In vitro antibacterial activities of silver nanoparticles synthesised using the seed extracts of three varieties of Phoenix dactylifera / Atividades antibacterianas in vitro de nanopartículas de prata sintetizadas usando extratos de sementes de três variedades de Phoenix dactylifera

Green synthesis of silver nanoparticles (AgNPs) is an ecofriendly, cost-effective and promising approach for discovery of novel therapeutics. The aim of the current work was to biogenic synthesize, characterize AgNPs using seed extracts of three economically important varieties of date palm (Iklas, Irziz and Shishi), and assess their anti-pathogenic bacterial activities. AgNPs were synthesised then characterised using electron microscopy and Fourier transform infrared analyses. The bactericidal activities of AgNPs against five different bacterial pathogens, Bacillus subtilis, Escherichia coli, Staphylococcus aureus, methicillin-resistant Staphylococcus aureus and Streptococcus pneumoniae, were determined in vitro. In particular, changes in membrane integrity of virulent bacterial strains in response to AgNPs were investigated. Results of lactate dehydrogenase, alkaline phosphatase activity assays, and measurement of membrane potential revealed that the cytotoxic effects of the AgNPs were mainly centred on the plasma membrane of bacterial cells, leading to loss of its integrity and eventually cell death. In conclusion, green synthesis of AgNPs is an efficient, cost-effective and promising strategy to combat virulent antibiotic-resistant strains.

A síntese verde de nanopartículas de prata (AgNPs) é uma abordagem ecologicamente correta, econômica e promissora para a descoberta de novas terapêuticas. O objetivo do presente trabalho foi sintetizar biogênica, caracterizar AgNPs usando extratos de sementes de três variedades economicamente importantes de tamareira (Iklas, Irziz e Shishi) e avaliar suas atividades bacterianas antipatogênicas. AgNPs foram sintetizados e caracterizados usando microscopia eletrônica e análise de infravermelho por transformada de Fourier. As atividades bactericidas de AgNPs contra cinco diferentes patógenos bacterianos, Bacillus subtilis, Escherichia coli, Staphylococcus aureus, Staphylococcus aureus resistente à meticilina e Streptococcus pneumoniae, foram determinadas in vitro. Em particular, foram investigadas alterações na integridade da membrana de cepas bacterianas virulentas em resposta a AgNPs. Os resultados da lactato desidrogenase, dos ensaios da atividade da fosfatase alcalina e da medição do potencial de membrana revelaram que os efeitos citotóxicos dos AgNPs estavam principalmente centrados na membrana plasmática das células bacterianas, levando à perda de sua integridade e, eventualmente, à morte celular. A síntese verde de AgNPs é uma estratégia eficiente, econômica e promissora para combater cepas virulentas resistentes a antibióticos.

Population parameters as a way to evaluate the risk of attack of Aphis craccivora (Hemiptera: Aphididae) on cowpea / Parâmetros populacionais como forma de avaliar os riscos de ataque de Aphis craccivora (Hemiptera: Aphididae) em feijão-caupi

The objective of this research was to develop fertility life tables in order to estimate the population parameters of black aphid (Aphis craccivora Koch) in cowpea (Vigna unguiculata) varieties, aiming to propose a risk scale for its use. The experiment consisted of six treatments and six replicates (five varieties plus the cultivar VITA 7 as a susceptible control). A cohort was formed with six adult females distributed in six replicates of each genotype, and the insects were observed daily. Based on the data, fertility life tables were drawn for each variety and the population parameters were estimated. The different values of the finite growth rate (λ) were considered to propose a risk scale for the use of the genotypes. The results obtained give the dimension of the variability of V. unguiculata in respect to the character, resistance to A. craccivora, with antibiosis as the main mechanism of resistance. Considering all the results, the varieties studied can be classified according to their suitability as a plant favorable to the development of the black aphid as follows: VITA 7 > CE-13 > CE-51 > CE-08 = CE-07. The proposal of a risk scale for the use of V. unguiculata genotypes against the A. craccivora population, based on the finite growth rate (λ) values, was adequate to discriminate the varieties studied.(AU)

