Albus. I. N2 fixation potential

This work aimed to evaluate physiological parameters, nodulation response and N2 fixation rate in mutants of Lupinus albus in comparison with the standard Multolupa cultivar. Two nitrate levels (0 and 5mM) and two evaluation periods (7 and 10 weeks) were used. Significant differences were observed among genotypes, in relation to fresh nodule weight, nitrate levels and growth stages. The overall average for nitrate level differed between them where 5mM severely inhibieted the number of nodules, reaching a 49.5 (per cent) reduction in relation to treatment without nitrate. There were no behaviour differences among genotypes, nor among evaluation periods. Although the level of nitrate did not influence the production of shoot dry matter in relation to the average among levels applied, the L-135 genotype, being an inefficient mutant, reached very low values. There were no significant differences in electron allocation coefficient (EAC) among nitrate levels, nor among genotypes studied. However , the evaluation periods revealed differences, where the EAC for the seventh week had a higher value than that for the tenth week, when a 5mM aplication was evaluated. The N2 fixation rate (N2FIX) showed the existence of the nitrate interference in fixation, given that the application of 5mM severely reduced. However, there were no differences among the genotypes and it was noted that the fixation rate was much higher in those that received nitrate. The L-88 and L-62 genotypes were the ones that have shown best adaptability in this experiment, thus being able to be recommended for new studies with higher nitrate levels and different evaluation periods. The nitrate (5mM) interferes in the nitrogen fixation rate, given that all the genotypes were affected by the level applied.

Nitrate levels and stages of growth in hypernodulating mutants of Lupinus Albus. II.Enzymatic activity and transport of N in the xylem SAP

The enzymatic study and transport of N in the xylem sap was carried out with a view to observing the influence of different nitrate levels and growth stages of the plant in chemically treated mutants of Lupinus albus. Several stresses induce a reduction in plant growth, resulting in the accumulation of free amino acids, amides or ureides, not only in the shoot, but also in the roots and nodules. Although enzyme activity is decisive in avoiding products that inhibit nitrogenase by ammonium, little is known about the mechanism by wich the xylem carries these products. However, this process may be the key to the function of avoiding the accumulation of amino acids in the cells of infected nodules. The behaviour of the enzymes nitrate reductase (NR), phosphoenolpyruvate carboxylase (PEPC), glutamine synthetase (GS) and nitrogen compounds derived from fixation, such as N-Ó-amino, N-ureides and N-amide in mutant genotypes were observed. The NR enzyme was highly influenced by the application of nitrate showing much higher values than those in the non-application of nitrate, independently of genotype, being that the NR, the best evaluation period was in the tenth week. The L-62 genotype characterized with nitrate-resistance, clearly showed that the enzyme PEPC is inhibited by presence of nitrate. The L-135 genotype (nor fix) showed GS activity extremely low, thus demonstrating that GS is an enzyme highly correlated with fixation. With regard to the best growth stage for GS, Lupinus albus should be evaluated in the seventh week.

Effect of nitrate on nodulation and n2-fixation of lupinus mutabilis mutants at different growth stages

Uma das principais técnicas usadas para estudar o efeito inibidor do NO3 na fixaçäo biológica do N2, tem sido o uso de mutantes de estirpes de Rhizobium ou de plantas hospedeiras que näo apresentam o padräo usual de nodulaçäo. Este estudo foi conduzido usando genótipos parentais e mutantes de Lupinus mutabilis, submetidos a dois níveis de NO3: 0 e 5mM, com duas épocas de colheita: sete e dez semanas após emergência. A concentraçäo de 5mM revelou-se da maior importância na inibiçäo da nodulaçäo inicial de Lupinus, apesar de os dados demonstrarem que os mutantes L-114 e L-105 foram mais eficientes quanto a nodulaçäo, N-total e atividade da nitrogenase que a cv. SCG-25, e podem permitir um aumento da fixaçäo de N2. Mas näo houve diferenças na atividade da glutamina sintetase (GS) dos nódulos entre os mutantes L-114, L-105 e a cv. SCG-25, quando as plantas näo receberam NO3, mas na presença de NO3 houve tendência para os mutantes, especialmente para os L-114, apresentarem maior atividade de GS. Isto indica que o mutante L-114 tem uma maior capacidade de assimilaçäo do N fixado, associada com um maior suprimento de carboidratos disponíveis, como indicado pela atividade da PEP-carboxilase. Portanto, a tolerância parcial ao NO3 apresentada pelos mutantes L-114 e L-105 está associada com o fenótipo hipernodulante. Esta sugestäo pode ser observada de reduçäo de acetileno, que foram maiores para o mutante L-114 que para a cv. SCG-25, na presença ou na ausência de NO3

Efetividade do inoculante com Bradyrhizobium sp em diferentes substratos / Effectivenees of Bradyrhizobium sp inoculants on different substrates

Resumo: Objetivando determinar as condiçöes ideais para a sobrevivência e efetividade do Bradyrhizobium sp em substratos alternativos com diferentes potenciais matriciais, foi conduzido u experimento utilizando-se como veículo de inoculaçäo os substratos: diatomia, vinhaça, seca, vermiculita e composto urbano em relaçäo à turfa. A inoculaçäo foi procedida com a estirpe UMKL-58, selecionada em trabalhos anteriores. Os substratos foram submetidos aos m de -0,33, -1,0 e -3,00 bar e incubados por 240dias. Para a qualificaçäo dos veiculos foram feitos testes de sobrevivência da estirpe nos materiais utilizando-se os métodos de diluiçäo em placas e de infecçäo em plantas coma leguminosa cunhä (Clitoria ternatea L.) nos intervalos de 0, 12, 30, 60, 90, 120, 150, 180 e 240dias após a inoculaçäo, como também, avaliaçäo da eficiência da fixaçäo biológica do N2 em funçäo do veículo de inoculaçäo. Os resultados demonstram que o Bradyrhizobium sobreviveu melhor na diatomita submetida a m -3,0 bar e nos demais potenciais foi comparável a turfa. Para todos os veículos, observou-se que altos valores de m reduzirama taxa de sobrevivência da bactéria como também afetaram a qualidade do inóculo diminuindo a efetividade da estirpe (au)