RESUMO
In 2015, sweetpotato producers in the United States experienced one of the worst outbreaks of black rot recorded in history, with up to 60% losses reported in the field and packing houses and at shipping ports. Host resistance remains the ideal management tool to decrease crop losses. Lack of knowledge of Ceratocystis fimbriata biology represents a critical barrier for the deployment of resistance to black rot in sweetpotato. In this study, we scanned the recent near chromosomal-level assembly for putative secreted effectors in the sweetpotato C. fimbriata isolate AS236 using a custom fungal effector annotation pipeline. We identified a set of 188 putative effectors on the basis of secretion signal and in silico prediction in EffectorP. We conducted a deep RNA time-course sequencing experiment to determine whether C. fimbriata modulates effectors in planta and to define a candidate list of effectors expressed during infection. We examined the expression profile of two C. fimbriata isolates, a pre-epidemic (1990s) isolate and a post-epidemic (2015) isolate. Our in planta expression profiling revealed clusters of co-expressed secreted effector candidates. Based on fold-change differences of putative effectors in both isolates and over the course of infection, we suggested prioritization of 31 effectors for functional characterization. Among this set, we identified several effectors that provide evidence for a marked biotrophic phase in C. fimbriata during infection of sweetpotato storage roots. Our study revealed a catalog of effector proteins that provide insight into C. fimbriata infection mechanisms and represent a core catalog to implement effector-assisted breeding in sweetpotato. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.
Assuntos
Ascomicetos , Ascomicetos/genética , Melhoramento Vegetal , Ceratocystis/genética , Sequência de BasesRESUMO
Ceratocystis fimbriata is a destructive fungal pathogen of sweetpotato (Ipomoea batatas (L.) Lam.) that leads to losses at all stages of sweetpotato production. Accurate detection of C. fimbriata would allow for more efficient deployment of management tactics in sweetpotato production. To develop a diagnostic assay, a hybrid genome assembly of C. fimbriata isolate AS236 was generated. The resulting 31.7 MB assembly was near-chromosome level, with 18 contigs, 6,481 predicted genes, and a BUSCO completion score of 98.4% when compared to the fungi-specific lineage database. Additional Illumina DNA reads from C. manginecans, C. platani, and a second C. fimbriata isolate (C1421) were then mapped to the assembled genome using BOWTIE2 and counted using HTSeq, which identified 148 genes present only within C. fimbriata as molecular diagnostic candidates; 6 single-copy and 35 highly multi-copy (>40 BLAST hits), as determined through a self-BLAST-P alignment. Primers for PCR were designed in the 200 bp flanking region of the first exon for each candidate, and the candidates were validated against a diverse DNA panel containing Ceratocystis species, sweetpotato pathogens, and plants. After validation, two diagnostic candidates amplified only C. fimbriata DNA and were considered to be highly specific to the species. These genetic markers will serve as valuable diagnostic tools with multiple applications including the detection of C. fimbriata in seed, soil, and wash water in sweetpotato production.
RESUMO
Black rot, caused by Ceratocystis fimbriata, is a devastating postharvest disease of sweetpotato that recently re-emerged in 2014. Although the disease is known to develop in storage and during export to overseas markets, little is known as to how pathogen dispersal occurs. This study was designed to investigate dump tank water as a means of dispersal through four different types of water treatments: inoculum concentration (0, 5, 5 × 101, 5 × 102, and 5 × 103 spores/ml), inoculum age (0, 24, 48, 96, and 144 h), water temperature (10°C, 23°C, 35°C, and 45°C), and presence of a water sanitizer (DryTec, SaniDate, FruitGard, and Selectrocide). Wounded and nonwounded sweetpotato storage roots were soaked in each water treatment for 20 min, stored at 29°C for a 14-day period, and rated for disease incidence every other day. Disease was observed in sweetpotato storage roots in all water treatments tested, except in the negative controls. Disease incidence decreased with both inoculum concentration and inoculum age, yet values of 16.26% and up to 50% were observed for roots exposed to 5 spores/ml and 144-h water treatments, respectively. Sanitizer products that contained a form of chlorine as the active ingredient significantly reduced disease incidence in storage roots when compared with control roots and roots exposed to a hydrogen-peroxide based product. Finally, no significant differences in final incidence were detected in wounded sweetpotato storage roots exposed to water treatments of any temperature, but a significant reduction in disease progression was observed in the 45°C treatment. These findings indicate that if packing line dump tanks are improperly managed, they can aid C. fimbriata dispersal through the build-up of inoculum as infected roots are unknowingly washed after storage. Chlorine-based sanitizers can reduce infection when applied after root washing and not in the presence of high organic matter typically found in dump tanks.
