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Abstract Background Horizontal gene transfer, also called lateral gene transfer, frequently occurs among prokaryotic organisms, and is considered an important force in their evolution. However, there are relatively few reports of transfer to or from fungi, with some notable exceptions in the acquisition of prokaryotic genes. Some fungal species have been found to contain sequences resembling those of bacterial genes, and with such sequences absent in other fungal species, this has been interpreted as horizontal gene transfer. Similarly, a few fungi have been found to contain genes absent in close relatives but present in more distantly related taxa, and horizontal gene transfer has been invoked as a parsimonious explanation. There is a paucity of direct experimental evidence demonstrating the occurrence of horizontal gene transfer in fungi. Results We found a fungal field isolate from rice (Oryzae sativa) that contains ribosomal DNA sequences from two species of fungal rice pathogens (Thanatephorus cucumeris and Ceratobasidium oryzae-sativae). This field isolate has four types of ribosomal DNA internal transcribed spacers (ITS), namely pure ITS of C. oryzae-sativae, which was dominant in this field isolate, pure ITS of T. cucumeris, and two chimeric ITS, with ITS1 derived from C. oryzae-sativae and ITS2 from T. cucumeris, or ITS1 from T. cucumeris and ITS2 from C. oryzae-sativae. The presence of chimeric forms indicates that the intergeneric hybrid was not merely composed of nuclei from the parental species, but that nuclear fusion and crossing over had taken place. Conclusion Hyphae of T. cucumeris and C. oryzae-sativae are vegetatively incompatible, and do not successfully anastomose. However, they parasitize the same host, and perhaps under the influence of host enzymes targeted to weaken pathogen cells or in dying host plant tissue, the fungal hyphae lost their integrity, and normal vegetative incompatibility mechanisms were overcome, allowing the hyphae to fuse. Based on the presence of other similarly anomalous isolates from the field, we speculate that these types of intergeneric hybridization events and occurrences of horizontal gene transfer may not be so rare in the field.

The deprotonation–reprotonation sequence introduces additional cyclization branches in terpene biosynthesis. However, the underlying mechanism remains poorly understood. In this study, we employed a combined approach of molecular dynamics (MD) simulations and site-directed mutagenesis on astellifadiene synthase EvAS from Emericella variecolor to investigate the role of a protonated S85 residue. This residue acts as a catalytic acid, previously unreported, that facilitates the reprotonation step in astellifadiene biosynthesis. Mutating S85 led to the production of a new tricyclic sesterterpene. [ABSTRACT FROM AUTHOR]

Terpene synthases (TPSs) play pivotal roles in generating diverse terpenoids through complex cyclization pathways. Protein engineering of TPSs offers a crucial approach to expanding terpene diversity. However, significant potential remains untapped due to limited understanding of the structure‐function relationships of TPSs. In this investigation, using a joint approach of molecular dynamics simulations‐assisted engineering and site‐directed mutagenesis, we manipulated the aromatic residue cluster (ARC) of a bifunctional terpene synthase (BFTPS), Pestalotiopsis fici nigtetraene synthase (PfNS). This led to the discovery of previously unreported catalytic functions yielding different cyclization patterns of sesterterpenes. Specifically, a quadruple variant (F89A/Y113F/W193L/T194W) completely altered PfNS′s function, converting it from producing the bicyclic sesterterpene nigtetraene to the tricyclic ophiobolin F. Additionally, analysis of catalytic profiles by double, triple, and quadruple variants demonstrated that the ARC functions as a switch, unprecedently redirecting the production of 5/11 bicyclic (Type B) sesterterpenes to 5/15 bicyclic (Type A) ones. Molecular dynamics simulations and theozyme calculations further elucidated that, in addition to cation‐π interactions, C−H⋅⋅⋅π interactions also play a key role in the cyclization patterns. This study offers a feasible strategy in protein engineering of TPSs for various industrial applications. [ABSTRACT FROM AUTHOR]