Objetivou-se nesta pesquisa elaborar tabelas de vida de fertilidade com o intuito de estimar os parâmetros populacionais de pulgão-preto (Aphis craccivora Koch) em variedades de feijão-caupi (Vigna unguiculata), visando propor uma escala de risco para o seu uso. O experimento constituiu-se em seis tratamentos (cinco variedades mais o cultivar VITA 7 como padrão de suscetibilidade) com seis repetições. Formou-se uma coorte com seis fêmeas adultas distribuídas em cada genótipo, sendo todos os indivíduos observados diariamente. De posse dos dados, foram elaboradas tabelas de vida de fertilidade para cada variedade e estimados os parâmetros populacionais. Para propor uma escala de risco de uso de genótipos, ponderou-se sob os diferentes valores da razão finita de crescimento (λ). Os resultados obtidos dão a dimensão da variabilidade de V. unguiculata com relação ao caráter da resistência a A. craccivora, sendo a antibiose o principal mecanismo de resistência associado. Considerando-se todos os resultados, foi possível hierarquizar as variedades estudadas segundo sua aptidão como planta favorável ao desenvolvimento ao pulgão-preto conforme segue: VITA 7 >> CE-13 > CE-51 > CE-08 = CE-07. A proposta de uma escala de risco do uso de genótipos de V. unguiculata frente à população de A. craccivora, baseada nos valores da razão finita de crescimento (λ), foi adequada para discriminar as variedades estudadas.(AU)

Antagonistic activity of volatile metabolites from Trichoderma asperellum / 生物工程学报

Trichoderma spp. is a kind of filamentous fungi with important biocontrol value. Twelve strains of Trichoderma spp. were isolated from the soils of different types of crops in Shaoxing, Zhejiang and Foshan, Guangdong. The antagonistic resistance to Fusarium oxysporum was compared by plate confrontation test. The further analysis of volatile secondary metabolites for two strains were carried out using HS-SPME-GC-MS analysis. The results showed that T. asperellum ZJSX5003 and GDFS1009 had fast growth ability, and the inhibition effects on F. oxysporum were 73% and 74% respectively. Six identical volatile metabolites were detected as follows 2-Methyl-1-propanol, 3-Methyl-1-butanol, 3-Methyl-3-buten-1-ol, Acetyl methyl carbinol, Butane-2,3-diol and 6-n-pentyl-2H-pyran-2-one (6-PAP). Among them, 6-PAP was validated to have a higher inhibitory effect on F. oxysporum in vitro. This study will provide basis for the development of biocontrol agents with metabolites of Trichoderma, such as 6-PAP.

In vitro assessment of the probiotic potential of lactobacilli isolated from Minas artisanal cheese produced in the Araxá region, Minas Gerais state, Brazil / Avaliação in vitro do potencial probiótico de lactobacilos isolados de queijo minas artesanal produzido na região de Araxá, estado de Minas Gerais, Brasil

Minas artisanal cheese is made from endogenous starter cultures, including lactic acid bacteria (LAB). Some LAB may possess probiotic potential. Thus, this study aimed to evaluate the in vitro probiotic properties of lactobacilli isolated from Minas artisanal cheeses produced in Minas Gerais. Ten samples of lactobacilli, formerly isolated from those cheeses, were submitted to the following assays: antimicrobial susceptibility, tolerance to artificial gastric juice and biliary salts, production of hydrogen peroxide and antagonism against pathogenic and non-pathogenic micro-organisms. Only L. plantarum (C0) was sensitive to all tested antimicrobials, while the other LAB samples were resistant to at least one drug. Six samples were tolerant to artificial gastric juice, and L. brevis (A6) even grew in that medium. Three samples were tolerant to biliary salts. Only L. brevis (E35) produced hydrogen peroxide. Difference (P< 0.05) was observed among the means of inhibition haloes of lactobacilli against Enterococcus faecalis ATCC 19433 and Lactobacillus plantarum C24 in spot-on-the-lawn assay. All samples of lactobacilli inhibited Escherichia coli ATCC 25922, Salmonella enterica var. Typhimurium ATCC 14028 in co-culture antagonism test (P< 0.0001). Most lactobacilli samples showed in vitro probiotic potential. From the tested samples, L. brevis (A6) presented the best results considering all in vitro probiotic tests.(AU)

O queijo minas artesanal é produzido por culturas starters endógenas, incluindo bactérias ácido-láticas (BAL). Algumas BAL podem possuir potencial probiótico. Com isso, este estudo teve como objetivo avaliar as propriedades probióticas in vitro de lactobacilos isolados de queijo minas artesanal produzido no estado de Minas Gerais. Dez amostras de lactobacilos, previamente isoladas desses queijos, foram submetidas aos seguintes testes: susceptibilidade aos antimicrobianos, tolerância ao suco gástrico artificial e aos sais biliares, produção de peróxido de hidrogênio e antagonismo contra micro-organismos patogênicos e não patogênicos. Apenas L. plantarum (C0) foi sensível a todos os antimicrobianos testados, enquanto as outras amostras de BAL foram resistentes a, pelo menos, uma droga testada. Seis amostras foram tolerantes ao suco gástrico artificial, e L. brevis (A6) apresentou crescimento nesse meio. Três amostras foram tolerantes aos sais biliares. Apenas L. brevis (E35) produziu peróxido de hidrogênio. Diferença (P<0,05) foi observada entre as médias dos halos de inibição de lactobacilos contra Enterococcus faecalis ATCC 19433 e Lactobacillus plantarum C24 no teste do spot-on-the-lawn. Todas as amostras de lactobacilos inibiram Escherichia coli ATCC 25922, Salmonella enterica var. Typhimurium ATCC 14028 no teste de antagonismo em cocultura (P<0,0001). A maioria das amostras de lactobacilos apresentou potencial probiótico in vitro. Com base nas amostras testadas, L. brevis (A6) apresentou os melhores resultados, considerando-se todos os testes probióticos in vitro.(AU)

