ABSTRACT Sugar and alcohol production in Brazil comes from planting hybrids from the crossing of species belonging to the Saccharum Complex. However, little is known about the growth response of these genera to inoculation using a diazotrophic bacteria Nitrospirillum viridazoti strain BR11145, currently recommended in Brazil as an inoculant for sugarcane growth promotion. This study aimed to evaluate the initial growth of 12 sugarcane genotypes inoculated or not with the strain BR11145. After 109 days, plant growth, biomass and macronutrient accumulation were evaluated. Fresh and dry mass at the end of the trial showed a positive response to inoculation for the genotypes US72-1319, CPDAU 849678, and NG77-122 of S. spontaneum and for Fiji 10 of Miscanthus. A negative response was observed for Q45416 of Saccharum sp. The growth of S. spontaneum US72-1319 is significantly improved by the inoculation with BR11145, which leads to a high accumulation of nutrients, especially N, standing out among the genotypes tested.

ABSTRACT Breeding of new cultivars of sugarcane uses several species belonging to the Saccharum complex genus. In this study, a set of 20 genotypes from the Saccharum complex was assessed for their growth promotion during germination using a mixed inoculant containing five selected diazotrophs and their resistance to short heat treatment (52°C / 30 min). The percentage of germination was evaluated, leading to the identification of three response groups. Saccharum officinarum (group I) showed the highest germination percentage, succeeded by Erianthus arundinaceus genotypes (group II) and the other genotypes (group III). The mean germination time (MGT) varied among the genotypes and segregated the growth response into two groups. Saccharum officinarum only required less than 15 days to reach the MGT, whereas the other genera required more than two or three days during the initial growth process. Bacterial counting also differed between the groups, being effective for S. officinarum and other genotypes, improving the colonization of the plants 100–1,000 times higher than the control and heat treatments. Further comparison was made among eight genotypes, including one Saccharum spontaneum(US72-1319), one S. officinarum (IN845), and six E. arundinaceus. This comparison measured biomass accumulation and root parameters. The E. arundinaceus group demonstrated less susceptibility to the heat treatment, resulting in stimulation of the root dry mass of IJ76-381 and root parameters of this genotype, as well as IJ76-364, although not associated to the inoculation treatment. Inoculation applied to US72-1319 and IN845 enhanced only the Shoot/Root ratio.

ABSTRACT Inoculating corn with diazotrophic bacteria as growth promoters has been demonstrated to be an efficient agricultural practice in Brazil, mainly due to the root stimulation they provide to plants. This study investigates the corn (Zea mays L.) root architecture in a greenhouse assay where A. baldaniorum Sp245 and H. seropedicae ZAE94 strains were inoculated and evaluated for 22 days under two N levels: 0.6 and 6 mmol L-1 of N. Short-term bioassays were conducted to assess the plant’s response to the addition of indole-acetic acid, two bacterial populations, and two corn genotypes, utilizing image capture software WinRhizo Pro. The growth and distribution of tips, crossing, and length of fine roots were determined to be the most sensitive aspects to inoculation and indole-acetic acid induction. These responses were influenced by the genotype and the number of bacterial cells present. Biomass accumulation analyses quantified these modifications after a 22-day period. Additionally, the growth response was found to be more significant when applying the Hs-ZAE94 strain to plants fertilized with a higher dose of nitrogen (6.0 mmol L-1), and this response was positively correlated with bioassay data. Selected strains used as an inoculant can improve root architecture and, consequently, the N use efficiency.

ABSTRACT Plant-growth-promoting bacteria of the genus Azospirillum and Herbaspirillum can improve crop yields of maize in tropical environments. The main mechanism proposed for its promotion is related to the auxins produced by these species. The aim of this study was to compare the maize growth response of inoculation using two species, Azospirillum baldaniorum Sp245 (Ab-Sp245) and Herbaspirillum seropedicae ZAE94 (Hs-ZAE94) during the initial growth under controlled conditions. A pot experiment was conducted with five harvests measuring bacterial counts, root morphology, biomass, and nitrogen content, and soluble metabolites for 38 days. Depending on the harvest period, a differential growth response between the two strains test was compared to the control. Plants use the bacterium applied to improve nitrogen acquisition and transform it into higher biomass and nitrogen accumulation, especially for Ab-Sp245. On the other side, Hs-ZAE94 altered the soluble metabolites resulting in higher NO3-, NH4+, and N-amino and lower soluble fractions compared to Ab-Sp245 and the control. The observed plant-bacteria interaction is closely fine-tuned and regulated by the strain used and differences in growth promotion effects cannot solely attributed to the root morphology.

