Optimization of oxalic acid production by fungi for biotechnological solubilization of rock phosphate

This study describes a biotechnological strategy for producing and applying oxalic acid to solubilize phosphorus (P) from rock phosphate (RP). We evaluated six fungal species (Aspergillus niger FS1, Penicillium islandicum FS41, Pleurotus ostreatus PO1, Rhizoctonia solani Rhiz555, Sclerotium rolfsii Sr25, and Sclerotinia sclerotiorum Scl134) and three culture media (potato dextrose broth, Tsao and Strasser media) to maximize oxalic acid production. Among the fungal isolates tested and culture media, S. rolfsii Sr25 and Tsao medium showed efficient oxalic acid production. Tsao medium was optimized following a response surface methodology after initial screening of factors affecting RP solubilization. The optimized concentrations were 1 g L-1 NaNO3, 100 g L-1 glucose, 2 g L-1 KH2PO4, 4.5 g L-1 yeast extract, and 25 mg L-1 MgSO4·7H2O used for 20 days of incubation. Under these conditions, 71 mmol L-1 oxalic acid was obtained, representing a three-fold increase over production under non-optimized conditions (20 mmol L-1). Under optimized conditions, oxalic acid produced by S. rolfsii Sr25 reacted with low-solubility RP and solubilized 100 % of the P contained in ore. Thus, using S. rolfsii Sr25 to produce oxalic acid seems a promising biotechnological alternative for P solubilization from RP.(AU)

Endophytic bacteria Bacillus safensis and Pseudomonas hibiscicola and their ability to increase rice seedling growth

Endophytic bacteria Bacillus safensis RS95 and Pseudomonas hibiscicola RS121 were evaluated for their ability to promote the growth of rice seedlings and produce indole-acetic acid (IAA) and siderophores and to solubilize phosphates. 'Guri' rice seeds were immersed in bacterial endophyte cell suspensions (separated and two-strain mixed), as well as in Escherichia coli DH5α, phosphate-buffered saline (PBS) and water treatments (negative controls). Seeds were sown on agar-water in Petri plates placed vertically at an angle of 65°. The ability of plant growth-promoting endophytic bacteria (PGPEB) to produce IAA and siderophores was determined by Salkowski colorimetric and chrome azurol S (CAS) assays, respectively. Mineral phosphate solubilization activity was calculated by inoculating the endophytes onto medium containing insoluble phosphate. PGPEB showed a positive effect on the growth of rice seedlings, causing a mean growth of shoots and primary-roots of 60 and 67%, respectively. Bacterial strains also showed positive traits for IAA and siderophore production, as well as phosphate-solubilization activity

Ornamental plant Pachystachys lutea as a source of promising endophytes for plant growth and phytoprotective activity

Endophytes are growth-promoting agents capable of synthesizing phytohormones, uptaking nutrients, and controlling pathogens. There is a strong potential to exploit them in the agriculture field like biofertilizers and biocontrol agents. In this work, we aimed to evaluate endophytic fungi isolated from Pachystachys lutea for their potential to solubilize phosphate, synthesise indole acetic acid (IAA), antagonize phytopathogens, and promote plant growth under greenhouse conditions. The phosphate solubilization efficiency was assessed on Pikovskayas agar medium. For analysis of IAA production, mycelia plugs of endophytes were cultured in Potato Dextrose Broth medium supplemented with L-tryptophan, with Salkowski Reagent, and the absorbance of the culture was measured. The antagonism evaluation of strain Alternaria sp. PL75 against phytopathogens was performed using the paired-culture technique. The promotion of plant growth provided by Alternaria sp. PL75 was evaluated in tomato plants. All strains evaluated were able to solubilize phosphate; however, the strain Alternaria sp. PL75 was the most effective (4.29). Two strains, Nemania sp. PL27 and Alternaria sp. PL75, produced 1.86 and 1.73 & 956;g mL-1 of IAA, respectively. In the antagonism assay, the endophyte Alternaria sp. PL75 and its fungal extract showed the best results against the pathogen Moniliophthora perniciosa. The greenhouse experiment result showed the endophyte Alternaria sp. PL75 increased the plantlets emergency speed index and the percentage of germination from 60 to 81.63%. It was also observed a statistical significance in the shoot length of the treated plants with the endophyte suspension (55.38 cm) compared to the control (41.67 cm).(AU)

Ornamental plant Pachystachys lutea as a source of promising endophytes for plant growth and phytoprotective activity

Endophytes are growth-promoting agents capable of synthesizing phytohormones, uptaking nutrients, and controlling pathogens. There is a strong potential to exploit them in the agriculture field like biofertilizers and biocontrol agents. In this work, we aimed to evaluate endophytic fungi isolated from Pachystachys lutea for their potential to solubilize phosphate, synthesise indole acetic acid (IAA), antagonize phytopathogens, and promote plant growth under greenhouse conditions. The phosphate solubilization efficiency was assessed on Pikovskaya’s agar medium. For analysis of IAA production, mycelia plugs of endophytes were cultured in Potato Dextrose Broth medium supplemented with L-tryptophan, with Salkowski Reagent, and the absorbance of the culture was measured. The antagonism evaluation of strain Alternaria sp. PL75 against phytopathogens was performed using the paired-culture technique. The promotion of plant growth provided by Alternaria sp. PL75 was evaluated in tomato plants. All strains evaluated were able to solubilize phosphate; however, the strain Alternaria sp. PL75 was the most effective (4.29). Two strains, Nemania sp. PL27 and Alternaria sp. PL75, produced 1.86 and 1.73 & 956;g mL-1 of IAA, respectively. In the antagonism assay, the endophyte Alternaria sp. PL75 and its fungal extract showed the best results against the pathogen Moniliophthora perniciosa. The greenhouse experiment result showed the endophyte Alternaria sp. PL75 increased the plantlets emergency speed index and the percentage of germination from 60 to 81.63%. It was also observed a statistical significance in the shoot length of the treated plants with the endophyte suspension (55.38 cm) compared to the control (41.67 cm).

