Genome-wide identification of the banana GLR gene family and its expression analysis in response to low temperature and abscisic acid/methyl jasmonate / 生物工程学报

Glutamate receptor-like (GLR) is an important class of Ca2+ channel proteins, playing important roles in plant growth and development as well as in response to biotic and abiotic stresses. In this paper, we performed genome-wide identification of banana GLR gene family based on banana genomic data. Moreover, we analyzed the basic physicochemical properties, gene structure, conserved motifs, promoter cis-acting elements, evolutionary relationships, and used real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) to verify the expression patterns of some GLR family members under low temperature of 4 ℃ and different hormone treatments. The results showed that there were 19 MaGLR family members in Musa acuminata, 16 MbGLR family members in Musa balbisiana and 14 MiGLR family members in Musa itinerans. Most of the members were stable proteins and had signal peptides, all of them had 3-6 transmembrane structures. Prediction of subcellular localization indicated that all of them were localized on the plasma membrane and irregularly distributed on the chromosome. Phylogenetic analysis revealed that banana GLRs could be divided into 3 subclades. The results of promoter cis-acting elements and transcription factor binding site prediction showed that there were multiple hormone- and stress-related response elements and 18 TFBS in banana GLR. RT-qPCR analysis showed that MaGLR1.1 and MaGLR3.5 responded positively to low temperature stress and were significantly expressed in abscisic acid/methyl jasmonate treatments. In conclusion, the results of this study suggest that GLR, a highly conserved family of ion channels, may play an important role in the growth and development process and stress resistance of banana.

Identification and verification of key salt-tolerant amino acid sites of banana MaNHX5 / 生物工程学报

In order to improve the salt tolerance of banana NHX genes, we cloned a MaNHX5 gene from Musa acuminata L. AAA group and predicted the key salt-tolerant amino acid sites and mutant protein structure changes of MaNHX5 by using bioinformatics tools. The 276-position serine (S) of MaNHX5 protein was successfully mutated to aspartic acid (D) by site-directed mutagenesis, and the AXT3 salt-sensitive mutant yeast was used for a functional complementation test. The results showed that after the mutated MaNHX5 gene was transferred to AXT3 salt-sensitive mutant yeast, the salt tolerance of the mutant yeast was significantly improved under 200 mmol/L NaCl treatment. It is hypothesized that Ser276 of MaNHX5 protein plays an important role in the transport of Na+ across the tonoplast.

El compostaje de cáscara de Musa paradisiaca L. para la producción de biogás y biometano como estrategia de preservación de la salud ambiental / Musa paradisiaca L. husk composting for biogas and biomethane production as a strategy for preserving environmental health

A medida que a se avanza en el siglo XXI, los sistemas de energía deben alejarse de los combustibles fósiles y aumentar la capacidad de las energías renovables si se quieren cumplir los objetivos de temperaturas máximas del Acuerdo de París. Sin embargo, debido a los desafíos en la adopción de tecnologías bajas en carbono, ciertas áreas de los sistemas energéticos globales son difíciles de controlar y descarburar. Por otra parte, el compostaje es una de las prácticas de gestión de residuos orgánicos más importantes que se puede utilizar para lograr la sostenibilidad del suelo y del medio ambiente. El compost tiene un mínimo impacto en algunas emisiones, y puede ayudar a controlar la huella de carbono y limitar los efectos ambientales negativos de los métodos de eliminación de desechos más deficientes. La investigación tuvo por objetivo determinar la calidad de producir biogás y biometano a partir de la cáscara de plátano (Musa paradisiaca L.). Metodológicamente se desarrolló una investigación aplicada, con nivel de investigación de tipo experimental. Las cáscaras de plátano se colectaron de la planta de compostaje de la Municipalidad Provincial de Leoncio Prado, Perú. De la muestra se prepararon cinco sub muestras para la producción de biogás y cinco muestras adicionales para la producción de biometano. Los sistemas mostraron una producción de 0,067 m3 BG/Kg ST de biogás y 0,059 m3CH4/Kg ST de biometano, que generó subproductos como el biol y biosol. Estos resultados presentaron una baja toxicidad al ser sometidos a pruebas germinativas, concluyéndose que solo el 11,5% de la cáscara introducida al biorreactor se degrado y de esta fracción solo el 2,8% se convirtió en biogás(AU)

