Colletotrichum acutatum sensu lato : from diversity study to genome analysis

Colletotrichum acutatum sensu lato includes a number of important pathogens that cause economically significant losses of various crops. The C. acutatum species complex has a wide host range in both domesticated and wild plant species, and its capability to infect different types of hosts such as insects has also been described. Members belonging to this complex are able to develop three different types of interaction with plant hosts including biotrophic, necrotrophic and hemibiotrophic infections and are also capable of surviving on weeds and non-hosts without causing visible symptoms. They are mainly asexual, but some have a teleomorphic state called Glomerella and can be either homothallic or heterothallic. The sexual behaviour in Glomerella is more complicated than in most ascomycetes, and strains within the same species do not show a typical MAT1-1/2 system. The overall aim of this study was to gain an improved understanding of the relationships between the genetic diversity of global populations, host association patterns, geographic distribution and biological and pathological attributes. A database (CaITSdb) containing more than 800 rRNA sequences deposited in GenBank was created along with key biogeographic information, and the data have been analysed in order to investigate genetic diversity and distribution of sub-populations and their evolutionary relationships. The combined evidence was used to assemble a core collection of 120 isolates that are representative of the diversity in host preference, geographic origin, mating behaviour and molecular genetic variation. A multi-locus sequencing approach (based on four neutral loci) has been used to evaluate phylogenetic relationships amongst the isolates in the core collection. A strong relationship was observed between various genetic groups distinguished and their mating behaviour, geographic distribution and host association patterns. Oceania has been identified as a likely geographic origin of this pathogen, as the highest level of variability and groups related to a hypothetical ancestral population are mainly distributed in these countries. All homothallic isolates capable of producing perithecia belonged to the same genetic group A7; whereas all self-sterile heterothallic isolates were classified as either A3 or A5. Isolates derived from the same host tend to cluster together into genetic groups or sub-populations. This evidence is generally in agreement with recent published work on taxonomic re-assessment of Colletotrichum acutatum sensu lato, which indicates at least fifteen new species. This study has provided the evidence for the occurrence of three distinct genetic groups on strawberry in the UK corresponding to three species reported in the literature namely, C. nymphaeae, C. fioriniae and C. godetiae. Isolates belonging to the genetic groups that correspond to C. nymphaeae and C. fioriniae appeared to be the most aggressive on strawberry, followed by C. godetiae, and C. simmondsii (not found in the UK). Representative isolates of other species were less aggressive. The first whole genome sequence an isolate (A9 = C. simmondsii) from the C. acutatum sensu species complex was assembled and analysed using a range of bioinformatics algorithms. An isolate of C. simmondsii was chosen based on its wide host range including strawberry and the phylogeographic position. Genome analyses enabled prediction and annotation of the whole gene set at 13549 including 6 % unique to this species. The data also suggested an interesting expansion of several gene families, such as those encoding carbohydrate-active enzymes, secondary metabolites pathways and effectors which could be associated with the wide host range. The new knowledge and resources developed with the genome analyses along with the results of the population level diversity studies provide a platform for future comparative and functional genomics investigations to advance this research

Insights on KP4 Killer Toxin-like Proteins of Fusarium Species in Interspecific Interactions

KP4 killer toxins are secreted proteins that inhibit cell growth and induce cell death in target organisms. In Fusarium graminearum, KP4-like (KP4L) proteins contribute to fungal virulence in wheat seedling rot and are expressed during Fusarium head blight development. However, fungal KP4L proteins are also hypothesized to support fungal antagonism by permeabilizing cell walls of competing fungi to enable penetration of toxic compounds. Here, we report the differential expression patterns of F. graminearum KP4L genes (Fgkp4l-1, -2, -3 and -4) in a competitive interaction, using Trichoderma gamsii as the antagonist. The results from dual cultures indicate that Fgkp4l-3 and Fgkp4l-4 could participate in the recognition at the distance of the antagonist, while all Fgkp4l genes were highly activated in the pathogen during the physical interaction of both fungi. Only Fgkp4l-4 was up-regulated during the interaction with T. gamsii in wheat spikes. This suggests the KP4L proteins could participate in supporting F. graminearum interspecific interactions, even in living plant tissues. The distribution of KP4L orthologous within the genus Fusarium revealed they are more represented in species with broad host-plant range than in host-specific species. Phylogeny inferred provides evidence that KP4L genes evolved through gene duplications, gene loss and sequence diversification in the genus Fusarium

