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Fusarium wilt of banana (FWB), caused by the soil-borne fungus Fusarium oxysporum f. sp. cubense (Foc), poses an undeniable threat to global banana production. This disease has intensified in recent years, with the tropical race 4 (TR4) strain spreading rapidly. Since 2018, the number of affected countries has increased from 16 to 23, presenting a significant challenge to researchers, producers, and National Plant Protection Organizations (NPPOs) worldwide. The potential impact of TR4 in Latin America and the Caribbean (LAC) is particularly concerning. This region boasts seven of the top ten banana-exporting countries, and bananas and plantains are crucial for food security and income generation. In Colombia, where TR4was detected in 2019, the disease has already spread fromLa Guajira to Magdalena, and it is currently affecting 20 large commercial export farms. In Peru, the disease was detected in 2021 and although still restricted to the northern region, flood irrigation and heavy rains associated with the Yaku cyclone, boosted pathogen spread, and more than 400 small organic banana farmers are currently affected. In Venezuela, TR4 detection occurred in 2023, with plantations across three states and five municipalities now affected. Worryingly, TR4 has also been confirmed in plantains, a staple food in the region. Current national responses in LAC primarily rely on preventive and reactive measures: preventing initial incursions and containing outbreaks to avoid further spread. However, the disease's relentless progression suggests that its eventual presence in all banana-producing areas is likely. Therefore, exploring alternative management approaches beyond pathogen exclusion becomes crucial, both in affected and disease-free regions. This paper examines the current spread of TR4, focusing on epidemiological aspects and recent research-based management options. Key epidemiological features were highlighted, drawing practical examples from various scales (plots to landscapes) and utilizing experiences from LAC's fight against TR4. The paper also reviews field-tested approaches in biosecurity, biological control, resistant varieties, soil health, and integrated disease management, acknowledging the specific challenges faced by smallholder settings. In each section research initiatives were analyzed, identifying gaps, and proposing directions to minimize TR4 impact and accelerate the development of sustainable solutions for managing this devastating disease. [ABSTRACT FROM AUTHOR]

Banana production in Australia is under threat from Fusarium wilt Tropical Race 4 (TR4), and the option of growing a disease-resistant variety is sought after by many farmers. Goldfinger is one such alternative; however, it was previously rejected by Australian consumers. In Phase I of this investigation, mutagenesis was employed as a practical method to generate large numbers of novel variants. In Phase II, 631 variants were established in a field trial where agronomic and postharvest assessments, including taste-testing, identified 20 improved selections. Phase III involved a replicated field evaluation of the selections with favourable mutations, where further taste-testing revealed five selections with the most promising eating characteristics. In Phase IV, the five selections underwent large-scale consumer and sensory evaluations where four of them performed as well as the industry standard controls 'Williams' Cavendish and Lady Finger. The sensory analysis revealed differences in the fruit flavour profile and textural attributes, and these features will play a key role in bringing a new banana variety to market and for promotion to consumers. Screening of the variants' resistance to yellow Sigatoka was integrated into the project, but further work is needed to ensure these four selections have retained their TR4 resistance. [ABSTRACT FROM AUTHOR]