Simple Summary: Ginseng replant disease is associated with root rot caused by the soil-borne fungus, Ilyonectria mors-panacis, which is believed to be due to the combination of the pathogen with unknown host-related factors found in soil after harvesting ginseng. One candidate for the host-related factor was described in a prior report where methanol extracts from previous ginseng soil increased root lesion sizes during Ilyonectria mors-panacis infections associated with suppression of ginseng disease defense gene expression. As treatment of ginseng with ginseng root extract did not have these effects, it was unclear what was the source of this bioactivity in soil. This study showed that a soil bacterium, Pseudomonas plecoglossicida, isolated from soil previously used for ginseng production, can convert ginseng root compounds in culture into compounds able to increase infections by Ilyonectria mors-panacis, demonstrating a possible way that a host factor of ginseng replant disease is produced by microbial conversion of root materials in previous ginseng soils. A prior report showed that soil previously planted with American ginseng (Panax quinquefolius) contained compound(s) which could reduce ginseng resistance to root infection by Ilyonectria mors-panacis, and this was not found in extracts from ginseng roots or soils not previously planted with ginseng. However, the origin of this ginseng-related factor in ginseng soils is unknown. An isolate of Pseudomonas plecoglossicida obtained from soil where P. quinquefolius had been harvested grew more in culture media when ginseng root extract was included, indicating the use of compounds in the extract as nutrients. Treatment with cell-free extracts from media containing ginseng root extracts where P. plecoglossicida had been cultured resulted in root lesions caused by I. mors-panacis being significantly larger than roots treated with fresh media containing root extract or with cell-free media inoculated with the same bacterial isolate without root extract. Levels of ginsenosides in the media decreased over time with incubation. Genome sequencing revealed that the bacterium had genes homologous to those reported for ginsenoside metabolism, which can release sugars for microbial growth. Thus, a ginseng soil bacterium, P. plecoglossicida, can create compound(s) suppressive to root rot resistance, similar to that found in soils previously planted with ginseng, indicating that the activity suppressing root rot resistance in soil previously planted with ginseng may be of microbial origin, utilizing compounds from ginseng roots. [ABSTRACT FROM AUTHOR]

Simple Summary: Picromerus lewisi (P. lewisi), a predatory stink bug, is widely acknowledged as a beneficial natural enemy for controlling Lepidopteran pests in agroforestry systems. This species paralyzes its prey and facilitates extra-oral digestion by releasing venom. Although these insects harbor symbiotic microorganisms that play crucial roles in various biological aspects of their host interaction, comprehensive knowledge regarding the microbiota within the venom glands of P. lewisi is lacking. This study explored the numerous bacterial and fungal species in the venom glands of P. lewisi and the capability of these microbes to metabolize carbon sources. The dominant bacterial genera included Wolbachia, Enterococcus, Serratia, and Lactococcus, while the chief fungal genera were Vishniacozyma, Cladosporium, Papiliotrema, Penicillium, Fusarium, and Aspergillus. En masse, the microbial community showed abilities to metabolize a variety of carbon sources. This is the first report of the analysis of the microbiome in the venom glands of P. lewisi, enhancing our understanding of the microbial composition of this natural enemy's venom. These findings will provide useful insights for researchers to further investigate the venom microbiota and their associated functionalities among predatory stink bugs. The predatory stink bug, Picromerus lewisi (Hemiptera: Pentatomidae), is an important and valuable natural enemy of insect pests in their ecosystems. While insects are known to harbor symbiotic microorganisms, and these microbial symbionts play a crucial role in various aspects of the host's biology, there is a paucity of knowledge regarding the microbiota present in the venom glands of P. lewisi. This study investigated the venom glands of adult bugs using both traditional in vitro isolation and cultural methods, as well as Illumina high-throughput sequencing technology. Additionally, the carbon metabolism of the venom gland's microorganisms was analyzed using Biolog ECO metabolic phenotyping technology. The results showed 10 different culturable bacteria where the dominant ones were Enterococcus spp. and Lactococcus lactis. With high-throughput sequencing, the main bacterial phyla in the microbial community of the venom glands of P. lewisi were Proteobacteria (78.1%) and Firmicutes (20.3%), with the dominant bacterial genera being Wolbachia, Enterococcus, Serratia, and Lactococcus. At the fungal community level, Ascomycota accounted for the largest proportion (64.1%), followed by Basidiomycota (27.6%), with Vishniacozyma, Cladosporium, Papiliotrema, Penicillium, Fusarium, and Aspergillus as the most highly represented fungal genera. The bacterial and fungal community structure of the venom glands of P. lewisi exhibited high species richness and diversity, along with a strong metabolism of 22 carbon sources. Functional prediction indicated that the primary dominant function of P. lewisi venom-gland bacteria was metabolism. The dominant eco-functional groups of the fungal community included undefined saprotroph, fungal parasite–undefined saprotroph, unassigned, endophyte–plant pathogen, plant pathogen–soil saprotroph–wood saprotroph, animal pathogen–endophyte–plant pathogen–wood saprotroph, plant pathogen, and animal pathogen–endophyte–epiphyte–plant pathogen–undefined saprotroph. These results provide a comprehensive characterization of the venom-gland microbiota of P. lewisi and demonstrate the stability (over one week) of the microbial community within the venom glands. This study represents the first report on the characterization of microbial composition from the venom glands of captive-reared P. lewisi individuals. The insights gained from this study are invaluable for future investigations into P. lewisi's development and the possible interactions between P. lewisi's microbiota and some Lepidopteran pests. [ABSTRACT FROM AUTHOR]

