Foc Tropical Race 4 Research Articles

Secreted in Xylem (SIX) genes in Fusarium oxysporum f.sp. cubense (Foc) unravels the potential biomarkers for early detection of Fusarium wilt disease.

Secreted in Xylem (SIX) are small effector proteins released by Fusarium oxysporum f.sp. cubense (Foc) into the plant's xylem sap disrupting the host's defence responses causing Fusarium wilt disease resulting in a significant decline in banana crop yields and economic losses. Notably, different races of Foc possess unique sets of SIX genes responsible for their virulence, however, these genes remain underutilized, despite their potential as biomarkers for early disease detection. Herein, we identified seven SIX genes i.e. SIX1, SIX2, SIX4, SIX6, SIX8a, SIX9a and SIX13 present in Foc Tropical Race 4 (FocTR4), while only SIX9b in Foc Race 1 (Foc1). Analysis of SIX gene expression in infected banana roots revealed differential patterns during infection providing valuable insights into host-pathogen interactions, virulence level, and early detection time points. Additionally, a comprehensive analysis of virulent Foc1_C2HIR and FocTR4_C1HIR isolates yielded informative genomic insights. Hence, these discoveries contribute to our comprehension of potential disease control targets in these plants, as well as enhancing plant diagnostics and breeding programs.

Behavior and Use of Quaternary Ammonium-Based Disinfectants in Biosafety Protocols Against Fusarium oxysporum f. sp. cubense Race 1 and Tropical Race 4.

The banana is one of Colombia's main export products. However, production is seriously affected by Fusarium wilt of banana, which is the most destructive disease caused by the fungus Fusarium oxysporum f. sp. cubense tropical race 4 (Foc TR4). Currently, management strategies focus on containment and biosecurity protocols to prevent its spread to territories that are free of this disease. This study aimed to evaluate nine quaternary ammonium-based disinfectants (i.e., quaternary ammonium compounds [QACs]) in vitro in Colombia on reproductive (microconidia and macroconidia) and resistance structures (chlamydospores) of Foc race 1 (R1) and tropical race 4 (TR4), with and without soil, to determine the influence of organic matter and soil texture on the action of QACs. A method for inhibiting the action of QACs was standardized and evaluated at 1,200 ppm with a contact time of ≤30 s while evaluating the soil-inoculum and soil-disinfectant interactions. In the soil-inoculum interaction, the efficacy of QACs was 100% in the reproductive and resistance structures of Foc R1 and TR4 without soil. However, in the soil-disinfectant interaction, only QAC4 controlled the pathogen at 100%. The presence of organic matter influenced the biocidal action of the QACs, and fine textures had a greater reducing effect on the concentration. The soil decreased the efficacy of the QACs and, therefore, must be removed from contaminated boots before treatments are applied.

First report of Fusarium wilt of Cavendish bananas caused by Fusarium oxysporum f. sp. cubense Tropical Race 4 in the Grande Comoros Island.

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.

Antifungal effects and biocontrol potential of lipopeptide-producing Streptomyces against banana Fusarium wilt fungus Fusarium oxysporum f. sp. cubense.

Fusarium wilt of banana (FWB), caused by Fusarium oxysporum f. sp. cubense (Foc), especially tropical race 4 (TR4), presents the foremost menace to the global banana production. Extensive efforts have been made to search for efficient biological control agents for disease management. Our previous study showed that Streptomyces sp. XY006 exhibited a strong inhibitory activity against several phytopathogenic fungi, including F. oxysporum. Here, the corresponding antifungal metabolites were purified and determined to be two cyclic lipopeptide homologs, lipopeptin A and lipopeptin B. Combined treatment with lipopeptin complex antagonized Foc TR4 by inhibiting mycelial growth and conidial sporulation, suppressing the synthesis of ergosterol and fatty acids and lowering the production of fusaric acid. Electron microscopy observation showed that lipopeptide treatment induced a severe disruption of the plasma membrane, leading to cell leakage. Lipopeptin A displayed a more pronounced antifungal activity against Foc TR4 than lipopeptin B. In pot experiments, strain XY006 successfully colonized banana plantlets and suppressed the incidence of FWB, with a biocontrol efficacy of up to 87.7%. Additionally, XY006 fermentation culture application improved plant growth parameters and induced peroxidase activity in treated plantlets, suggesting a possible role in induced resistance. Our findings highlight the potential of strain XY006 as a biological agent for FWB, and further research is needed to enhance its efficacy and mode of action in planta.