Characterization of Pseudomonas bacteria of Piper tuberculatum regarding the production of potentially bio-stimulating compounds for plant growth / Caracterização de bactérias Pseudomonas de Piper tuberculatum quanto à produção de compostos potencialmente bioestimulantes para o crescimento de plantas

Despite advances in the identification and characterization of endophytic bacteria in various plant species worldwide, little is known about such microorganisms in plants from the Amazon region. Previous studies reported that Piper tuberculatum endophytic Pseudomonas (isolates Pt12 and Pt13, identified as Pseudomonas putida and Pseudomonas sp., respectively) were able to inhibit the in vitro growth of Fusarium solani f. sp. piperis, which causes root rot in black pepper (Piper nigrum), and that Pt13 promoted the growth of P. nigrum. Therefore, the aim here was to characterize these bacteria regarding their ability to produce plant growth-promoting substances [siderophores, indol acetic acid (IAA) and soluble phosphate]. Chrome azurol S assays were performed for the detection of siderophores. For qualitative and quantitative assays of IAA production and phosphate solubilization, Salkowski´s reagent and NBRIP medium with molybdenum blue reagent, respectively, were used. Results revealed that Pt12 and Pt13 were able to synthesize IAA, mainly under a high concentration of L-tryptophan, indicating that they are IAA-producing bacteria, probably through a tryptophan-dependent biosynthesis pathway. The presence of P. nigrum extract positively influenced the IAA production by Pt12 and Pt13, with highest values of 125 and 90 µg mL-1, respectively. In addition, Pt12 was positive for the production of siderophores and produced 56.56 µg mL-1 of soluble phosphate. In contrast, Pt13 showed no ability to produce siderophores or to solubilize phosphate. Besides their potential in controlling plant diseases, Pt12 and Pt13 have potential as biofertilizers, favoring sustainable agriculture.(AU)

Apesar dos avanços na identificação e caracterização de bactérias endofíticas em espécies vegetais em todo o mundo, pouco se sabe sobre esses microrganismos em plantas da região amazônica. Estudos anteriores mostraram que Pseudomonas de Piper tuberculatum (isolados Pt12 e Pt13, identificados como Pseudomonas putida e Pseudomonas sp., respectivamente) são capazes de inibir Fusarium solani f. sp. piperis, que causa a podridão das raízes da pimenteira-do-reino (Piper nigrum), e que Pt13 promoveu o crescimento de P. nigrum. Portanto, o objetivo do presente trabalho foi caracterizar essas bactérias quanto à capacidade de produzir substâncias potencialmente bioestimulantes para o crescimento vegetal [sideróforos, ácido indol acético (AIA) e fosfato solúvel]. Ensaios de Cromo Azurol S foram realizados para detecção de sideróforos. Para os ensaios qualitativos e quantitativos de produção de AIA e solubilização de fosfato, foram utilizados o reagente de Salkowski e o meio NBRIP com azul de molibdênio, respectivamente. Os resultados revelaram que Pt12 e Pt13 sintetizaram AIA, principalmente sob alta concentração de L-triptofano, indicando que provavelmente utilizam uma via de biossíntese dependente deste aminoácido. A presença do extrato de P. nigrum influenciou positivamente a produção de AIA por Pt12 e Pt13, com valores máximos de 125 e 90 µg mL-1, respectivamente. Além disso, Pt12 foi positiva para produção de sideróforos e produziu 56.56 µg.mL-1 de fosfato solúvel. Em contraste, Pt13 não produziu sideróforos, nem solubilizou fosfato. Além do potencial de controle de doenças de plantas, Pt12 e Pt13 têm potencial como biofertilizantes, favorecendo a agricultura sustentável.(AU)

Diversity of bacterial strains in biochar-enhanced Amazon soil and their potential for growth promotion and biological disease control in tomato / Diversidade de isolados bacterianos em solo amazônico enriquecido com biocarvão e seu potencial para a promoção de crescimento e controle biológico de doenças em tomate

The use of bacteria in growth promotion and biological control of plant diseases can minimize environmental contamination caused by the indiscriminate use of pesticides and chemical fertilizers. We aimed to evaluate growth promotion and biological control of Corynespora cassiicola in tomato seedlings mediated by beneficial bacteria isolated from a non-rhizospheric Amazon soil containing different amounts of biochar, and to identify to which groups of bacteria the strains belong. We obtained 200 strains of bacteria from experimental plots containing biochar doses of 0, 40, 80 and 120 t ha-1. Of these, 53 strains were selected by root colonization tests. Based on growth promotion parameters, 25 strains were screened, identified by molecular characterization and evaluated for indoleacetic acid (IAA) production, phosphate solubilization and biological control. The best dose of biochar for colony formation was 40 t ha-1, and a regression model indicated 34 t ha-1 as the optimal dose. The production of IAA was observed in 18 (75%) strains, and two (8%) strains were able to solubilize phosphate. The efficiency in root growth promotion was up to 125%, and the percentage of plant protection ranged from 50 to 59%. Molecular characterization showed that the bacteria used in this study belong to the genera Bacillus and Lysinibacillus.(AU)

