Foc TR4 Research Articles

Soil Predisposing Factors to Fusarium oxysporum f.sp Cubense Tropical Race 4 on Banana Crops of La Guajira, Colombia

Fusarium wilt of banana caused by the soil-borne fungi Fusarium oxysporum f. sp. cubense, Tropical Race 4 (Foc TR4) (Syn. Fusarium odoratissimum), is a major threat to the global banana industry. Aiming to identify predisposing soil factors for Fusarium wilt of banana (FWB) TR4, the 23 physical and chemical soil properties were studied in three commercial banana farms in La Guajira, Colombia. Disturbed and undisturbed soil samples were collected from areas affected by the disease (affected plots) and disease-free areas (healthy plots). Five repetitions per farm were considered, with a total sample of n = 30. The data were analyzed using one-way analysis of variance (ANOVA). Subsequently, the debiased sparse partial correlation (DSPC) algorithm was applied. Organic matter (OM), pH, calcium (Ca), magnesium (Mg), zinc (Zn), and cation-exchange capacity on the exchange complex (ECEC), showed significant differences between the affected and healthy plots. In addition, the bulk density and saturated hydraulic conductivity (HC) were associated suggesting that physical attributes, such as soil compaction and poor drainage, create favorable conditions for FWB. According to the DSPC algorithm, the HC variable presented a grade of 5 and an intermediation of 14.67, which indicates that it has significant associations with BD, sand, porosity, ECEC, and OM and plays a critical role in the connection of other variables in the network and the differentiation of healthy and affected plots. These findings establish a baseline of information under field conditions in Colombia, which can be used to design soil management strategies to mitigate the detrimental effects of Foc TR4 by creating less favorable conditions for the pathogen.

First report of Fusarium wilt of Cavendish bananas caused by Fusarium oxysporum f. sp. cubense Tropical Race 4 in Venezuela.

Fusarium wilt of banana (Musa spp.), caused by the soil-borne fungus Fusarium oxysporum f. sp. cubense (Foc), is a major constraint to banana production worldwide (Dita et al., 2018). A strain of Foc that affects Cavendish (AAA) bananas in the tropics, called Foc tropical race 4 (TR4; VCG 01213), is of particular concern. Foc TR4 was first detected in Malaysia and Indonesia around 1990 but was restricted to Southeast Asia and northern Australia until 2012. The fungus has since been reported from Africa, the Indian subcontinent, and the Middle East (Viljoen et al., 2020). Foc TR4 was detected in Colombia in 2019 and in Perú in 2021 (Reyes-Herrera et al., 2020). The incursions into Latin America and the Caribbean (LAC) triggered global concerns, as 75% of international export bananas are produced in the region. Banana production in Venezuela, however, is primarily intended for domestic consumption (Aular and Casares, 2011). In 2021 the country produced 533,190 metric tons of banana on an area of 35,896 ha, with an approximate yield of 14,853 kg/ha (FAOSTAT, 2023). In July 2022, severe leaf-yellowing, and wilting, along with internal vascular discoloration of the pseudostem, were noted in Cavendish banana plants cultivar 'Valery' in the states of Aragua (10°11'8″N; 67°34'51″W), Carabobo (10º14'24″N; 67º48'51″W), and Cojedes (9°37'44″N; 68°55'4″W). Necrotic strands from the pseudostems of diseased plants were collected for identification of the causal agent using DNA-based techniques, vegetative compatibility group (VCG) analysis and pathogenicity testing. The samples were first surface disinfected and plated onto potato dextrose agar medium. Single-spored isolates were identified as F. oxysporum based on cultural and morphological characteristics, including white colonies with purple centres, infrequent macroconidia, abundant microconidia on short monophialides, and terminal or intercalary chlamydospores (Leslie and Summerell, 2006). Foc TR4 was identified from five isolates by endpoint and quantitative-PCR using four different primer sets (Li et al. 2013; Dita et al. 2010; Aguayo et al. 2017; Matthews et al. 2020). The same isolates were identified as VCG 01213 by successfully pairing nitrate non-utilizing (nit-1) mutants of the unknown strains with Nit-M testers of Foc TR4 available at Stellenbosch University (Leslie and Summerell, 2006). For pathogenicity testing, 3-month-old Cavendish banana plants cultivar 'Williams' were inoculated with isolates from Venezuela grown on sterile millet seed (Viljoen et al., 2017). Plants developed typical Fusarium wilt symptoms 60 days after inoculation, including yellowing of leaves that progressed from the older to the younger leaves, wilting, and internal discoloration of the pseudostem. Koch's postulates were fulfilled by reisolating and identifying Foc TR4 from the plants by qPCR (Matthews et al., 2020). These results provide scientific proof of the presence of Foc TR4 in Venezuela. The Venezuelan Plant Protection Organization (INSAI) has declared Foc TR4 as a newly introduced pest (January 19, 2023), and infested banana fields were placed under quarantine. Comprehensive surveys are now conducted in all production areas in Venezuela to assess the presence and impact of Foc TR4, and information campaigns were started to make farmers aware of biosecurity protocols. Collaborative initiatives and coordinated actions among all stakeholders are needed to prevent the spread of Foc TR4 to other countries in Latin America, and to develop Foc TR4-resistant bananas (Figueiredo et al. 2023).

