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Conditioning and Coating of Urochloa Brizantha Seeds Associated with Inoculation of Bacillus Subtilis1

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Conditioning and Coating of Urochloa Brizantha Seeds Associated with Inoculation of Bacillus Subtilis1 e-ISSN 1983-4063 - www.agro.ufg.br/pat - Pesq. Agropec. Trop., Goiânia, v. 49, e55536, 2019 Research Article Conditioning and coating of Urochloa brizantha seeds associated with inoculation of Bacillus subtilis1 Antônio Emilio Rodrigues Manrique2, Rita de Cássia Lima Mazzuchelli2, Ademir Sergio Ferreira Araujo3, Fabio Fernando de Araujo2 ABSTRACT RESUMO Condicionamento e revestimento de sementes de Urochloa Technological advances for improving the sowing stage brizanta associados à inoculação com Bacillus subtilis of forage plants are important because, besides increasing the operation efficiency, they may also positively affect the Avanços tecnológicos para melhorar a etapa de semeadura plant growth. This study aimed to evaluate the inoculation de forrageiras são importantes, pois, além de aumentarem a of Bacillus subtilis in Urochloa brizantha seeds, using eficiência na operação, podem proporcionar ganhos no crescimento coating, physiological conditioning and storage procedures, da planta. Objetivou-se avaliar a inoculação de Bacillus subtilis as well as to observe its effects on seed storage and plant em sementes de Urochloa brizantha, utilizando-se procedimentos growth. A completely randomized design, in a 2 × 3 × 5 + 1 de revestimento, condicionamento fisiológico e armazenamento, factorial scheme [two inoculation procedures (coating and bem como seus efeitos na germinação e no crescimento das conditioning), three inoculants (B. subtilis AP-3, B. subtilis plantas. Utilizou-se delineamento inteiramente casualizado, em PRBS-1 and without bacteria), five storage periods (0, 30, esquema fatorial 2 × 3 × 5 + 1 [dois procedimentos de inoculação 60, 90 and 120 days) and control (bare seeds)], with five (revestimento e condicionamento), três inoculantes (B. subtilis replicates, was used. Two lots of U. brizantha seeds were used, AP-3, B. subtilis PRBS-1 e sem bactérias), cinco períodos de characterized as being of higher and lower viability. After the armazenamento (0, 30, 60, 90 e 120 dias) e controle (sementes treatments, part of the seeds was employed to evaluate the plant nuas)], com cinco repetições. Foram utilizados dois lotes de sementes growth promotion in a greenhouse, during 180 days, while de U. brizantha, caracterizados como de maior e menor viabilidade. the remaining portion was stored at a temperature of 25 ºC, Após os tratamentos, parte das sementes foi utilizada para avaliar a with the seed germination being evaluated every 30 days, for promoção de crescimento de plantas em casa-de-vegetação, durante 120 days. The physiological conditioning showed the best 180 dias, e o restante foi armazenado sob temperatura de 15 ºC, performance in the evaluations carried out for U. brizantha com avaliação da germinação a cada 30 dias, durante 120 dias. O seeds with B. subtilis. The coating and conditioning of the condicionamento fisiológico apresentou melhor desempenho nas U. brizantha seeds with lower viability, both with and without avaliações efetuadas para sementes de U. brizantha com B. subtilis. the inoculation of bacteria, increases the shoot mass of plants O revestimento e o condicionamento nas sementes de U. brizantha after 180 days of growth. com menor viabilidade, com e sem inoculação de bactérias, aumenta a massa da parte aérea nas plantas após 180 dias de crescimento. KEYWORDS: Seed inoculation, rhizobacteria, germination. PALAVRAS-CHAVE: Inoculação de sementes, rizobactérias, germinação. INTRODUCTION high germination, what results in a vigorous seedling growth, and, in turn, this provides a suitable soil cover In Brazil, the area cultivated with forages and inhibits weed species. occupies about 150 million hectares (IBGE 2017), Adding value to Urochloa spp. seeds by using where the genus Urochloa spp. (Brachiaria) technology for seed processing, such as seed coating is present. However, agricultural practices for and conditioning, can provide a unique selling point improving the quantity and quality of forages are in an increasingly competitive market. Seed coating necessary to increase the animal productivity (Silva is the deposition of inert and adhesive materials on et al. 2008). Thus, grasses seeds should exhibit a the seed surface, with or without modification of 1. Received: Oct. 22, 2018. Accepted: June 06, 2019. Published: Oct. 10, 2019. DOI: 10.1590/1983-40632019v4955536. 2. Universidade do Oeste Paulista, Presidente Prudente, SP, Brasil. E-mail/ORCID: [email protected]/ 0000-0003-1455-6854, [email protected]/0000-0002-6299-9628, [email protected]/0000-0002-4614-9260. 3. Universidade Federal do Piauí, Teresina, PI, Brasil. E-mail/ORCID: [email protected]/0000-0002-3212-3852. 