The recent incursion of Fusarium wilt of banana (FWB) caused by TR4 threatens banana production in the Chira Valley of Peru. To develop a management strategy, we mapped the entire production area from the air. During 12 flights at an altitude of 610 m (2,000 ft), we gathered 133,700 images in a timeframe of 2 weeks and constructed an orthomosaic map of 73,000 ha. This unveiled the complex logistic network across the banana-producing region, comprising 150 km of primary roads, as well as numerous secondary and tertiary unpaved paths through and bordering banana plantations. Moreover, the Poechos reservoir—with a total length of 450 km—feeds the entire Chira Valley irrigation system, which could further exacerbate TR4 expansion. Using georeferenced landscape details (digital terrain model), we determined areas prone to flooding, which is also relevant for disseminating TR4. Analyses of the maps resulted in the identification of many suspect areas for direct sampling and subsequent analyses to study the expansion of TR4. At four locations, we confirmed two cases, and by June 8, 2023, a total of 207 cases were reported. We conclude that TR4 is a serious threat to the entire region, which exports approximately 25% of the global organic bananas. [Figure: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Fusarium wilt of banana (FWB), caused by the soil-borne fungus Fusarium oxysporum f. sp. cubense (Foc), poses an undeniable threat to global banana production. This disease has intensified in recent years, with the tropical race 4 (TR4) strain spreading rapidly. Since 2018, the number of affected countries has increased from 16 to 23, presenting a significant challenge to researchers, producers, and National Plant Protection Organizations (NPPOs) worldwide. The potential impact of TR4 in Latin America and the Caribbean (LAC) is particularly concerning. This region boasts seven of the top ten banana-exporting countries, and bananas and plantains are crucial for food security and income generation. In Colombia, where TR4 was detected in 2019, the disease has already spread from La Guajira to Magdalena, and it is currently affecting 20 large commercial export farms. In Peru, the disease was detected in 2021 and although still restricted to the northern region, flood irrigation and heavy rains associated with the Yaku cyclone, boosted pathogen spread, and more than 400 small organic banana farmers are currently affected. In Venezuela, TR4 detection occurred in 2023, with plantations across three states and five municipalities now affected. Worryingly, TR4 has also been confirmed in plantains, a staple food in the region. Current national responses in LAC primarily rely on preventive and reactive measures: preventing initial incursions and containing outbreaks to avoid further spread. However, the disease’s relentless progression suggests that its eventual presence in all banana-producing areas is likely. Therefore, exploring alternative management approaches beyond pathogen exclusion becomes crucial, both in affected and disease-free regions. This paper examines the current spread of TR4, focusing on epidemiological aspects and recent research-based management options. Key epidemiological features were highlighted, drawing practical examples from various scales (plots to landscapes) and utilizing experiences from LAC’s fight against TR4. The paper also reviews field-tested approaches in biosecurity, biological control, resistant varieties, soil health, and integrated disease management, acknowledging the specific challenges faced by smallholder settings. In each section research initiatives were analyzed, identifying gaps, and proposing directions to minimize TR4 impact and accelerate the development of sustainable solutions for managing this devastating disease.

This editorial titled "Microbial interaction with banana: mechanisms, symbiosis, and integrated diseases control" explores the significance of bananas as a global fruit and the need for new methods to combat diseases that affect banana crops. The document emphasizes the role of microbial interactions in mitigating soil-borne diseases and presents original research articles on various aspects of this topic. It discusses the development of a compound microbial agent for controlling Fusarium wilt disease in heavily infected banana fields, achieving a disease control effect of 57.14%. The text also suggests that manipulating banana holobionts by introducing novel endophytes into plantlets could be a future direction for enhancing biocontrol agents. The authors stress the importance of understanding host-pathogen-endophyte interactions for sustainable development in the banana industry. [Extracted from the article]

Banana production in Australia is under threat from Fusarium wilt Tropical Race 4 (TR4), and the option of growing a disease-resistant variety is sought after by many farmers. Goldfinger is one such alternative; however, it was previously rejected by Australian consumers. In Phase I of this investigation, mutagenesis was employed as a practical method to generate large numbers of novel variants. In Phase II, 631 variants were established in a field trial where agronomic and postharvest assessments, including taste-testing, identified 20 improved selections. Phase III involved a replicated field evaluation of the selections with favourable mutations, where further taste-testing revealed five selections with the most promising eating characteristics. In Phase IV, the five selections underwent large-scale consumer and sensory evaluations where four of them performed as well as the industry standard controls ‘Williams’ Cavendish and Lady Finger. The sensory analysis revealed differences in the fruit flavour profile and textural attributes, and these features will play a key role in bringing a new banana variety to market and for promotion to consumers. Screening of the variants’ resistance to yellow Sigatoka was integrated into the project, but further work is needed to ensure these four selections have retained their TR4 resistance.

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Fusarium wilt of banana is a devastating disease that has decimated banana production worldwide. Host resistance to Fusarium oxysporum f. sp. Cubense (Foc), the causal agent of this disease, is genetically dissected in this study using two Musa acuminata ssp. Malaccensis segregating populations, segregating for Foc Tropical (TR4) and Subtropical (STR4) race 4 resistance. Marker loci and trait association using 11 SNP-based PCR markers allowed the candidate region to be delimited to a 12.9 cM genetic interval corresponding to a 959 kb region on chromosome 3 of 'DH-Pahang' reference assembly v4. Within this region, there was a cluster of pattern recognition receptors, namely leucine-rich repeat ectodomain containing receptor-like protein kinases, cysteine-rich cell-wall-associated protein kinases, and leaf rust 10 disease-resistance locus receptor-like proteins, positioned in an interspersed arrangement. Their transcript levels were rapidly upregulated in the resistant progenies but not in the susceptible F2 progenies at the onset of infection. This suggests that one or several of these genes may control resistance at this locus. To confirm the segregation of single-gene resistance, we generated an inter-cross between the resistant parent 'Ma850' and a susceptible line 'Ma848', to show that the STR4 resistance co-segregated with marker '28820' at this locus. Finally, an informative SNP marker 29730 allowed the locus-specific resistance to be assessed in a collection of diploid and polyploid banana plants. Of the 60 lines screened, 22 lines were predicted to carry resistance at this locus, including lines known to be TR4-resistant, such as 'Pahang', 'SH-3362', 'SH-3217', 'Ma-ITC0250', and 'DH-Pahang/CIRAD 930'. Additional screening in the International Institute for Tropical Agriculture's collection suggests that the dominant allele is common among the elite 'Matooke' NARITA hybrids, as well as in other triploid or tetraploid hybrids derived from East African highland bananas. Fine mapping and candidate gene identification will allow characterization of molecular mechanisms underlying the TR4 resistance. The markers developed in this study can now aid the marker-assisted selection of TR4 resistance in breeding programs around the world. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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.