Fungal endophytes as potential biocontrol agent of Panama disease of banana

BackgroundFusarium odoratissimum (Foc TR4) is a devastating fungal pathogen that causes Panama wilt in Cavendish banana, a high value crop that generates significant revenue. The excessive use of synthetic fungicides for disease control poses risks to both human health and the environment. Consequently, there is a pressing need for eco-friendly alternatives to manage this disease. One potential approach is the utilization of biocontrol agents, which have shown promise in disease control. This study aimed to evaluate the biocontrol potential of fungal endophytes isolated from the naturally-resistant cultivar Cardaba banana against Foc TR4. These fungal isolates were subjected to a dual culture assay, characterized based on morpho-cultural characteristics, and confirmed molecularly using the internal transcribed spacer (ITS) region of rDNA.ResultsOf the 15 fungal endophytes isolated from Cardaba; Blakeslea trispora, Pseudopestalotiopsis theae, Xylaria badia, Nigrospora hainanensis, Colletotrichum gloeosporioides, Xylaria feejeensis, Gymnoascus reessii, Fusarium subglutinans, Rigidoporus vinctus, and Macrophomina phaseolina showed potential antagonistic activity against F. odoratissimum. Isolates X. feejeensis (USMJMTBL10), G. reesii (USMJMTBL11), and M. phaseolina (USMJMTBR1) had the highest antagonistic activity of 87.8–96.6% against F. odoratissimum in vitro.ConclusionThis study presents evidence of the potential of Cardaba-associated endophytes isolated and identified in this research, as effective biocontrol agents against F. odoratissimum, the causal agent of Panama disease in Cavendish banana. Our findings suggest that these endophytes hold promise as a biocontrol agent for managing Foc TR4-infected banana plants. Future research will focus on in planta testing, validation in greenhouse and field trials, as well as development of formulations and application protocols to optimize the utilization of these biocontrol agents in an integrated management approach for Foc TR4-infected bananas.

Banana disease-suppressive soil drives Bacillus assembled to defense Fusarium wilt of banana.

Fusarium wilt of banana (FWB) caused by Fusarium oxysporum f. sp. cubense tropical race 4 (Foc TR4), poses a serious problem for sustainable banana production. Biological control is one of the effective measures to control this destructive disease. High-throughput sequencing of soil microorganisms could significantly improve the efficiency and accuracy of biocontrol strain screening. In this study, the soil microbial diversity of six main banana-producing areas in Yunnan was sequenced by Illumina Miseq platform. The outcome of this study showed the genus of Chujaibacter, Bacillus, and Sphingomonas were significantly enriched in microorganism community composition. Further correlation analysis with soil pathogen (Foc TR4) content showed that Bacillus was significantly negatively correlated with pathogen content. Therefore, we isolated and identified Bacillus from the disease-suppressive soils, and obtained a B. velezensis strain YN1910. In vitro and pot experiments showed that YN1910 had a significant control effect (78.43-81.76%) on banana Fusarium wilt and had a significant growth promotion effect on banana plants.

