Assessing the functional diversity of rhizobacteria from cacao by partitioning root and shoot biomasses.

Plant-microbe interactions are critical for the sustainability of agricultural production. In this study, our aims were to characterize the genetic and functional diversity of the culturable bacterial community associated with the cacao rhizosphere and access their potential for growth promotion of cacao seedling. Culture-dependent and molecular methods were used to characterize the population densities and diversity of bacterial communities from soil and cacao plants at two locations and two plant ages. A total of 63 strains were identified through hsp60 sequencing. Pseudomonas and Enterobacter were the most abundant genera in association with the cacao rhizosphere, whereas Bacillus was more numerous in soil. Parameters of seedling growth promotion were evaluated 60 days after inoculation of seeds, with partition of the assessments into root and shoot weight. Each isolate showed beneficial, neutral or deleterious effects on plant growth, depending on the isolate and on the parts of plant assessed. Interestingly, although an apparent overall decrease in total biomass of seedlings (roots + shoots dry matters) was observed for the majority of isolates (89%), 94% of all isolates, in fact, revealed an increase in plant roots/shoots dry biomass ratio. Despite that part of the isolates (35%) appeared to significantly decrease plant height, and that 65% did not influence plant height (neutral effect), 18 had significantly increased root dry biomass; nevertheless, seven of these root growth-increasing isolates simultaneously decreased shoots-related growth parameters. The results of this study evidentiated the functional diversity of culturable cacao rhizobacteria and how the partitioning of roots and shoots in the assessment of plant growth parameters could reveal the biotechnological potential of these isolates for promoting growth of clones for rehabilitation of commercial cacao plantations. KEY POINTS: • The most common culturable bacteria in cacao roots were Pseudomonas and Enterobacter • Most culturable bacteria from cacao roots increased the root/shoot ratio • Roots and shoots should be examined separately to detect cacao beneficial bacteria.

Phylogenomics and gene selection in Aspergillus welwitschiae: Possible implications in the pathogenicity in Agave sisalana.

Aspergillus welwitschiae causes bole rot disease in sisal (Agave sisalana and related species) which affects the production of natural fibers in Brazil, the main worldwide producer of sisal fibers. This fungus is a saprotroph with a broad host range. Previous research established A. welwitschiae as the only causative agent of bole rot in the field, but little is known about the evolution of this species and its strains. In this work, we performed a comparative genomics analysis of 40 Aspergillus strains. We show the conflicting molecular identity of this species, with one sisal-infecting strain sharing its last common ancestor with Aspergillus niger, having diverged only 833 thousand years ago. Furthermore, our analysis of positive selection reveals sites under selection in genes coding for siderophore transporters, Sodium­calcium exchangers, and Phosphatidylethanolamine-binding proteins (PEBPs). Herein, we discuss the possible impacts of these gene functions on the pathogenicity in sisal.

Infestation dynamics of Brevipalpus phoenicis (Geijskes) (Acari: Tenuipalpidae) in citrus orchards as affected by edaphic and climatic variables.

Brevipalpus phoenicis (Geijskes) is a cosmopolitan and polyphagous mite that transmits important phytoviruses, such as coffee ringspot virus, passion fruit green spot virus and Citrus leprosis virus C. To characterise the dynamics of the probability and the rate of B. phoenicis infestation in response to edaphic and climatic factors, monthly inspections were performed in nine orchards in a citrus region of the State of Bahia, Brazil, for 35 months. Three fruits per plant were examined using a magnifying glass (10×) on 21 plants distributed along a "W"-shaped path in each orchard. Meteorological data were collected from a conventional station. To determine the correlations among the climatic variables, the data were analysed using Spearman correlations. Variables were selected by principal component analysis, and those that contributed the most to differentiate the groups were evaluated via a Mann-Whitney test. Using the quantile-quantile method, the limit values for the following climatic variables were determined: temperature (24.5 °C), photoperiod (12 h), relative humidity (83%), evapotranspiration (71 mm) and rainy days (14 days). The combination of longer days, higher temperatures, lower relative humidity levels and lower evapotranspiration increased the probability of B. phoenicis infestation, whereas successive rain events decreased that risk. Infestation rates were negatively affected by relative humidity levels above 83% and were positively affected by a decreasing available soil-water fraction and increasing insolation and photoperiod.