As progress is being made in the 21st century, energy systems must move away from fossil fuels and increase the capacity of renewable energies if you want to meet the maximum temperatures objectives of the Paris Agreement. However, due to the challenges in the adoption of low carbon technologies, certain areas of global energy systems are difficult to control and decarbure. On the other hand, composting is one of the most important organic waste management practices that can be used to achieve soil and environmental sustainability. The compost has a minimum impact on some emissions, and can help control the carbon footprint and limit the negative environmental effects of the most deficient waste removal methods. The research aimed to determine the quality of producing biogas and biomethane from the banana peel (Musa paradisiaca L.). Methodologically, an applied investigation was developed, with experimental research level. The banana peels were collected from the composting plant of the Provincial Municipality of Leoncio Prado, Peru. From the sample, five sub samples were prepared for the production of biogas and five additional samples for biomethane production. The systems showed a production of 0.067 m3 bg/kg ST of biogas and 0.059 m3ch4/kg ST of biomethane, which generated by -products such as biol and biosol. These results presented a low toxicity when they were subjected to germinative evidence, concluding that only 11.5% of the shell introduced into the bioreactor was degraded and of this fraction only 2.8% became biogas(AU)

A degradação do amido da banana depende da ação combinada de α-amilase e ß-amilase em regiões de diferentes graus de cristalinidade do grânulo / The banana starch degradation depends on the combined action of α-amylase and ß-amylase in regions of different degrees of crystallinity of the granules

A banana é considerada um bom modelo de estudo para a transformação amido-sacarose, já que acumula um teor alto de amido durante o desenvolvimento, que é degradado durante o amadurecimento. Já foram detectadas em polpa de banana atividade e proteína relativa a várias enzimas supostamente envolvidas no processo de degradação do amido. Entre elas, a α-amilase, a ß-amilase, a amido fosforilase e as glucano-água-diquinases (GWD). Estas enzimas estão envolvidas em dois processos distintos de degradação de amido em plantas: o dependente da ação inicial da α-amilase e o dependente da fosforilação do grânulo pela GWD e PWD e posterior ação da ß-amilase. A dificuldade do estabelecimento da participação efetiva de cada enzima no processo de degradação do amido está associada a muitos fatores, entre eles a não-correlação entre atividade e real envolvimento em um processo, e a acessibilidade da enzima ao seu substrato. Aliado ao estudo da morfologia do grânulo de amido e suas modificações sofridas durante o processo de degradação que ocorre durante o amadurecimento do fruto, estudos in vitro que simulem a ação da enzima sobre o seu substrato poderiam ser mais efetivos no estabelecimento da real ação de dada enzima sobre o suposto substrato. Tentativas no sentido de obter as proteínas relativa à degradação não foram bem sucedidas. Assim, os ensaios de grânulos de amido isolados versus enzimas foram feitos com α-amilase e ß-amilase comerciais. O grau de fosforilação da amilopectina nas posições Glic-6 e Glic-3 foi determinado, condição necessária para o início da degradação do grânulo pela ß-amilase. Os resultados mostraram que os grânulos de amido isolados de bananas recém colhidas, ou verdes, já estão fosforilados e as enzimas responsáveis por esta fosforilação estão associadas aos grânulos. Após 72 h de incubação dos grânulos de amido com as enzimas hidrolítica, os grânulos foram separados do tampão contendo as enzimas e os produtos de hidrólise. Os sobrenadantes foram analisados por cromatografia líquida acoplada a detector amperométrico e os grânulos por Microscopia Eletrônica de Varredura (MEV) e microscopia de força atômica (MFA). Os resultados mostraram que a α-amilase hidrolisa preferencialmente regiões amorfas dos grânulos, com predominância de amilose, expondo as regiões mais cristalinas dos anéis de crescimento, enquanto que a ß-amilase parece atuar preferencialmente nas regiões cristalinas dos grânulos, degradando os bloquetes, que são formados por amilopectina. Pode-se concluir que ambas as enzimas parecem ser importantes no processo de degradação do amido da banana, com diferentes especificidades