Probiotics study with Streptococcus salivarius and its ability to produce bacteriocins and adherence to KB cells / Estudo de probióticos Streptococcus salivarius e sua habilidade de produzir bacteriocinas e aderência à células KB

Abstract Introduction Streptococcus salivarius is a dominant oral species and the best suitable candidate for probiotic of the oral cavity. Since Streptococcus salivarius is able to produce bacteriocins against Streptococcus pyogenes interest has been focused on the use of it as a probiotic to avoid sore throats by Streptococcus pyogenes. Objective This study is for selecting Streptococcus salivarius strains for potential use as probiotics for the oral mucosa, that is, production of bacteriocin against Streptococcus pyogenes and the ability to bind to KB cells. Material and method Tongue material from 45 students was collected and seeded on Mitis Salivarius Agar plaques. The strains were tested by the production of bacteriocin-like substances (BLIS) against S. pyogenes, biochemically and PCR for identification of S. salivarius. The best strains were tested for adherence to KB cells. Briefly, S. salivarius strains were cultured in broth, washed and suspended at 108cells/ml. KB cells were inoculated into plaques, washed and incubated with the bacteria, for adhesion. These were washed for lysis of the KB cells and release bacteria for determination of CFU. Result The bacteriocin test showed that 133 strains presented inhibition of S. pyogenes. The samples tested for adhesion to KB cells, presented different profiles and only three strains presenting high adhesion capacity. Conclusion The selection of strains of Streptococcus salivarius with high inhibitory activity against Streptococcus pyogenes, as well as adherence to KB cells leads us to the next future step, that is, to use the best strains for in vivo colonization tests

Resumo Introdução Streptococcus salivarius é uma espécie dominante na cavidade bucal e tem sido indicada como um ótimo candidato para uso como probiótico. Visto que a espécie Streptococcus salivarius é capaz de produzir bacteriocinas contra Streptococcus pyogenes, desenvolveu-se interesse no uso desse microrganismo como probiótico, para evitar amigdalites causadas por Streptococcus pyogenes. Objetivo A pesquisa em questão tem o objetivo de selecionar cepas de Streptococcus salivarius para seu uso potencial como probióticos na cavidade bucal, ou seja, produção de bacteriocinas contra Streptococcus pyogenes e habilidade de aderência à células KB. Material e método Coletou-se material de língua de 45 estudantes e semeou-se em placas de ágar Mitis Salivarius. As amostras foram testadas para verificar a produção de substâncias semelhantes à bacteriocina (BLIS) contra S. pyogenes, bioquimicamente e através de PCR para identificação de S. salivarius. As melhores cepas foram testadas quanto aderência à células KB. Resumidamente, as cepas de S. salivarius foram cultivadas em caldo, lavadas e suspensas à correspondência de 108 cels/ml. As células KB foram inoculadas em placas, lavadas e incubadas com as bactérias, para adesão. Estas foram lavadas para lise das células KB e liberação das bactérias para determinação de UFC. Resultado O teste de bacteriocina, mostrou que 133 cepas apresentaram atividade inibitória contra Streptococcus pyogenes. As cepas testadas para aderência à células KB, apresentaram diferentes perfis e somente três com alta capacidade de adesão. Conclusão: A seleção de cepas de Streptococcus salivarius com alta atividade inibitória contra Streptococcus pyogenes, bem como aderência a células KB, pode nos levar ao próximo passo, ou seja, o uso das melhores cepas para o estudo de colonização in vivo.

Isolation and identification of endophytic fungi from Chelidonium majus and their antifungal activity / 中国中药杂志

In order to find new source of antifungal agents, eleven cultivable endophytic fungi were isolated from the roots,stems and leaves of Chelidonium majus by traditional method. Seven of them were identified as Colletotrichum(L1, L2, L3, S1, S3, S4, S5), and three of them were identified as Fusarium(R1,R2,R3) by morphological features and molecular biological technology. The antifungal activity test showed that all the tested fungi displayed some inhibitory activity against five common plant pathogens(C. gloeosporioides, Curvularia lunata, Pyricularia oryza, Alternaria alternate and A. brassicae), and their inhibition rate of some test items were over 60%. Among them, R1, S2, S3 and S4 were more potent than others. This study enriches the understanding of endophytes from Ch. majus and provides a basis for the study of new microbial fungicides.