The Sisal Virome: Uncovering the Viral Diversity of Agave Varieties Reveals New and Organ-Specific Viruses.

Sisal is a common name for different plant varieties in the genus Agave (especially Agave sisalana) used for high-quality natural leaf fiber extraction. Despite the economic value of these plants, we still lack information about the diversity of viruses (virome) in non-tequilana species from the genus Agave. In this work, by associating RNA and DNA deep sequencing we were able to identify 25 putative viral species infecting A. sisalana, A. fourcroydes, and Agave hybrid 11648, including one strain of Cowpea Mild Mottle Virus (CPMMV) and 24 elements likely representing new viruses. Phylogenetic analysis indicated they belong to at least six viral families: Alphaflexiviridae, Betaflexiviridae, Botourmiaviridae, Closteroviridae, Partitiviridae, Virgaviridae, and three distinct unclassified groups. We observed higher viral taxa richness in roots when compared to leaves and stems. Furthermore, leaves and stems are very similar diversity-wise, with a lower number of taxa and dominance of a single viral species. Finally, approximately 50% of the identified viruses were found in all Agave organs investigated, which suggests that they likely produce a systemic infection. This is the first metatranscriptomics study focused on viral identification in species from the genus Agave. Despite having analyzed symptomless individuals, we identified several viruses supposedly infecting Agave species, including organ-specific and systemic species. Surprisingly, some of these putative viruses are probably infecting microorganisms composing the plant microbiota. Altogether, our results reinforce the importance of unbiased strategies for the identification and monitoring of viruses in plant species, including those with asymptomatic phenotypes.

Calm Before the Storm: A Glimpse into the Secondary Metabolism of Aspergillus welwitschiae, the Etiologic Agent of the Sisal Bole Rot.

Aspergillus welwitschiae is a species of the Nigri section of the genus Aspergillus. In nature, it is usually a saprotroph, decomposing plant material. However, it causes the bole rot disease of Agave sisalana (sisal), a plant species used for the extraction of hard natural fibers, causing great economic loss to this culture. In this study, we isolated and sequenced one genome of A. welwitschiae (isolate CCMB 674 (Collection of Cultures of Microorganisms of Bahia)) from the stem tissues of sisal and performed in silico and wet lab experimental strategies to describe its ability to produce mycotoxins. CCMB 674 possesses 64 secondary metabolite gene clusters (SMGCs) and, under normal conditions, it produces secondary metabolism compounds that could disturb the cellular cycle of sisal or induce abnormalities in plant growth, such as malformin C. This isolate also produces a pigment that might explain the characteristic red color of the affected tissues. Additionally, this isolate is defective for the production of fumonisin B1, and, despite bearing the full cluster for the synthesis of this compound, it did not produce ochratoxin A. Altogether, these results provide new information on possible strategies used by the fungi during the sisal bole rot, helping to better understand this disease and how to control it.

Biological efficiency and nutritional value of Pleurotus ostreatus cultivated in agroindustrial wastes of palm oil fruits and cocoa almonds / Eficiência biológica e valor nutricional de Pleurotus ostreatus cultivados em resíduos agroindustriais de dendê e amêndoa de cacau