Abstract The objective of this work was to evaluate inoculation response to Azospirillum baldaniorum strain Sp245 in 14 Urochloa cultivars. The evaluated cultivars were: Marandú, Xaraés, Paiaguás, Piatã, Basilisk, B140, Comum, Tupi, H47, Llanero, Ruziziensis, Ipyporã, Mulato, and Convert HD364. Three experiments were conducted to measure root traits: two using soil and one a sterile substrate. Compared with the control, a single application of A. baldaniorum Sp245 increases dry matter accumulation and number of roots in cultivars Marandú, Llanero, Ruziziensis, and Convert HD364. However, after inoculation, cultivars Xaraés and Ipyporã show some root traits with lower values.

Resumo O objetivo deste trabalho foi avaliar a resposta à inoculação da estirpe Sp245 de Azospirillum baldaniorum em 14 cultivares de Urochloa. As cultivares avaliadas foram: Marandú, Xaraés, Paiaguás, Piatã, Basilisk, B140, Comum, Tupi, H47, Llanero, Ruziziensis, Ipyporã, Mulato e Convert HD364. Foram conduzidos três experimentos para mensuração de parâmetros radiculares: dois em solo e um em substrato estéril. Em comparação ao controle, a aplicação única de A. baldaniorum Sp245 aumenta o acúmulo de massa seca e a quantidade de raízes das cultivares Marandú, Llanero, Ruziziensis e Convert HD364. No entanto, após a inoculação, as cultivares Xaraés e Ipyporã apresentam alguns parâmetros radiculares com valores menores.

ABSTRACT Little is known about the interaction between the inoculation of diazotrophic bacteria, the variety, and the substrate used in inoculated sugarcane seedling production. Bearing this in mind, the aim of this study was to evaluate the effects of diazotrophic bacteria inoculation, four sugarcane varieties and four different substrates on the growth and nitrogen accumulation of sugarcane seedlings. Parameters related to sprouting, height, and root growth, as well as nitrogen accumulation, were evaluated. The results indicate that inoculating sugarcane seedlings belonging to the following varieties: RB867515, RB92579, RB966928 and RB975932 with bacteria may increase sprouting speed index, height, root length, fresh and dry matter weight, as well as nitrogen accumulation, which has resulted in a more uniform stem diameter. However, the responses to inoculation differ from variety to variety and are also dependent on the substrate used with better results observed in the commercial substrate and the substrate mixtures containing a higher proportion of organic compost. Understanding the interaction between the variety and the substrate with the bacterial inoculation is essential to the success of the production of inoculated sugarcane seedlings.

RESUMO Aumentar a produtividade de milho em condições de clima e solo do nordeste brasileiro podem ser atingida com a utilização de insumos biológicos acoplado a aplicação de menores doses de N -fertilizante. O objetivo deste trabalho foi avaliar o rendimento de grãos de milho inoculado com duas espécies de bactérias diazotróficas combinadas com doses de N-fertilizante durante três anos consecutivos em um Cambissolo no estado de Sergipe, Brasil. Duas espécies de bactérias foram utilizadas: Azospirillum brasilen se estirpe BR11005 (Sp245) e Herbaspirillum seropedicae BR11147 (ZAE94) e as doses crescentes de ureia de 0, 100, 200 e 300 kg ha-1. Sob condições de campo houve limitação do rendimento dependendo do ano agrícola e regime hídrico. A inoculação, associada a uma dose de 100 kg ha-1 N com as estirpes BR11147 e BR11005, aumentou a produtividade em 1.230 e 614 kg ha-1, respectivamente; sendo 37% e 19% maior que o controle não inoculado. Na ausência do fertilizante também foi observado incremento da produtividade no milho inoculado com BR11147 de 18% e no teor de N na folha índice pela inoculação da BR11145 em 10% comparado ao controle.