α-Pheromone Precursor Protein Foc4-PP1 Is Essential for the Full Virulence of Fusarium oxysporum f. sp. cubense Tropical Race 4.

Fusarium oxysporum f. sp. cubense (Foc), which causes Fusarium wilt of bananas, is considered one of the most destructive fungal pathogens of banana crops worldwide. During infection, Foc secretes many different proteins which promote its colonization of plant tissues. Although F. oxysporum has no sexual cycle, it has been reported to secrete an α-pheromone, which acts as a growth regulator, chemoattractant, and quorum-sensing signaling molecule; and to encode a putative protein with the hallmarks of fungal α-pheromone precursors. In this study, we identified an ortholog of the α-pheromone precursor gene, Foc4-PP1, in Foc tropical race 4 (TR4), and showed that it was necessary for the growth and virulence of Foc TR4. Foc4-PP1 deletion from the Foc TR4 genome resulted in decreased fungal growth, increased sensitivity to oxidative stress and cell-wall-damaging agents, and attenuation of pathogen virulence towards banana plantlets. Subcellular localization analysis revealed that Foc4-PP1 was concentrated in the nuclei and cytoplasm of Nicotiana benthamiana cells, where it could suppress BAX-induced programmed cell death. In conclusion, these findings suggest that Foc4-PP1 contributes to Foc TR4 virulence by promoting hyphal growth and abiotic stress resistance and inhibiting the immune defense responses of host plants.

Genome Sequence Data Reveal at Least Two Distinct Incursions of the Tropical Race 4 Variant of Fusarium Wilt into South America.

The global banana industry is threatened by one of the most devastating diseases: Fusarium wilt of banana. Fusarium wilt of banana is caused by the soilborne fungus Fusarium oxysporum f. sp. cubense (Foc), which almost annihilated the banana production in the late 1950s. A new strain of Foc, known as tropical race 4 (TR4), attacks a wide range of banana varieties, including Cavendish clones, which are the source of 99% of banana exports. In 2019, Foc TR4 was reported in Colombia, and more recently (2021) in Peru. In this study, we sequenced three fungal isolates identified as Foc TR4 from La Guajira (Colombia) and compared them against 19 whole-genome sequences of Foc TR4 publicly available, including four genome sequences recently released from Peru. To understand the genetic relatedness of the Colombian Foc TR4 isolates and those from Peru, we conducted a phylogenetic analysis based on a genome-wide set of single nucleotide polymorphisms (SNPs). Additionally, we compared the genomes of the 22 available Foc TR4 isolates, looking for the presence-absence of gene polymorphisms and genomic regions. Our results reveal that (i) the Colombian and Peruvian isolates are genetically distant, which could be better explained by independent incursions of the pathogen to the continent, and (ii) there is a high correspondence between the genetic relatedness and geographic origin of Foc TR4. The profile of present/absent genes and the distribution of missing genomic regions showed a high correspondence to the clades recovered in the phylogenetic analysis, supporting the results obtained by SNP-based phylogeny.

First Report of Fusarium Wilt of Iholena Banana (Musa spp.) Caused by Fusarium oxysporum f. sp. cubense Tropical Race 4 in China.

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 theITS 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.

Designing a synthetic microbial community devoted to biological control: The case study of Fusarium wilt of banana.