Isolation,screening and identification of endophytic fungi and detection of its antifungal effects against Alternaria panax / 中国中药杂志

To obtain biocontrol fungus for Alternaria panax,the antifungal effects of one strain of endophytic fungi isolated from leaves of healthy ginseng were screened and evaluated by using dual-culture method,and the taxonomic assignment of the screened strain was identified based on the morphological characters and ITS sequence analysis. The results of dual-culture showed that one of the endophytes marked as FS-01 had good antifungal effects and the inhibitory rates of FS-01 strain to A. panax was( 60. 21±0. 12) %.The hyphae junction of the both strains,A. panax dissolved,broke and winded,while the hyphae of FS-01 strain remained normal. The inhibitory rates of non-sterilized FS-01 strain fermentation liqud was( 13. 94±0. 21) %. Strain FS-01 identified as Chaetomium globosum.

Silicon as resistance inducer in to control black aphid Aphis craccivora Koch, 1854 in Phaseolus lunatus lima beans / Silício como indutor de resistência no controle do pulgão preto Aphis craccivora Koch, 1854 em fava Phaseolus lunatus

In Brazil, there are few records of insects associated with the cultivation of lima beans; among them, there is the black aphid Aphis craccivora Koch, 1854. The objective of this study was to evaluate the effects of silicon application on the resistance induction of lima bean plants, Phaseolus lunatus, to the black aphid A. craccivora. The experiment was conducted in the Entomology Laboratory of the Phytosanitary Sector of Centro de Ciências Agrárias, Universidade Federal do Piauí (UFPI), Brazil. The effects of the following treatments on biological aspects of the insect were evaluated: silicon applied to soil; silicon applied to soil + leaf; silicon applied to leaf; and control, without silicon application. The following biological variables were evaluated: generation period, reproductive period, and the fertility and daily average of produced nymphs per female. Plant silicon and lignin content were also evaluated. A 1% solution of silicic acid (2.0 g of product diluted in 200 mL of water) was applied around the plant stem (on soil), 15 days after emergence. Leaf application was performed with a 1-L spray, 5 days after the soil application. The non-preference of A. craccivora on lima beans was also evaluated. The evaluations were performed after 48 and 72 hours of infestation by counting nymphs and adults at each leaf section. Silicon application reduces nymph production, thereby interfering in the biological aspects of A. craccivora. Therefore, it can be used in cowpea pest management programs.(AU)

São poucos os registros no Brasil de insetos associados à cultura da fava; entre eles, destaca-se o pulgão preto Aphis craccivora Koch, 1854. O objetivo deste estudo foi avaliar os efeitos da aplicação de silício na indução de resistência de plantas de feijão-fava Phaseolus lunatus a esse inseto. O experimento foi conduzido no Laboratório de Entomologia do setor de Fitossanidade do Centro de Ciências Agrárias da Universidade Federal do Piauí (UFPI). Os efeitos da aplicação do silício sobre os aspectos biológicos do inseto foram avaliados utilizando os tratamentos: silício aplicado no solo, silício aplicado no solo + folha, silício aplicado à folha; e controle, sem aplicação de silício. Foram avaliadas as variáveis biológicas: duração do período pré-reprodutivo, período reprodutivo e fertilidade, e média diárias de ninfas produzidas por fêmea. Os teores de silício e de lignina nas plantas também foram avaliados. O ácido silícico foi aplicado em uma solução a 1% ao redor do caule das plantas (no solo), 15 dias após a emergência, diluindo-se 2,0 g do produto em 200 mL de água. Já a aplicação foliar foi realizada com um pulverizador de 1 L, 5 dias após a aplicação em solo. A não preferência de A. craccivora em feijão também foi avaliada. As avaliações foram realizadas após 48 e 72 horas da infestação, por meio da contagem de ninfas e adultos em cada secção foliar. A aplicação de silício promove a redução da produção de ninfas, interferindo nos aspectos biológicos de A. craccivora, podendo ser utilizado em programas de manejo de pragas do feijão-fava.(AU)

Antagonistic endophytic bacteria associated with nodules of soybean (Glycine max L. ) and plant growth-promoting properties

