Antifungal activity of volatile compounds generated by endophytic fungi Sarocladium brachiariae HND5 against Fusarium oxysporum f. sp. cubense.

The soil-born filamentous fungal pathogen Fusarium oxysporum f. sp. cubense (FOC), which causes vascular wilt disease in banana plants, is one of the most economically important Fusarium species. Biocontrol using endophytic microorganisms is among the most effective methods for controlling banana Fusarium wilt. In this study, volatile organic compounds (VOCs) showed strong antifungal activity against FOC. Seventeen compounds were identified from the VOCs produced by endophytic fungi Sarocladium brachiariae HND5, and three (2-methoxy-4-vinylphenol, 3,4-dimethoxystyrol and caryophyllene) showed antifungal activity against FOC with 50% effective concentrations of 36, 60 and 2900 µL/L headspace, respectively. Transmission electron microscopy (TEM) and double fluorescence staining revealed that 2-methoxy-4-vinylphenol and 3,4-dimethoxystyrol damaged the plasma membranes, resulting in cell death. 3,4-dimethoxystyrol also could induce expression of chitin synthases genes and altered the cell walls of FOC hyphae. Dichloro-dihydro-fluorescein diacetate staining indicated the caryophyllene induced accumulation of reactive oxygen species (ROS) in FOC hyphae. FOC secondary metabolism also responded to active VOC challenge by producing less fusaric acid and expressions of genes related to fusaric acid production were interrupted at sublethal concentrations. These findings indicate the potential of S. brachiariae HND5 as a biocontrol agent against FOC and the antifungal VOCs as fumigants.

Total phenolics, flavonoids and antioxidant activity following simulated gastro-intestinal digestion and dialysis of banana (Musa acuminata, AAB) as affected by induced ripening agents.

The present study was conducted to evaluate effect of ethephon and acetylene treatments on phenolics, flavonoids and antioxidant activity of banana flesh and their bioaccessibility. Total phenolics, flavonoids and antioxidant activity (DPPH, ABTS, FRAP) were measured at different phases of simulated gastrointestinal digestion of banana treated with 1000 ppm ethephon and 1000 ppm acetylene against natural ripening. The results revealed that inducing ripening lowers the content of phenolics, flavonoids and antioxidant activity considerably in the fresh fruit. Bioavailability of phenolics, flavonoids and FRAP activity were increased significantly (p < 0.05) after gastric digestion regardless of the treatment. The release of polyphenols and flavonoids during gastric digestion in treated banana was more significant than in naturally ripened banana. Recovery of polyphenols after dialysis was significantly high in naturally ripened banana. Dialyzable flavonoids, DPPH and ABTS activities of dialyzed fractions were not significantly affected by ethephon or acetylene treatments.

Predicting Virulence of Fusarium Oxysporum f. sp. Cubense Based on the Production of Mycotoxin Using a Linear Regression Model.

Fusarium wilt caused by Fusarium oxysporum f.sp. cubense (Foc) is one of the most destructive diseases for banana. For their risk assessment and hazard characterization, it is vital to quickly determine the virulence of Foc isolates. However, this usually takes weeks or months using banana plant assays, which demands a better approach to speed up the process with reliable results. Foc produces various mycotoxins, such as fusaric acid (FSA), beauvericin (BEA), and enniatins (ENs) to facilitate their infection. In this study, we developed a linear regression model to predict Foc virulence using the production levels of the three mycotoxins. We collected data of 40 Foc isolates from 20 vegetative compatibility groups (VCGs), including their mycotoxin profiles (LC-MS) and their plant disease index (PDI) values on Pisang Awak plantlets in greenhouse. A linear regression model was trained from the collected data using FSA, BEA and ENs as predictor variables and PDI values as the response variable. Linearity test statistics showed this model meets all linearity assumptions. We used all data to predict PDI with high fitness of the model (coefficient of determination (R2 = 0.906) and adjust coefficient (R2adj = 0.898)) indicating a strong predictive power of the model. In summary, we developed a linear regression model useful for the prediction of Foc virulence on banana plants from the quantification of mycotoxins in Foc strains, which will facilitate quick determination of virulence in newly isolated Foc emerging Fusarium wilt of banana epidemics threatening banana plantations worldwide.