Fusarium oxysporum f. sp. cubense (Foc) tropical race 4 (TR4) is threatening banana production because of its increasing spread. Biological control approaches have been widely studied and constitute interesting complementary measures to integrated disease management strategies. They have been based mainly on the use of single biological control agents (BCAs). In this study, we moved a step forward by designing a synthetic microbial community (SynCom) for the control of Fusarium wilt of banana (FWB). Ninety-six isolates of Pseudomonas spp., Bacillus spp., Streptomyces spp., and Trichoderma spp. were obtained from the banana rhizosphere and selected in vitro for the antagonism against Foc TR4. In pot experiments, a large community such as SynCom 1.0 (44 isolates with moderate to high antagonistic activity) or a small one such as SynCom 1.1 (seven highly effective isolates) provided similar disease control (35% symptom severity reduction). An in vitro study of the interactions among SynCom 1.1 isolates and between them and Foc revealed that beneficial microorganisms not only antagonized the pathogen but also some of the SynCom constituents. Furthermore, Foc defended itself by antagonizing the beneficial microbes. We also demonstrated that fusaric acid, known as one of the secondary metabolites of Fusarium species, might be involved in such an interaction. With this knowledge, SynCom 1.2 was then designed with three isolates: Pseudomonas chlororaphis subsp. piscium PS5, Bacillus velezensis BN8.2, and Trichoderma virens T2C1.4. A non-simultaneous soil application of these isolates (to diminish cross-inhibition) delayed FWB progress over time, with significant reductions in incidence and severity. SynCom 1.2 also performed better than two commercial BCAs, BioPak® and T-Gro. Eventually, SynCom 1.2 isolates were characterized for several biocontrol traits and their genome was sequenced. Our data showed that assembling a SynCom for biocontrol is not an easy task. The mere mixtures of antagonists (e.g., SynCom 1.0 and 1.1) might provide effective biocontrol, but an accurate investigation of the interactions among beneficial microorganisms is needed to improve the results (e.g., SynCom 1.2). SynCom 1.2 is a valuable tool to be further developed for the biological control of FWB.

Biocontrol potential and antifungal mechanism of a novel Streptomyces sichuanensis against Fusarium oxysporum f. sp. cubense tropical race 4 in vitro and in vivo.

Most commercial banana cultivars are highly susceptible to Fusarium wilt caused by soilborne fungus Fusarium oxysporum f. sp. cubense (Foc), especially tropical race 4 (TR4). Biological control using antagonistic microorganism has been considered as an alternative method to fungicide. Our previous study showed that Streptomyces sp. SCA3-4T had a broad-spectrum antifungal activity from the rhizosphere soil of Opuntia stricta in a dry hot valley. Here, the sequenced genome of strain SCA3-4T contained 6614 predicted genes with 72.38% of G + C content. A polymorphic tree was constructed using the multilocus sequence analysis (MLSA) of five house-keeping gene alleles (atpD, gyrB, recA, rpoB, and trpB). Strain SCA3-4T formed a distinct clade with Streptomyces mobaraensis NBRC 13819T with 71% of bootstrap. Average nucleotide identity (ANI) values between genomes of strain SCA3-4T and S. mobaraensis NBRC 13819T was 85.83% below 95-96% of the novel species threshold, and named after Streptomyces sichuanensis sp. nov. The type strain is SCA3-4T (= GDMCC 4.214T = JCM 34964T). Genomic analysis revealed that strain SCA3-4T contained 36 known biosynthetic gene clusters of secondary metabolites. Antifungal activity of strain SCA3-4T was closely associated with the production of siderophore and its extracts induced the apoptosis of Foc TR4 cells. A total of 12 potential antifungal metabolites including terpenoids, esters, acid, macrolides etc. were obtained by the gas chromatography-mass spectrometry (GC-MS). Greenhouse experiment indicated that strain SCA3-4T could significantly inhibit infection of Foc TR4 in the roots and corms of banana seedlings and reduce disease index. Therefore, strain SCA3-4T is an important microbial resource for exploring novel natural compounds and developing biopesticides to manage Foc TR4. KEY POINTS: • Strain SCA3-4T was identified as a novel species of Streptomyces. • Siderophore participates in the antifungal regulation. • Secondary metabolites of strain SCA3-4T improves the plant resistance to Foc TR4.

Geographical Distribution and Genetic Diversity of the Banana Fusarium Wilt Fungus in Laos and Vietnam.