ABSTRACT Increasing the crop yield of maize planted in the climate and soil conditions of the northeast region of Brazil can be accomplished with the use of biological inputs coupled with the application of lower doses of nitrogen (N) fertilizer. The objective of this work was to evaluate the yield of maize inoculated with two diazotrophic bacterial species under different N rates, during three consecutive years in an Inseptisol of Sergipe state, Brazil. The two bacterial species used were Azospirillum brasilense BR11005 (Sp245) and Herbaspirillum seropedicae BR11147 (ZAE94). N was applied in the form of urea at 0, 100, 200, and 300 kg ha-1. Under field conditions and depending on the year, the crop yield was limited by the rainfall regime. The inoculation associated with the 100 kg ha-1 N treatment with BR11147 and BR11005, increased productivity by 1,230 kg ha-1 and 614 hg ha-1, respectively. This corresponded to a 37% and 19% productivity increase, respectively, compared to the productivity of the uninoculated control. At zero N, inoculation increased grain yield of maize plants inoculated with BR11147 by 18%. Additionally, the N content of the index leaf inoculated with BR11005 increased by 10% over that of the uninoculated control.

Resumo O objetivo deste trabalho foi avaliar a produtividade e a diluição do isótopo 15N de duas variedades de cana-de-açúcar, inoculadas ou não com bactérias diazotróficas. Dois experimentos foram realizados nas regiões Sudeste e Nordeste do Brasil, durante três safras (2007/2008, 2008/2009 e 2009/2010): o primeiro em Argissolo Vermelho eutrófico, de textura argilosa, e o segundo em Planossolo Háplico distrófico, de textura média. Utilizou-se o delineamento de blocos ao acaso, com quatro repetições. As variedades testadas foram a RB867515 e a RB72454, cultivadas com inoculação com bactérias diazotróficas (cinco estirpes combinadas no inoculante para cana-de-açúcar) ou com adubação de 120 kg ha-1 de N, além do controle experimental sem inoculação e sem adubação. Avaliaram-se a produtividade de colmos e a diluição isotópica de 15N nas folhas-bandeira. A variedade RB72454 foi mais responsiva aos tratamentos, nos ciclos de cana-planta e na segunda soca. A abundância natural de 15N nas folhas-bandeira diminui ao longo dos anos, o que indica aumento da fixação biológica de N na medida em que a disponibilidade de N do solo vem sendo reduzida.

Abstract The objective of this work was to evaluate the productivity and 15N isotope dilution of two sugarcane varieties inoculated or not with diazotrophic bacteria. Two experiments were carried out in the Southeast and Northeast regions of Brazil, during three cropping seasons (2007/2008, 2008/2009 e 2009/2010): the first one on a clayey Acrisol, and the second one on a medium-textured Planosol. A randomized complete block design, with four replicates was used. The varieties RB867515 and RB72454 were tested with inoculation of diazotrophic bacteria (five strains combined in the inoculant for sugarcane) or fertilization with 120 kg ha-1 N, besides the control treatment without inoculation and nitrogen fertilization. Stalk yield and 15N isotope dilution of the flag-leaves were evaluated. The RB72454 variety showed greater response to the treatments during the plant-cane cycle and the second ratoon. 15N natural abundance in the flag-leaves decreases over the years, which suggests the increase of biological N fixation when N availability in soil reduces.

Resumo: O objetivo deste trabalho foi avaliar o efeito de nitrogênio e da inoculação de Herbaspirillum seropedicae sobre a produtividade de milho (Zea mays) e os teores de nutrientes nos grãos. Os híbridos simples BRS 1030 e BRS 1060 receberam inoculação da estirpe BR 11417, na presença ou não de doses de adubação nitrogenada, em Argissolo Vermelho-Amarelo distrófico, durante os períodos de entressafra (plantio em maio de 2012) e safra (plantio em outubro de 2012). Em cada época, os seguintes tratamentos foram avaliados: controle absoluto, sem adubação nitrogenada nem inoculação; controle com inoculação; doses de 50 e 100 kg ha-1 de N, sem inoculação; e dose de 50 kg ha-1 de N mais inoculação. BRS 1030 produziu 1.157 kg ha-1 a mais de grãos que BRS 1060, na análise conjunta dos dois cultivos. Para o primeiro genótipo, a dose de 50 kg ha-1 de N, com inoculação, incrementou em 2% a produtividade na entressafra e em 4,5% na safra. A inoculação de H. seropedicae, estirpe BR 11417, favorece o acúmulo de P nos grãos, mas tem efeito positivo sobre a produtividade somente em combinação com doses de N mineral, o que indica que seu efeito é mais destacado na promoção do crescimento do milho do que na fixação biológica de N.