O uso de bactérias na promoção do crescimento e no controle biológico de doenças em plantas pode minimizar a contaminação ambiental causada pela aplicação indiscriminada de pesticidas e fertilizantes químicos. Objetivamos avaliar a promoção do crescimento e o controle biológico de Corynespora cassiicola em mudas de tomate mediadas por bactérias benéficas isoladas de solo amazônico não-rizosférico contendo diferentes dosagens de biocarvão, e identificar a quais grupos de bactérias os isolados pertencem. Obtivemos 200 isolados de parcelas experimentais contendo doses de biocarvão de 0, 40, 80 e 120 t ha-1. Destes, 53 foram selecionados por testes de colonização radicular. Com base nos parâmetros de promoção do crescimento, 25 isolados foram selecionados, identificados através de análise molecular e avaliados para produção de ácido indolacético (AIA), solubilização de fosfato e controle biológico. A melhor dose de biocarvão para a formação de colônias foi 40 t ha-1, e um modelo de regressão indicou 34 t ha-1como dose ótima. A produção de AIA foi observada em 18 (75%) isolados e dois (8%) isolados foram capazes de solubilizar fosfato. A eficiência na promoção do crescimento das raízes foi de até 125%, e a porcentagem de proteção das plantas variou de 50,3 a 59,0%. A caracterização molecular indicou que as bactérias utilizadas nesse estudo pertencem aos gêneros Bacillus e Lysinibacillus.(AU)

Characterization of associative diazotrophic bacteria in torch ginger / Caracterização de bactérias diazotróficas associativas em bastão-do-imperador

Associative diazotrophic bacteria perform several processes that promote increased plant development and production, allowing a reduction in the use of agricultural inputs and costs. However, for some species, such as torch ginger, there are still no reports of studies aimed at identifying diazotrophic bacteria associated with this species. On this basis, this study proposes to isolate and characterize associative diazotrophic bacteria in rhizospheric soils and roots of torch ginger as well as analyze the potential of these isolates in solubilizing phosphorus (P) and potassium (K) and producing indole-3- acetic acid (IAA). Soil and roots samples of torch ginger were inoculated into five different semi-solid and semi-selective culture media, namely, NFb, JNFb, LGI, JMV and FAM, where bacterial growth was diagnosed by the formation of a characteristic film on the surface of the media. Subsequently, the bacterial isolates were analyzed for their ability to solubilize P and K in liquid medium, using phosphate rock powder (AO-15) and potassium rock powder (phonolite) as sources of P and K, respectively. All culture media showed bacterial growth, making this the first report of isolation of diazotrophic bacterial strains in this species. Eight of the obtained strains originated from rhizospheric soils and four from roots of torch ginger. Of these, 10 solubilized P, with the UNIFENAS 100-340, UNIFENAS 100-342 and UNIFENAS 100-348 strains standing out. Six strains showed K solubilizing ability, UNIFENAS 100-346 being the most efficient. All strains were able to produce the IAA phytohormone, both in the presence and absence of tryptophan, with superior results obtained by UNIFENAS 100-344 and UNIFENAS 100-351.(AU)

As bactérias diazotróficas associativas realizam diversos processos que promovem o maior desenvolvimento e produção vegetal, reduzindo o uso de insumos agrícolas e custos. No entanto, para algumas espécies, como o bastão-do-imperador, ainda não existem relatos de estudos visando identificar as bactérias diazotróficas associadas à esta espécie. Objetivou-se isolar e caracterizar bactérias diazotróficas associativas em solos rizosféricos e raízes de bastão-do-imperador e analisar o potencial destes isolados em solubilizar fósforo (P) e potássio (K) e produzir ácido 3-indol acético (AIA). As amostras de solos e raízes do bastão-do-imperador foram inoculadas em cinco diferentes meios de cultura semi-sólidos e semisseletivos, NFb, JNFb, LGI, JMV e FAM, em que o crescimento bacteriano foi diagnosticado pela formação de película característica na superfície dos meios. Posteriormente, os isolados bacterianos foram analisados quanto à capacidade de solubilizar P e potássio K em meio líquido, utilizando pó de rocha fosfatada, AO-15 e potássica, fonolito, como fontes de P e K, respectivamente. Todos os meios de cultivo utilizados apresentaram crescimento bacteriano, sendo este o primeiro relato de isolamento de estirpes bacterianas diazotróficas desta espécie. Das estirpes obtidas, oito foram de solos rizosféricos e quatro das raízes do bastão-do-imperador. Destas, 10 solubilizaram P, sendo as estirpes UNIFENAS 100-340 UNIFENAS 100-342 e UNIFENAS 100-348 as que se destacaram. Quanto a solubilização de K, seis estipes apresentaram a capacidade de solubilização, sendo a estirpe UNIFENAS 100-346 a mais eficiente. Em relação à produção de AIA, todas foram capazes de produzir esse fitormônio, tanto na presença quanto ausência do triptofano, com destaque para as estirpes UNIFENAS 100-344 e UNIFENAS 100-351.(AU)

Characterization of associative diazotrophic bacteria in torch ginger / Caracterização de bactérias diazotróficas associativas em bastão-do-imperador