Banana is considered a good model to study the starch-sucrose metabolism, since it accumulates a high starch content during development, which is degraded during fruit ripening. It have been detected in banana pulp some proteins and activities of several enzymes supposedly involved in starch degradation process. Among them, α-amylase, ß-amylase, starch phosphorylase and glucan-water-diquinases (GWD). These enzymes are involved in two separate processes of starch degradation in plants: the initial action of α-amylase dependent, and the starch granule phosphorylation by GWD and PWD enzymes and subsequent action of ß-amylase. The difficulty of establishing the effective participation of each enzyme in the starch degradation process is associated with many factors, including the lack of correlation between real activity and involvement in the process, and accessibility of the enzyme to its substrate. Allied to study the morphology of the starch granule and its modifications suffered during the process of degradation, which occurs during the fruit ripening, in vitro studies that simulate the action of the enzyme on its substrate could be more effective in establishing the real action of a given enzyme on the argued substrate. However, attempts to obtain the proteins related to the degradation process were unsuccessful. Thus, assays of isolated starch granules versus enzymes were made with commercial α-amylase and ß-amylase enzymes. The degree of phosphorylation of amylopectin in the Gluc-6 and Gluc-3 positions was determined, a necessary condition for the start of degradation by ß-amylase enzyme. The results showed that the starch granules isolated from freshly harvested bananas, or green, are already phosphorylated and the enzymes responsible for this phosphorylation is associated with the starch granules surface. After 72 h incubation of the starch granules with the hydrolytic enzymes, the granules were separated from the buffer containing the enzymes and the hydrolysis products. The supernatants were analyzed by liquid chromatography coupled with amperometric detector and the granules were visualized by scanning electron microscopy (SEM) and atomic force microscopy (AFM). The results showed that the α-amylase preferentially hydrolyzes amorphous regions of the granule, especially amylose, exposing more crystalline regions of the growth rings, whereas ß-amylase appears to act preferentially on crystalline regions of the granule, degrading blocklets that consist of amylopectin. It can be concluded that both enzymes appear to be important in the banana starch degradation process, with different specificities

O efeito da biodiversidade nativa da Mata Atlântica sobre o metabolismo de amadurecimento da banana (Musa acuminata AAA, Cavendish, cv. Nanicão) / he effect of the Atlantic Coastal Forest´s native biodiversity on the metabolism of banana ripening (Musa acuminata AAA, Cavendish, cv. Nanicão).

A produção de banana no Vale do Ribeira, em São Paulo, está inserida numa região que abriga um fragmento remanescente da Mata Atlântica, considerada um dos principais repositórios de biodiversidade e a floresta mais devastada entre os biomas brasileiros. Medidas sustentáveis, como a adoção de práticas agrícolas alternativas, tem sido o caminho para a conservação da fauna e flora, como também para a redução do uso inapropriado de terras que consequentemente pode aumentar o índice de doenças que atacam as plantas. Apesar, do razoável conhecimento científico sobre a fisiologia pós-colheita da banana, ainda não foram elucidadas quais são as respostas fisiológicas do fruto diante das alterações do meio ambiente decorrentes das práticas agroecológicas. Principalmente quais são os mecanismos de ação de compostos específicos, relacionados com a resistência da planta contra os estresses bióticos e abióticos. Também, não há uma abordagem analítica integrada que identifique os reguladores das vias metabólicas e possibilite um estudo holístico a nível molecular. Neste trabalho, foi avaliado o efeito da proximidade da biodiversidade nativa da Mata Atlântica sobre o perfil de metabólitos da banana (Musa acuminata AAA, Cavendish, cv. Nanicão). Foram comparados os frutos da parcela Biodiversidade, o qual apresenta 60% de seu perímetro limítrofe à floresta com a parcela Controle, inserida em uma área de produção de banana convencional. Neste estudo, foram determinados os perfis de etileno, vida verde, amido, açúcares, textura, cor, metabólitos semi-voláteis e não-voláteis e poliaminas. Como resultado, ambas as parcelas avaliadas apresentaram diferenças significativas no metabolismo primário e secundário, nos frutos verdes e maduros, respectivamente. Os metabólitos com diferença significativa entre as parcelas experimentais apresentaram maior envolvimento no metabolismo primário, sobretudo na via de biossíntese de aminoácidos, compostos precursores de uma ampla faixa de metabólitos secundários. As alterações quanto à abundância destes compostos são uma referência de que as mudanças do meio modificam as respostas da planta a estresses e estímulos

