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

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

Bacterial secondary metabolites: possible mechanism for weed suppression in wheat.

Chemical weed control is an effective method, but has proved hazardous for humans, environment, and soil biodiversity. Use of allelopathic bacteria may be more efficient and sustainable weed control measure. The bacterial inoculants have never been studied in context of their interaction with weed root exudates and precursor-dependent production of the natural phytotoxins (cyanide, cytolytic enzymes and auxin) by these strains to understand their weed suppression and wheat growth promotion abilities. Therefore, root exudates of Avena fatua, Phalaris minor, Rumex dentatus, and wheat were quantified and their role in microbial root colonization and secondary metabolite production, i.e., cyanide, cytolytic enzymes, phenolics, and elevated auxin concentration, was studied. The results depicted l-tryptophan and glycine as major contributors of elevated cyanide and elevated levels in weed rhizosphere by the studied Pseudomonas strains, through their higher root colonization ability in weeds as compared with wheat. Furthermore, the higher root colonization also enhanced p-coumaric acid (photosynthesis inhibitor by impairing cytochrome c oxidase activity in plants) and cytolytic enzyme (root cell wall degradation) concentration in weed rhizosphere. In conclusion, the differential root colonization of wheat and weeds by these strains is responsible for enhancing weed suppression (enhancing phytotoxic effect) and wheat growth promotion (lowering phytotoxic effect).

De novo synthesis of ferrichrome by Fusarium oxysporum f. sp. cubense TR4 in response to iron starvation.

Manipulation of iron bioavailability in the banana rhizosphere may suppress Fusarium wilt, caused by Fusarium oxysporum f. sp. cubense (Foc). However, iron starvation induced by application of synthetic iron chelators does not effectively suppress Fusarium wilt. It is unclear whether Foc can subvert iron chelators and thereby evade iron starvation through the synthesis of iron-scavenging secondary metabolites, called siderophores. In vitro studies were conducted using iron-deficient growth medium and medium supplemented with a synthetic iron chelator, 2,2'-dipyridyl, to mimic iron starvation in Foc Tropical Race 4 (Foc TR4). Concentration of extracellular siderophores increased three-fold (p < 0.05) in the absence of iron. Liquid chromatography-mass spectrometry analysis detected the hydroxamate siderophore, ferrichrome, only in the mycelia of iron-starved cultures. Moreover, iron-starved cultures exhibited a reduction in total cellular protein concentration. In contrast, out of the 20 proteinogenic amino acids, only arginine increased (p < 0.05) under iron starvation. Our findings suggest that iron starvation does not cause a remodelling of amino acid metabolism in Foc TR4, except for arginine, which is required for biosynthesis of ornithine, the precursor for siderophore biosynthesis. Collectively, our findings suggest that biosynthesis of siderophores, particularly ferrichrome, could be a counteractive mechanism for Foc TR4 to evade iron starvation.

Spent Pleurotus ostreatus Substrate Has Potential for Managing Fusarium Wilt of Banana.

A range of basidiomycetes including the edible mushroom Pleurotus ostreatus (Po) can suppress plant pathogens such as Fusarium spp. With the current increase in production and consumption of Po in Uganda, the spent Po substrate (SPoS) could be an alternative to manage Fusarium wilt of banana (FWB), caused by the soil borne pathogen Fusarium oxysporum f. sp. cubense, race 1 (Foc). This study determined the potential of SPoS to inhibit Foc in vitro and in potted plants. In vitro studies confirmed suppression of Foc in pure co-culture (Po vs. Foc) assays and media amended with different concentrations (0% to 50% w/v) of un-sterilized SPoS filtrates. Foc growth in the sterile SPoS filtrate was comparable to the water control, suggesting possible roles of biotic or thermolabile components of the SPoS. To further verify the suppressive effects of SPoS, pot experiments were carried out with a resistant ('Mbwazirume', AAA) and susceptible ('Sukali Ndizi', AAB) banana cultivar using both artificially and naturally infested soils. Independent of the inoculation method, SPoS significantly reduced the severity of FWB in pot experiments. Susceptible cultivar 'Sukali Ndizi' growing in substrates amended with SPoS showed lower (1.25) corm damage (Scale 0-5) than the un-amended control (3.75). No corm damage was observed in uninoculated controls. The resistant cultivar 'Mbwazirume', showed slight (0.25) corm damage only in the Foc-inoculated plants without SPoS. These findings suggest that SPoS could be used as part of the management practices to reduce the impact of FWB.

Sources of resistance to Pseudocercospora fijiensis, the cause of black Sigatoka in banana.