Abstract: The objective of this work was to evaluate the effect of nitrogen and inoculation with Herbaspirullum seropedicae on maize (Zea mays) yield and nutrient contents in grains. The BRS 1030 and BRS 1060 simple hybrids received inoculation with the BR 11417 strain in the presence or absence of nitrogen-fertilizer levels in a Typic Hapladult, during the periods of off-season (sowing in May 2012) and season (sowing in October 2012). In each season, the following treatments were evaluated: absolute control, without nitrogen fertilization or inoculation; inoculated control; doses of 50 and 100 kg ha-1 N, without inoculation; and dose of 50 kg ha-1 N plus inoculation. BRS 1030 produced 1,157 kg ha-1 more grains than BRS 1060, in the joint analysis of both crop seasons. For the first genotype, the dose of 50 kg ha-1 N with inoculation increased crop productivity by 2% in the off-season and by 4.5% in the season. The inoculation with H. seropedicae, BR 11417 strain, favors P accumulation in grains, but it has positive effect on yield only in combination with mineral N levels, which indicates that its effect is more important for maize growth promotion than for biological N fixation.

RESUMO A cana-de-açúcar é uma cultura de grande destaque na economia, em razão da produção de açúcar, etanol e energia. Tecnologias que possam contribuir para o aumento da produtividade e qualidade da cultura com mínimos danos ao meio ambiente são necessárias. Objetivou-se avaliar a produção de ácido indol acético de cinco estirpes de bactérias diazotróficas e o efeito da inoculação delas na brotação de duas variedades de cana-de-açúcar, RB867515 e IACSP95-5000. A produção de auxina foi determinada pelo teste colorimétrico, usando o reagente de Salkowski. Para avaliar a germinação, foi conduzido um experimento em casa de vegetação, utilizando-se delineamento experimental em blocos ao acaso com quatro repetições e sete tratamentos: controle não inoculado; inoculação mista com as cinco estirpes e inoculação individual com Gluconacetobacter diazotrophicus (Gd) estirpe BR11281T(PAL-5T), Herbaspirillum seropedicae (Hs - BR11335 = HRC54), Herbaspirillum rubrisubalbicans (Hr - BR11504 = HCC103), Burkholderia tropica (Bt - BR11366T = PPe 8 T) e Azospirillum amazonense (Aa - BR11145 = CBAMc). As bactérias mais eficientes na produção de auxina foram Hs e Hr, declinando 48 h após o crescimento. Hr, Aa e Bt aumentaram o índice de velocidade de germinação e o número de brotações nas duas variedades. Na var. RB867515, a velocidade de germinação ainda foi positivamente influenciada pela inoculação mista, sendo o mesmo observado pela inoculação de Gd na var. IACSP95-5000.

ABSTRACT Sugarcane is one of the most important crops in the economy due to production of sugar, ethanol, and energy. Technologies are necessary to improve its yield and quality with minimum damage to the environment. We evaluated the indole acetic acid production and germination in the sugarcane varieties RB867515 and IACSP95-5000 after inoculation with five strains of diazotrophic bacteria. The production of indole acetic acid was assayed through the colorimetric method using the Salkowski reagent. To evaluate germination, an experiment was conducted in a greenhouse using a randomized block experimental design with four replications and seven treatments: uninoculated control; mixed inoculation with the five strains; and individual inoculation with Gluconacetobacter diazotrophicus (Gd) estirpe BR11281T (PAL-5T), Herbaspirillum seropedicae (Hs - BR11335 = HRC54), Herbaspirillum rubrisubalbicans (Hr - BR11504 = HCC103), Burkholderia tropica (Bt - BR11366T = PPe 8 T) e Azospirillum amazonense (Aa - BR11145 = CBAMc). The most efficient bacteria for production of indol acetic acid were Hs and Hr, and production declined after 48 h of growth. Hr, Aa, and Bt also improved the germination speed index and number of stalks sprouted in both varieties. Furthermore, in the var. RB867515, germination speed was positively affected by the mixed inoculation, and the same effect was observed from Gd in the var. IACSP95-5000.