Physiological and transcriptomic analysis reveals the roles of 1-MCP in the ripening and fruit aroma quality of banana fruit (Fenjiao).

Fenjiao (Musa ABB Pisang Awak) is a popular banana cultivar due to its good taste and stress resistance, but it has a short shelf-life and deteriorates rapidly post-harvest. The effects of 1-methylcyclopropene (1-MCP) treatment on fruit physiology and quality and transcriptomic profiles are investigated in this study. The results showed that 1-MCP significantly delayed fruit ripening by repressing fruit softening and inhibiting the respiratory rate and ethylene production. The 1-MCP treatment delayed sugar accumulation and influenced the content of the precursors of the biosynthesis of aroma volatiles. 1-MCP reduced the production of flavor-contributing volatile esters isoamyl isobutyrate, isoamyl acetate and trans-2-hexenal and hexanal, but dramatically increased the hexyl acetate production at the full-ripening stage. The transcriptomic analysis showed that 1-MCP dramatically affected the transcript profiles during fruit ripening, especially the KEGG pathways involved in amino acid metabolism, biosynthesis of other secondary metabolites, carbohydrate metabolism, lipid metabolism, signal transduction, and translation classes. The key genes and the corresponding enzyme activities involved in the volatile and ethylene synthesis were severely repressed due to the 1-MCP treatment. The 1-MCP treatment effectively delayed Fenjiao fruit ripening, but affected volatile production by reducing the precursor production and expression level of genes involved in the metabolism pathways of ethylene, auxin and volatiles.

Genome-Wide Identification and Characterization of the NAC Transcription Factor Family in Musa Acuminata and Expression Analysis during Fruit Ripening.

Banana (Musa acuminata, AAA group) is a representative climacteric fruit with essential nutrients and pleasant flavors. Control of its ripening determines both the fruit quality and the shelf life. NAC (NAM, ATAF, CUC2) proteins, as one of the largest superfamilies of transcription factors, play crucial roles in various functions, especially developmental processes. Thus, it is important to conduct a comprehensive identification and characterization of the NAC transcription factor family at the genomic level in M. acuminata. In this article, a total of 181 banana NAC genes were identified. Phylogenetic analysis indicated that NAC genes in M. acuminata, Arabidopsis, and rice were clustered into 18 groups (S1-S18), and MCScanX analysis disclosed that the evolution of MaNAC genes was promoted by segmental duplication events. Expression patterns of NAC genes during banana fruit ripening induced by ethylene were investigated using RNA-Seq data, and 10 MaNAC genes were identified as related to fruit ripening. A subcellular localization assay of selected MaNACs revealed that they were all localized to the nucleus. These results lay a good foundation for the investigation of NAC genes in banana toward the biological functions and evolution.

What are the Main Sensor Methods for Quantifying Pesticides in Agricultural Activities? A Review.

In recent years, there has been an increase in pesticide use to improve crop production due to the growth of agricultural activities. Consequently, various pesticides have been present in the environment for an extended period of time. This review presents a general description of recent advances in the development of methods for the quantification of pesticides used in agricultural activities. Current advances focus on improving sensitivity and selectivity through the use of nanomaterials in both sensor assemblies and new biosensors. In this study, we summarize the electrochemical, optical, nano-colorimetric, piezoelectric, chemo-luminescent and fluorescent techniques related to the determination of agricultural pesticides. A brief description of each method and its applications, detection limit, purpose-which is to efficiently determine pesticides-cost and precision are considered. The main crops that are assessed in this study are bananas, although other fruits and vegetables contaminated with pesticides are also mentioned. While many studies have assessed biosensors for the determination of pesticides, the research in this area needs to be expanded to allow for a balance between agricultural activities and environmental protection.

Ripening of bananas using Bowdichia virgilioides Kunth leaves.

Bananas are usually ripened with calcium carbide (CaC2), a dangerous substance that can cause food poisoning. The objective was to test the empirical ripening banana method using Bowdichia virgilioides leaves compared to carbide. Ripening tests were carried out using 'Pacovan' banana fruits with B. virgilioides leaves and carbide following the empirical method used by Borborema farmers, Paraíba, Brazil. Bowdichia virgilioides leaves induced increased respiration and ascorbic acid production and reduced acidity, chlorophyll and pH in banana fruits like CaC2. Leaves of B. virgilioides induce ripening of 'Pacovan' banana with safer and same results than with CaC2.