Abstract A total of 276 endophytic bacteria were isolated from the root nodules of soybean (Glycine max L.) grown in 14 sites in Henan Province, China. The inhibitory activity of these bacteria against pathogenic fungus Phytophthora sojae 01 was screened in vitro. Six strains with more than 63% inhibitory activities were further characterized through optical epifluorescence microscopic observation, sequencing, and phylogenetic analysis of 16S rRNA gene, potential plant growth-promoting properties analysis, and plant inoculation assay. On the basis of the phylogeny of 16S rRNA genes, the six endophytic antagonists were identified as belonging to five genera: Enterobacter, Acinetobacter, Pseudomonas, Ochrobactrum, and Bacillus. The strain Acinetobacter calcoaceticus DD161 had the strongest inhibitory activity (71.14%) against the P. sojae 01, which caused morphological abnormal changes of fungal mycelia; such changes include fracture, lysis, formation of a protoplast ball at the end of hyphae, and split ends. Except for Ochrobactrum haematophilum DD234, other antagonistic strains showed the capacity to produce siderophore, indole acetic acid, and nitrogen fixation activity. Regression analysis suggested a significant positive correlation between siderophore production and inhibition ratio against P. sojae 01. This study demonstrated that nodule endophytic bacteria are important resources for searching for inhibitors specific to the fungi and for promoting effects for soybean seedlings.

Analysis of bacterial communities and characterization of antimicrobial strains from cave microbiota

Abstract In this study for the first-time microbial communities in the caves located in the mountain range of Hindu Kush were evaluated. The samples were analyzed using culture-independent (16S rRNA gene amplicon sequencing) and culture-dependent methods. The amplicon sequencing results revealed a broad taxonomic diversity, including 21 phyla and 20 candidate phyla. Proteobacteria were dominant in both caves, followed by Bacteroidetes, Actinobacteria, Firmicutes, Verrucomicrobia, Planctomycetes, and the archaeal phylum Euryarchaeota. Representative operational taxonomic units from Koat Maqbari Ghaar and Smasse-Rawo Ghaar were grouped into 235 and 445 different genera, respectively. Comparative analysis of the cultured bacterial isolates revealed distinct bacterial taxonomic profiles in the studied caves dominated by Proteobacteria in Koat Maqbari Ghaar and Firmicutes in Smasse-Rawo Ghaar. Majority of those isolates were associated with the genera Pseudomonas and Bacillus. Thirty strains among the identified isolates from both caves showed antimicrobial activity. Overall, the present study gave insight into the great bacterial taxonomic diversity and antimicrobial potential of the isolates from the previously uncharacterized caves located in the world's highest mountains range in the Indian sub-continent.

Polyphasic characterization of bacteria obtained from upland rice cultivated in Cerrado soil

ABSTRACT This work aimed to characterize 20 isolates obtained from upland rice plants, based on phenotypic (morphology, enzymatic activity, inorganic phosphate solubilization, carbon source use, antagonism), genotypic assays (16S rRNA sequencing) and plant growth promotion. Results showed a great morphological, metabolic and genetic variability among bacterial isolates. All isolates showed positive activity for catalase and protease enzymes and, 90% of the isolates showed positive activity for amylase, catalase and, nitrogenase. All isolates were able to metabolize sucrose and malic acid in contrast with mannitol, which was metabolized only by one isolate. For the other carbon sources, we observed a great variability in its use by the isolates. Most isolates showed antibiosis against Rhizoctonia solani (75%) and Sclerotinia sclerotiorum (55%) and, 50% of them showed antibiosis against both pathogens. Six isolates showed simultaneous ability of antibiosis, inorganic phosphate solubilization and protease activity. Based on phylogenetic analysis of the 16S rRNA gene all the isolates belong to Bacillus genus. Under greenhouse conditions, two isolates (S4 and S22) improved to about 24%, 25%, 30% and 31% the Total N, leaf area, shoot dry weight and root dry weight, respectively, of rice plants, indicating that they should be tested for this ability under field conditions.

Antibiosis and antixenosis resistance to oviposition by Bemisia tabaci (Gennadius) B biotype (Hemiptera: Aleyrodidae) in soybean genotypes / Antibiose e antixenose para oviposição da Bemisia tabaci (Gennadius) Biótipo B (Hemiptera: Aleyrodidae) em genótipos de soja