2 A. E. R. Manrique et al. (2019) the seed shape and size (Silva et al. 2002), while beneficial microorganisms to the seed through in conditioning seeds are soaked with a chemical either conditioning or coating may further enhance or biological solution for a certain period of time the establishment of the crop, particularly if the (Batista et al. 2016). Both techniques may be used applied microorganisms settle in the rhizosphere, to introduce chemical or biological materials that and subsequently contribute to the long-term plant can benefit seeds (Santos et al. 2010). The use of health or to the promotion of plant growth (Bennett & coated or conditioned seeds facilitates sowing by Whipps 2008). increasing the sanity of the seeds, what, in turn, leads Seed storage is a very important stage in to an improved seedling establishment, allowing the the seed production process and, in the case of incorporation of nutrients, growth regulators and U. brizantha, other aspects should also be explored, other inputs (Silva et al. 2002, Bonome et al. 2017). such as dormancy and seed viability (Novembre et However, Santos et al. (2011) state that techniques al. 2006). In addition, it is necessary to evaluate if the such as seed coating, despite being a method used introduction of chemical and biological agents may to increase the pasture productivity, may cause affect the germination and emergence speed of the inhibition in the germination of Urochloa brizantha. U. brizantha seedlings (Santos et al. 2010). The plant growth-promoting rhizobacteria that Thus, this study aimed to evaluate the effect of inhabit the soil are isolated from the rhizosphere of the inoculation of B. subtilis on U. brizantha seeds several cultivated plants. Among the most studied using either coating or osmotic conditioning, as well genera are Bacillus, Pseudomonas, Azospirillum as the effects of the inoculation on germination, plant and Rhizobium. The effects of these microorganisms development and seed storage. on the plant development are broad and include beneficial effects on seed germination, seedling MATERIAL AND METHODS emergence and plant growth (Figueiredo et al. 2010). There are few studies about the inoculation The experiments were conducted from of Brachiaria seeds with plant growth-promoting September 2016 to June 2017. Non-scarified seeds of rhizobacteria. For instance, Araujo et al. (2012a) U. brizantha (Hochst ex A. Rich.) Stapf. ‘Marandu’ concluded that the inoculation of Bacillus sp. in were used from two lots (1 and 2) with lower and seeds of U. brizantha promotes the growth of higher viability, respectively, based on a tetrazolium plants. In a study carried out with other grasses, the test carried out previously (Novembre 2006). Two production of hormones by rhizobacteria changed strains of B. subtilis (PRBS-1 and AP-3) isolated the root morphology and increased the biomass, from an agricultural soil and described in Araujo et thus increasing the soil exploration capacity (Malik al. (2005) were used in this study. et al. 1997). Araujo (2008) evaluated the inoculation For the seed treatment, initially, B. subtilis of Bacillus subtilis in different plant species and was multiplied in a culture medium for five days. showed that the inoculation significantly increased Afterwards, the cells were separated by centrifugation the nitrogen content in maize leaf tissue, even though (5,000 × g) and resuspended in water. All suspensions the introduced bacterial species is not known as were adjusted to 108 colony forming units (CFU) per diazotrophic. In addition to effects on plant growth, mL. Initially, the seed coating was carried out using other benefits related to the inoculation ofB. subtilis 200 g of U. brizantha seeds and 50 mL of bacterial may be found in disease management, such as cell suspension with 0.75 g of methyl cellulose, which nematode control (Morgado et al. 2015). were mixed in a rotary drum. During mixing, 200 g of When inoculating microorganisms in seeds, it inert talc were added to impart the uniformity in the is important to ensure that they have enough bacteria coating. To perform the osmotic conditioning of the for a greater process efficiency. Following this, a seeds, 200 g of seeds were packed in an Erlenmeyer study was conducted, wherein several techniques flask and immersed with 50 mL of bacterial cell for bacterial inoculation in seeds were evaluated, suspension with 0.425 g of sodium chloride. The vials such as the use of suspensions, pastes, peat or were stirred (120 rpm) for 12 h, at 25 ºC. Finally, encapsulation (Walker et al. 2004). Seed coating the seeds were dried for 24 h, at 30 ºC. In parallel, or conditioning techniques can ensure the highest treatments with seed coating and conditioning were concentrations of bacteria in the seed. Applying conducted using sterilized water, instead of bacterial e-ISSN 1983-4063 - www.agro.ufg.br/pat - Pesq. Agropec. Trop., Goiânia, v. 49, e55536, 2019 3 Conditioning and coating of Urochloa brizantha seeds associated with inoculation of Bacillus subtilis 3 suspensions, and these were used as control. All final cutting of the plants. Roots were also collected treated seeds were stored under a temperature of and washed, and both were oven dried at 65 ºC, to 15 ºC, for 120 days. obtain the dry mass. The moisture in the seed environment was Analyses of variance and F-tests at 5 % of controlled, and 15 % of humidity for conditioning significance were used to estimate and test the effects and 12 % for coating (Brasil 2009) were maintained.
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