Mapping of the Susceptibility of Colombian Musaceae Lands to a Deadly Disease: Fusarium oxysporum f. sp. cubense Tropical Race 4

Fusarium oxysporum f. sp. cubense Tropical Race 4 (Foc TR4) (Syn. Fusarium odoratissimum) is a devastating soil-borne pathogen that infects the roots of banana plants and causes Fusarium wilt disease. Colombia is one of the world’s leading banana producers; therefore, new uncontrolled outbreaks could have serious consequences. Despite this, little is known about the susceptibility of Musaceae lands in Colombia to Foc TR4. This work presents a pioneering study on the susceptibility of Colombian soils to Foc TR4. For this, a study was carried out to characterize climatic, edaphic, and density factors of Musaceae productive systems at the Colombian level, articulated with expert criteria to map and define areas with different levels of susceptibility to Foc R4T. These criteria are typically selected based on the existing scientific literature, consultation with domain experts, and consideration of established methods for assessing soil health and disease susceptibility in Musaceae plantations. By joining the analyzed susceptibility factors, differentiated areas were generated that imply a greater or lesser predisposition to the disease. Subsequently, a validation of the classification was made with Random Forest. The results indicate that at the level of climate, soil, and farm density as a fit factor, practically 50% of the cultivated territory of Musaceae are areas high and very highly susceptible to the pathogen (572,000 km2). The results showed that from the total Musaceae area, Antioquia, Bolívar, Chocó, and Santander turned out to be the departments with the highest proportion of very high susceptibility class of the production farms. The analysis of Random Forest classification performance shows that the model has a relatively low out-of-bag (OOB) error rate (0.023). The study on the susceptibility is highly novel and original, as it represents the first systematic investigation of Foc TR4 susceptibility in Colombian soils. This paper provides important insights into the susceptibility of Musaceae lands in Colombia to Foc TR4. The study highlights the need for ongoing monitoring, containment, and control measures to prevent the spread of this deadly pathogen and protect Colombia’s important banana industry.

Iron necessity for chlamydospore germination in Fusarium oxysporum f. sp. cubense TR4.

Fusarium wilt disease of banana, caused by the notorious soil-borne pathogen Fusarium oxysporum f. sp. cubense Tropical Race 4 (Foc TR4), is extremely difficult to manage. Manipulation of soil pH or application of synthetic iron chelators can suppress the disease through iron starvation, which inhibits the germination of pathogen propagules called chlamydospores. However, the effect of iron starvation on chlamydospore germination is largely unknown. In this study, scanning electron microscopy was used to assemble the developmental sequence of chlamydospore germination and to assess the effect of iron starvation and pH in vitro. Germination occurs in three distinct phenotypic transitions (swelling, polarized growth, outgrowth). Outgrowth, characterized by formation of a single protrusion (germ tube), occurred at 2 to 3h, and a maximum value of 69.3% to 76.7% outgrowth was observed at 8 to 10h after germination induction. Germination exhibited plasticity with pH as over 60% of the chlamydospores formed a germ tube between pH 3 and pH 11. Iron-starved chlamydospores exhibited polarized-growth arrest, characterized by the inability to form a germ tube. Gene expression analysis of rnr1 and rnr2, which encode the iron-dependent enzyme ribonucleotide reductase, showed that rnr2 was upregulated (p < 0.0001) in iron-starved chlamydospores compared to the control. Collectively, these findings suggest that iron andextracellular pH arecrucial for chlamydospore germination in Foc TR4. Moreover, inhibition of germination by iron starvation may be linked to a different mechanism, rather than repression of the function of ribonucleotide reductase, the enzyme that controls growth by regulation of DNA synthesis.

Antimicrobial mechanisms and secondary metabolite profiles of Streptomyces hygroscopicus subsp. hygroscopicus 5-4 against banana fusarium wilt disease using metabolomics.