This study was aimed at assessing some biological parameters of Bemisia tabaci B biotype at immature stages to examine antibiosis in six soybean genotypes, besides antixenosis resistance to oviposition in choice and non-choice tests, under greenhouse and laboratory conditions. Between April 2016 and August 2016, bioassays were carried out at the Laboratory of Applied Entomology at College of Agricultural Sciences at Federal University of Grande Dourados (UFGD) Dourados (MS), Brazil. The following variables were evaluated: egg incubation period, nymphal and egg-to-adult duration, egg and nymph viability, and egg-to-adult survival on the soybean genotypes 68i70 RSF IPRO, M6210 IPRO, MS 947 IPRO, BMX Potência RR, M6410 IPRO, and ANTA 82 RR. Antixenosis resistance to oviposition by the silverleaf whitefly was found for MS 947 IPRO, while oviposition was highest for ANTA 82 RR, M6210 IPRO, and M6410 IPRO. The genotype MS 947 IPRO adversely affected egg and nymph viability, and egg-to-adult survival rates was the lowest among genotypes (59.75%), suggesting antibiosis resistance to this pest.(AU)

Este trabalho teve por objetivo avaliar a antibiose em seis genótipos de soja da Bemisia tabaci biótipo B mediante alguns parâmetros biológicos das fases imaturas do inseto, assim como analisar a resistência do tipo antixenose para oviposição em testes com e sem chance de escolha, em laboratório e casa de vegetação. Os ensaios foram conduzidos no Laboratório de Entomologia Aplicada da Faculdade de Ciências Agrárias da Universidade Federal da Grande Dourados (UFGD), Dourados (MS), entre os meses de abril e agosto de 2016. As variáveis averiguadas foram: período de incubação dos ovos; período ninfal; duração de ovo a adulto; viabilidade de ovo; viabilidade de ninfa e sobrevivência de ovo a adulto. Os genótipos de soja avaliados foram: 68i70 RSF IPRO, M6210 IPRO, MS 947 IPRO, BMX Potência RR, M6410 IPRO e ANTA 82 RR. O genótipo que apresentou antixenose para oviposição da mosca-branca foi o MS 947 IPRO, e constatou-se que o maior número de postura foi realizado nos genótipos ANTA 82 RR, M6210 IPRO e M6410 IPRO. O genótipo MS 947 IPRO também afetou negativamente a viabilidade de ovos e ninfas, bem como a sobrevivência de ovo a adulto, com as menores porcentagens de sobrevivência (59,75%), indicando possível resistência do tipo antibiose à mosca-branca.(AU)

Biocontrol potential of saline- or alkaline-tolerant Trichoderma asperellum mutants against three pathogenic fungi under saline or alkaline stress conditions

ABSTRACT Salinity and alkalinity are major abiotic stresses that limit growth and development of poplar. We investigated biocontrol potential of saline- and alkaline-tolerant mutants of Trichoderma asperellum to mediate the effects of salinity or alkalinity stresses on Populus davidiana × P. alba var. pyramidalis (PdPap poplar) seedlings. A T-DNA insertion mutant library of T. asperellum was constructed using an Agrobacterium tumefaciens mediated transformation system; this process yielded sixty five positive transformants (T1-T65). The salinity tolerant mutant, T59, grew in Potato Dextrose Agar (PDA) containing up to 10% (1709.40 mM) NaCl. Under NaCl-rich conditions, T59 was most effective in inhibiting Alternaria alternata (52.00%). The alkalinity tolerant mutants, T3 and T5, grew in PDA containing up to 0.4% (47.62 mM) NaHCO3. The ability of the T3 and T5 mutants to inhibit Fusarium oxysporum declined as NaHCO3 concentrations increased. NaHCO3 tolerance of the PdPap seedlings improved following treatment with the spores of the WT, T3, and T5 strains. The salinity tolerant mutant (T59) and two alkalinity tolerant mutants (T3 and T5) generated in this study can be applied to decrease the incidence of pathogenic fungi infection under saline or alkaline stress.

Isolation, identification, and biocontrol of antagonistic bacterium against Botrytis cinerea after tomato harvest

ABSTRACT Tomato is one of the most important vegetables in the world. Decay after harvest is a major issue in the development of tomato industry. Currently, the most effective method for controlling decay after harvest is storage of tomato at low temperature combined with usage of chemical bactericide; however, long-term usage of chemical bactericide not only causes pathogen resistance but also is harmful for human health and environment. Biocontrol method for the management of disease after tomato harvest has great practical significance. In this study, antagonistic bacterium B-6-1 strain was isolated from the surface of tomato and identified as Enterobacter cowanii based on morphological characteristics and physiological and biochemical features combined with sequence analysis of 16SrDNA and ropB gene and construction of dendrogram. Effects of different concentrations of antagonistic bacterium E. cowanii suspension on antifungal activity after tomato harvest were analyzed by mycelium growth rate method. Results revealed that antifungal activity was also enhanced with increasing concentrations of antagonistic bacterium; inhibitory rates of 1 × 105 colony-forming units (cfu)/mL antagonistic bacterial solution on Fusarium verticillioides, Alternaria tenuissima, and Botrytis cinerea were 46.31%, 67.48%, and 75.67%, respectively. By using in vivo inoculation method, it was further confirmed that antagonistic bacterium could effectively inhibit the occurrence of B. cinerae after tomato harvest, biocontrol effect of 1 × 109 cfu/mL zymotic fluid reached up to 95.24%, and antagonistic bacterium E. cowanii has biocontrol potential against B. cinerea after harvest of fruits and vegetables.