The cocoa and palm oil agro-industries active in the state of Bahia, Brazil, generate high quantities of lignocellulosic wastes that could be recycled through their use in the formulation of substrates to cultivate edible mushrooms. Pleurotus ostreatus, also known as oyster mushroom, is the second most cultivated mushroom in the world due to its highly appreciated gastronomic, nutritional, and medicinal characteristics. This work evaluated the vertical mycelium growth, biological efficiency, mushroom yield, and nutritional composition of P. ostreatus produced in substrates formulated with a combination of palm oil fruit mesocarp (POFM) and cocoa almond peels (CAP) processing wastes. The substrates were formulated with the following POFM/CAP proportions (%/%): S1 ­ 86.4/9.6; S2 ­ 76.8/19.2; S3 ­ 67.2/28.8; S4 ­ 57.6/38.4, and S5 ­ 48.0/48.0. Substrates also received 3% powdered charcoal and 1% calcium carbonate. Substrates S1, S2, S3, and S4 were superior for vertical mycelium growth. S2 promoted the best biological efficiency (148.8%) and yield (560.5g·kg-1).The mushrooms produced in all substrates presented good nutritional values, although mushrooms produced using the S2 presented the highest crude protein content. Overall, S1 is the recommended substrate as it results in higher yields of nutrient rich mushrooms. Production of P. ostreatus in substrates composed of POFM and CAP represents a good alternative for recycling these wastes with potential economic and ecological benefits to regions where palm oil and cocoa are grown.(AU)

As indústrias de cacau e óleo de dendê no estado da Bahia, Brasil, geram grandes quantidades de resíduos lignocelulósicos que podem ser reciclados na formulação de substratos para o cultivo de cogumelos comestíveis. Pleurotus ostreatus ou cogumelo ostra é o segundo cogumelo mais cultivado no mundo por apresentar características gastronômicas, nutricionais e medicinais muito apreciadas. Este estudo avaliou o crescimento micelial vertical, a eficiência biológica, a produção e a composição nutricional de P. ostreatus produzido em substratos formulados com a combinação de resíduos do processamento de frutos de dendê (mesocarpo do fruto de dendê ­ MFD) e de amêndoas de cacau (tegumento de amêndoas de cacau ­ TAC). Os substratos foram formulados com as seguintes proporções de MFD e TAC (%/%): S1: 86,4/9,6; S2: 76,8/19,2; S3: 67,2/28,8; S4: 57,6/38,4 e S5: 48,0/48,0. Os substratos também receberam 3% de carvão e 1% de carbonato de cálcio. Os substratos S1, S2, S3 e S4 foram superiores quanto ao crescimento micelial vertical. S2 promoveu os melhores resultados para eficiência biológica (148,8%) e produção (560.5 g·kg-1). Os cogumelos produzidos em todos os substratos apresentaram valores nutricionais promissores. Entretanto, os cogumelos produzidos com o substrato S2 apresentaram o maior conteúdo de proteína bruta. De modo geral, S1 é o substrato recomendado por resultar na maior produção de cogumelos ricos em nutrientes. A produção de P. ostreatus em substratos compostos por MFD e TAC representa uma boa alternativa para a reciclagem desses resíduos com potenciais benefícios econômicos e ecológicos para as regiões produtoras de dendê e cacau.(AU)

Five egg-laying queens in a single colony of brazilian stingless bees (Melipona scutellaris Latreille / Presença de cinco rainhas fisogástricas em colônia de abelha sem ferrão (Melipona scutellaris Latreille)

Polygyny, characterized by the presence of several egg-laying queens, is considered as a temporary colony status. In stingless bees it is rarely observed. This paper reports the first case of natural polygyny in Melipona scutellaris colony, with five egg-laying queens.

Poliginia, caracterizada pela presença de mais de uma rainha poedeira, é considerada como uma condição temporária em colônias. Em abelhas sem ferrão isso é raramente observado. Este artigo registra o primeiro caso de poliginia natural em colônia de Melipona scutellaris, com cinco rainhas poedeiras.