Fusarium wilt of bananas (FWB) is seriously affecting the sustainable development of the banana industry and is caused by the devastating soil-borne fungus Fusarium oxysporum f. sp. cubense tropical race 4 (Foc TR4). Biological control is a promising strategy for controlling Fusarium wilt in bananas. We previously identified Streptomyces hygroscopicus subsp. hygroscopicus 5-4 with strong antifungal activity against the FWB. The most possible antimicrobial mechanism of strain 5-4 was explored using the metabolomics approach, light microscopy imaging, and transmission electron microscopy (TEM). The membrane integrity and ultrastructure of Foc TR4 was damaged after extract treatment, which was supported by the degradation of mycelium, soluble protein content, extracellular reducing sugar content, NADH oxidase activity, malondialdehyde content, mitochondrial membrane potential, and mitochondrial respiratory chain complex enzyme activity. The extracts of strain 5-4 cultivated at different times were characterized by a liquid chromatography-mass spectrometer (LC-MS). 647 known metabolites were detected in the extracts of strains 5-4. Hygromycin B, gluten exorphin B4, torvoside G, (z)-8-tetradecenal, piperitoside, sarmentosin, pubescenol, and other compounds were the main differential metabolites on fermentation culture for 7 days. Compared with strain 5-4 extracts, hygromycin B inhibited the mycelial growth of Foc TR4, and the EC50 concentration was 7.4 μg/mL. These results showed that strain 5-4 could destroy the cell membrane of Foc TR4 to inhibit the mycelial growth, and hygromycin B may be the key antimicrobial active metabolite. Streptomyces hygroscopicus subsp. hygroscopicus 5-4 might be a promising candidate strain to control the FWB and provide a scientific basis for the practical application of hygromycin B as a biological control agent.

Antifungal Potential of Melaleuca alternifolia against Fungal Pathogen Fusarium oxysporum f. sp. cubense Tropical Race 4.

Fusarium wilt of bananas caused by Fusarium oxysporum f. sp. cubense Tropical Race 4 (Foc TR4) poses the most serious threat to banana production globally. The disease has been managed using chemical fungicides, yet the control levels are still unsatisfactory. This study investigated the antifungal activities of tea tree (Melaleuca alternifolia) essential oil (TTO) and hydrosol (TTH) against Foc TR4 and their bioactive components. The potential of TTO and TTH in inhibiting the growth of Foc TR4 was evaluated in vitro using agar well diffusion and spore germination assays. Compared to the chemical fungicide, TTO effectively suppressed the mycelial growth of Foc TR4 at 69%. Both the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of TTO and TTH were established at 0.2 µg/µL and 50% v/v, respectively, suggesting the fungicidal nature of the plant extracts. The disease control efficacies were also demonstrated by a (p ≤ 0.05) delayed Fusarium wilt symptom development in the susceptible banana plants with reduced LSI dan RDI scores from 70% to around 20-30%. A GC/MS analysis of TTO identified terpinen-4-ol, eucalyptol, and α-terpineol as the major components. In contrast, an LC/MS analysis of TTH identified different compounds, including dihydro-jasmonic acid and methyl ester. Our findings indicate the potential of tea tree extracts as natural alternatives to chemical fungicides to control Foc TR4.

Loss of Phosphomannose Isomerase Impairs Growth, Perturbs Cell Wall Integrity, and Reduces Virulence of Fusarium oxysporum f. sp. cubense on Banana Plants.

Fusarium oxysporum f. sp. cubense tropical race 4 (Foc TR4) causes Fusarium wilt of banana, necessitating urgent measures to control this disease. However, the molecular mechanisms underlying Foc TR4 virulence remain elusive. Phosphomannose isomerase is a key enzyme involved in the biosynthesis of GDP mannose, an important precursor of fungal cell walls. In this study, two phosphomannose isomerases were identified in the Foc TR4 genome, of which only Focpmi1 was highly expressed throughout all developmental stages. Generated null mutants in Foc TR4 showed that only the ΔFocpmi1 mutant required exogenous mannose for growth, indicating that Focpmi1 is the key enzyme involved in GDP mannose biosynthesis. The Focpmi1 deficient strain was unable to grow without exogenous mannose and exhibited impaired growth under stress conditions. The mutant had reduced chitin content in its cell wall, rendering it vulnerable to cell wall stresses. Transcriptomic analysis revealed up- and down-regulation of several genes involved in host cell wall degradation and physiological processes due to the loss of Focpmi1. Furthermore, Focpmi1 was also found to be crucial for Foc TR4 infection and virulence, making it a potential antifungal target to address the threats posed by Foc TR4.