Exogenous progesterone treatment alleviates chilling injury in postharvest banana fruit associated with induction of alternative oxidase and antioxidant defense.

The effects of exogenous progesterone (PROG) on chilling injury (CI) in postharvest banana fruit were investigated. Concentration screening tests showed that 10-5 mol/l PROG was most effective in reducing CI in banana fruit stored for 25 d at 5 ±â€¯1 °C, but did not markly increase PROG content of pulps. This PROG treatment significantly reduced the electrolyte leakage, levels of malondialdehyde, O2- production rate and H2O2 contents in banana compared with control fruit. The PROG treatment caused an early induction of alternative oxidase (AOX) at the transcript and protein level to reduce the generation of O2- and H2O2. PROG treatment also enhanced the transcript levels and activities of antioxidant enzymes and maintained higher levels of reduced glutathione and ascorbic acid than the control fruit. These results suggested that PROG attenuating CI in banana fruit may be attributed to the induction of AOX and the improvement of enzyme and non-enzymatic antioxidant defenses.

Effect of morin on the degradation of water-soluble polysaccharides in banana during softening.

The degradation of cell wall polysaccharides is highly associated with the softening process of banana. In this work, banana was treated by morin to delay softening during storage. Water-soluble polysaccharides were extracted from banana pulp at four storage stages. Their levels increased when the banana was green, but decreased when turned to yellow. Three types of polysaccharides were identified by nuclear magnetic resonance spectroscopy, including starches, homogalacturonans and arabinogalactan proteins. (1 → 4)-α-D-galacturonan constructed the backbone of homogalacturonans, which was the leading water-soluble polysaccharide in the last stage. Starch was the dominant polysaccharide at all stages in morin-treated banana. Deassembly of insoluble starch granules and degradation of soluble starches were responsible for its high level in banana. Arabinogalactan proteins were only detected in late stages, and could be a marker for banana deterioration. The inhibited degradation of cell wall polysaccharides contributed to the preservative effect of morin.

Risk assessment and copper geochemistry of an orchard irrigated with mine water: a case study in the semiarid region of Brazil.

This study aimed to evaluate mine water reuse, elucidating the potential problems related to trace metal biogeochemistry focusing on Cu dynamics in water, soil, and plants. Water samples were collected from a Cu mine and a reservoir used to store mine water. Additional samples were taken from soils from an uncultivated area and a banana orchard (irrigated with mine water for at least 10 years) and plant from the irrigated area. The following parameters were analyzed: pH, redox potential, dissolved ions in water samples (e.g., Ca2+, Mg2+, Na+, K+, Cu2+, SO 4 2- , and Cl-), bioavailable Cu and Cu solid-phase fractionation (in soils and reservoir sediments samples), as well as Cu content in banana plants. Mine water presents high dissolved Cu concentration (mean 2.3 ± 0.0 mg L-1), limiting its use for irrigation. Water storage at the reservoir increased water quality, reducing dissolved Cu concentration (mean 0.2 ± 0.0 mg L-1), due to adsorption/precipitation as carbonates (mean 131.8 ± 24.6 mg kg-1), organic matter (mean 1526.2 ± 4.7 mg kg-1) and sulfides (mean 158.4 ± 56.9 mg kg-1). Despite higher water quality at the reservoir, the use of mine water increased the amount of bioavailable Cu in soils, which was primarily associated with organic matter. Increased bioavailable Cu in the soil did not increase the Cu content of banana leaves but resulted in high Cu content of roots and fruit, increasing the risk of toxicity for the population.

Independent and combined abiotic stresses affect the physiology and expression patterns of DREB genes differently in stress-susceptible and resistant genotypes of banana.