Atividade antimicrobiana in vitro dos extratos aquosos, hidroalcoólicos e alcoólicos das folhas de espécies da família Myrtaceae frente à cepas de bactérias de interesse / Antimicrobial activity in vitro of aqueous extracts, hydroalcoholic and spirits of Myrtaceae family species leaves front of the strains of interest of bacteria

Introdução: a Organização Mundial da Saúde (OMS) tem incentivado o estudo de plantas tradicionalmente conhecidas como medicinais, tendo como objetivo avaliar cientificamente os benefícios da utilização de medicamentos fitoterápicos e de conhecer, ao mesmo tempo, a segurança de seu uso indevido. Objetivos: este trabalho teve como objetivo avaliar a atividade antimicrobiana do extrato aquoso, hidroalcoólico e alcoólico de folhas de espécies da família Myrtaceae frente às cepas de bactérias de interesse. Metodologia: a análise da atividade antibacteriana dos extratos das folhas das plantas, jambo, azeitona preta, pitanga e goiaba foram verificados a partir do teste de difusão em Ágar Mueller-Hinton. As cepas utilizadas, após reativação, foram diluídas em solução salina estéril a 0,85%. Sendo consideradas com potencial antimicrobiano aqueles extratos que geraram halos ≥ 7 mm (sete milímetros). Resultados: os resultados encontrados na análise dos dados do extrato aquoso demonstraram que Eugenia uniflora, Syzygium cumini e Psidium guajava apresentaram atividade antimicrobiana sobre Staphylococcus aureus, na concentração de 100 mg/mL. Na análise dos dados dos extratos hidroalcoólico, Syzygium cumini demonstrou atividade antimicrobiana na concentração de 100 mg/mL sobre Pseudomonas aeruginosa, diante de Staphylococcus aureus, Eugenia uniflora, Syzygium malaccense, Syzygium cumini e Psidium guajava demonstraram atividade na concentração de 100 mg/mL cada. Observando os resultados dos extratos alcoólicos, Psidium guajava e Syzygium cumini demonstraram atividade antimicrobiana diante das quatro cepas estudadas. Conclusão: o presente estudo demonstrou que os extratos alcoólicos das espécies em estudo apresentaram melhor atividade antimicrobiana e maior espectro de ação quando comparado ao extrato aquoso e hidroalcoólico.

Introduction: this study aimed to evaluate the antimicrobial activity of aqueous extract, hydro-alcoholic and alcoholic leaf front Myrtaceae family species to strains of bacteria of interest. Objective: this study aimed to evaluate the antimicrobial activity of aqueous extract, hydro alcoholic and alcoholic leaves Myrtaceae front family species to strains of bacteria of interest. Methodology: the antibacterial activity of plant leaves extracts, rose apple, black olive, cherry and guava were verified from the diffusion test on Mueller-Hinton agar. Where the strains used after reactivation were diluted in 0.85% sterile saline. Being considered with antimicrobial potential that statement that generated halos ≥ 7 mm (seven millimeters). Results: the results of the analysis of the aqueous extract data demonstrated that Eugenia uniflora, Syzygium cumini and Psidium guajava showed antimicrobial activity against Staphylococcus aureus at a concentration of 100 mg/mL. In the data analysis of hydro-alcoholic extracts, Syzygium cumini demonstrated antimicrobial activity at a concentration of 100 mg/mL of Pseudomonas aeruginosa, before Staphylococcus aureus, Eugenia uniflora, Syzygium malaccense, Syzygium cumini and Psidium guajava showed activity at a concentration of 100 mg/mL each. Noting the results of alcoholic extracts, Psidium guajava and Syzygium cumini demonstrated antimicrobial activity against the four strains studied. Conclusion: the present study demonstrated that the alcoholic extracts of the species under study presented better antimicrobial activity and a greater spectrum of action when compared to the aqueous and hydroalcoholic extract.

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

Abstract 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.