Associative diazotrophic bacteria perform several processes that promote increased plant development and production, allowing a reduction in the use of agricultural inputs and costs. However, for some species, such as torch ginger, there are still no reports of studies aimed at identifying diazotrophic bacteria associated with this species. On this basis, this study proposes to isolate and characterize associative diazotrophic bacteria in rhizospheric soils and roots of torch ginger as well as analyze the potential of these isolates in solubilizing phosphorus (P) and potassium (K) and producing indole-3- acetic acid (IAA). Soil and roots samples of torch ginger were inoculated into five different semi-solid and semi-selective culture media, namely, NFb, JNFb, LGI, JMV and FAM, where bacterial growth was diagnosed by the formation of a characteristic film on the surface of the media. Subsequently, the bacterial isolates were analyzed for their ability to solubilize P and K in liquid medium, using phosphate rock powder (AO-15) and potassium rock powder (phonolite) as sources of P and K, respectively. All culture media showed bacterial growth, making this the first report of isolation of diazotrophic bacterial strains in this species. Eight of the obtained strains originated from rhizospheric soils and four from roots of torch ginger. Of these, 10 solubilized P, with the UNIFENAS 100-340, UNIFENAS 100-342 and UNIFENAS 100-348 strains standing out. Six strains showed K solubilizing ability, UNIFENAS 100-346 being the most efficient. All strains were able to produce the IAA phytohormone, both in the presence and absence of tryptophan, with superior results obtained by UNIFENAS 100-344 and UNIFENAS 100-351.

As bactérias diazotróficas associativas realizam diversos processos que promovem o maior desenvolvimento e produção vegetal, reduzindo o uso de insumos agrícolas e custos. No entanto, para algumas espécies, como o bastão-do-imperador, ainda não existem relatos de estudos visando identificar as bactérias diazotróficas associadas à esta espécie. Objetivou-se isolar e caracterizar bactérias diazotróficas associativas em solos rizosféricos e raízes de bastão-do-imperador e analisar o potencial destes isolados em solubilizar fósforo (P) e potássio (K) e produzir ácido 3-indol acético (AIA). As amostras de solos e raízes do bastão-do-imperador foram inoculadas em cinco diferentes meios de cultura semi-sólidos e semisseletivos, NFb, JNFb, LGI, JMV e FAM, em que o crescimento bacteriano foi diagnosticado pela formação de película característica na superfície dos meios. Posteriormente, os isolados bacterianos foram analisados quanto à capacidade de solubilizar P e potássio K em meio líquido, utilizando pó de rocha fosfatada, AO-15 e potássica, fonolito, como fontes de P e K, respectivamente. Todos os meios de cultivo utilizados apresentaram crescimento bacteriano, sendo este o primeiro relato de isolamento de estirpes bacterianas diazotróficas desta espécie. Das estirpes obtidas, oito foram de solos rizosféricos e quatro das raízes do bastão-do-imperador. Destas, 10 solubilizaram P, sendo as estirpes UNIFENAS 100-340 UNIFENAS 100-342 e UNIFENAS 100-348 as que se destacaram. Quanto a solubilização de K, seis estipes apresentaram a capacidade de solubilização, sendo a estirpe UNIFENAS 100-346 a mais eficiente. Em relação à produção de AIA, todas foram capazes de produzir esse fitormônio, tanto na presença quanto ausência do triptofano, com destaque para as estirpes UNIFENAS 100-344 e UNIFENAS 100-351.

Characterization of bacterial endophytes from the roots of native and cultivated Brazil nut trees (Bertholletia excelsa) / Caracterização de bactérias endofíticas de raízes de castanha-do-Brasil (Bertholletia excelsa) em habitats nativos e cultivados

Brazil nut is a very important nontimber forest product in the Amazon region. Propagation of this tree still represents a challenge due to slow and uneven seed germination. In this context, plant growth-promoting bacteria can facilitate the process of propagation. The aims of this study were to isolate and characterize endophytic bacteria from the roots of Brazil nut trees in native terra firme forest and cultivation areas in northern Brazil, and to identify mechanisms by which bacteria act in plant growth promotion. Overall, 90 bacterial isolates were obtained from the roots of Brazil nut trees in monoculture, agroforestry and native forest areas by using different semisolid media. The isolates were characterized by sequencing the 16S rRNA gene. Plant growth-promoting characteristics were evaluated by the presence of the nifH gene, aluminum phosphate solubilization and the production of indole compounds. The isolates were affiliated with 18 genera belonging to 5 different classes (α-Proteobacteria, β-Proteobacteria, γ-Proteobacteria, Bacilli and Actinobacteria). The genus Bacillus was predominant in the forest and monoculture areas. Fourteen isolates presented the nifH gene. Most of the bacteria were able to solubilize aluminum phosphate and synthetize indole compounds. The results indicated high diversity of endophytic bacteria present among the roots of Brazil nut trees, mainly in the agroforestry area, which could be related to soil attributes. Among the 90 isolates, the 22 that presented the best results regarding plant growth promotion traits were good candidates for testing in seedling production of Brazil nut trees.(AU)

A castanha-do-brasil é um produto florestal não madeireiro muito importante na região amazônica. A propagação desta árvore ainda representa um desafio, devido ao lento e irregular processo de germinação das sementes. Neste contexto, bactérias promotoras do crescimento vegetal podem facilitar o processo de propagação. O objetivo deste estudo foi isolar e caracterizar bactérias endofíticas em raízes de castanha-do-Brasil em floresta de terra firme e em áreas cultivadas no norte do Brasil, e identificar alguns mecanismos de promoção do crescimento vegetal executados por essas bactérias. No total, 90 isolados bacterianos foram obtidos de raízes de castanha-do-Brasil em monocultura, agrofloresta e floresta nativa, usando diferentes meios de cultivo semi-sólidos. Os isolados foram caracterizados pelo sequenciamento do gene 16S rRNA. As características de promoção do crescimento vegetal foram avaliadas através da presença do gene nifH, solubilização de fosfato de alumínio e produção de compostos indólicos. Os isolados foram afiliados a 18 gêneros, pertencentes a cinco diferentes classes (α-Proteobacteria, β-Proteobacteria, γ-Proteobacteria, Bacilli e Actinobacteria). O gênero Bacillus foi predominante, principalmente nas áreas de floresta e monocultura. Quatorze isolados apresentaram o gene nifH. A maioria dos isolados foi capaz de solubilizar fosfato de alumínio e sintetizar compostos indólicos. Os resultados indicam uma elevada diversidade de bactérias endofíticas presente em raízes de castanha-do-Brasil, principalmente em área de agrofloresta, que pode estar relacionado aos atributos do solo. Entre os 90 isolados, 22 apresentaram os melhores resultados relacionados às características de promoção do crescimento vegetal, e são bons candidatos para testes em produção de mudas de castanha-do-Brasil.(AU)