Black Sigatoka, caused by Pseudocercospora fijiensis, is one of the most devastating diseases of banana. In commercial banana-growing systems, black Sigatoka is primarily managed by fungicides. This mode of disease management is not feasible for resource-limited smallholder farmers. Therefore, bananas resistant to P. fijiensis provide a practical solution for managing the disease, especially under smallholder farming systems. Most banana and plantain hybrids with resistance to P. fijiensis were developed using few sources of resistance, which include Calcutta 4 and Pisang Lilin. To broaden the pool of resistance sources to P. fijiensis, 95 banana accessions were evaluated under field conditions in Sendusu, Uganda. Eleven accessions were resistant to P. fijiensis. Black Sigatoka symptoms did not progress past Stage 2 (narrow brown streaks) in the diploid accessions Pahang (AA), Pisang KRA (AA), Malaccensis 0074 (AA), Long Tavoy (AA), M.A. Truncata (AA), Tani (BB), and Balbisiana (BB), a response similar to the resistant control Calcutta 4. These accessions are potential sources of P. fijiensis resistance and banana breeding programmes can use them to broaden the genetic base for resistance to P. fijiensis.

SNP-based genotyping and whole-genome sequencing reveal previously unknown genetic diversity in Xanthomonas vasicola pv. musacearum, causal agent of banana xanthomonas wilt, in its presumed Ethiopian origin.

For decades, Xanthomonas vasicola pv. musacearum (Xvm) has been an economically important bacterial pathogen on enset in Ethiopia. Since 2001, Xvm has also been responsible for significant losses to banana crops in several East and Central African countries, with devastating consequences for smallholder farmers. Understanding the genetic diversity within Xvm populations is essential for the smart design of transnationally reasoned, durable, and effective management practices. Previous studies have revealed limited genetic diversity in Xvm, with East African isolates from banana each falling into one of two closely related clades previously designated as sublineages SL 1 and SL 2, the former of which had also been detected on banana and enset in Ethiopia. Given the presumed origin of Xvm in Ethiopia, we hypothesized that both clades might be found in that country, along with additional genotypes not seen in Central and East African bananas. Genotyping of 97 isolates and whole-genome sequencing of 15 isolates revealed not only the presence of SL 2 in Ethiopia, but additional diversity beyond SL 1 and SL 2 in four new clades. Moreover, SL 2 was detected in the Democratic Republic of Congo, where previously SL 1 was the only clade reported. These results demonstrate a greater range of genetic diversity among Xvm isolates than previously reported, especially in Ethiopia, and further support the hypothesis that the East/Central Africa xanthomonas wilt epidemic has been caused by a restricted set of genotypes drawn from a highly diverse pathogen pool in Ethiopia.

Risks Posed by Intercrops and Weeds as Alternative Hosts to Xanthomonas campestris pv. musacearum in Banana Fields.

Alternative host plants are important in the survival and perpetuation of several crop pathogens and have been suspected to play a role in the survival of Xanthomonas campestris pv. musacearum (Xcm) and perpetuation of Xanthomonas wilt (XW) disease of banana and enset. This study determined the potential risk posed by two weeds (Canna spp. and wild sorghum) and common banana intercrops (maize, millet, sorghum, taro, and sugarcane) as alternative hosts to Xcm. The study employed screenhouse experiments, laboratory procedures and diagnosis of banana fields in XW-affected landscapes. Typical XW symptoms were only observed in artificially inoculated Canna sp., with an incidence of 96%. Leaf lesions characteristic of xanthomonads occurred on millet (50%) and sorghum (35%), though the plants recovered. No symptoms occurred in maize, sugarcane, taro or wild sorghum. However, Xcm was recovered from all these plant species, with higher recoveries in Canna sp. (47%), millet (27%), sugarcane (27%), and wild sorghum (25%). Only isolates recovered from Canna sp., millet, sorghum and wild sorghum caused disease in banana plantlets. The presence and incidence of XW on-farm was positively associated with the presence of susceptible ABB Musa genotypes and negatively with number of banana cultivars on farm and household access to training on XW management. Only 0.02% of field sampled Canna spp. plants had Xcm. Risk posed by Canna spp. on-farm could be limited to tool transmission as it has persistent floral bracts that prevent insect-mediated infections. Given the high susceptibility, perennial nature and propagation through rhizomes of Canna sp., it could pose a moderate-high risk, thus warranting some attention in the management of XW disease. Sugarcane could offer a low-moderate risk due to its perennial nature and propagation through rhizomes while risk from maize, millet, and sorghum was deemed zero-low due to their annual nature, wind-mediated mode of pollination and propagation through seed. Understanding the interactions of a crop pathogen with other plants is thus important when diversifying agroecosystems. The study findings also suggest other factors such as cultivar composition and management of the disease at farm and landscape level to be important in the perpetuation of XW disease.