Fusarium oxysporum f.sp. cubense, causal agent of Panama disease, is one of the biggest threats to global banana production, particularly the Cavendish competent tropical race 4 (Foc TR4). It continues to spread globally with detections occurring in regions of the Middle East and new continents such as Africa and South America in the last decade. As the search was on for new management strategies and resistant cultivars to combat the disease, a banana cultivar-screening trial took place in the Northern Territory of Australia, which examined the responses of 24 banana cultivars to the soil borne fungus. These cultivars included material from TBRI, FHIA and selections from Thailand, Indonesia and Australia and evaluated for their resistance to tropical race 4 for two cropping cycles. Several cultivars displayed considerable resistance to Foc TR4, including several FHIA parental lines and hybrids, the Cavendish (AAA) selections GCTCV 215 and GCTCV 247 from TBRI and an Indonesian selection CJ19 showed either very little to no plant death due to the disease. [ABSTRACT FROM AUTHOR]

The socio‐economic impact of Fusarium odoratissimum, which is colloquially called tropical race 4 (TR4), is escalating as this fungal pathogen spreads to new banana‐growing areas. Hence, the development of simple, reliable and rapid detection technologies is indispensable for implementing quarantine measures. Here, a versatile loop‐mediated isothermal amplification (LAMP) assay has been developed that is applicable under field and laboratory conditions. DNA markers unique to TR4 isolates were obtained by diversity arrays technology sequencing (DArTseq), a genotyping by sequencing technology that was conducted on 27 genotypes, comprising 24 previously reported vegetative compatibility groups (VCGs) and three TR4 isolates. The developed LAMP TR4 assay was successfully tested using 22 TR4 isolates and 45 non‐target fungal and bacterial isolates, as well as on infected plants under greenhouse and field conditions. The detection limit was 1 pg µL−1 pure TR4 DNA or 102 copies plasmid‐localized TR4 unique sequence (SeqA) per reaction, which was not affected by background DNA in complex samples. The LAMP TR4 assay offers a powerful tool for the routine and unambiguous identification of banana plants infected with TR4, contributing to advanced diagnosis in field situations and monitoring of fusarium wilt. [ABSTRACT FROM AUTHOR]

Fusarium oxysporum f.sp. cubense, causal agent of Panama disease, is one of the biggest threats to global banana production, particularly the Cavendish competent tropical race 4 (Foc TR4). It continues to spread globally with detections occurring in regions of the Middle East and new continents such as Africa and South America in the last decade. As the search was on for new management strategies and resistant cultivars to combat the disease, a banana cultivar-screening trial took place in the Northern Territory of Australia, which examined the responses of 24 banana cultivars to the soil borne fungus. These cultivars included material from TBRI, FHIA and selections from Thailand, Indonesia and Australia and evaluated for their resistance to tropical race 4 for two cropping cycles. Several cultivars displayed considerable resistance to Foc TR4, including several FHIA parental lines and hybrids, the Cavendish (AAA) selections GCTCV 215 and GCTCV 247 from TBRI and an Indonesian selection CJ19 showed either very little to no plant death due to the disease.