Fusarium wilt, caused by the fungus Fusarium oxysporum f. sp. cubense (Foc), poses a major threat to global banana production. The tropical race 4 (TR4) variant of Foc is a highly virulent form with a large host range, and severely affects Cavendish bananas. Foc TR4 was recently observed within the Greater Mekong Subregion, after Chinese private companies expanded Cavendish production to the region. In this study, extensive surveys conducted across Laos and Vietnam show that Foc TR4 is still mainly constricted to the northern regions of these countries and is limited to Cavendish cultivation settings. In Laos, Foc TR4 is associated with large-scale Cavendish plantations owned by or involved with Chinese companies through which infected planting material could have been imported. In Vietnam, mostly small-holder Cavendish farmers and backyard gardens were affected by Foc TR4. In Vietnam, no direct link is found with Chinese growers, and it is expected the pathogen mainly spreads through local and regional movement of infected planting materials. Foc TR4 was not recorded on banana cultivars other than Cavendish. The extensively cultivated ‘Pisang Awak’ cultivar was solely infected by VCGs belonging to Foc race 1 and 2, with a high occurrence of VCG 0123 across Laos, and of VCG 0124/5 in Vietnam. Substantial diversity of Foc VCGs was recorded (VCGs 0123, 0124/5, 01218 and 01221) from northern to southern regions in both countries, suggesting that Fusarium wilt is well established in the region. Interviews with farmers indicated that the local knowledge of Fusarium wilt epidemiology and options for disease management was limited. Clear communication efforts on disease epidemiology and management with emphasis on biosecurity practices need to be improved in order to prevent further spread of Foc TR4 to mixed variety smallholder settings.

Phenolics mediate suppression of Fusarium oxysporum f. sp. cubense TR4 by legume root exudates

Fusarium oxysporum Schlecht f. sp. cubense (E.F. Smith) Snyder & H.N. Hansen (Foc) is a notorious soil-borne fungal pathogen that causes Fusarium wilt disease of bananas (Musa spp. L.). Foc, especially Tropical Race 4 (TR4), is particularly devastating due its ability to infect diverse banana cultivars including Cavendish bananas. Fusarium wilt is difficult to manage, but intercropping has been reported to efficiently suppress Fusarium wilt through exudation of root metabolites with antagonistic effects on Foc. Yet, the metabolites that inhibit Foc and the underlying mode of action remain unclear. Hydroponic culture and targeted metabolite profiling were used to investigate the potential of root-exuded phenolic acids and flavonoids of two legume species, Desmodium uncinatum (Jacq.) DC. and Mucuna pruriens (L.) DC., to inhibit growth and the biosynthesis of the phytotoxins beauvericin and fusaric acid (FA) in Foc TR4. Of the twelve metabolites targeted, four phenolic acids (benzoic, t-cinnamic, p-coumaric, p-hydroxybenzoic) were common in root exudates of both legumes, whereas vanillin and the flavonoid quercetin were only detected in root exudates of M. pruriens. Bioassays using synthetic benzoic-, t-cinnamic-, or p-hydroxybenzoic acid, or a mixture thereof, showed a concentration-dependent suppressive effect on Foc TR4. Low concentrations of phenolic acids (0.01 and 0.1 mM) inhibited chlamydospore germination and the production of macro- and micro-conidia, as well as the biosynthesis of FA. Mycelial growth of Foc TR4 was only inhibited at high concentrations (1 mM) of benzoic acid, t-cinnamic acid, and a mixture of the three phenolics. This study demonstrates that phenolics in root exudates of D. uncinatum and M. pruriens suppress Foc TR4 by directly inhibiting the early stages of pathogen development. Intercropping banana with D. uncinatum and M. pruriens may thus be considered as a valuable option for suppression of Foc TR4.

Identification and characterization of early Fusarium wilt responsive mRNAs and long non-coding RNAs in banana root using high-throughput sequencing