Streptomycetes antagonism against Cladosporium fulvum Cooke and Fusarium oxysporium f.sp. lycopersici / Antagonismo de estreptomicetos a Cladosporium fulvum Cooke e Fusarium oxysporium f.sp. lycopersici

This research aimed to evaluate the secondary effects of secondary metabolites produced by streptomycetes on spore germination and mycelial growth of the phytopathogenic fungi Cladosporium fulvum Cooke and Fusarium oxysporium f. sp. lycopersici from tomato plants. Metabolites produced by streptomycete isolates codified as AC-147 and AC-92 caused 94.1 percent inhibition of C. fulvum while AC-95 isolate caused 33.9 percent inhibition. AC-92 was the most efficient for F. oxysporum f. sp. lycopersici, causing 94.2 percent inhibition of spore germination. For mycelial growth, AC-26 and AC-92 were the most efficient in inhibiting C. fulvum growth by 46.6 percent and F. oxysporum f. sp. lycopersici by 29.9 percent. These streptomycetes are potential agents for biocontrol development methods of these tomato plant pathogenic fungi.

Este trabalho teve como objetivo avaliar o efeito de metabólitos secundários produzidos por estreptomicetos na germinação de esporos e no crescimento micelial dos fungos Cladosporium fulvum Cooke e Fusarium oxysporum sp. f. lycopersici da cultura do tomateiro. Metabólitos produzidos pelos isolados AC-147 e AC-92 causaram 94,1 por cento de inibição da germinação de esporos de C. fluvum, enquanto que o isolado AC-95 causou 33,9 por cento de inibição. O AC-92 foi o mais eficiente para F. oxysporum f. sp. lycopersici, causando 94,2 por cento de inibição na germinação de esporos. Para o crescimento micelial, AC-26 e AC-92 foram os mais eficientes na inibição dos fungos C. fulvum, em 46,6 por cento, e F. oxysporum f. sp. Lycopersici, em 29,9 por cento. Esses estreptomicetos são potenciais agentes para o desenvolvimento de métodos de controle biológico desses fungos fitopatogênicos do tomateiro.

Amostragem, caracterização de sintomas e escala diagramática da mancha graxa dos citros (Mycosphaerella citri) no Recôncavo Baiano / Sampling, characterization of citrus greasy spot (Mycosphaerella citri) symptoms in Recôncavo Baiano and development of a diagrammatic scale for assessment of severity

O Recôncavo da Bahia é uma região de clima propício à mancha graxa dos citros (Mycosphaerella citri), doença nunca antes estudada no Brasil. Os objetivos deste trabalho foram definir o tamanho mínimo de amostra para quantificar a incidência, caracterizar os sintomas da doença e elaborar uma escala diagramática para a avaliação de sua severidade. A partir de uma amostragem piloto realizada por avaliação de cinco folhas por quadrante, quatro quadrantes por planta e 30 plantas em cada um dos 10 pomares visitados no município de Cruz das Almas, foi determinado que o tamanho mínimo da amostra em quadrantes e folhas seria de quatro e 16 plantas, respectivamente. De 320 folhas coletadas de diferentes plantas em 11 pomares, foi constatada uma grande quantidade de lesões (média de 131) por folha sintomática, em sua maioria diminutas (média de 0,014cm²). A severidade variou de 0,15 por cento a 35,85 por cento, com média de 7,3 por cento. Com base na severidade real em campo, foi elaborada uma escala diagramática com seis níveis: 1 por cento, 2 por cento, 5 por cento, 9 por cento, 18 por cento e 36 por cento de área foliar lesionada. Cinqüenta imagens de folhas foram submetidas a avaliadores por duas vezes, os quais estimaram a severidade com e sem a utilização da escala, constatando-se um discreto benefício no grau de precisão e de acurácia das estimativas com o uso da escala.

