Efficacy of disinfectants against Tropical Race 4 causing Fusarium wilt in Cavendish bananas

Fusarium wilt is one of the most damaging diseases of banana that currently puts the global production and food security at risk. The causal agents belong to the genus Fusarium and the soilborne species that is currently threatening Cavendish varieties and many locally important cultivars is F. odoratissimum, colloquially called Tropical Race 4 (TR4), which spreads easily and rapidly within and between farms. Even though limited information exists on the efficacy of disinfectants, they are promoted as a front-line defense against the spread of TR4 in the banana production areas by equipment and human activity. In this study, 13 disinfectants, the majority marketed in the Philippines, were tested for efficacy on TR4 mycelium, conidia, chlamydospores suspensions and chlamydospore-infested soil. They encompass five chemical groups, quaternary ammonium, halogens, alcohols, diamidines and aldehydes, and were tested at a range of concentrations and exposure times. Conidiospores were sensitive to all tested disinfectants at the manufacturer’s suggested rates. Ten disinfectants controlled mycelial growth with the exception of the quaternary ammonium-based disinfectant GUAA. Eleven disinfectants showed adequate efficacy at all tested concentrations towards chlamydospores in suspension, whereas a calcium hypochlorite-based disinfectant lacked efficacy at all concentrations tested. Despite in vitro efficacy, all disinfectants, except the diamidine-based disinfectant Formo, were largely ineffective in treating chlamydospores present in soil (in situ). However, Formo is corrosive to metals, making it less suitable for various applications. The in vitro results illustrate that overall efficacy of disinfectants depends on the fungal propagule, exposure time, and the environment in which they are exposed. We also determined the exposure times of footwear and vehicles in sanitation baths under field conditions and conclude that those significantly deviate from the minimal required contact time under in vitro conditions. Suboptimal quarantine measures, therefore, are likely to contribute to continued local, regional, and international TR4 dissemination.

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Induced mutations for generating bananas resistant to Fusarium wilt tropical race 4.

10.1079/9781789249095.0038 ◽

2021 ◽

pp. 366-378

Author(s):

Joanna Jankowicz-Cieslak ◽

Florian Goessnitzer ◽

Sneha Datta ◽

Altus Viljoen ◽

Ivan Ingelbrecht ◽

...

Keyword(s):

Genetic Diversity ◽

Fusarium Wilt ◽

Point Mutations ◽

Crop Productivity ◽

Banana Production ◽

Biotic Stresses ◽

Large Deletions ◽

Forward Genetic Screens ◽

Race 4

Abstract Bananas are a staple for more than 400 million people. Additionally, more than 16.5 million tonnes are exported, making it both an important food security and a cash crop. Productivity of Cavendish-type bananas is threatened by both abiotic and biotic stresses. The fact that triploid bananas are sterile, parthenocarpic and obligate vegetatively propagated makes them particularly susceptible to diseases, including Fusarium wilt caused by Fusarium oxysporum f. sp. cubense (Foc) tropical race 4 (Foc TR4). This is because continual clonal propagation has led to loss of genetic diversity. Additionally, lack of meiosis limits methods for breeding. Foc TR4 has been devastating Cavendish bananas in South-east Asia but has recently also been reported from Queensland in Australia, the Middle East and Mozambique, thus threatening global banana production. To address this, we are performing mutagenesis of in vitro propagated bananas to broaden the genetic diversity in order to find new alleles conferring disease resistance. We have developed methods for efficient induction of mutations in isolated apical meristems from shoot tips using chemical mutagens and ionizing radiation. Mutation discovery methods have been adapted to recover mutations including single point mutations and large deletions spanning millions of base pairs. We have created approximately 5000 mutated lines for forward-genetic screens to identify TR4 resistance in greenhouse- evaluated material. A subset of ca. 500 in vitro plantlets was subjected to glasshouse-based screening using a virulent F. oxysporum isolate. To date, 23 lines showing altered resistance responses to Foc TR4 have been identified.