Fusarium wilt is currently spreading in banana growing regions around the world leading to substantial losses. The disease is caused by the fungus Fusarium oxysporum f. sp. cubense (Foc), which is further classified into distinct races according to the banana varieties that they infect. Cavendish banana is resistant to Foc race 1, to which the popular Gros Michel subgroup succumbed last century. Cavendish effectively saved the banana industry, and became the most cultivated commercial subgroup worldwide. However, Foc tropical race 4 (TR4) subsequently emerged in Southeast Asia, causing significant yield losses due to its high level of aggressiveness to cultivars of Cavendish, and other commonly grown cultivars. Preventing further spread is crucially important in the absence of effective control methods or resistant market-acceptable banana cultivars. Implementation of quarantine and containment measures depends on early detection of the pathogen through reliable diagnostics. In this study, we tested the hypothesis that secreted in xylem (SIX) genes, which currently comprise the only known family of effectors in F. oxysporum, contain polymorphisms to allow the design of molecular diagnostic assays that distinguish races and relevant VCGs of Foc. We present specific and reproducible diagnostic assays based on conventional PCR targeting SIX genes, using as templates DNA extracted from pure Foc cultures. Sets of primers specifically amplify regions of: SIX6 in Foc race 1, SIX1 gene in TR4, SIX8 in subtropical race 4, SIX9/SIX10 in Foc VCG 0121, and SIX13 in Foc VCG 0122. These assays include simplex and duplex PCRs, with additional restriction digestion steps applied to amplification products of genes SIX1 and SIX13. Assay validations were conducted to a high international standard including the use of 250 Fusarium spp. isolates representing 16 distinct Fusarium species, 59 isolates of F. oxysporum, and 21 different vegetative compatibility groups (VCGs). Tested parameters included inter and intraspecific analytical specificity, sensitivity, robustness, repeatability, and reproducibility. The resulting suite of assays is able to reliably and accurately detect R1, STR4, and TR4 as well as two VCGs (0121 and 0122) causing Fusarium wilt in bananas.

Fusarium wilt is currently spreading in banana growing regions around the world leading to substantial losses. The disease is caused by the fungus Fusarium oxysporum f. sp. cubense (Foc), which is further classified into distinct races according to the banana varieties that they infect. Cavendish banana is resistant to Foc race 1, to which the popular Gros Michel subgroup succumbed last century. Cavendish effectively saved the banana industry, and became the most cultivated commercial subgroup worldwide. However, Foc tropical race 4 (TR4) subsequently emerged in Southeast Asia, causing significant yield losses due to its high level of aggressiveness to cultivars of Cavendish, and other commonly grown cultivars. Preventing further spread is crucially important in the absence of effective control methods or resistant market-acceptable banana cultivars. Implementation of quarantine and containment measures depends on early detection of the pathogen through reliable diagnostics. In this study, we tested the hypothesis that secreted in xylem (SIX) genes, which currently comprise the only known family of effectors in F. oxysporum , contain polymorphisms to allow the design of molecular diagnostic assays that distinguish races and relevant VCGs of Foc. We present specific and reproducible diagnostic assays based on conventional PCR targeting SIX genes, using as templates DNA extracted from pure Foc cultures. Sets of primers specifically amplify regions of: SIX6 in Foc race 1, SIX1 gene in TR4, SIX8 in subtropical race 4, SIX9 / SIX10 in Foc VCG 0121, and SIX13 in Foc VCG 0122. These assays include simplex and duplex PCRs, with additional restriction digestion steps applied to amplification products of genes SIX1 and SIX13. Assay validations were conducted to a high international standard including the use of 250 Fusarium spp. isolates representing 16 distinct Fusarium species, 59 isolates of F. oxysporum , and 21 different vegetative compatibility groups (VCGs). Tested parameters included inter and intraspecific analytical specificity, sensitivity, robustness, repeatability, and reproducibility. The resulting suite of assays is able to reliably and accurately detect R1, STR4, and TR4 as well as two VCGs (0121 and 0122) causing Fusarium wilt in bananas. [ABSTRACT FROM AUTHOR]

The expression profile of pathogenesis related genes are signatures of an infection response in plant cells. Pathogenic infections can increase or reduce gene expression in a plant system in a relatively specific pattern. These expression patterns can be used as standards in pathogenicity studies and, where phenotypic expression is normally used to gauge a plants response to infection, it could additionally present a more rapid and early screening reference tool. Three genes: catalase (CAT), pathogen related protein (PR10), and phenylalanine ammonia (PAL) all implicitly implicated in the plant disease response pathway were targeted for analysis during the infection of Fusarium oxysporum f. sp. cubense tropical race 4 (FOCR4) in banana Musa acuminata cv. Berangan seedlings after a standard challenge under growth room conditions. Distinct patterns of gene expression were observed at three infection time points by real time expression analysis. There was a sequential 10-fold reduction in expression for the PR gene while, the PAL and CAT genes were both upregulated. These results present a set of reference genes that could be used for screening of a plant's response to Fusarium before the onset of symptoms. [ABSTRACT FROM AUTHOR]