Fusarium wilt disease, caused by Fusarium oxysporum f.sp. cubense (Foc), has been recognized as the most devastating disease to banana. The regulatory role of long non-coding RNAs (lncRNAs) in plant defense has been verified in many plant species. However, the understanding of their role during early FocTR4 (Foc tropical race 4) infection stage is very limited. In this study, lncRNA sequencing was used to reveal banana root transcriptome profile changes during early FocTR4 infection stages. Quantitative real time PCR (qRT-PCR) was performed to confirm the expression of eight differentially expressed (DE) lncRNAs (DELs) and their predicted target genes (DETs), and three DE genes (DEGs). Totally, 12,109 lncRNAs, 36,519 mRNAs and 2642 novel genes were obtained, of which 1398 (including 78 DELs, 1220 DE known genes and 100 DE novel genes) were identified as FocTR4 responsive DE transcripts. Gene function analysis revealed that most DEGs were involved in biosynthesis of secondary metabolites, plant–pathogen interaction, plant hormone signal transduction, phenylalanine metabolism, phenylpropanoid biosynthesis, alpha-linolenic acid metabolism and so on. Coincidently, many DETs have been identified as DEGs in previous transcriptome studies. Moreover, many DETs were found to be involved in ribosome, oxidative phosphorylation, lipoic acid metabolism, ubiquitin mediated proteolysis, N-glycan biosynthesis, protein processing in endoplasmic reticulum and DNA damage response pathways. QRT-PCR result showed the expression patterns of the selected transcripts were mostly consistent with our lncRNA sequencing data. Our present study showed the regulatory role of lncRNAs on known biotic and abiotic stress responsive genes and some new-found FocTR4 responsive genes, which can provide new insights into FocTR4-induced changes in the banana root transcriptome during the early pathogen infection stage.

Fusarium Wilt of Bananas: A Review of Agro-Environmental Factors in the Venezuelan Production System Affecting Its Development

Bananas and plantains (Musa spp.) are among the main staple of millions of people in the world. Among the main Musaceae diseases that may limit its productivity, Fusarium wilt (FW), caused by Fusarium oxysporum f. sp. cubense (Foc), has been threatening the banana industry for many years, with devastating effects on the economy of many tropical countries, becoming the leading cause of changes in the land use on severely affected areas. In this article, an updated, reflective and practical review of the current state of knowledge concerning the main agro-environmental factors that may affect disease progression and dissemination of this dangerous pathogen has been carried out, focusing on the Venezuelan Musaceae production systems. Environmental variables together with soil management and sustainable cultural practices are important factors affecting FW incidence and severity, excluding that the widespread dissemination of Foc, especially of its highly virulent tropical race 4 (TR4), is mainly caused by human activities. Additionally, risk analysis and climatic suitability maps for Foc TR4 in Venezuela have been developed. Although currently there are no effective management solutions available for FW control, this perspective provides an overview on the influence that environmental and agricultural variables would have on FW incidence and severity, giving some insight into management factors that can contribute to reducing its detrimental effects on banana production and how climate change may affect its development.

Efficacy of Disinfectants against Fusarium oxysporum f. sp. cubense Tropical Race 4 Isolated from La Guajira, Colombia.

Banana, the main export fruit for Colombia, is threatened by Fusarium wilt (FWB), caused by Fusarium oxysporum f. sp. cubense (Foc), tropical race 4 (TR4). Pathogen containment through disinfecting tools, machinery, shoes, and any means that may carry contaminated soil particles with proper disinfectants is at the forefront of disease management. In this study, the biocide efficacy of 10 commercial quaternary ammonium compounds (QACs) products and one based on glutaraldehyde (GA) were evaluated on both reproductive structures (microconidia and macroconidia) and survival spores (chlamydospores) of Foc TR4 (strain 140038) isolated from La Guajira, Colombia. QACs were evaluated at 1200 ppm and two exposure times: <1 and 15 min in the absence or presence of soil. For GA disinfectant, four different concentrations (500, 800, 1200, and 2000 ppm) were evaluated at both contact times in the presence of soil. In the absence of soil, all QACs showed 100% biocidal efficiency against microconidia, macroconidia, and chlamydospores at both <1 and 15 min. The presence of soil decreased the efficacy of disinfectants, but some of them, such as QAC3_1st, QAC7_4th, and QAC5_4th, showed 98%, 98%, and 100% efficacy against Foc TR4 chlamydospores, respectively, after <1 min of contact time. For instance, the GA-based disinfectant was able to eliminate all Foc TR4 propagules after 15 min for all concentrations tested.