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Seleksi In Vitro dan Pengujian Mutan Tanaman Pisang Ambon Kuning untuk Ketahanan terhadap Penyakit Layu Fusarium

Jurnal AgroBiogen ◽

10.21082/jbio.v9n2.2013.p66-76 ◽

2016 ◽

Vol 9 (2) ◽

pp. 66

Author(s):

Deden Sukmadjaja ◽

Ragapadmi Purnamaningsih ◽

Tri P. Priyatno

Keyword(s):

Fusarium Wilt ◽

In Vitro Selection ◽

Fusaric Acid ◽

Mutation Breeding ◽

Induce Mutagenesis ◽

Fusarium Wilt Of Banana ◽

Basal Media ◽

Generative Phase ◽

Race 4

Fusarium wilt of banana (Musa spp.) caused by Fusarium oxysporum f. sp. cubense (Foc) is the most serious problem faced in banana cultivation in terms of plant productivity and fruit quality. Mutation breeding is one of the alternative method that can be applied in producing new banana cultivar. Mutants can be induced by chemical mutagen such as ethyl methane sulfonate (EMS) followed by in vitro selection and then evaluation of the mutants to fusarium wilt disease in glasshouse and Foc infected field. The aim of this research was obtained EMS induced and in vitro selected mutants of banana var. Ambon Kuning and evaluated Foc disease resistant clones in glasshouse and Foc infected field. The first step to obtain the explants for this research was initiation and formation of multiple bud clumps (MBC) using MS basal media supplemented with 5, 10, and 20 mg/l of benzyladenin. Plant regeneration of MBC was also studied by using MS media containing 0, 0.2, and 1 mg/l of benzyladenin. To induce mutagenesis, MBC was soaked in 0.1, 0.3, and 0.5% (v/v) EMS for 1, 2, and 3 hours. The assesment of resistant MBC mutants to Fusarium phytotoxin was conducted by using fusaric acid (FA) as selection agent in concentration of 30, 45, and 60 ppm. Putative mutant plants produced by in vitro selection were further tested using spore solution of Foc race 4 in glasshouse. Meanwhile, Foc resistance assesment in the infected field was conducted in Pasirkuda Experimental Station, Bogor Agricultural University. The results showed that MBC can be formed in MS basal media supplemented with 10 or 20 mg/l benzyladenin. The EMS played a role in obtaining mutants by producing 68 MBC putative mutants tolerant to Foc based on FA selection. Further evaluation in the glasshouse was obtained 64 Foc resistant plants from 391 putative mutants produced by in vitro selection. Evaluation in the Foc infected field showed six clones survived until generative phase (12 month of age).

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Effect of in Planta Treatment of ‘Cavendish’ Banana with Herbicides and Fungicides on the Colonisation and Sporulation by Fusarium oxysporum f.sp. cubense Subtropical Race 4

Journal of Fungi ◽

10.3390/jof7030184 ◽

2021 ◽

Vol 7 (3) ◽

pp. 184

Author(s):

Jay Anderson ◽

Elizabeth Aitken

Keyword(s):

Fusarium Oxysporum ◽

Fusarium Wilt ◽

Effective Control ◽

In Planta ◽

Banana Production ◽

Cavendish Banana ◽

Green Fluorescent ◽

Race 4

Fusarium wilt caused by the soil-borne fungus Fusarium oxysporum f.sp. cubense (Foc) is a significant constraint to banana production worldwide, with the recent expansion of banana growing regions impacted by Foc Tropical Race 4 (TR4). The lack of commercially acceptable Cavendish cultivars with Foc resistance means the only current means of effective control is through strict quarantine and inoculum management. One method of control that is currently advocated includes the removal of infected plants which have been killed using herbicide injections. The aim of this work was to examine the effect of herbicide and fungicide treatments on sporulation of the fungus. In glasshouse studies using a green fluorescent transformed Foc Subtropical Race 4 isolate, we found treatments with herbicide hastened colonisation of the banana tissue and the production of micro- and macroconidia. The use of a fungicide did not prevent sporulation of the fungus in such tissue. This study demonstrates that herbicide treated plants are a source of potential inoculum for infection of nearby plants.

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Evaluation of disinfectant products for microbial decontamination of imported used footwear

New Zealand Plant Protection ◽

10.30843/nzpp.2009.62.4782 ◽

2009 ◽

Vol 62 ◽

pp. 130-135 ◽

Cited By ~ 1

Author(s):

L-H. Cheah ◽

A.T. Marsh ◽

M.R. McNeill ◽

D.I. Hedderley

Keyword(s):