The continued dispersal of Fusarium oxysporum f. sp. cubense Tropical race 4 (FocTR4), a quarantine soil-borne pathogen that kills banana, has placed this worldwide industry on alert and triggered enormous pressure on National Plant Protection (NPOs) agencies to limit new incursions. Accordingly, biosecurity plays an important role while long-term control strategies are developed. Aiming to strengthen the contingency response plan of Ecuador against FocTR4, a population biology study—including phylogenetics, mating type, vegetative compatibility group (VCG), and pathogenicity testing—was performed on isolates affecting local bananas, presumably associated with race 1 of F. oxysporum f. sp. cubense (Foc). Our results revealed that Foc populations in Ecuador comprise a single clonal lineage, associated with VCG0120. The lack of diversity observed in Foc populations is consistent with a single introduction event from which secondary outbreaks originated. The predominance of VCG0120, together with previous reports of its presence in Latin America countries, suggests this group as the main cause of the devastating Fusarium wilt epidemics that occurred in the 1950s associated to the demise of ‘Gros Michel’ bananas in the region. The isolates sampled from Ecuador caused disease in cultivars that are susceptible to races 1 and 2 under greenhouse experiments, although Fusarium wilt symptoms in the field were only found in ‘Gros Michel’. Isolates belonging to the same VCG0120 have historically caused disease on Cavendish cultivars in the subtropics. Overall, this study shows how Foc can be easily dispersed to other areas if restriction of contaminated materials is not well enforced. We highlight the need of major efforts on awareness and monitoring campaigns to analyze suspected cases and to contain potential first introduction events of FocTR4 in Ecuador.

Bananas are Australia’s number-one selling supermarket product (in volume), with over fivemillion of them eaten daily [...]

Panama wilt, caused by Fusarium oxysporum f. sp. cubense (Foc) is considered one of the most devastating banana diseases in recorded history. The disease threatens the banana industry due to Tropical Race 4 (TR4) infecting the Cavendish cultivar. Forty-two of the 45 representative isolates from Luzon were pathogenic, based on leaf symptom index and vascular discoloration rating. Accurate, fast and reliable identification are pre-requisites for effective management considering there are yet no proven effective chemicals to control the disease, thus the confirmation by a PCR-based diagnostic tool is essential. Using race-specific primers, FocTr4-F/FocTr4-R and Foc-1/Foc-2, the absence of TR4 in Luzon has been confirmed, however, the occurrence of Race 4 has been reported, which should also be taken in consideration as the latter can also cause severe damage under favorable conditions. Furthermore, to examine genetic diversity of Foc in bananas, 55 of the 164 isolates collected from Regions I, II, III, IV and Cordillera Administrative Region (CAR) were analyzed by fingerprinting techniques using M13, ERIC and REP primers. Twenty-two reference isolates from Mindanao were also analyzed using the same primers. Foc isolates were differentiated into two clades at 25% similarity level, classifying all Mindanao isolates to clade A. Consistently high genetic variation was obtained from Luzon isolates using M13, an arbitrarily primed fingerprinting technique and repetitive elements, REP and ERIC-PCR, while low genetic variation was obtained from Mindanao isolates. ERIC-PCR was the most informative and predictive fingerprinting method as the TR4 isolates from Mindanao were grouped together. No grouping of Foc isolates was observed with respect to geographical origin, except isolates from Mindanao. In addition, grouping of Foc4 is also regardless of host variety in all analyses conducted. Overall, high genetic variability was recorded in Foc Philippine population for the three primers used, which might render host resistance vulnerable.