In tropics, combined stresses of drought and heat often reduce crop productivity in plants like Musa acuminata L. We compared responses of two contrasting banana genotypes, namely the drought-sensitive Grand Nain (GN; AAA genome) and drought tolerant Hill banana (HB; AAB genome) to individual drought, heat and their combination under controlled and field conditions. Drought and combined drought and heat treatments caused greater reduction in leaf relative water content and greater increase in ion leakage and H2 O2 content in GN plants, especially in early stages, while the responses were more pronounced in HB at later stages. A combination of drought and heat increased the severity of responses. Real-time expression patterns of the A-1 and A-2 group DEHYDRATION-RESPONSIVE ELEMENT BINDING (DREB) genes revealed greater changes in expression in leaves of HB plants for both the individual stresses under controlled conditions compared to GN plants. A combination of heat and drought, however, activated most DREB genes in GN but surprisingly suppressed their expression in HB in controlled and field conditions. Its response seems correlated to a better stomatal control over transpiration in HB and a DREB-independent pathway for the more severe combined stresses unlike in GN. Most of the DREB genes had abscisic acid (ABA)-responsive elements in their promoters and were also activated by ABA suggesting at least partial dependence on ABA. This study provides valuable information on physiological and molecular responses of the two genotypes to individual and combined drought and heat stresses.

Identification of Ethylene Responsive miRNAs and Their Targets from Newly Harvested Banana Fruits Using High-Throughput Sequencing.

The roles of microRNAs (miRNAs) related to ethylene response in banana fruits remain unknown because many miRNAs are differentially expressed as the fruit ripens, making the identification of ethylene-responsive miRNAs difficult. Using newly harvested banana fruits (within 5 h after harvest) as material, we found that these fruit did not ripen when treated with 5 µL/L of ethylene for 12 h at 22 °C. Two miRNA libraries were generated from newly harvested banana fruits with and without ethylene treatment and sequenced. In total, 128 known miRNAs belonging to 42 miRNA families were obtained, and 12 novel miRNAs were identified. Among them, 22 were differentially expressed in response to ethylene treatment, among which 6 known miRNAs and their putative targets were validated using qRT-PCR. These putative targets encoded proteins including GATA, ARF, DLC, and AGO, etc. KEGG and GO analyses showed that miRNAs differentially expressed in response to ethylene mainly function in the molecular and biological processes.

Morin as a Preservative for Delaying Senescence of Banana.

Banana is a climacteric fruit with desirable palatability and high nutritional value. It ripens rapidly accompanied with metabolite changes during postharvest storage. In this work, morin was applied to treat banana to delay senescence. Nuclear magnetic resonance (NMR) spectroscopy was used to monitor the changes of metabolite composition and levels in banana. The results showed that morin significantly delayed the changes of color and firmness. 1D and 2D NMR spectra reflected that the levels and composition of metabolites were changed with the senescence initiation. The principal component analysis revealed that the first principal components responsible for banana senescence were carbohydrates, amino acids, lipids and phenolics. Morin treatment delayed the transformation of starch to glucose, fructose and sucrose, accelerated the accumulations of alanine and γ-Amino-butyrate (GABA), postponed the generations of valine and l-aspartic acid, suppressed the degradation of saponin a. It indicated that morin was effective in delaying banana senescence.

Genome-wide transcriptome analysis and identification of benzothiadiazole-induced genes and pathways potentially associated with defense response in banana.

BACKGROUND: Bananas (Musa spp.) are the most important fruit crops worldwide due to their high nutrition value. Fusarium wilt of banana, caused by fungal pathogen Fusarium oxysporum f. sp. cubense tropical race 4 (Foc 4), is considered as the most destructive disease in the world and results in extensive damage leading to productivity loss. The widespread use of plant resistance inducers (PRIs), such as benzothiadiazole (BTH), is a novel strategy to stimulate defense responses in banana plants to protect against pathogens infection. The recent focus on the crop defense against fungal infections has led to a renewed interest on understanding the molecular mechanisms of specific PRIs-mediated resistance. This transcriptome study aimed to identify genes that are associated with BTH-induced resistance. Patterns of gene expression in the leaves and roots of BTH-sprayed banana plants were studied using RNA-Seq. RESULTS: In this study, 18 RNA-Seq libraries from BTH-sprayed and untreated leaves and roots of the Cavendish plants, the most widely grown banana cultivar, were used for studying the transcriptional basis of BTH-related resistance. Comparative analyses have revealed that 6689 and 3624 differentially expressed genes were identified in leaves and roots, respectively, as compared to the control. Approximately 80% of these genes were differentially expressed in a tissue-specific manner. Further analysis showed that signaling perception and transduction, transcription factors, disease resistant proteins, plant hormones and cell wall organization-related genes were stimulated by BTH treatment, especially in roots. Interestingly, the ethylene and auxin biosynthesis and response genes were found to be up-regulated in leaves and roots, respectively, suggesting a choice among BTH-responsive phytohormone regulation. CONCLUSIONS: Our data suggests a role for BTH in enhancing banana plant defense responses to Foc 4 infection, and demonstrates that BTH selectively affect biological processes associated with plant defenses. The genes identified in the study could be further studied and exploited to develop Foc 4-resistant banana varieties.