Sodium Hypochlorite ◽

Quaternary Ammonium ◽

Hypochlorous Acid ◽

Field Tests ◽

In Vitro Tests ◽

Calcium Hypochlorite ◽

Hypobromous Acid ◽

Soil Bacteria And Fungi ◽

Bacteria And Fungi

Fourteen disinfectants were tested in vitro for activity against soil bacteria and fungi attached to a boot worn in New Zealand and then eight were further evaluated on three types of fieldworn footwear From in vitro tests sodium hypochlorite (Janola) calcium hypochlorite (HazTab) and a quaternary ammonium product (Trimove) gave almost complete kill of bacteria Four disinfectants (sodium hypochlorite phenols (Prephen) hypochlorous acid plus hypobromous acid (Nylate) and calcium hypochlorite) also gave complete kill of fungi Field tests showed that five disinfectants (sodium hypochlorite calcium hypochlorite formaldehyde hypochlorous acid plus hypobromous acid and the quaternary ammonium product) were as effective as dipotassium peroxodisulphate (Virkon a standard biosecurity treatment for disinfecting footwear) Sodium hypochlorite was found to be significantly more effective than dipotassium peroxodisulphate for disinfecting shoes of soil fungi The disinfectants caused no visible changes in colour or damage to footwear materials

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Field resistance of Cavendish somaclonal variants and local banana cultivars to tropical race 4 of Fusarium wilt in the Philippines

Acta Horticulturae ◽

10.17660/actahortic.2016.1114.31 ◽

2016 ◽

pp. 227-230 ◽

Cited By ~ 8

Author(s):

A.B. Molina ◽

V.O. Sinohin ◽

E.G. Fabregar ◽

E.B. Ramillete ◽

M.M. Loayan ◽

...

Keyword(s):

Fusarium Wilt ◽

The Philippines ◽

Field Resistance ◽

Somaclonal Variants ◽

Race 4

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Ex Ante Assessment of Returns on Research Investments to Address the Impact of Fusarium Wilt Tropical Race 4 on Global Banana Production

Frontiers in Plant Science ◽

10.3389/fpls.2020.00844 ◽

2020 ◽

Vol 11 ◽

Author(s):

Charles Staver ◽

Diemuth E. Pemsl ◽

Lars Scheerer ◽

Luis Perez Vicente ◽

Miguel Dita

Keyword(s):

Fusarium Wilt ◽

Banana Production ◽

Ex Ante ◽

Research Investments ◽

Race 4 ◽

The Impact

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

Journal of Fungi ◽

10.3390/jof7040297 ◽

2021 ◽

Vol 7 (4) ◽

pp. 297

Author(s):

Luisa F. Izquierdo-García ◽

Sandra L. Carmona ◽

Paola Zuluaga ◽

Gustavo Rodríguez ◽

Miguel Dita ◽

...

Keyword(s):

Disease Management ◽

Fusarium Oxysporum ◽

Fusarium Wilt ◽

Contaminated Soil ◽

Quaternary Ammonium ◽

Contact Time ◽

Soil Particles ◽

Reproductive Structures ◽

Race 4 ◽

Ammonium Compounds

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.

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An in vitro co-culture system for rapid differential response to Fusarium oxysporum f. sp. vasinfectum race 4 in three cotton cultivars

Plant Disease ◽

10.1094/pdis-08-21-1743-re ◽

2021 ◽

Author(s):

Stephen Parris ◽

Steven N. Jeffers ◽

James Olvey ◽

Jeffrey Adelberg ◽

...

Keyword(s):

Fusarium Oxysporum ◽

Fusarium Wilt ◽

Culture System ◽

Biotic Interactions ◽

In Vitro System ◽

Pima Cotton ◽

Cotton Species ◽

Race 4 ◽

Cotton Cultivar

Fusarium oxysporum f. sp. vasinfectum race 4 (FOV4) is a devastating fungus pathogen that causes Fusarium wilt in both domesticated cotton species, Gossypium hirsutum (Upland) and G. barbadense (Pima). Greenhouse and field-based pathogenicity assays can be a challenge due to non-uniform inoculum levels, the presence of endophytes, and varying environmental factors. Therefore, an in vitro co-culture system was designed to support the growth of both domesticated cotton species and FOV4 using an inert polyphenolic foam substrate with a liquid medium. A Fusarium wilt-susceptible Pima cotton cultivar, G. barbadense ‘GB1031’, a highly resistant Pima cotton cultivar, G. barbadense ‘DP348RF’, and a susceptible Upland cotton cultivar, G. hirsutum ‘TM-1’, were evaluated for 30 days during co-culture with FOV4 in this foam-based system. Thirty days after inoculation, disease symptoms were more severe in both the susceptible cultivars, which displayed higher percentages of foliar damage, and greater plant mortality, than observed in ‘DP348RF’, the resistant Pima cotton cultivar. This foam-based in vitro system may be useful for screening cotton germplasm for resistance to a variety of fungus pathogens and to facilitate the study of biotic interactions in domesticated cotton species under controlled environmental conditions.