Endophytic Bacillus strains enhance pearl millet growth and nutrient uptake under low-P

Bacterial endophytes are considered to have a beneficial effect on host plants, improving their growth by different mechanisms. The objective of this study was to investigate the capacity of four endophytic Bacillus strains to solubilize iron phosphate (Fe-P), produce siderophores and indole-acetic acid (IAA) in vitro, and to evaluate their plant growth promotion ability in greenhouse conditions by inoculation into pearl millet cultivated in a P-deficient soils without P fertilization, with Araxá rock phosphate or soluble triple superphosphate. All strains solubilized Fe-P and three of them produced carboxylate-type siderophores and high levels of IAA in the presence of tryptophan. Positive effect of inoculation of some of these strains on shoot and root dry weight and the N P K content of plants cultivated in soil with no P fertilization might result from the synergistic combination of multiple plant growth promoting (PGP) traits. Specifically, while B1923 enhanced shoot and root dry weight and root N P content of plants cultivated with no P added, B2084 and B2088 strains showed positive performance on biomass production and accumulation of N P K in the shoot, indicating that they have higher potential to be microbial biofertilizer candidates for commercial applications in the absence of fertilization.(AU)

Endophytic Bacillus strains enhance pearl millet growth and nutrient uptake under low-P

Abstract Bacterial endophytes are considered to have a beneficial effect on host plants, improving their growth by different mechanisms. The objective of this study was to investigate the capacity of four endophytic Bacillus strains to solubilize iron phosphate (Fe-P), produce siderophores and indole-acetic acid (IAA) in vitro, and to evaluate their plant growth promotion ability in greenhouse conditions by inoculation into pearl millet cultivated in a P-deficient soils without P fertilization, with Araxá rock phosphate or soluble triple superphosphate. All strains solubilized Fe-P and three of them produced carboxylate-type siderophores and high levels of IAA in the presence of tryptophan. Positive effect of inoculation of some of these strains on shoot and root dry weight and the N P K content of plants cultivated in soil with no P fertilization might result from the synergistic combination of multiple plant growth promoting (PGP) traits. Specifically, while B1923 enhanced shoot and root dry weight and root N P content of plants cultivated with no P added, B2084 and B2088 strains showed positive performance on biomass production and accumulation of N P K in the shoot, indicating that they have higher potential to be microbial biofertilizer candidates for commercial applications in the absence of fertilization.

Mangrove endophyte promotes reforestation tree (Acacia polyphylla) growth

Mangroves are ecosystems located in the transition zone between land and sea that serve as a potential source of biotechnological resources. Brazil's extensive coast contains one of the largest mangrove forests in the world (encompassing an area of 25,000 km2 along all the coast). Endophytic bacteria were isolated from the following three plant species: Rhizophora mangle, Laguncularia racemosa and Avicennia nitida. A large number of these isolates, 115 in total, were evaluated for their ability to fix nitrogen and solubilize phosphorous. Bacteria that tested positive for both of these tests were examined further to determine their level of indole acetic acid production. Two strains with high indole acetic acid production were selected for use as inoculants for reforestation trees, and then the growth of the plants was evaluated under field conditions. The bacterium Pseudomonas fluorescens (strain MCR1.10) had a low phosphorus solubilization index, while this index was higher in the other strain used, Enterobacter sp. (strain MCR1.48). We used the reforestation tree Acacia polyphylla. The results indicate that inoculation with the MCR1.48 endophyte increases Acacia polyphylla shoot dry mass, demonstrating that this strain effectively promotes the plant's growth and fitness, which can be used in the seedling production of this tree. Therefore, we successfully screened the biotechnological potential of endophyte isolates from mangrove, with a focus on plant growth promotion, and selected a strain able to provide limited nutrients and hormones for in plant growth.(AU)

Tricalcium phosphate solubilization and nitrogen fixation by newly isolated Aneurinibacillus aneurinilyticus CKMV1 from rhizosphere of Valeriana jatamansi and its growth promotional effect

Aneurinibacillus aneurinilyticus strain CKMV1 was isolated from rhizosphere of Valeriana jatamansi and possessed multiple plant growth promoting traits like production of phosphate solubilization (260 mg/L), nitrogen fixation (202.91 nmol ethylene mL-1 h-1), indole-3-acetic acid (IAA) (8.1 µg/mL), siderophores (61.60%), HCN (hydrogen cyanide) production and antifungal activity. We investigated the ability of isolate CKMV1 to solubilize insoluble P via mechanism of organic acid production. High-performance liquid chromatography (HPLC) study showed that isolate CKMV1 produced mainly gluconic (1.34%) and oxalic acids. However, genetic evidences for nitrogen fixation and phosphate solubilization by organic acid production have been reported first time for A. aneurinilyticus strain CKMV1. A unique combination of glucose dehydrogenase (gdh) gene and pyrroloquinoline quinone synthase (pqq) gene, a cofactor of gdh involved in phosphate solubilization has been elucidated. Nitrogenase (nif H) gene for nitrogen fixation was reported from A. aneurinilyticus. It was notable that isolate CKMV1 exhibited highest antifungal against Sclerotium rolfsii (93.58%) followed by Fusarium oxysporum (64.3%), Dematophora necatrix (52.71%), Rhizoctonia solani (91.58%), Alternaria sp. (71.08%) and Phytophthora sp. (71.37%). Remarkable increase was observed in seed germination (27.07%), shoot length (42.33%), root length (52.6%), shoot dry weight (62.01%) and root dry weight (45.7%) along with NPK (0.74, 0.36, 1.82%) content of tomato under net house condition. Isolate CKMV1 possessed traits related to plant growth promotion, therefore, could be a potential candidate for the development of biofertiliser or biocontrol agent and this is the first study to include the Aneurinibacillus as PGPR.(AU)