A Novel Effector, FSE1, Regulates the Pathogenicity of Fusarium oxysporum f. sp. cubense Tropical Race 4 to Banana by Targeting the MYB Transcription Factor MaEFM-Like.

Phytopathogenic fungi secretes a range of effectors to manipulate plant defenses. Fusarium oxysporum f. sp. cubense tropical race 4 (Foc TR4) is a soil-borne pathogen that causes destructive banana wilt disease. Understanding the molecular mechanisms behind Foc TR4 effectors and their regulation of pathogenicity is helpful for developing disease control strategies. In the present study, we identified a novel effector, Fusarium special effector 1 (FSE1), in Foc TR4. We constructed FSE1 knock-out and overexpression mutants and investigated the functions of this effector. In vitro assays revealed that FSE1 was not required for vegetative growth and conidiation of Foc TR4. However, inoculation analysis of banana plantlets demonstrated that knock-out of FSE1 increased the disease index, while overexpression of FSE1 decreased it. Microscope analysis suggested that FSE1 was distributed in the cytoplasm and nuclei of plant cells. Furthermore, we identified an MYB transcription factor, MaEFM-like, as the target of FSE1, and the two proteins physically interacted in the nuclei of plant cells. In addition, Transient expression of MaEFM-like induced cell death in tobacco leaves. Our findings suggest that FSE1 is involved in the pathogenicity of Foc TR4 by targeting MaEFM-like.

Plant pathogen resistance is mediated by recruitment of specific rhizosphere fungi.

Beneficial interactions between plants and rhizosphere microorganisms are key determinants of plant health with the potential to enhance the sustainability of agricultural practices. However, pinpointing the mechanisms that determine plant disease protection is often difficult due to the complexity of microbial and plant-microbe interactions and their links with the plant's own defense systems. Here, we found that the resistance level of different banana varieties was correlated with the plant's ability to stimulate specific fungal taxa in the rhizosphere that are able to inhibit the Foc TR4 pathogen. These fungal taxa included members of the genera Trichoderma and Penicillium, and their growth was stimulated by plant exudates such as shikimic acid, D-(-)-ribofuranose, and propylene glycol. Furthermore, amending soils with these metabolites enhanced the resistance of a susceptible variety to Foc TR4, with no effect observed for the resistant variety. In total, our findings suggest that the ability to recruit pathogen-suppressive fungal taxa may be an important component in determining the level of pathogen resistance exhibited by plant varieties. This perspective opens up new avenues for improving plant health, in which both plant and associated microbial properties are considered.

The Advance of Fusarium Wilt Tropical Race 4 in Musaceae of Latin America and the Caribbean: Current Situation

The fungus Fusarium oxysporum f. sp. cubense tropical race 4 (syn. Fusarium odoratissimum) (Foc TR4) causes vascular wilt in Musaceae plants and is considered the most lethal for these crops. In Latin America and the Caribbean (LAC), it was reported for the first time in Colombia (2019), later in Peru (2021), and recently declared in Venezuela (2023). This work aimed to analyze the evolution of Foc TR4 in Musaceae in LAC between 2018 and 2022. This perspective contains a selection of topics related to Foc TR4 in LAC that address and describe (i) the threat of Foc TR4 in LAC, (ii) a bibliometric analysis of the scientific production of Foc TR4 in LAC, (iii) the current situation of Foc TR4 in Colombia, Peru, and Venezuela, (iv) medium-term prospects in LAC member countries, and (v) export trade and local food security. In this study, the presence of Foc TR4 in Venezuela and the possible consequences of the production of Musaceae in the long term were reported for the first time. In conclusion, TR4 is a major threat to banana production in Latin America and the world, and it is important to take measures to control the spread of the fungus and minimize its impact on the banana industry. It is important to keep working on the control of Foc TR4, which requires the participation of the local and international industry, researchers, and consumers, among others, to prevent the disappearance of bananas.