Banana production in the Ribeira Valley, in São Paulo, is set in a region that embraces a remaining fragment of the Atlantic Coastal Forest (Mata Atlântica), considered one of the main biodiversity repositories and the most devastated forest among the Brazilians´ biomes. Sustainable measures such as the adoption of alternative farming practices has been the way for the conservation of fauna and flora, but also to reduce the inappropriate use of land which in turn may increase the disease index that attack plants. Although, reasonable scientific knowledge about banana postharvest physiology is available, the fruit´s physiologic response given the environment changes as a result agroecological practices has not been elucidated yet. Primarily what are the mechanisms of action of specific compounds, related to plant resistance against biotic and abiotic stresses. In addition, there is not an integrated analytical approach to identify regulators of metabolic pathways and enable a holistic study at the molecular level. In this study, we evaluated the effect of proximity to the native biodiversity of the Atlantic Forest on the banana metabolites profile (Musa acuminata AAA, Cavendish, cv. Nanicão). The fruits of Biodiversity group were compared, which has 60% of its perimeter adjacent to the forest with the Control group set in a conventional banana production area. In this study, we determined the ethylene profiles, green life, starch, sugars, texture, colour, semi- volatile and non- volatile metabolites and polyamines. As a result, both evaluated groups showed significant differences in primary and secondary metabolism, green and ripen fruits respectively. The metabolites with significant differences between treatments showed greater involvement in primary metabolism, especially in amino acid biosynthesis, precursors of secondary metabolites. Changes in the abundance of these compounds are indicators that alterations in the environment modify the plant responses to stresses and stimuli

Production of plant cell wall degrading enzymes by monoculture and co-culture of Aspergillus niger and Aspergillus terreus under SSF of banana peels

Filamentous fungi are considered to be the most important group of microorganisms for the production of plant cell wall degrading enzymes (CWDE), in solid state fermentations. In this study, two fungal strains Aspergillus niger MS23 and Aspergillus terreus MS105 were screened for plant CWDE such as amylase, pectinase, xylanase and cellulases (β-glucosidase, endoglucanase and filterpaperase) using a novel substrate, Banana Peels (BP) for SSF process. This is the first study, to the best of our knowledge, to use BP as SSF substrate for plant CWDE production by co-culture of fungal strains. The titers of pectinase were significantly improved in co-culture compared to mono-culture. Furthermore, the enzyme preparations obtained from monoculture and co-culture were used to study the hydrolysis of BP along with some crude and purified substrates. It was observed that the enzymatic hydrolysis of different crude and purified substrates accomplished after 26 h of incubation, where pectin was maximally hydrolyzed by the enzyme preparations of mono and co-culture. Along with purified substrates, crude materials were also proved to be efficiently degraded by the cocktail of the CWDE. These results demonstrated that banana peels may be a potential substrate in solid-state fermentation for the production of plant cell wall degrading enzymes to be used for improving various biotechnological and industrial processes.

Production of bioethanol using agricultural waste: banana pseudo stem

India is amongst the largest banana (Musa acuminata) producing countries and thus banana pseudo stem is commonly available agricultural waste to be used as lignocellulosic substrate. Present study focuses on exploitation of banana pseudo stem as a source for bioethanol production from the sugars released due to different chemical and biological pretreatments. Two fungal strains Aspergillus ellipticus and Aspergillus fumigatus reported to be producing cellulolytic enzymes on sugarcane bagasse were used under co-culture fermentation on banana pseudo stem to degrade holocellulose and facilitate maximum release of reducing sugars. The hydrolysate obtained after alkali and microbial treatments was fermented by Saccharomyces cerevisiae NCIM 3570 to produce ethanol. Fermentation of cellulosic hydrolysate (4.1 g%) gave maximum ethanol (17.1 g/L) with yield (84%) and productivity (0.024 g%/h) after 72 h. Some critical aspects of fungal pretreatment for saccharification of cellulosic substrate using A. ellipticus and A. fumigatus for ethanol production by S. cerevisiae NCIM 3570 have been explored in this study. It was observed that pretreated banana pseudo stem can be economically utilized as a cheaper substrate for ethanol production.