Diversity and capacity to promote maize growth of bacteria isolated from the Amazon region / Diversidade e capacidade de promoção de crescimento de milho de bactérias isoladas da região Amazônica

Maize plants can establish beneficial associations with plant growth-promoting bacteria. However, few studies have been conducted on the characterization and inoculation of these bacteria in the Amazon region. This study aimed to characterize endophytic bacteria isolated from maize in the Amazon region and to assess their capacity to promote plant growth. Fifty-five bacterial isolates were obtained from maize grown in two types of ecosystems, i.e., a cerrado (savanna) and a forest area. The isolates were characterized by the presence of the nifH gene, their ability to synthesize indole-3-acetic acid (IAA) and solubilize calcium phosphate (CaHPO4), and 16S rRNA partial gene sequencing. Twenty-four bacteria contained the nifH gene, of which seven were isolated from maize plants cultivated in a cerrado area and seventeen from a forest area. Fourteen samples showed the capacity to synthesize IAA and only four solubilized calcium phosphate. The following genera were found among these isolates: Pseudomonas; Acinetobacter; Enterobacter; Pantoea; Burkholderia and Bacillus. In addition, eight isolates with plant growth-promoting capacity were selected for a glasshouse experiment involving the inoculation of two maize genotypes (a hybrid and a variety) grown in pots containing soil. Inoculation promoted the development of the maize plants but no significant interaction between maize cultivar and bacterial inoculation was found. A high diversity of endophytic bacteria is present in the Amazon region and these bacteria have potential to promote the development of maize plants.

Plantas de milho podem estabelecer associações benéficas com bactérias promotoras do crescimento vegetal. No entanto, poucos estudos de caracterização e inoculação com essas bactérias foram realizados na região Amazônica. O objetivo deste estudo foi caracterizar bactérias endofíticas isoladas de milho na região Amazônica e avaliar sua capacidade de promover o crescimento. Cinquenta e cinco isolados bacterianos foram obtidos de plantas de milho cultivadas em diferentes tipos de ecossistemas, cerrado (savana) e área de floresta. Estes foram caracterizados pela presença do gene nifH, capacidade de sintetizar ácido indol-3-acético (AIA), solubilizar fosfato de cálcio (CaHPO4) e sequenciamento parcial do gene 16S rRNA. Vinte e quatro bactérias apresentaram o gene nifH. Destas, sete são oriundas de plantas cultivadas em uma área de cerrado e 17 em área de floresta. Quatorze bactérias demonstraram a capacidade de solubilizar AIA e quatro de solubilizar fosfato de cálcio. Entre estas bactérias, foram encontrados os seguintes gêneros: Pseudomonas; Acinetobacter; Enterobacter; Pantoea; Burkholderia e Bacillus. Oito isolados com capacidade de promoção do crescimento vegetal foram selecionados para experimento em casa de vegetação com a inoculação em dois diferentes genótipos de milho (um híbrido e uma variedade) em vasos com solo. Quanto à promoção de crescimento vegetal, a inoculação promoveu o desenvolvimento de plantas de milho, mas não foi encontrada interação significativa entre a cultivar de milho e a inoculação bacteriana. Na região Amazônica há uma diversidade de bactérias endofíticas e estas possuem potencial para promover o desenvolvimento de plantas de milho.

Isolation and identification by 16S rRNA sequence analysis of plant growth-promoting azospirilla from the rhizosphere of wheat

The main objective of the present study was to isolate phytohormone-producing, phosphate-solubilizing strains of Azospirillum from wheat to be used as inoculants for plant growth promotion. Five Azospirillum strains were isolated from the rhizosphere of field-grown wheat (Triticum aestivum L.), and it was confirmed by BOX-polymerase chain reaction (PCR) that the isolates were different and not re-isolates of the same strain. Sequence analysis of the PCR-amplified 16S rRNA gene indicated that four isolates showed maximum similarity to Azospirillum brasilense and one isolate showed maximum similarity to Azospirillum zeae. This is the first report indicating the presence of an A. zeae like isolate in the wheat rhizosphere in Pakistan. The bacterial isolates were characterized for their plant growth-promoting traits, phosphate solubilization, and indole-3-acetic acid (IAA) production. None of the isolates showed phosphate solubilization activity in the commonly used Pikovskaya medium. However, all strains (except AzoK4) exhibited ability to solubilize tricalcium phosphate (TCP) in modified Pikovskaya medium in which sucrose was replaced by Na-malate, as well as in TCP-supplemented Luria-Bertani (LB) medium. Organic acids, such as acetic, citric, lactic, malic, and succinic acids, were detected in culture supernatants of the tested Azospirillum strains. All strains exhibited ability to produce IAA in the growth medium, except Azospirillum sp. AzoK1. Among the strains tested, the maximum IAA production (30.49 ± 1.04 mg L-1) and phosphate solubilization (105.50 ± 4.93 mg L-1) were shown by a pure culture of Azospirillum sp. AzoK2. In pot experiments, single-strain inocula of Azospirillum sp. AzoK1 and AzoK2 improved wheat plant growth.(AU)