15N tracing reveals preference for different nitrogen forms of Fusarium oxysporum f. sp. cubense tropical race 4.

Plant uptake of nitrogen is often associated with increased incidence of banana Fusarium wilt, a disease caused by the soil-borne fungus Fusarium oxysporum f. sp. cubense tropical race 4 (Foc TR4). However, the nitrogen metabolic preferences of Foc TR4 pathogens remain unknown. In this study, we investigated the ecophysiological patterns of Foc TR4 grown on different combinations of organic and inorganic nitrogen. Potato Dextrose Agar (PDA) and Rose Bengal Medium (RBM) were used as an organic nitrogen source, which was sequentially replaced with inorganic N (0, 50% or 90%) in the form 15NH4NO3 or NH4 15NO3 to reveal preferential assimilation of ammonium or nitrate. The results showed that mycelium biomass and nitrogen content decreased significantly, while the carbon content and C:N ratio increased in Foc TR4 grown on media containing inorganic nitrogen sources. Mycelium biomass was negatively correlated with C:N ratio. Mycelium 15N abundance increased significantly between the PDA50 + A50/RBM50 + A50 treatments (50% organic nitrogen+50%15NH4NO3) and the PDA10 + A90/RBM10 + A90 treatments (10% organic nitrogen+90%15NH4NO3). These results indicate that the higher C:N ratio reduced mycelium growth by reducing its biomass and diameter and showed that Foc TR4 preferred to use ammonium nitrogen to promote the growth. These findings suggest that treating banana crops with a combination of organic and inorganic (i.e., nitrate) nitrogen could be a better way to defend against Fusarium wilt of banana.

Extracellular DNA of Fusarium oxysporum f. sp. cubense as a Priming Agent for Inducing the Resistance of Banana Plantlets

Fusarium wilt is one of the major causes of global losses in the banana industry. The application of extracellular DNA (eDNA) is explored as a natural fungicide. eDNA is categorized on the basis of the receiving cell’s perception, namely self and non-self. The application of self-eDNA in agriculture presents the potential for limiting the growth of pathogens, while non-self-eDNA, as a vaccine for plants. This study evaluated whether the eDNA from Fusarium oxysporum f. sp. cubense (Foc) could limit the growth of Foc itself (self-inhibition test) while increasing the resistance of banana plant (priming test). A self-inhibition test showed that the administration of 400 and 800 μg mL−1 eDNA inhibited Foc TR4 spore germination. A priming test was carried out on banana plantlets in the interaction medium containing eDNA Foc TR4 suspension with final concentrations of 40, 80, and 200 μg mL−1. After 24 h, the plantlets were infected with a suspension of 106 spore mL−1. Increased resistance was observed in plantlets treated with 200 μg mL−1 of Foc TR4 eDNA, as indicated by the decrease in leaf symptoms and rhizome discoloration. The accumulation of O2- and H2O2 was observed 24 h after priming as was a significant increase in the relative expression of CAT, PR1, and chitinase 1 genes on day 9 post-infection. In conclusion, eDNA Foc TR4 as a growth inhibitor of pathogen and a priming agent to the banana plantlets could be considered as a biofungicide candidate to induce the resistance of banana plants.

Spatiotemporal biocontrol and rhizosphere microbiome analysis of Fusarium wilt of banana

The soil-borne fungus Fusarium oxysporum f. sp. cubense tropical race 4 (Foc TR4) causes Fusarium wilt of banana (FWB), which devastates banana production worldwide. Biocontrol is considered to be the most efficient approach to reducing FWB. Here we introduce an approach that spatiotemporally applies Piriformospore indica and Streptomyces morookaensis strains according to their respective strength to increase biocontrol efficacy of FWB. P. indica successfully colonizes banana roots, promotes lateral root formation, inhibits Foc TR4 growth inside the banana plants and reduces FWB. S. morookaensis strain Sm4-1986 secretes different secondary compounds, of which xerucitrinin A (XcA) and 6-pentyl-α-pyrone (6-PP) show the strongest anti-Foc TR4 activity. XcA chelates iron, an essential nutrient in pathogen-plant interaction that determines the output of FWB. 6-PP, a volatile organic compound, inhibits Foc TR4 germination and promotes banana growth. Biocontrol trials in the field demonstrated that application of S. morookaensis lead to improvement of soil properties and increase of rhizosphere-associated microbes that are beneficial to banana growth, which significantly reduces disease incidence of FWB. Our study suggests that optimal utilization of the two biocontrol strains increases efficacy of biocontrol and that regulating iron accessibility in the rhizosphere is a promising strategy to control FWB.