Ocorrência e biossíntese de frutooligossacarídeos em banana / Occurrence and biosynthesis of fructooligosaccharides in banana

A banana tem sido comumente indicada como uma boa fonte de frutooligossacarídeos (FOS), que são considerados componentes funcionais de alimentos. Contudo, diferenças significantes em suas quantidades têm sido referidas na literatura. Portanto, uma parte do trabalho foi destinada à identificação e quantificação de FOS durante o amadurecimento de cultivares de bananas pertencentes aos grupos genômicos mais comumente cultivados no Brasil. Considerando as diferencas de cultivar, estágio do amadurecimento e metodologia usada para análise de FOS, os conteúdos dos acúcares foram analisados por cromatografia líquida de alta performance (HPAEC-PAD) e cromatografia a gás (CG-MS). Uma pesquisa inicial entre oito cultivares no estágio maduro, mostrou acúmulo de 1-cestose, primeiro membro da série de FOS, em todas elas (quantidades entre 297 e 1600 ´MUg/g M. S´). A nistose, o segundo membro, foi detectado somente no cultivar Prata. Com bases nestes dados, foram escolhidas cinco cultivares, para que fossem analisadas durantes todo o amadurecimento. Os resultados mostraram uma forte correlação entre a síntese de 1-cestose e um nível específico de sacarose (~200mg/g M.S)...

Caracterização funcional do gene maBMY que codifica para uma `BETA´-amilase endereçada a plastídeos e expressa durante o amadurecimento da banana / Functional characterization of maBMY gene that encodes a `BETA´-amylase targeted to plastids and expressed during banana ripening

O amadurecimento dos frutos é um processo caracterizado pela ocorrência de diversas alterações bioquímicas que ocorrem em um curto intervalo de tempo e que são importantes para a qualidade desses alimentos. Na banana uma das características mais importantes é o adoçamento do fruto, que ocorre como resultado da degradação do amido e acúmulo de sacarose. Resultados do nosso grupo apontam a ´BETA` amilase como uma enzima importante no processo de mobilização do amido, o que também é visto em estudos recentes utilizando Arabidopsis thaliana como modelo, os quais mostram que a principal via de degradação do amido transitório presente nas folhas ocorre pela ação da ´BETA`-amilase. Entretanto, em bananas, faltam evidências quanto à funcionalidade de um gene de ´BETA`amilase, parcialmente isolado da polpa do fruto, e que é expresso durante o amadurecimento e que parece ser modulado por hormônios vegetais. Em vista disso, esse trabalho objetivou realizar a caracterização funcional desse gene, a qual permitiu constatar que esse gene codifica, de fato, para uma proteína capaz de ser endereçada aos cloroplastos. Também foi observado que o promotor desse gene contém motivos regulatórios para os mesmos hormônios previamente relacionados com a modulação da expressão desse gene em bananas. Essas novas evidências reforçam a idéia de que o produto desse gene de ´BETA`amilase tem um importante papel no processo de degradação do amido durante o amadurecimento da banana...

Infra-red spectroscopic analyses of banana waste degraded by oyster mushroom.

Carbon, hydrogen and nitrogen analyses of banana leaf and pseudostem biomass revealed their potentiality as substrates for microorganisms. Infra-red (IR) spectra of both biomass show presence of cellulose, xylan and lignin. IR spectra of leaf and pseudostem biomass degraded in solid state fermentation (SSF) by two Pleurotus species (P. sajor-caju and P. ostreatus) for 40 days showed the utilization of cellulose, xylan and lignin by these microbes. Dynamics of various lignocellulolytic enzymes of Pleurotus species and analyses of carbon, hydrogen and nitrogen contents of degraded biomass supported the same. Both the Pleurotus species exhibited lignin consumption ability on both the substrates.

Biotransformation of banana waste into protein by pleurotus sajor-caju.

Pseudostems and leaves from banana waste were used for biotransformation into protein by using P. sajor-caju, an oyster mushroom. Treatment of formalin (500 ppm) + carbendazim (12.5 ppm) of these substrates was found to favour relatively high percentage biological efficiency (BE) of P. sajor-caju.. Steam sterilization also exhibited comparable yield performance by P. sajor-caju. Fruiting bodies harvested from all the treatments had relatively higher protein contents. The spent substrate (steam sterilized) was found to be suitable as an ideal animal feed because of its rich nutritive composition.