O objetivo deste trabalho foi avaliar os efeitos da inoculação de bactérias promotoras de crescimento sobre a formação de mudas pré-brotadas de cana-de-açúcar, oriundas de gemas individualizadas, e quantificar o crescimento inicial dessas mudas, em associação à aplicação de nitrogênio, em solo de baixa fertilidade. Foram conduzidos dois experimentos: um em casa de vegetação, com duração de 50 dias, e o outro, em vasos no campo, com duração de 180 dias. Em ambos os experimentos, utilizou-se o delineamento de blocos ao acaso, em arranjo fatorial 2x3, no primeiro experimento - com ou sem inoculante, e com três quantidades de reserva nas gemas -, e 2x2x4, no segundo - com ou sem inoculante, com ou sem nitrogênio, avaliados em quatro épocas: aos 45, 90, 135 e 180 dias. O inoculante produziu efeito na fase inicial de crescimento das mudas pré-brotadas, com aumento na velocidade de brotação e no acúmulo da matéria seca de raízes e da parte aérea, independentemente da quantidade de reserva da gema. No segundo experimento, o inoculante promoveu ganhos no crescimento inicial da parte aérea e do sistema radicular, até os 180 dias após o transplantio, com aumento em altura, perfilhamento, diâmetro do colmo, produção da matéria seca de colmos e de palha e do comprimento radicular, independentemente da aplicação de nitrogênio. O inoculante tem efeito fisiológico positivo sobre o crescimento das plantas.

The objective of this work was to evaluate the effects of the inoculation with plant growth-promoting bacteria on the formation of sugarcane pre-sprouted seedlings, originated from individual buds, and to quantify the initial growth of these seedlings in association with nitrogen application, in a low fertility soil. Two experiments were carried out: one in greenhouse conditions, with a time span of 50 days, and the other in vases under field conditions, with a time span of 180 days. In both experiments, a randomized complete block design was used with a factorial arrangement of 2x3, in the first experiment - with or without inoculation, and with three quantities of bud reserve -, and of 2x2x4, in the second one - with or without inoculation, with or without nitrogen, evaluated in four times: at 45, 90, 135, and 180 days. The inoculant had effect on the initial growth of the pre-sprouted seedlings, increasing sprouting speed and dry matter accumulation on roots and shoots, regardless of the bud reserve amount. In the second experiment, the inoculant increased the initial growth of shoots and root system up to 180 days after transplantation, increasing height, tillering, stalk diameter, dry matter production of stalks and straw, and root length, regardless of nitrogen application. The inoculant has a positive physiological effect on plant growth.

O aumento da eficiência produtiva da cana-de-açúcar com tecnologias que possibilitem aumentar seu balanço energético é uma necessidade do setor sucroenergético, por se tratar de uma cultura estratégica para o país. Entre as diversas alternativas atualmente estudadas, o inoculante com bactérias diazotróficas vem se evidenciando uma tecnologia com elevado potencial. Diante desse contexto, este estudo teve como objetivo avaliar a eficiência do inoculante bacteriano no desempenho agronômico e na nutrição nitrogenada da cana-de-açúcar. O ensaio foi realizado no campo experimental da Embrapa Agrobiologia, em Seropédica, RJ. O delineamento experimental foi em bloco ao acaso, em esquema fatorial 2 × 3 (duas variedades e três tratamentos), com quatro repetições, totalizando 24 parcelas. As variedades utilizadas foram a RB867515 e RB72454. Os tratamentos foram: inoculação com bactérias diazotróficas, controle nitrogenado com 120 kg ha-1 de N e controle absoluto (sem inoculação e sem N). O inoculante foi composto por cinco estirpes de cinco espécies de bactérias diazotróficas. As variáveis avaliadas foram a produtividade de colmos, o acúmulo de matéria seca e N total da parte aérea das plantas e a contribuição da fixação biológica de N (FBN), utilizando como referência a abundância natural de 15N da própria cana-de-açúcar não inoculada. O inoculante bacteriano aumentou a produtividade de colmos na variedade RB72454 de maneira similar à adubação com 120 kg ha-1 de N, nas colheitas de cana-planta e primeira soqueira; no entanto, não houve modificação na contribuição da FBN advinda da inoculação, dando evidências de que os benefícios do inoculante para a cana-de-açúcar podem ser provenientes do efeito da promoção de crescimento de plantas.