Effect of plant growth-promoting bacteria on the growth and fructan production of Agave americana L

The effect of plant growth-promoting bacteria inoculation on plant growth and the sugar content in Agave americana was assessed. The bacterial strains ACO-34A, ACO-40, and ACO-140, isolated from the A. americana rhizosphere, were selected for this study to evaluate their phenotypic and genotypic characteristics. The three bacterial strains were evaluated via plant inoculation assays, and Azospirillum brasilense Cd served as a control strain. Phylogenetic analysis based on the 16S rRNA gene showed that strains ACO-34A, ACO-40 and ACO-140 were Rhizobium daejeonense, Acinetobacter calcoaceticus and Pseudomonas mosselii, respectively. All of the strains were able to synthesize indole-3-acetic acid (IAA), solubilize phosphate, and had nitrogenase activity. Inoculation using the plant growth-promoting bacteria strains had a significant effect (p 0.05) on plant growth and the sugar content of A. americana, showing that these native plant growth-promoting bacteria are a practical, simple, and efficient alternative to promote the growth of agave plants with proper biological characteristics for agroindustrial and biotechnological use and to increase the sugar content in this agave species.(AU)

Diversity and capacity to promote maize growth of bacteria isolated from the Amazon region / Diversidade e capacidade de promoção de crescimento de milho de bactérias isoladas da região Amazônica

Maize plants can establish beneficial associations with plant growth-promoting bacteria. However, few studies have been conducted on the characterization and inoculation of these bacteria in the Amazon region. This study aimed to characterize endophytic bacteria isolated from maize in the Amazon region and to assess their capacity to promote plant growth. Fifty-five bacterial isolates were obtained from maize grown in two types of ecosystems, i.e., a cerrado (savanna) and a forest area. The isolates were characterized by the presence of the nifH gene, their ability to synthesize indole-3-acetic acid (IAA) and solubilize calcium phosphate (CaHPO4), and 16S rRNA partial gene sequencing. Twenty-four bacteria contained the nifH gene, of which seven were isolated from maize plants cultivated in a cerrado area and seventeen from a forest area. Fourteen samples showed the capacity to synthesize IAA and only four solubilized calcium phosphate. The following genera were found among these isolates: Pseudomonas; Acinetobacter; Enterobacter; Pantoea; Burkholderia and Bacillus. In addition, eight isolates with plant growth-promoting capacity were selected for a glasshouse experiment involving the inoculation of two maize genotypes (a hybrid and a variety) grown in pots containing soil. Inoculation promoted the development of the maize plants but no significant interaction between maize cultivar and bacterial inoculation was found. A high diversity of endophytic bacteria is present in the Amazon region and these bacteria have potential to promote the development of maize plants.(AU)

Plantas de milho podem estabelecer associações benéficas com bactérias promotoras do crescimento vegetal. No entanto, poucos estudos de caracterização e inoculação com essas bactérias foram realizados na região Amazônica. O objetivo deste estudo foi caracterizar bactérias endofíticas isoladas de milho na região Amazônica e avaliar sua capacidade de promover o crescimento. Cinquenta e cinco isolados bacterianos foram obtidos de plantas de milho cultivadas em diferentes tipos de ecossistemas, cerrado (savana) e área de floresta. Estes foram caracterizados pela presença do gene nifH, capacidade de sintetizar ácido indol-3-acético (AIA), solubilizar fosfato de cálcio (CaHPO4) e sequenciamento parcial do gene 16S rRNA. Vinte e quatro bactérias apresentaram o gene nifH. Destas, sete são oriundas de plantas cultivadas em uma área de cerrado e 17 em área de floresta. Quatorze bactérias demonstraram a capacidade de solubilizar AIA e quatro de solubilizar fosfato de cálcio. Entre estas bactérias, foram encontrados os seguintes gêneros: Pseudomonas; Acinetobacter; Enterobacter; Pantoea; Burkholderia e Bacillus. Oito isolados com capacidade de promoção do crescimento vegetal foram selecionados para experimento em casa de vegetação com a inoculação em dois diferentes genótipos de milho (um híbrido e uma variedade) em vasos com solo. Quanto à promoção de crescimento vegetal, a inoculação promoveu o desenvolvimento de plantas de milho, mas não foi encontrada interação significativa entre a cultivar de milho e a inoculação bacteriana. Na região Amazônica há uma diversidade de bactérias endofíticas e estas possuem potencial para promover o desenvolvimento de plantas de milho.(AU)

HIDRÓLISE ENZIMÁTICA DAS VÍSCERAS DE Macrobrachium rosenbergii PROVENIENTES DE PROCESSAMENTO