Development of an in vitro hydroponic system for studying the interaction between banana plantlet and Fusarium oxysporum f. sp. cubense

Fusarium wilt of banana, caused by Fusarium oxysporum f. sp. cubense (Foc), is one of the most destructive diseases of banana. Several experimental systems have been used to study banana—Foc interaction, including pot system and hydroponic system. In this study, an in vitro hydroponic system was developed for the growth of rooted banana plantlet. It was characterized by a filter-paper holder, which consisted of a cone cup and a round tube. Besides being used as supporting material and providing nutrients to the banana plantlet, the filter-paper holder was designed to create a humid and aerobic environment for root growth. It was observed that rooted plantlet could grow normally in inoculum-free nutrient solution for at least 8 weeks. In the following rejuvenation experiment, each plantlet was back-inoculated by adding spore suspension of a degenerated strain of Foc tropical race 4 (TR4) into nutrient solution. Three to four weeks after in vitro inoculation, the rejuvenated strain was isolated from the rhizome of the plantlet showing symptoms under sterile conditions. The results of pathogenicity test indicated that the third-generation rejuvenated strain had partially regained virulence, which was characterized by lower radial growth rate and higher level of sporulation compared with the degenerated strain. In contrast to previously reported hydroponic systems which applied to studies in relation to Fusarium wilt of banana, the one described here is a totally closed system. It is demonstrated that the in vitro hydroponic system can be used for studying the interaction between banana plantlet and Foc TR4. A novel in vitro hydroponic system is described, in which a rooted banana plantlet can grow normally for at least 8 weeks without an air pump. In vitro inoculation technique allows banana plantlet—Foc interaction without interference of other microorganisms.

Biological control of banana wilt disease caused by Fusarium oxyspoum f. sp. Cubense using Streptomyces sp. H4

Banana Fusarium wilt caused by Fusarium oxyspoum f. sp. cubense (Foc) is one of the most destructive diseases. Foc tropical race 4 (TR4) is threatening the global banana industry and can infect almost all banana cultivars. Until now, there is still a lack of an effective method for controlling fusarium wilt. Application of beneficial microbes is considered as an environmentally sound option to manage phytopathogenic fungi. We previously identified Streptomyces sp. H4 with a strong antifungal activity against Foc TR4. In this study, strain H4 extracts obviously inhibited mycelial growth, spore germination and tube elongation of Foc TR4. Spore germination was completely suppressed by 8 × EC50 of extracts. Cell membrane integrity and ultrastructure of Foc TR4 were destroyed, which was supported by the increase of extracellular conductivity and nucleic acid releasing. In a pot experiment, the fermentation broth of strain H4 significantly decreased the disease index of Foc TR4 and improved the growth of banana seedlings. Hence, Streptomyces sp. H4 will be a potential microbial resource for the biocontrol of banana Fusarium wilt and the promotion of plant growth.

Occurrence and spread of the banana fungus Fusarium oxysporum f. sp. cubense TR4 in Mozambique

Fusarium wilt, caused by the soil-borne fungus Fusarium oxysporum f. sp. cubense (Foc), poses a major threat to banana production globally. A variant of Foc that originated in Southeast Asia, called tropical race 4 (TR4), was detected on a Cavendish banana export plantation (Metocheria) in northern Mozambique in 2013. Foc TR4 was rapidly disseminated on the farm, and affected approximately half a million plants within 3 years. The fungus was also detected on a second commercial property approximately 200 km away (Lurio farm) a year later, and on a small-grower’s property near Metocheria farm in 2015. Surveys in Mozambique showed that non-Cavendish banana varieties were only affected by Foc race 1 and race 2 strains. The testing of Cavendish banana somaclones in northern Mozambique revealed that GCTCV-119 was most resistant to Foc TR4, but that GCTCV-218 produced better bunches. The occurrence of Foc TR4 in northern Mozambique poses a potential threat to food security on the African continent, where banana is considered a staple food and source of income to millions of people. Cavendish somaclones can be used, in combination with integrated disease management practices, to replace susceptible Cavendish cultivars in southern Africa. The comprehensive testing of African cooking bananas for resistance to Foc TR4 is required, along with the improvement of biosecurity and preparedness of growers on the African continent.&#x0D; Significance:&#x0D; &#x0D; This paper presents the first official report of the invasive pest Foc TR4 in Africa.&#x0D; The spread of Foc TR4 on Cavendish banana farms in Mozambique was documented.&#x0D; Banana varieties that could replace susceptible Cavendish bananas were identified.&#x0D;

