Isolation and characterization of extracellular vesicles from Fusarium oxysporum f. sp. cubense, a banana wilt pathogen.

Fusarium wilt of banana is a destructive widespread disease caused by Fusarium oxysporum f. sp. cubense (Foc) that ravaged banana plantations globally, incurring huge economic losses. Current knowledge demonstrates the involvement of several transcription factors, effector proteins, and small RNAs in the Foc-banana interaction. However, the precise mode of communication at the interface remains elusive. Cutting-edge research has emphasized the significance of extracellular vesicles (EVs) in trafficking the virulent factors modulating the host physiology and defence system. EVs are ubiquitous inter- and intra-cellular communicators across kingdoms. This study focuses on the isolation and characterization of Foc EVs from methods that make use of sodium acetate, polyethylene glycol, ethyl acetate, and high-speed centrifugation. Isolated EVs were microscopically visualized using Nile red staining. Further, the EVs were characterized using transmission electron microscopy, which revealed the presence of spherical, double-membrane, vesicular structures ranging in size from 50 to 200 nm (diameter). The size was also determined using the principle based on Dynamic Light Scattering. The Foc EVs contained proteins that were separated using SDS-PAGE and ranged between 10 and 315 kDa. Mass spectrometry analysis revealed the presence of EV-specific marker proteins, toxic peptides, and effectors. The Foc EVs were found to be cytotoxic, whose toxicity increased with EVs isolated from the co-culture preparation. Taken together, a better understanding of Foc EVs and their cargo will aid in deciphering the molecular crosstalk between banana and Foc.

Molecular insights into the jasmonate signaling and associated defense responses against wilt caused by Fusarium oxysporum.

Biotic and abiotic stress factors drastically limit plant growth and development as well as alter the physiological, biochemical and cellular processes. This negatively impacts plant productivity, ultimately leading to agricultural and economical loss. Plant defense mechanisms elicited in response to these stressors are crucially regulated by the intricate crosstalk between defense hormones such as jasmonic acid (JA), salicylic acid and ethylene. These hormones orchestrate adaptive responses by modulating the gene regulatory networks leading to sequential changes in the root architecture, cell wall composition, secondary metabolite production and expression of defense-related genes. Fusarium wilt is a widespread vascular disease in plants caused by the soil-borne ascomycete Fusarium oxysporum and is known to attack several economically important plant cultivars. JA along with its conjugated forms methyl jasmonate and jasmonic acid isoleucine critically tunes plant defense mechanisms by regulating the expression of JA-associated genes imparting resistance phenotype. However, it should be noted that some members of F. oxysporum utilize the JA signaling pathway for disease development leading to susceptibility in plants. Therefore, JA signaling pathway becomes one of the important targets amenable for modulation to develop resistance response against Fusarium wilt in plants. In this review, we have emphasized on the physiological and molecular aspects of JA and its significant role in mounting an early defense response against Fusarium wilt disease. Further, utilization of the inherent JA signaling pathway and/or exogenous application of JA in generating Fusarium wilt resistant plants is discussed.