A aquicultura está em constante ascensão, e um dos setores que mais destaca-se é a carcinicultura, por sua ampla difusão no mundo e expansão na produção. Aliado a essa produção, a geração de resíduos é inevitável e, consequentemente, buscam-se formas de tratar e aproveitar os mesmos. Os resíduos da carcinicultura são ricos em componentes como proteínas, lipídeos e minerais que podem ser utilizados na formulação de rações, o que permite sua valorização e aproveitamento. O objetivo desse trabalho foi avaliar a hidrólise enzimática proteica das vísceras de camarões Macrobrachium rosenbergii oriundas de seu processamento, como alternativa para aproveitamento desse material. Para o preparo das dispersões foram utilizadas as vísceras do camarão com teor de proteína bruta de 19,43% e as enzimas Alcalase® 2.4L e Flavourzyme® 1000L. Foram realizados 2 planejamentos experimentais a fim de otimizar o processo. No primeiro foi feito um delineamento fatorial 2³ sobre o efeito de três fatores, cada um em 3 níveis: temperatura (45; 55; 65 °C) e concentração das enzimas (0,75; 1,50 e 2,25% proteína da enzima/proteína do substrato). Em seguida foi feito um delineamento composto central rotacional sobre o efeito de três fatores, cada um em 5 níveis: temperatura (34,2; 41,0; 51,0; 61,0 e 67,8 °C) e concentração das enzimas (0,08; 0,25; 0,50; 0,75 e 0,92%). Para ambos os delineamentos foram conduzidas reações controles, que são reações nas mesmas condições das reações enzimáticas, mas sem a adição das enzimas. Foi demonstrado que a temperatura teve uma maior influência na hidrólise enzimática proteica das vísceras de camarão M. rosenbergii, obtendo-se valores de GH de 2,11%±0,25 na temperatura de 67,8 ºC, e concentração das enzimas Alcalase® 2.4L e Flavourzyme® 1000L 0,50%. No entanto, uma das reações controle apresentou um GH de 2,06%±0,11 em uma temperatura de 61,0 °C, sendo estatisticamente igual ao valor encontrado para a reação enzimática. Além disso, a variável temperatura em 51,0 °C atingiu um rendimento otimizado. Dessa forma, utilizando essa temperatura, foi possível solubilizar a proteína presente nas vísceras de camarão M. rosenbergii, podendo tornar o hidrolisado uma fonte de proteína para a aquicultura.

Aquaculture is constantly on the rise, and one of the sectors that stands out the most is carciniculture, for its wide spread in the world and expansion in production. Allied to this production, the generation of waste is inevitable and, consequently, ways of treating and taking advantage of it are sought. Carciniculture residues are rich in components such as proteins, lipids and minerals that can be used in the formulation of rations, which allows their valorization and utilization. The objective of this work was to evaluate the protein enzymatic hydrolysis of the viscera of Macrobrachium rosenbergii shrimps from their processing, as an alternative for the use of this material. For the preparation of the dispersions, the shrimp viscera with raw protein content of 19.43% and the enzymes Alcalase® 2.4L and Flavourzyme® 1000L were used. Two experimental plans were made in order to optimize the process. In the first one, a 2³ factorial design was made on the effect of three factors, each in 3 levels: temperature (45; 55; 65 °C) and enzyme concentration (0.75; 1.50 and 2.25% enzyme/protein protein of the substrate). Then a rotational central compound delineat was made on the effect of three factors, each in 5 levels: temperature (34.2; 41.0; 51.0; 61.0 and 67.8 °C) and enzyme concentration (0.08; 0.25; 0.50; 0.75 and 0.92%). For both delineations, control reactions were conducted, which are reactions under the same conditions as enzymatic reactions, but without the addition of enzymes. It was demonstrated that the temperature had a greater influence on the proteinic enzymatic hydrolysis of M. rosenbergii shrimp viscera, obtaining GH values of 2.11%±0.25 at a temperature of 67.8 ºC, and concentration of the enzymes Alkalase® 2.4L and Flavourzyme® 1000L 0.50%. However, one of the control reactions presented a GH of 2.06%±0.11 at a temperature of 61.0 ºC, being close to the value found for the enzymatic reaction. In addition, the temperature variable at 51.0 °C achieved an optimum yield. Thus, using this temperature, it was possible to solubilize the protein present in the viscera of M. rosenbergii shrimps, which could make hydrolysate a source of protein for aquaculture.

Poly R decolorization and APPL production by Streptomyces violaceoruber and Streptomyces spiroverticillatus

Two mesophilic streptomycetes (S. violaceoruber and S. spiroverticillatus) were selected to study their Poly R-478 decolorization ability and lignocellulose solubilizing activity. Both strains were able to degrade Poly R-478 dye and ferulic acid during growth on a minimal salts medium. The Poly R-478 decolorizing activities of both strains were induced by adding different carbon sources to the culture media. S. violaceoruber could decolorize 63% of Poly R-478 after 24 h. Both strains could solubilize straw and produce acid-precipitable polymeric lignin (APPL) with different efficiency. From the major extracellular enzymes recovery of both strains on rice and wheat straw, we can predicate that the biodegradation process was partial indicating a possible utilization in biological delignification.

Bioinformatics based structural characterization of glucose dehydrogenase (gdh) gene and growth promoting activity of Leclercia sp. QAU-66

Glucose dehydrogenase (GDH; EC 1.1. 5.2) is the member of quinoproteins group that use the redox cofactor pyrroloquinoline quinoine, calcium ions and glucose as substrate for its activity. In present study, Leclercia sp. QAU-66, isolated from rhizosphere of Vigna mungo, was characterized for phosphate solubilization and the role of GDH in plant growth promotion of Phaseolus vulgaris. The strain QAU-66 had ability to solubilize phosphorus and significantly (p < 0.05) promoted the shoot and root lengths of Phaseolus vulgaris. The structural determination of GDH protein was carried out using bioinformatics tools like Pfam, InterProScan, I-TASSER and COFACTOR. These tools predicted the structural based functional homology of pyrroloquinoline quinone domains in GDH. GDH of Leclercia sp. QAU-66 is one of the main factor that involved in plant growth promotion and provides a solid background for further research in plant growth promoting activities.