Pathogenic potential of Beauveria pseudobassiana as bioinsecticide in protein baits for the control of the medfly Ceratitis capitata

The medfly, Ceratitis capitata (Wiedemann) (Diptera Tephritidae), is a major insect pest affecting fruit production worldwide whose control is mainly based on the use of protein baits laced with chemical insecticides. Entomopathogenic fungi are well- known to be effective against a wide spectrum of insect pests and are commonly utilized in integrated pest management and bio- logical control programs. Here, we assess the feasibility of using the recently described entomopathogenic species Beauveria pseudobassiana Rehner et Humber (Hypocreales Cordycipitaceae) as a biological insecticide in protein bait sprays for the control of the medfly. Firstly, we evaluated the pathogenicity of B. pseudobassiana against eggs, larvae, pupae and adults of the medfly. Secondly, we tested its efficacy as bioinsecticide in protein bait sprays. The results of the pathogenicity tests showed that B. pseu- dobassiana is able to infect, and lead to the death, all instar of the medfly. The efficacy of B. pseudobassiana was confirmed also when used as bioinsecticide in protein baits. In planta tests, the survival probability (Kaplan-Meier estimates) of flies in contact with the B. pseudobassiana-laced protein bait was significantly lower respect to control. Median survival time of flies treated with B. pseudobassiana-laced protein (6 ± 1.422 d) was at least three times shorter than in control (> 20 d). Based on our results, we confirmed the potential of B. pseudobassiana as bioinsecticide in entomopathogenic fungi-laced protein baits for the control of tephritid fruit flies

Draft Whole-Genome Sequence of Trichoderma gamsii T6085, a Promising Biocontrol Agent of Fusarium Head Blight on Wheat

Trichoderma gamsii T6085 is a promising beneficial isolate whose effects consist of growth inhibition of the main agents causing Fusarium head blight, reduction of mycotoxin accumulation, competition for wheat debris, and reduction of the disease in both the lab and the field. Here, we present the first genome assembly of a T. gamsii isolate, providing a useful platform for the scientific community

Comparative genomics of the genus Pseudomonas reveals host- and environment-specific evolution

[EN]Each Earth ecosystem has unique microbial communities. Pseudomonas bacteria have evolved to occupy a plethora of different ecological niches, including living hosts, such as animals and plants. Many genes necessary for the Pseudomonas-niche interaction and their encoded functions remain unknown. Here, we describe a comparative genomic study of 3,274 genomes with 19,056,667 protein-coding sequences from Pseudomonas strains isolated from diverse environments. We detected functional divergence of Pseudomonas that depends on the niche. Each group of strains from a certain environment harbored a distinctive set of metabolic pathways or functions. The horizontal transfer of genes, which mainly proceeded between closely related taxa, was dependent on the isolation source. Finally, we detected thousands of undescribed proteins and functions associated with each Pseudomonas lifestyle. This research represents an effort to reveal the mechanisms underlying the ecology, pathogenicity, and evolution of Pseudomonas, and it will enable clinical, ecological, and biotechnological advances

Genome sequence of the mycotoxigenic crop pathogen Fusarium proliferatum strain ITEM 2341 from date palm

Fusarium proliferatum is a widely distributed fungal pathogen associated with more than 26 crop species important in global food security. Its strong mycotoxigenic capability with potential impacts on human and animal health is well recognized. In this work, we report the draft genome sequence of F. proliferatum strain ITEM 2341, originally isolated from date palm, providing a platform for further comparative and functional genomic investigations