Bacterial selection for biological control of plant disease: criterion determination and validation

Abstract This study aimed to evaluate the biocontrol potential of bacteria isolated from different plant species and soils. The production of compounds related to phytopathogen biocontrol and/or promotion of plant growth in bacterial isolates was evaluated by measuring the production of antimicrobial compounds (ammonia and antibiosis) and hydrolytic enzymes (amylases, lipases, proteases, and chitinases) and phosphate solubilization. Of the 1219 bacterial isolates, 92% produced one or more of the eight compounds evaluated, but only 1% of the isolates produced all the compounds. Proteolytic activity was most frequently observed among the bacterial isolates. Among the compounds which often determine the success of biocontrol, 43% produced compounds which inhibit mycelial growth of Monilinia fructicola, but only 11% hydrolyzed chitin. Bacteria from different plant species (rhizosphere or phylloplane) exhibited differences in the ability to produce the compounds evaluated. Most bacterial isolates with biocontrol potential were isolated from rhizospheric soil. The most efficient bacteria (producing at least five compounds related to phytopathogen biocontrol and/or plant growth), 86 in total, were evaluated for their biocontrol potential by observing their ability to kill juvenile Mesocriconema xenoplax. Thus, we clearly observed that bacteria that produced more compounds related to phytopathogen biocontrol and/or plant growth had a higher efficacy for nematode biocontrol, which validated the selection strategy used.

Potential for biocontrol of melanized fungi by actinobacteria isolated from intertidal region of Ilha Do Mel, Paraná, Brazil

Abstract Actinobacteria occur in many environments and have the capacity to produce secondary metabolites with antibiotic potential. Identification and taxonomy of actinobacteria that produce antimicrobial substances is essential for the screening of new compounds, and sequencing of the 16S region of ribosomal DNA (rDNA), which is conserved and present in all bacteria, is an important method of identification. Melanized fungi are free-living organisms, which can also be pathogens of clinical importance. This work aimed to evaluate growth inhibition of melanized fungi by actinobacteria and to identify the latter to the species level. In this study, antimicrobial activity of 13 actinobacterial isolates from the genus Streptomyces was evaluated against seven melanized fungi of the genera Exophiala, Cladosporium, and Rhinocladiella. In all tests, all actinobacterial isolates showed inhibitory activity against all isolates of melanized fungi, and only one actinobacterial isolate had less efficient inhibitory activity. The 16S rDNA region of five previously unidentified actinobacterial isolates from Ilha do Mel, Paraná, Brazil, was sequenced; four of the isolates were identified as Streptomyces globisporus subsp. globisporus, and one isolate was identified as Streptomyces aureus. This work highlights the potential of actinobacteria with antifungal activity and their role in the pursuit of novel antimicrobial substances.

Evaluación del crecimiento de cuatro especies del género Bacillus sp, primer paso para entender su efecto biocontrolador sobre Fusarium sp / Evaluation of growth of four species of the genus Bacillus sp, the first step to understand their biocontrol effect on Fusarium sp

Evaluar las condiciones de crecimiento de cuatro especies de Bacillus sp. nativas a escala de 10ml en Medio Mínimo de Sales (MMS) como primer paso para entender su acción biocontroladora contra Fusarium sp. Métodos. El procedimiento para evaluar el crecimiento de los aislamientos UCMC-TB1, UCMC-TB2, UCMC-TB3 y UCMC-TB4 se realizó utilizando espectrofotometría y recuento directo en placa y pruebas de antagonismo dual en placa para evaluar el efecto controlador contra Fusarium sp. Resultados. Se confirmó la identificación por pruebas bioquímicas de los cuatro aislamientos: Bacillus licheniformis, Bacillus subtilis, Bacillus pumilusy Bacillus cereus; todas las cepas presentaron antagonismo in vitro. El Bacillus subtilis fue la especie que demostró mayor capacidad antagónica (79,73%PICR) y las características más destacadas de esta cepa fueron su velocidad de crecimiento. El género Bacillus es uno de los más reportados para usar en el control biológico de hongos como Fusarium sp. el cual ataca un gran número de cultivos de interés económico para el sector agrícola en Colombia.

Objetive. Evaluate the conditions of growth of four species of native Bacillus sp. on the scale of in minimal medium 10ml Sales (MMS) as a first step to understand their biocontrol action against Fusarium sp. Methods. The procedure for evaluating the growth of UCMC-TB1, UCMC-TB2, UCMC-TB3 and TB4 UCMC-isolates was performed using spectrophotometry and direct plate count and dual antagonism tests to evaluate the effect against Fusarium sp. Results. Identification by biochemical tests of the four isolates were confirmed: Bacillus licheniformis, Bacillus subtilis, Bacillus pumilus and Bacillus cereus; all showed antagonism in vitro. Bacillus subtilis was the species that showed increased antagonistic capacity (79.73% PICR) and the main features of this strain were the speed of growth and death. The genus Bacillus is one of the most reported for use in the biological control of fungi such as Fusarium sp. It is attacking a large number of crops of economic interest to the agricultural sector in Colombia.