Proteomic profiling of 24-epibrassinolide-induced chilling tolerance in harvested banana fruit.

The mechanism of 24-epibrassinolide (EBR)-induced chilling tolerance in harvested banana fruit was investigated. Results showed that EBR pretreatment remarkably suppressed the development of chilling injury (CI) in harvested banana fruit during 12 days of cold storage at 8 °C, as indicated by lower CI index in treated fruit. Physiological measurements exhibited that EBR treatment reduced the relative electrolyte leakage and malondialdehyde (MDA) content while increased the chlorophyll fluorescence (Fv/Fm), total soluble solids (TSS) and ratio of TSS and titratable acidity. Furthermore, the differentially accumulated proteins of banana fruit in response to EBR and cold treatment were investigated by employing gel-based proteomic in combination with MALDI-TOF-TOF MS and LC-ESI-MS/MS analyses. There were fifty five protein spots to be successfully identified. Notably, most of up-regulated proteins by EBR treatment were related to energy biosynthesis, stress response and cell wall modification. In contrast, proteins involved in protein degradation and energy consumption were down-regulated by EBR treatment. These results suggest that EBR treatment could enhance the defense ability, promote the synthesis and utilization of energy, as well as maintain the protein function via enhancing protein biosynthesis and inhibiting protein degradation, consequently contributing to improvement of cold tolerance in harvested banana fruit. SIGNIFICANCE: To extend our understanding of chilling injury (CI) of harvested banana fruit, we reported the effect of 24-epibrassinolide (EBR) on CI of banana fruit when stored at 8 °C. It was the first report on the comprehensive proteomic analysis of banana fruit in response to EBR treatment at low temperature. EBR pretreatment significantly reduced CI in harvested banana fruit. Fifty five protein spots were successfully identified. Notably, the most of up-regulated proteins by EBR treatment were related to energy biosynthesis, stress response and cell wall modification. In contrast, proteins involved in protein degradation and energy consumption were down-regulated. These results suggest that exogenous EBR treatment could enhance the defense ability and maintain high energy status. Meanwhile, EBR treatment maintained protein function via enhancing protein biosynthesis and inhibiting protein degradation. These results may help us to understand the molecular mechanism of the chilling tolerance induced by EBR treatment and broaden the current knowledge of the mechanism of CI of harvested banana fruit.

Comparison of tissue deterioration of ripening banana fruit (Musa spp., AAA group, Cavendish subgroup) under chilling and non-chilling temperatures.

BACKGROUND: In fleshy fruits, induced programmed cell death (PCD) has been observed in heat-treated tomato, and in ethylene-treated and low-temperature exposure in immature cucumber. No other fleshy fruit has been evaluated for chilling-injury-induced PCD, especially mature fruit with full ripening capacity. The purpose of this research was to identify and evaluate the presence of PCD processes during the development of low-temperature-induced physiopathy of banana fruit. RESULTS: Exposure of fruit to 5 °C for 4 days induced degradative processes similar to those occurring during ripening and overripening of non-chilled fruit. Nuclease from banana peel showed activity in both DNA substrates and RNA substrates. No exclusive low-temperature-induced proteases and nucleases were observed. DNA of chilled peel showed earlier signs of degradation and higher levels of DNA tailing during overripening. CONCLUSION: This study shows that exposure to low temperatures did not induce a pattern of degradative processes that differed from that occurring during ripening and overripening of non-chilled fruit. DNA showed earlier signs of degradation and higher levels of DNA tailing. Nuclease activity analysis showed bifunctionality in both chilled and non-chilled tissue and no chilling-exclusive protease and nuclease. Fleshy fruit might use their available resources on degradative processes and adjust them depending on environmental conditions. © 2018 Society of Chemical Industry.