The sugarcane industry, a strategic crop in Brazil, requires technological improvements in production efficiency to increase the crop energy balance. Among the various currently studied alternatives, inoculation with diazotrophic bacteria proved to be a technology with great potential. In this context, the efficiency of a mixture of bacterial inoculant was evaluated with regard to the agronomic performance and N nutrition of sugarcane. The experiment was carried out on an experimental field of Embrapa Agrobiologia, in Seropédica, Rio de Janeiro, using a randomized block, 2 × 3 factorial design (two varieties and three treatments) with four replications, totaling 24 plots. The varieties RB867515 and RB72454 were tested in treatments consisting of: inoculation with diazotrophic bacteria, N-fertilized control with 120 kg ha-1 N and absolute control (no inoculation and no N fertilizer). The inoculum was composed of five strains of five diazotrophic species. The yield, dry matter accumulation, total N in the shoot dry matter and the contribution of N by biological fixation were evaluated, using the natural 15N abundance in non-inoculated sugarcane as reference. The bacterial inoculant increased the stalk yield of variety RB72454 similarly to fertilization with 120 kg ha-1 N in the harvests of plant-cane and first ratoon crops, however the contribution of biological N fixation was unchanged by inoculation, indicating that the benefits of the inoculant in sugarcane may have resulted from plant growth promotion.

O objetivo do estudo foi avaliar a contribuição da inoculação com bactérias diazotróficas, aplicadas individualmente e em mistura, em variedades comerciais de cana-de-açúcar. O experimento f oi conduzido no Campo Experimental da Embrapa Agrobiologia, em Seropédica-RJ, Brasil. O delineamento experimental foi em blocos casualizados, com parcelas subdivididas e com seis repetições. Foram utilizadas seis variedades de cana-de-açúcar e seis tratamentos: controle absoluto, controle nitrogenado, inoculação individual das estipes: BR 11512, BR 11724 e BR 11411; além de inoculação com um coquetel composto por cinco estirpes de bactérias diazotróficas. Foi observada diferença entre variedades, entre tratamentos e uma interação entre tratamentos e variedades. Para todos os parâmetros avaliados não foi observado resposta nas variedades de cana-de-açúcar RB855536 e RB92606. Na variedade RB918639, houve apenas diferença no acúmulo de biomassa verde. Não foi observada nenhuma diferença entre tratamentos no acúmulo de nitrogênio total do ponteiro. A variedade mais responsiva à inoculação foi a RB867515, diferenciando no acúmulo de biomassa seca e fresca. Este estudo mostrou que a inoculação promove o acúmulo de biomassa, sendo a contribuição diferente entre variedades e estirpes de bactérias, sugerindo uma interação entre os fatores estudados. A variedade RB867515 é promissora para os estudos de inoculação com bactérias diazotróficas.

The objective of this study was to evaluate the contribution of inoculation with diazotrophs, applied both individually and in combination, in commercial varieties of sugarcane. The experiment was carried out at the experimental grounds of Embrapa Agrobiologia, in Seropédica, Rio de Janeiro, Brazil. The experimental design was of randomised blocks, with sub-divided plots and six replications. Six varieties of sugarcane with six treatments were used: control, control with nitrogen, inoculation of the individual strains: BR 11512, BR 11724 and BR 11411, in addition to inoculation with a cocktail consisting of five strains of diazotrophs. Differences were observed between varieties and between treatments, and an interaction observed between the treatments and varieties. For all the parameters evaluated, no response was observed in the sugarcane varieties RB855536 and RB92606. In the variety RB918639, the only difference was in the accumulation of green biomass. No difference was observed between the treatments for total nitrogen accumulation. The most responsive variety to inoculation was RB867515, differing in the accumulation of fresh and dry biomass. This study showed that inoculation promotes the accumulation of biomass, the contribution being different for the varieties and strains of bacteria, suggesting an interaction between the factors studied. The variety RB867515 is promising for use in studies of inoculation with diazotrophs.