Transcriptome and metabolome profiling provide insights into hormone-mediated enhanced growth in autotetraploid seedlings of banana (Musa spp.)

IntroductionReconstructive breeding based on autotetraploids to generate triploid varieties is a promising breeding strategy in banana (Musa spp.). Therefore understanding the molecular mechanisms underlying the phenotypic differences between the original diploid and its autopolyploid derivatives is of significant importance in such breeding programs of banana.MethodsIn this study, a number of non-chimeric autotetraploid plants, confirmed by flow cytometry and chromosome counting were obtained using colchicine treatment of ‘Pisang Berlin' (AA Group), a diploid banana cultivar highly resistant to Fusarium wilt Tropical Race 4 (Foc TR4) and widely cultivated in Asia.Results and discussionThe autotetraploids showed significant increase in plant height, pseudostem diameter, root length, leaf thickness, leaf area, and leaf chlorophyll content. Transcriptomic analysis indicated that differentially expressed genes were mainly enriched in plant hormone signal transduction, mitogen-activated protein kinase (MAPK) signaling pathway, and carbon fixation in photosynthetic organelles. The genes related to the metabolism, transport or signaling of auxin, abscisic acid (ABA), cytokinin (CTK) and gibberellin (GA), as well as the genes encoding essential enzymes in photosynthetic CO2 fixation were differentially expressed in leaves of autotetraploids and most of them were up-regulated. Metabolomic analysis revealed that the differentially accumulated metabolites were mainly involved in plant hormone signal transduction, porphyrin and chlorophyll metabolism, indole alkaloid biosynthesis, and carbon fixation in photosynthetic organelles. The results therefore, demonstrate that the hormones IAA, ABA, and photosynthetic regulation may play a vital role in the observed enhancement in the autotetraploids. These could be used as molecular and biochemical markers to facilitate the generation of triploid progenies as suitable new varieties for cultivation.

The backbone of agrifood value chain resilience: Innovation in the Ecuadorian banana value chain from a historical perspective

Given the accelerated global shifts and disruptions to agrifood value chains, an approach is required that reveals multiple drivers, recognizable patterns, and behaviors that can be understood and navigated towards value chain resilience. This research shares roots with and builds further on resilience approaches, the concept of upgrading in global value chains and innovation to contribute to the study of agrifood value chain resilience. The analysis is based on narratives extracted from interviews with local actors in the chain. Narrative, as formed from a combination of temporal succession and causality, helps us track the chain’s historical trajectory, identify critical historical and contemporary drivers of change and actors’ adaptations to historical events, the latest being the Covid-19 pandemic. Our study finds that main drivers, such as natural hazards, market dynamics, actors’ organization, and digital and technological innovation, have been key to Covid-19 adaptation. These constitute crucial areas for policy action and management of systemic risk. For example, previous experience with the fungal disease Foc TR4 one year before the Covid-19 pandemic has been perceived as a key factor in easing Covid-19 adaptation. The analysis shows that some shocks may be challenging to predict, while others may result from the chronic build-up of vulnerabilities, such as the impacts of climate change or the spread of transboundary diseases. As observed in this study, the backbone of overall agrifood value chain resilience and Covid-19 adaptation lie in experiential learning and build on the past; hence, identifying lessons from past innovations is crucial to finding solutions in novel contexts and to deriving opportunities for agrifood value chain development.

Antifungal Mechanism of Metabolites from Newly Isolated Streptomyces sp. Y1-14 against Banana Fusarium Wilt Disease Using Metabolomics.