Cryopreservation of banana's cv Grand Naine in vitro rhizomes

ABSTRACT The preservation of banana genetic material is usually performed through seedlings. However, most banana cultivars do not produce seed and are propagated vegetatively. Therefore, cryopreservation is a feasible technique that allows the preservation of banana genotypes indefinitely. For the success of cryopreservation protocols, the selection of cryoprotectants and pre-freezing techniques are important factor. Therefore, the objective of this study was to verify the effects of different cryoprotectants with and without 1% phloroglucinol and pre-cooling periods on the development of a protocol for cryopreservation of in vitro rhizomes ofMusa accuminata(AAA) cv Grand Naine banana. The addition of 1% phloroglucinol to the cryoprotective solutions, such as PVS2 enhanced recovery of cryopreserved banana rhizomes. In addition, pre-cooling of explants in ice for 3 hours in PVS2 + 1% of phloroglucinol allowed efficient cryopreservation of banana rhizomes, followed by successful recovery and regeneration of in vitro shoots of banana cv Grand Naine.

Thap Maeo bananas: Fast ripening and full ethylene perception at low doses.

Brazil is a major producer and consumer of various banana types. Thap Maeo is a promising cultivar for the market due to its resistance to Black and Yellow Sigatoka disease. However, a lack of information and postharvest technologies concerning Thap Maeo physiology seems to be a significant problem limiting its expansion in the market. Thus, this study aims to establish Thap Maeo fruit's physical, biochemical, and physiological aspects, defining the best ethylene dosage for treatment considering fruit ripening parameters. Bananas were harvested and monitored during both natural and ethylene-induced ripening processes. Assessments of pulp firmness, peel color and endogenous ethylene production showed different profiles between ethylene-treated and non-treated fruits, whereas the ethylene responses concerning the carbohydrates and hormones profiles, as well as the ethylene receptors expression, were observed in all ethylene-treated fruits, even applying low concentrations of the hormone. It thus indicated the high ethylene-sensitivity of Thap Maeo cultivar. Such postharvest behaviors reverberate in lower ethylene requirements for treatment, which was established at 10µLL-1. Ethylene-inducible changes in fruit volatile compounds throughout ripening are also discussed.

Cryopreservation of banana's cv Grand Naine in vitro rhizomes.

The preservation of banana genetic material is usually performed through seedlings. However, most banana cultivars do not produce seed and are propagated vegetatively. Therefore, cryopreservation is a feasible technique that allows the preservation of banana genotypes indefinitely. For the success of cryopreservation protocols, the selection of cryoprotectants and pre-freezing techniques are important factor. Therefore, the objective of this study was to verify the effects of different cryoprotectants with and without 1% phloroglucinol and pre-cooling periods on the development of a protocol for cryopreservation of in vitro rhizomes ofMusa accuminata(AAA) cv Grand Naine banana. The addition of 1% phloroglucinol to the cryoprotective solutions, such as PVS2 enhanced recovery of cryopreserved banana rhizomes. In addition, pre-cooling of explants in ice for 3 hours in PVS2 + 1% of phloroglucinol allowed efficient cryopreservation of banana rhizomes, followed by successful recovery and regeneration of in vitro shoots of banana cv Grand Naine.

Involvement of WRKY Transcription Factors in Abscisic-Acid-Induced Cold Tolerance of Banana Fruit.

Phytohormone abscisic acid (ABA) and plant-specific WRKY transcription factors (TFs) have been implicated to play important roles in various stress responses. The involvement of WRKY TFs in ABA-mediated cold tolerance of economical fruits, such as banana fruit, however remains largely unknown. Here, we reported that ABA application could induce expressions of ABA biosynthesis-related genes MaNCED1 and MaNCED2, increase endogenous ABA contents, and thereby enhance cold tolerance in banana fruit. Four banana fruit WRKY TFs, designated as MaWRKY31, MaWRKY33, MaWRKY60, and MaWRKY71, were identified and characterized. All four of these MaWRKYs were nuclear-localized and displayed transactivation activities. Their expressions were induced by ABA treatment during cold storage. More importantly, the gel mobility shift assay and transient expression analysis revealed that MaWRKY31, MaWRKY33, MaWRKY60, and MaWRKY71 directly bound to the W-box elements in MaNCED1 and MaNCED2 promoters and activated their expressions. Taken together, our findings demonstrate that banana fruit WRKY TFs are involved in ABA-induced cold tolerance by, at least in part, increasing ABA levels via directly activating NECD expressions.