Draft Whole-Genome Sequence of the Biocontrol Agent Trichoderma harzianum T6776

Trichoderma harzianum T6776 is a promising beneficial isolate whose effects consist of growth promotion, positive response of photosynthetic activity, hormonal signaling, and carbon partitioning in tomato, coupled with biocontrol of plant pathogens. Here, we present the first genome assembly of T6776, providing a useful platform for the scientific community

Identification and Comparison of Colletotrichum Secreted Effector Candidates Reveal Two Independent Lineages Pathogenic to Soybean

Colletotrichum is one of the most important plant pathogenic genus of fungi due to its scientific and economic impact. A wide range of hosts can be infected by Colletotrichum spp., which causes losses in crops of major importance worldwide, such as soybean. Soybean anthracnose is mainly caused by C. truncatum, but other species have been identified at an increasing rate during the last decade, becoming one of the most important limiting factors to soybean production in several regions. To gain a better understanding of the evolutionary origin of soybean anthracnose, we compared the repertoire of effector candidates of four Colletotrichum species pathogenic to soybean and eight species not pathogenic. Our results show that the four species infecting soybean belong to two lineages and do not share any effector candidates. These results strongly suggest that two Colletotrichum lineages have acquired the capability to infect soybean independently. This study also provides, for each lineage, a set of candidate effectors encoding genes that may have important roles in pathogenicity towards soybean offering a new resource useful for further research on soybean anthracnose management

Molecular diversity of anthracnose pathogen populations associated with UK strawberry production suggests multiple introductions of three different Colletotrichum species.

Fragaria × ananassa (common name: strawberry) is a globally cultivated hybrid species belonging to Rosaceae family. Colletotrichum acutatum sensu lato (s.l.) is considered to be the second most economically important pathogen worldwide affecting strawberries. A collection of 148 Colletotrichum spp. isolates including 67 C. acutatum s.l. isolates associated with the phytosanitary history of UK strawberry production were used to characterize multi-locus genetic variation of this pathogen in the UK, relative to additional reference isolates that represent a worldwide sampling of the diversity of the fungus. The evidence indicates that three different species C. nymphaeae, C. godetiae and C. fioriniae are associated with strawberry production in the UK, which correspond to previously designated genetic groups A2, A4 and A3, respectively. Among these species, 12 distinct haplotypes were identified suggesting multiple introductions into the country. A subset of isolates was also used to compare aggressiveness in causing disease on strawberry plants and fruits. Isolates belonging to C. nymphaeae, C. godetiae and C. fioriniae representative of the UK anthracnose pathogen populations showed variation in their aggressiveness. Among the three species, C. nymphaeae and C. fioriniae appeared to be more aggressive compared to C. godetiae. This study highlights the genetic and pathogenic heterogeneity of the C. acutatum s.l. populations introduced into the UK linked to strawberry production

Straw competition and wheat root endophytism of Trichoderma gamsii T6085 as useful traits in the biocontrol of Fusarium head blight

Trichoderma gamsii T6085 has been investigated for many years as a beneficial isolate for use in the biocontrol of Fusarium head blight (FHB) of wheat caused primarily by Fusarium graminearum. Previous work focused on application of T6085 to wheat spikes at anthesis, whereas application to soil before and/or at sowing has received limited attention. In the present study, the competitive ability of T6085 on plant residues against F. graminearum was investigated. Results showed a significant reduction of wheat straw colonization by the pathogen and of the development of perithecia, not only when T6085 was applied alone but also in the presence of a Fusarium oxysporum isolate (7121), well known as a natural competitor on wheat plant residues. T6085 was able to endophytically colonize wheat roots, resulting in internal colonization of the radical cortex area, without reaching the vascular system, as confirmed by confocal microscopy. This intimate interaction with the plant resulted in a significant increase of the expression of the plant defense-related genes PAL1 and PR1. Taken together, competitive ability, endophytic behavior, and host resistance induction represent three important traits that can be of great use in the application of T6085 against FHB, not only on spikes at anthesis but potentially also in soil before and/or at sowing