Fusarium wilt of banana, caused by the soil-borne pathogen Fusarium oxysporum f. sp. cubense (Foc), is a major constraint to banana production worldwide (Viljoen et al ., 2020). Currently, Cavendish bananas are severely affected by Foc Tropical Race 4 (TR4) globally. In Africa, Foc TR4 was first detected in northern Mozambique in 2013 (Viljoen et al ., 2020), and has since been found on the island of Mayotte in the Mozambique Channel off the coast of southeastern Africa (Aguayo et al ., 2021). In early 2023, severe leaf-yellowing and wilting of Cavendish banana plants was observed on Cavendish banana plants at several small holder farmer properties in Grande Comoros (Ngazidja) including in Ntsinimoipanga (-11,790054 S, 43°25'47,04384 E), Batou (-11,499716 S, 43°21'51,71976''E), Madjeweni (-11,8217 S , 43°16'41''E) and Mdé (-11°41'54'' S, 43°15'20''E). When the pseudostems of these plants were split open, a reddish-brown internal discoloration of the vascular tissue became apparent. Discolored strands of diseased plants were collected, and the causal agent identified using DNA-based techniques, vegetative compatibility group (VCG) analysis and pathogenicity testing. The samples were plated onto potato dextrose agar and single-spored and isolated from individual diseased plants identified as F. oxysporum based on cultural and morphological characteristics. These include the production of white fungal colonies with a purple center, infrequent production of macroconidia, but an abundance of microconidia on short monophialides, and terminal or intercalary chlamydospores (Leslie and Summerell, 2006). Foc TR4 was identified from seven isolates by conventional (Dita et al ., 2010) and quantitative-PCR (Matthews et al ., 2020), and with loop-mediated isothermal amplification (LAMP) (Ordóñez et al ., 2021). All seven isolates were confirmed as members of the VCG 01213/16 complex when nit -1 mutants of the unknown Foc isolates were compatible with Nit-M mutants of the Foc VCG 01213 and VCG 01216 tester strains. Two isolates were then selected for pathogenicity testing, and 2-month-old tissue culture-derived Cavendish plants (cv. Williams) inoculated by using the method described by Ndayihanzamaso et al . (2022). After 4 weeks, the Foc TR4-inoculated plants produced wilting symptoms and internal rhizome discoloration typical of Fusarium wilt. Foc TR4 was reisolated from the inoculated plants and identified by qPCR (Matthews et al . 2020), thereby fulfilling Koch's postulates. These results provide scientific proof of the presence of Foc TR4 in a second island in the Comoros archipelago. Comprehensive surveys will be conducted in all three of the Comoros Islands to assess the presence and impact of Foc TR4 to implement containment strategies. Collaborative initiatives and coordinated actions among growers and other stakeholders are needed to prevent the spread of Foc TR4 to more Southwest Indian Ocean islands and countries on the East African coasts. The importance of banana for food security and livelihoods, and the unique genetic diversity of bananas found on the Comoros islands, requires the eradication and isolation of diseased bananas on the short term, and the screening of local banana varieties for Foc TR4 resistance on the longer term.

Fusarium wilt or Panama disease poses a great risk to banana production worldwide and is caused by Fusarium oxysporum f. sp. Cubense (Foc). Its tropical race 4 (TR4) is a major problem for Cavendish banana growing areas. Basrai (Dwarf Cavendish) is the major cultivated variety in Pakistan and now recently been reported to be affected by Foc TR4 in certain areas in the Sindh province. To genetically characterize, three independent isolates (P.TH1, P.TH2 and P.Th3) were sampled from different fields in district Thatta and fungus was isolated on potato dextrose agar. The pure fungal isolates were used to establish Koch's postulates by testing their pathogenicity on tissue cultured Cavendish banana. A diagnostic PCR assay was optimized and used to sequence a reported molecular marker for specific detection of Foc TR4 and to sequence 28S-18S ribosomal RNA intergenic spacer region (IGS) from these local isolates. The phylogenetic of Pakistani isolates suggests that they belong to previously characterized lineage V of tropical race 4 of F. oxysporum cubense with 99% bootstrap support and show close relationship with Indonesian and Malaysian Foc TR4 isolates. The rapid spread of this disease in different banana growing areas of the Sindh is alarming and necessitates effective control of this through chemical and biocontrol, if remains unchecked, Foc TR4 can raise havoc to banana industry in Pakistan and would be devastating economically. [ABSTRACT FROM AUTHOR]

This study is the first to adopt a hypothetical approach to establish the influence of the complex endophyte-host interaction on endophyte survival and antifungal expression. Three key interactions were evaluated; (I) influence of host-induced enzymes on endophyte growth (biomass) and colonization, (II) link between endophyte-produced cellulase, their growth and colonization, and (III) the influence of host environment on antifungal expression of endophytes. The interactions with the host were performed using plant slurry (PS) to mimic in planta (host) environment with analysis on interactions evaluated using Pearson correlation coefficients (r). Results revealed that host induced enzymes may be a limiting factor to colonization of endophytes with inverse correlations observed (-0.046 = r = -0.7164). These enzymes may also limit growth of endophyte although, PAL (r = 0.536) and TPC (r = 0.8894) appeared contrary. Results were also suggestive that endophytes produced cellulase to aid in colonization in host plants (r = 0.7073 in PS), and cellulase activities are continuously produced even when growth of endophytes are limited (r = -0.314 in PS). Endophytes are presumed to produce antifungal compounds in planta (r = 0.2760 in PS), and these compounds may be secondary metabolites, which are primarily produced under nutrient-depleted conditions where growth is poor (in host plant). The superior growth of endophytes in synthetic PDB media has an inverse correlation to antifungal activity (r = -0.5129), confirming that secondary metabolites are involved in antifungal activities. This study clearly presents that success of inoculated endophytes in colonizing, growing and expressing antifungal activities is dependent on the host plant. [ABSTRACT FROM AUTHOR]