O objetivo deste trabalho foi avaliar a eficiência da inoculação de bactérias diazotróficas e da adubação nitrogenada, em duas variedades de cana-de-açúcar, cultivadas nas mesmas condições edafoclimáticas. O experimento foi conduzido durante os anos agrícolas de 2006/2007 e 2008/2009, em delineamento experimental de blocos ao acaso, com quatro repetições, instalado em março de 2006 em área de cultivo comercial, no Município de Campos dos Goytacazes, RJ. Os tratamentos foram: inoculação com bactérias diazotróficas, adubação com 120 kg ha-1 de N, e o controle sem inoculação e sem adubação com nitrogênio. As variedades de cana-de-açúcar avaliadas foram RB72454 e RB867515. O inoculante continha estirpes de cinco espécies de bactérias diazotróficas. Foram feitas avaliações quanto à produtividade de colmos frescos, ao acúmulo de matéria seca total, ao N total da parte aérea e quanto à abundância natural de 15N do N disponível no solo e na cana-de-açúcar. As variedades apresentaram comportamentos distintos com os tratamentos, em que a RB867515 foi responsiva e a RB72454 não responsiva à inoculação e à adubação nitrogenada. Na variedade RB867515, o crescimento e o acúmulo de N total na parte aérea das plantas, promovidos pela inoculação, foram similares aos do tratamento com adubação nitrogenada.

The objective of this work was to evaluate the efficiency of inoculation with diazotrophic bacteria and nitrogen fertilization, in two varieties of sugarcane grown under the same edaphic and climatic conditions. The experiment was carried out during the agricultural years 2006/2007 and 2008/2009, in a randomized block design with four replicates, installed in March 2006 in an area of commercial cultivation in Campos dos Goytacazes city, in Rio de Janeiro state, Brazil. The treatments were: inoculation with diazotrophic bacteria, fertilization with 120 kg ha-1 N, and a control without inoculation and without nitrogen fertilization. The sugarcane varieties evaluated were RB72454 and RB867515. The inoculant was composed of five strains of diazotrophic bacteria. Evaluations were done for fresh stalk yield, total dry matter accumulation, total N of shoots, and 15N natural abundance from available N in the soil and in sugarcane. The varieties performed differently to treatments, in which RB867515 was responsive and RB72454 unresponsive to inoculation and nitrogen fertilization. In the variety RB867515, growth and accumulation of total N in the shoots, promoted by inoculation, were similar to the ones in the treatment with N fertilization.

The species Gluconacetobacterdiazotrophicus, Herbaspirillum seropedicae and H. rubrisubalbicans are endophytic N2-fixing [diazotrophic] bacteria which colonise not only roots, but also the aerial tissue of sugar cane. However, the technique most commonly used to quantify the populations of these microbes in plants is by culturing serial dilutions of macerates of plant tissues in N free semi-solid media which are only semi-selective for the species/genera [the Most Probable Number (MPN) Technique] and each culture must be further subjected to several tests to identify the isolates at the species level. The use of species-specific polyclonal antibodies with the indirect ELISA (enzyme-linked immunosorbent assay) can be an alternative which is rapid and specific to quantify these populations of bacteria. This study was performed to investigate the viability of adapting the indirect ELISA technique to quantify individually the populations of these three species of diazotroph within the root and shoot tissues of sugarcane. The results showed that species-specific polyclonal antibodies could be obtained by purifying sera in protein-A columns which removed non-specific immuno-globulins. It was possible to quantify the three bacterial species in the Brazilian sugarcane variety SP 70-1143 in numbers above 10(5) cells per g fresh weight in roots, rhizomes and leaves. The numbers of the different bacterial species evaluated using the ELISA technique were found to be higher than when the same populations were evaluated using the MPN technique, reaching 1400 times greater for G. diazotrophicus and 225 times greater for Herbaspirillum spp. These results constitute the first quantification of Herbaspirillum using immunological techniques.