As an evergreen shrub, Euonymus japonicus plays a crucial role in urban landscape construction, and its growth is affected by severe foliar anthracnose caused by Colletotrichum spp. However, the biodiversity of Colletotrichum species associated with anthracnose on E. japonicus remains undetermined. This study involved a two-year collection of E. japonicus leaf samples with typical anthracnose symptoms from 9 districts in Beijing, China. A total of 194 Colletotrichum isolates were obtained, and eight Colletotrichum species were subsequently identified using morphological characteristics and molecular identification with the ACT, GADPH, CHS, TUB2, and CAL genes, as well as the rDNA-ITS region. These species included Colletotrichum aenigma, C. fructicola, C. gloeosporioides, C. grossum, C. hebeiense, C. karstii, C. siamense, and C. theobromicola with C. siamense being the most prevalent (57%), followed by C. aenigma and C. theobromicola. Furthermore, C. fructicola, C. grossum and C. hebeiense are reported for the first time as causal agents of anthracnose on E. japonicus worldwide, and C. karstii is newly reported to be associated with E. japonicus anthracnose in China. Pathogenicity tests revealed that all tested isolates exhibited pathogenicity in the presence of wounds, emphasizing the need to avoid artificial or mechanical wounds to prevent infection in E. japonicus management. The EC50 values of five fungicides, namely difenoconazole, flusilazole, tebuconazole, hexaconazole, and prochloraz, were found to be less than 10 mg/L, indicating their strong potential for application. Notably, the EC50 of prochloraz was less than 0.05 mg/L for C. theobromicola. These findings offer valuable insights for the management of anthracnose on E. japonicus. [ABSTRACT FROM AUTHOR]

Interspecies hybrid sterility has been extensively studied, especially in the genus Drosophila. Hybrid sterility is more often found in the heterogametic (XY or ZW) sex, a trend called Haldane's rule. Although this phenomenon is pervasive, identification of a common genetic mechanism remains elusive, with modest support found for a range of potential theories. Here, we identify a single precise morphological phenotype, which we call 'needle-eye sperm', that is associated with hybrid sterility in three separate species pairs that span the Drosophila genus. The nature of the phenotype indicates a common point of meiotic failure in sterile hybrid males. We used 10 generations of backcross selection paired with whole-genome pooled sequencing to genetically map the regions underlying the needle-eye (NE) sperm phenotype. Surprisingly, the sterility phenotype was present in ~50% of males even after 10 generations of backcrossing, and only a single region of the X chromosome was associated with sterility in one direction of backcross. Owing to the common phenotype among sterile male hybrids, and the strong effect of individual loci, further exploration of these findings may identify a universal mechanism for the evolution of hybrid sterility. [ABSTRACT FROM AUTHOR]