Fusarium wilt, caused by Fusarium oxysporum f. sp. cubense (Foc), has been considered as the most devastating disease affecting bananas (Musa spp.) worldwide. A highly virulent strain of Foc, known as tropical race 4 (TR4), has been detected in the southeast Asia in the 1990s, and has since spread to western Asia, Australia, the Middle East, southern Africa, and South America (Viljoen et al. 2020). Foc TR4 can cause severe yield losses in Cavendish (AAA), Gros Michel (AAA), Silk (AAB), Pisang Awak (ABB) and Bluggoe (ABB) bananas (Ploetz et al. 2006). However, cooking bananas such as plantain (AAB) and Matooke (AAA) bananas, appear to be resistant (Zuo et al. 2017). Iholena bananas (AAB), a subgroup of varieties related to plantains (also known as Pacific plantains), is an important staple food in the Pacific Islands where it was domesticated. It is also popular in Peru, probably due to its nutritional value (Kepler et al. 2011) and is wildly cultivated in other South American countries (Dita et al. 2013). In December 2019, typical symptoms of banana Fusarium wilt were observed on Iholena accession 'Pacific Plantain' (ITC0210) in experimental fields located in Dongguan, Guangdong Province of China. The symptoms included leaf yellowing and pseudostem splitting. The vascular tissue inside the pseudostems was dark red to brown, and the inner rhizomes yellowish-brown. Vascular tissues from three diseased plants were sampled aseptically and placed on potato dextrose agar (PDA) containing 0.05 g/liter kanamycin. Fungal colonies typical of F. oxysporum developed rapidly, with purple-tinged white aerial mycelia and an abundance of microconidia borne in false heads on short microconidia (Nelson et al. 1983). Chlamydospores were produced singly or in pairs in hyphae and macroconidia. Molecular identification was performed using Foc race 4-specific primers (Lin et al. 2009), Foc TR4-specific primers (Dita et al. 2010), Foc race 1 and Foc STR4-specific primers (Ndayihanzamaso et al. 2020). Amplicons of expected sizes were obtained for Foc TR4 and race 4, but not for Foc race 1 and STR4. Sequencing of the ITS and 18S rDNA from the three Iholena isolates and BLAST result showed a 100% similarity to the Foc TR4 reference sequences in GenBank (Foc II5, PRJNA73539 and PRJNA56513) to prove that the isolates were Foc TR4. Pathogenicity of the three isolates from Iholena bananas was determined by infecting 4-month-old Cavendish cv. 'Grand Nain' bananas and three Iholena accessions, 'Pacific Plantain' 'Tigua' and 'Uzakan', under greenhouse conditions by root immersion in a Foc conidial suspension and soil drenching at 106 conidia/ml (Dita, 2010). Control plants were treated with sterile distilled water. Three replications of five plantlets were used for each accession. After 35 days, the inoculated plantlets developed typical Fusarium wilt symptoms such as yellowing of the older leaves and discoloration of the inner rhizome. The control plants did not develop symptoms. To complete Koch's postulates, the fungus was re-isolated from inoculated plants and identified as Foc TR4 by PCR (Dita et al, 2010). The susceptibility of 'Tigua' and 'Uzakan' was also confirmed in Foc TR4-infested field trials, with both accessions developing severe Fusarium wilt symptoms. The susceptibility of Iholena bananas to Foc TR4 is of significant concern to all countries where this subgroup is cultivated for major food source, including Peru and other South American countries.