RESEARCH PROGRESS, CHALLENGES, FUTURE PERSPECTIVES ON THE MANAGEMENT OF FUSARIUM WILT OF BANANA IN MALAYSIA: A REVIEW

The global banana production is threatened by Fusarium wilt a fungal soil-borne disease caused by Fusarium oxysporum f. sp. cubense (Foc). In Malaysia, Foc Tropical Race 4 (Foc-TR4) is the most devastating disease that has completely wiped out the Cavendish plantations in 1990s. The TR4 strain attacks a diverse range of bananas and plantains which makes disease management challenging. In this review, we have summarized the current status of Fusarium wilt in Malaysia, current knowledge on the epidemiology of Foc and recommended control measures outlined by the Department of Agriculture (DOA) in preventing and/or avoiding disease spread. Research progress on the control of Foc-TR4 in Malaysia was also presented to highlight current findings and the knowledge gaps in each approaches. Recommendations for future studies were included to improve current disease management approaches while possible challenges were also provided for researchers, farmers and policy makers to consider.

Effect of bioformulations on the biocontrol efficacy, microbial viability and storage stability of a consortium of biocontrol agents against Fusarium wilt of banana.

This study sought to investigate the effect of bioformulation on the biocontrol efficacy, microbial viability and storage stability of a consortium of Pseudomonas aeruginosa DRB1 and Trichoderma harzianum CBF2 against Foc Tropical Race 4 (Foc-TR4). Four bioformulations consisting of dry (pesta granules, talc powder and alginate beads) and liquid formulations were evaluated for their ability to control Foc-TR4, sustain microbial populations after application and maintain microbial stability during storage. All tested bioformulations reduced disease severity (DS) by more than 43·00% with pesta granules producing the highest reduction in DS by 66·67% and the lowest area under the disease progress curve value (468·75) in a glasshouse trial. Microbial populations of DRB1 and CBF2 were abundant in the rhizosphere, rhizoplane and within the roots of bananas after pesta granules application as compared to talc powder, alginate beads and liquid formulations 84days after inoculation (DAI). The stability of both microbial populations after 180days of storage at 4°C was the greatest in the pesta granule formulation. The pesta granule formulation was a suitable carrier of biological control agents (BCA) without compromising biocontrol efficacy, microbial population and storage stability as compared to other bioformulations used in this study. Pesta granules could be utilized to formulate BCA consortia into biofertilizers. This formulation could be further investigated for possible applications under agricultural field settings.

Comparative transcriptome analysis reveals resistance-related genes and pathways in Musa acuminata banana 'Guijiao 9' in response to Fusarium wilt

Fusarium wilt caused by Fusarium oxysporum f. sp. cubense (Foc), is one of the most devastating diseases in bananas resulting in significant loss of Cavendish bananas production worldwide. Here we show the agronomic traits and the resistance of 'Guijiao 9′ in the field trials from 2012 to 2017. And then we dissect and compare the transcriptome response from these two cultivars (cv. 'Guijiao 9′ and cv. Williams) in an attempt to understand the molecular basis that contribute to the enhanced Foc tropical race 4 (Foc-TR4) resistance.'Guijiao 9′ is a Cavendish cultivar with strong resistance to Foc-TR4, which was reflected in a lower disease severity and incidence in glasshouse and field trails, when compared to the susceptible cultivar Williams. Gene expression profiles of 'Guijiao 9′ and Williams were captured by performing RNA-Seq analysis on 16 biological samples collected over a six day period post inoculation with Foc-TR4. Transcriptional reprogramming in response to Foc-TR4 was detected in both genotypes but the response was more drastic in 'Guijiao 9′ than in Williams. Specific genes involved in plant-pathogen interaction and defense signaling including MAPK, calcium, salicylic acid, jasmonic acid and ethylene pathways were analyzed and compared between 'Guijiao 9′ and Williams. Genes associated with defense-related metabolites synthesis such as NB-LRR proteins, calmodulin-binding protein and phenylpropanoids biosynthesis genes were significantly up-regulated in 'Guijiao 9′ resistant to Foc-TR4 infection. Taken together, this study highlights the important roles of plant hormone regulation and defense gene activation in mediating resistance in 'Guijiao 9'.