Weather conditions make the Recôncavo Region in the State of Bahia, a favorable region to the occurrence of citrus greasy spot (Mycosphaerella citri), a disease that hadn't been studied before in Brazil. Thus, this research aimed to develop a sampling method to quantify its incidence, to characterize its symptoms, and to develop a diagrammatic scale for assessing disease severity. The minimum sample size to estimate greasy spot incidence in leaves (16 plants) was calculated based on a prospective sampling of five leaves per quadrant, four quadrants per tree, and 30 trees per grove, in ten groves. From 320 mature leaves collected in different plants of 11 groves, it was observed a high number of lesions per symptomatic leaf (mean of 131±102), most of them small (mean of 0.014±.,011cm²). Greasy spot severity varied from 0,15 percent to 35,85 percent, with a mean of 7,3 percent. Based on those severity values, a six level (1 percent, 2 percent, 5 percent, 9 percent, 18 percent e 36 percent of lesioned leaf area) diagrammatic scale was developed. 50 images of symptomatic leaves were showed to evaluators, which estimated the severity with and without the use of the scale. The diagrammatic scale use, slightly increased the estimates accuracy and precision.

Soil streptomycetes with in vitro activity against the yam pathogens Curvularia eragrostides and Colletotrichum gloeosporioides

Yam (Dioscorea cayennensis Lam), an important crop in the Northeastern region of Brazil, is susceptible to the attack of several phytopathogenic fungi, such as Curvularia eragrostides (Henn.) Meyer and Colletotrichum gloeosporioides (Penz.), which cause leaf spot diseases. This work aimed to evaluate the effect of six actinomycete isolates (S. thermotolerans, S. griseus subsp. griseus, Streptomyces sp. N0035, S. purpurascens, and two isolates identified as Streptomyces sp.), as well as the effect of their secondary metabolites, on mycelium growth and spore germination of C. eragrostides and C. gloeosporioides, from yam plants, cultivar da Costa. Additionally, the chitinolytic activity of these actinomycete strains was determined in a minimum salt growth medium with chitin as the sole carbon source. There was significant interaction between the actinomycete isolates and the phytopathogenic fungi for spore germination and mycelium growth inhibition. Streptomyces sp. codified as AC 26 was more efficient in inhibiting spore germination of C. eragrostides, while S. thermotolerans and Streptomyces sp. N0035 were more efficient for C. gloeosporioides. Streptomyces sp. (AC 26) was the most efficient in inhibiting mycelium growth of both fungi, and the inhibitory effect was positively correlated to metabolite concentration. S. griseus subsp. griseus did not show chitinolytic activity and did not control mycelium growth of both fungi. The actinomycete strains evaluated in this study can be considered potential biological agents for controlling yam phytopathogenic fungi associated with leaf spot diseases.

O inhame da Costa (Dioscorea cayennensis Lam.), uma cultura importante na região do nordeste Brasileiro, é suscetível ao ataque de diversos fungos fitopatogênicos, como Curvularia eragrostides (Henn.) Meyer e Colletotrichum gloeosporioides (Penz.), que causam manchas foliares. Este trabalho teve o objetivo de avaliar o efeito de seis isolados de actinomicetos (S. thermotolerans, S. griseus subsp. griseus, Streptomyces sp. N0035, S. purpurascen e dois isolados identificados como Streptomyces sp.) e de seus metabólitos secundários no crescimento micelial e na germinação de esporos de C. gloeosporioides e C. eragrostides, da cultura do inhame da Costa. Adicionalmente, foi avaliada a atividade quitinolítica desses microrganismos, em meio de sais minerais e ágar, contendo quitina coloidal como única fonte de carbono. Houve interação significativa entre os isolados de actinomicetos e os fungos fitopatogênicos para inibição da germinação de esporos e do crescimento micelial. Streptomyces sp., codificado como AC 26, foi eficiente na inibição da germinação de esporos de C. eragrostides, enquanto que S. thermotolerans e Streptomyces sp. N0035 foram mais eficientes para C. gloeosporioides. Streptomyces sp. (AC 26) foi o mais eficiente na inibição do crescimento micelial de ambos os fungos e o efeito inibitório aumentou com o aumento na concentração dos metabólitos. S. griseus subsp. griseus não apresentou atividade quitinolítica e não inibiu o crescimento micelial de ambos os fungos. Os actinomicetos testados neste trabalho podem ser considerados potenciais agentes de biocontrole de doenças foliares do inhame.