The continued dispersal of Fusarium oxysporum f. sp. cubense Tropical race 4 (FocTR4), a quarantine soil-borne pathogen that kills banana, has placed this worldwide industry on alert and triggered enormous pressure on National Plant Protection (NPOs) agencies to limit new incursions. Accordingly, biosecurity plays an important role while long-term control strategies are developed. Aiming to strengthen the contingency response plan of Ecuador against FocTR4, a population biology study—including phylogenetics, mating type, vegetative compatibility group (VCG), and pathogenicity testing—was performed on isolates affecting local bananas, presumably associated with race 1 of F. oxysporum f. sp. cubense (Foc). Our results revealed that Foc populations in Ecuador comprise a single clonal lineage, associated with VCG0120. The lack of diversity observed in Foc populations is consistent with a single introduction event from which secondary outbreaks originated. The predominance of VCG0120, together with previous reports of its presence in Latin America countries, suggests this group as the main cause of the devastating Fusarium wilt epidemics that occurred in the 1950s associated to the demise of 'Gros Michel' bananas in the region. The isolates sampled from Ecuador caused disease in cultivars that are susceptible to races 1 and 2 under greenhouse experiments, although Fusarium wilt symptoms in the field were only found in 'Gros Michel'. Isolates belonging to the same VCG0120 have historically caused disease on Cavendish cultivars in the subtropics. Overall, this study shows how Foc can be easily dispersed to other areas if restriction of contaminated materials is not well enforced. We highlight the need of major efforts on awareness and monitoring campaigns to analyze suspected cases and to contain potential first introduction events of FocTR4 in Ecuador. [ABSTRACT FROM AUTHOR]

Panama wilt, caused by Fusarium oxysporum f. sp. cubense (Foc) is considered one of the most devastating banana diseases in recorded history. The disease threatens the banana industry due to Tropical Race 4 (TR4) infecting the Cavendish cultivar. Forty-two of the 45 representative isolates from Luzon were pathogenic, based on leaf symptom index and vascular discoloration rating. Accurate, fast and reliable identification are pre-requisites for effective management considering there are yet no proven effective chemicals to control the disease, thus the confirmation by a PCR-based diagnostic tool is essential. Using race-specific primers, FocTr4-F/FocTr4-R and Foc-1/Foc-2, the absence of TR4 in Luzon has been confirmed, however, the occurrence of Race 4 has been reported, which should also be taken in consideration as the latter can also cause severe damage under favorable conditions. Furthermore, to examine genetic diversity of Foc in bananas, 55 of the 164 isolates collected from Regions I, II, III, IV and Cordillera Administrative Region (CAR) were analyzed by fingerprinting techniques using M13, ERIC and REP primers. Twenty-two reference isolates from Mindanao were also analyzed using the same primers. Foc isolates were differentiated into two clades at 25% similarity level, classifying all Mindanao isolates to clade A. Consistently high genetic variation was obtained from Luzon isolates using M13, an arbitrarily primed fingerprinting technique and repetitive elements, REP and ERIC-PCR, while low genetic variation was obtained from Mindanao isolates. ERIC-PCR was the most informative and predictive fingerprinting method as the TR4 isolates from Mindanao were grouped together. No grouping of Foc isolates was observed with respect to geographical origin, except isolates from Mindanao. In addition, grouping of Foc4 is also regardless of host variety in all analyses conducted. Overall, high genetic variability was recorded in Foc Philippine population for the three primers used, which might render host resistance vulnerable. [ABSTRACT FROM AUTHOR]

Fusarium oxysporum f. sp. cubense (Foc), the causal agent of Fusarium wilt or Panama disease on banana, is one of the major constraints in banana production worldwide. Indonesia is the centre of origin for wild and cultivated bananas, which likely co-evolved with Foc. This study explored the widest possible genetic diversity of Foc by sampling across Indonesia at 34 geographically and environmentally different locations in 15 provinces at six islands. This resulted in a comprehensive collection of ∼200 isolates from 40 different local banana varieties. Isolates were identified and assessed using sequence analysis of the translation elongation factor-1alpha ( tef1 ), the RNA polymerase II largest subunit ( rpb1 ), and the RNA polymerase II second largest subunit ( rpb2 ). Phylogenetic analyses of these genes allowed the identification of 180 isolates of Fusarium oxysporum f. sp. cubense (Foc), and 20 isolates of the Fusarium fujikuroi species complex (FFSC), the Fusarium incarnatum-equiseti species complex (FIESC), and the Fusarium sambucinum species complex (FSSC). Further analyses, incorporating a worldwide collection of Foc strains, revealed nine independent genetic lineages for Foc, and one novel clade in the Fusarium oxysporum species complex (FOSC). Selected isolates from each lineage were tested on the banana varieties Gros Michel and Cavendish to characterise their pathogenicity profiles. More than 65 % of the isolates were diagnosed as Tropical Race 4 (Foc-TR4) due to their pathogenicity to Cavendish banana, which supports the hypothesis that Foc-TR4 is of Indonesian origin. Nine independent genetic lineages for Foc are formally described in this study. This biodiversity has not been studied since the initial description of Foc in 1919. This study provides a detailed overview of the complexity of Fusarium wilt on banana and its diversity and distribution across Indonesia.