Banana Fusarium wilt caused by Fusarium oxysporum f. sp. cubense tropical race 4 (Foc TR4) is one of the most destructive banana diseases in the world, which limits the development of the banana industry. Compared with traditional physical and chemical practices, biological control becomes a promising safe and efficient strategy. In this study, strain Y1-14 with strong antagonistic activity against Foc TR4 was isolated from the rhizosphere soil of a banana plantation, where no disease symptom was detected for more than ten years. The strain was identified as Streptomyces according to the morphological, physiological, and biochemical characteristics and the phylogenetic tree of 16S rRNA. Streptomyces sp. Y1-14 also showed a broad-spectrum antifungal activity against the selected 12 plant pathogenic fungi. Its extracts inhibited the growth and spore germination of Foc TR4 by destroying the integrity of the cell membrane and the ultrastructure of mycelia. Twenty-three compounds were identified by gas chromatography-mass spectrometry (GC-MS). The antifungal mechanism was investigated further by metabolomic analysis. Strain Y1-14 extracts significantly affect the carbohydrate metabolism pathway of Foc TR4 by disrupting energy metabolism.

Comparative Transcriptome Analysis Unravels the Response Mechanisms of Fusarium oxysporum f.sp. cubense to a Biocontrol Agent, Pseudomonas aeruginosa Gxun-2

Banana Fusarium wilt, which is caused by Fusarium oxysporum f.sp. cubense Tropical Race 4 (FOC TR4), is one of the most serious fungal diseases in the banana-producing regions in east Asia. Pseudomonas aeruginosa Gxun-2 could significantly inhibit the growth of FOC TR4. Strain Gxun-2 strongly inhibited the mycelial growth of FOC TR4 on dual culture plates and caused hyphal wrinkles, ruptures, and deformities on in vitro cultures. Banana seedlings under pot experiment treatment with Gxun-2 in a greenhouse resulted in an 84.21% reduction in the disease. Comparative transcriptome analysis was applied to reveal the response and resistance of FOC TR4 to Gxun-2 stress. The RNA-seq analysis of FOC TR4 during dual-culture with P. aeruginosa Gxun-2 revealed 3075 differentially expressed genes (DEGs) compared with the control. Among the genes, 1158 genes were up-regulated, and 1917 genes were down-regulated. Further analysis of gene function and the pathway of DEGs revealed that genes related to the cell membrane, cell wall formation, peroxidase, ABC transporter, and autophagy were up-regulated, while down-regulated DEGs were enriched in the sphingolipid metabolism and chitinase. These results indicated that FOC TR4 upregulates a large number of genes in order to maintain cell functions. The results of qRT-PCR conducted on a subset of 13 genes were consistent with the results of RNA-seq data. Thus, this study serves as a valuable resource regarding the mechanisms of fungal pathogen resistance to biocontrol agents.

Evaluation of Resistance of Banana Genotypes with AAB Genome to Fusarium Wilt Tropical Race 4 in China.

Banana cultivars with the AAB genome group comprise diverse subgroups, such as Plantain, Silk, Iholena, and Pisang Raja, among others, which play an important role in food security in many developing countries. Some of these cultivars are susceptible to Fusarium oxysporum f. sp. cubense tropical race 4 (Foc TR4), the most destructive pathogen threatening banana production worldwide, and some of them are still largely unknown. We evaluated the resistance of 37 banana genotypes, including Plantain, Silk, Iholena, Maia Maoli/Popoulu, Pisang Raja, Pome, and Mysore, to Foc TR4 under both greenhouse and field conditions. Genotypes from the Silk and Iholena subgroups were highly susceptible to Foc TR4. Pome and Mysore showed resistance and intermediate resistance, respectively. However, Pisang Raja ranged from susceptible to intermediate resistance. One cultivar from the Maia Maoli/Popoulu subgroup was highly susceptible, while the other displayed significant resistance. Most Plantain cultivars exhibited high resistance to Foc TR4, except two French types of cultivar, 'Uganda Plantain' and 'Njombe N°2', which were susceptible. The susceptibility to Foc TR4 of some of the AAB genotypes evaluated, especially Plantain and other cooking bananas, indicates that growers dependent on these varieties need to be included as part of the prevention and integrated Foc TR4 management strategies, as these genotypes play a crucial role in food security and livelihoods.