Can the banana weevil Cosmopolites sordidus be a vector of Fusarium oxysporum f.sp. cubense race 1? Unravelling the internal and external acquisition of effective inoculum.

BACKGROUND: With the undergoing world outbreak of Fusarium wilt of bananas, it is essential to unravel all the possible process of dissemination of this disease. The host-pest interactions of the banana weevil with banana plants make this insect an important potential vector. This study, carried out in controlled conditions, explores the interaction between the banana weevil and Fusarium oxysporum f.sp. cubense race 1 (Foc), with a focus on the external and internal transport of viable fungal propagules. RESULTS: Viable inoculum of Foc was detected very rapidly on external teguments and in the digestive tract of the insect, i.e. at the lowest time studied of 5 min after contact with infected pseudostems. Maximal inoculum acquisition occurred after 1 h contact with an inoculum source. External inoculum was higher than the inoculum present in the digestive tract, but external and internal inoculum had the same dynamics. After a contact of an infected source, external and internal inoculum decreased exponentially within 50 h, but weevils remained infested for a long time, as long as 2 or 3 days that would be enough for inoculum dispersal. Viable inoculum was also detected in feces. Foc strains isolated were pathogenic when inoculated to banana plants of the Gros Michel variety but did not provoke any symptom on Cavendish banana plants. CONCLUSION: These results demonstrate that the infective structures of Foc remain externally viable in the digestive system and the excreta of the banana weevil. Such excreta are capable of making healthy banana plants of the Gros Michel variety. © 2021 Society of Chemical Industry.

Efficient transformation of Mycosphaerella fijiensis by underwater shock waves.

Black leaf streak disease, also known as black Sigatoka, causes dramatic losses in production of banana and plantains fruits. The disease is caused by the pathogenic fungus Mycosphaerella fijiensis (anamorph Pseudocercospora fijiensis; Mycosphaerellaceae). Genetic transformation of M. fijiensis would allow a better understanding of molecular basis of pathogenicity and design novel approaches to control the infection caused by this pathogen. However, transformation of this fungus has not been easy. We report here a protocol for genetic transformation of M. fijiensis employing underwater shock waves and intact conidia. The recombinant strains recovered showed genetic stability over >10 generations. The frequency of transformation obtained was between 75 and 150 times higher than the efficiency reported in the only article published on transformation of M. fijiensis using spheroplasts. This improvement allowed the use of a thousand times less cells than the amount employed before, avoiding the need for cumbersome successive batch cultures. Our protocol is simple, highly efficient, fast and reproducible and together with the available genomes of M. fijiensis and Musa acuminata, it offers new possibilities to study the diverse mechanisms of pathogenesis of the fungus.

Comparative analysis of the in vitro and in planta secretomes from Mycosphaerella fijiensis isolates.

Black Sigatoka, a devastating disease of bananas and plantains worldwide, is caused by the fungus Mycosphaerella fijiensis. Several banana cultivars such as 'Yangambi Km 5' and Calcutta IV, have been known to be resistant to the fungus, but the resistance has been broken in 'Yangambi Km 5' in Costa Rica. Since the resistance of this variety still persists in Mexico, the aim of this study was to compare the in vitro and in planta secretomes from two avirulent and virulent M. fijiensis isolates using proteomics and bioinformatics approaches. We aimed to identify differentially expressed proteins in fungal isolates that differ in pathogenicity and that might be responsible for breaking the resistance in 'Yangambi Km 5'. We were able to identify 90 protein spots in the secretomes of fungal isolates encoding 42 unique proteins and 35 differential spots between them. Proteins involved in carbohydrate transport and metabolism were more prevalent. Several proteases, pathogenicity-related, ROS detoxification and unknown proteins were also highly or specifically expressed by the virulent isolate in vitro or during in planta infection. An unknown protein representing a virulence factor candidate was also identified. These results demonstrated that the secretome reflects major differences between both M. fijiensis isolates.