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Beneficial Microbes and Basal Fertilization in Antagonism of Banana Fusarium Wilt

Agronomy ◽

10.3390/agronomy11102043 ◽

2021 ◽

Vol 11 (10) ◽

pp. 2043

Author(s):

Chung-Pao Lin ◽

Yi-Cheng Ho

Keyword(s):

Fusarium Wilt ◽

Metal Ion ◽

Seedling Survival ◽

Survival Rates ◽

Severity Index ◽

Banana Production ◽

Beneficial Microbes ◽

Iron Copper ◽

Banana Leaves ◽

Race 4

Fusarium Wilt Tropical Race 4 (TR4) is spreading rapidly all over the world and threatens banana production, especially the Cavendish variety, which is the main commercial cultivar in Asia. This work aims to use beneficial microbes and fertilizers to control TR4. Both Bacillus mycoides NP02 (BM) and Bacillus amyloliquefaciens BaPD1 (BA) antagonized TR4 growth in plate tests. In addition, basal nutrients (BN) in combination with BM and BA were used to treat 1-month-old banana seedlings infected by Fusarium Wilt; plant growth and nutrients of leaves were investigated after 6-month cultivation in the field. The seedling survival rates for uninfected and infected controls without the microbes were 89.4 ± 14.8% and 50 ± 10.7%, respectively. In contrast, the corresponding rates for 2BM, 5BM, 1BA, F1 (5BM + 4BN + 1BA) and G1 (1BM + 4BN + 1BA) in the infected plants were 80 ± 9.4%, 83 ± 4.3%, 85 ± 14.2%, 96.7 ± 1.9% and 96.7 ± 3.3%, respectively. The 2BM treatment promoted the growth of both uninfected and infected banana plants and the 5BA treatment significantly reduced the severity index by 1.45-fold. Plants infected with TR4 showed significantly reduced contents of nitrogen and potassium, but the contents of iron, copper and zinc were higher than those of healthy plants. Additionally, 1BM treatment stimulated the accumulation of nitrogen and zinc ions in the leaves of uninfected plants. Both the 1BA and 2BN treatments increased the iron (Fe) and nickel (Ni) metal ion levels of TR4 infected plants. The F2 (5BM + 2BN + 1BA) treatment significantly maintained the growth of banana plants under TR4 stress with increased contents of nickel and zinc in banana leaves, suggesting that these ions may play a key role in stimulating the growth of banana plants under the threat of TR4. This work shows the potential of applying BM, BN and BA in the control of Fusarium wilt in field conditions.

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An Additional Threat to ‘Cavendish’ Banana Growers and Traders: The Infection of Banana Peduncles by Fusarium oxysporum f. sp. cubense Tropical Race 4 (Foc TR4)

Plant Health Progress ◽

10.1094/php-08-20-0067-rs ◽

2020 ◽

Vol 21 (4) ◽

pp. 312-316

Author(s):

Tingting Bai ◽

Meng Qin ◽

Xundong Li ◽

Huacai Fan ◽

Shengtao Xu ◽

...

Keyword(s):

Fusarium Oxysporum ◽

Fusarium Wilt ◽

Vascular Bundle ◽

Fungal Colonization ◽

Banana Production ◽

Specific Pcr ◽

Planting Material ◽

Cavendish Banana ◽

Race 4 ◽

Inoculation Assay

Fusarium oxysporum f. sp. cubense tropical race 4 (Foc TR4) is one of the most destructive banana pathogens worldwide. Until now, it has been generally accepted that Foc TR4 mainly colonizes the roots, rhizomes, pseudostems, and leaves of banana plants and spreads among plantations with infected planting material, soil residues, and water. Because banana Fusarium wilt is a typical vascular bundle disease, the pathogen is accepted to possess the ability to enter the peduncle. However, no scientific proof has been presented. Recently, more and more signs of peduncle Foc TR4 in ‘Cavendish’ plants in different banana production provinces of China suggest that further investigations ought to be made regarding fungal colonization. We observed discoloration inside some peduncles from the Foc TR4-infected plants. The fungus isolated from these symptoms was identified using Foc TR4-specific PCR, sequencing, and a host inoculation assay. In a greenhouse bioassay, the Foc TR4 isolates from the peduncle showed the capacity for reinfecting Brazilian plants (Cavendish, AAA) up into the leaves. Collectively, we isolated the Foc TR4 from the peduncle and directly testified that this pathogen can invade the peduncle via xylem vascular bundle.

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