The complete mitochondrial genome of the nematophagous fungus Acremonium implicatum is reported for the first time. The genome is concatenated with 22,367 bp in length, encoding 13 protein-coding genes, 2 ribosomal RNA genes and a set of 17 transfer RNA genes. The synteny analysis reveals that 50.35% of A. implicatum mitochondrial sequences matched to 48.21% of Acremonium chrysogenum mitochondrial sequences with 85.68% identity. Two proteins of cox3 and nad6, as well as seven tRNAs are lost in A. implicatum mitogenome compared to A. chrysogenum mitogenome. The gene orders in A. implicatum and A. chrysogenum mitogenome is different, which is mainly due to the location of nad4 and cox2. In addition, one transposition event related to tRNAs is identified in these two mitogenomes. This study may provide valuable mitochondrial information for research on A. implicatum and facilitate the study of mitochondrial evolution.
Introduction Root-knot nematode (RKN; Meloidogyne spp.) is one of the most infamous soilborne plant diseases, causing severe crop losses every year. Effector proteins secreted by RKNs play crucial roles during plant-nematode interaction. However, less is known about whether RKN effector proteins can impact the rhizosphere microbial environment. Methods In this study, we investigated the rhizosphere microbiome community of MiMIF-2 (a plant immunity-modulating effector) transgenic Arabidopsis thaliana with or without nematode infection using the Illumina high-throughput sequencing analysis. Results and discussion The results showed that the bacterial species richness index increased, while the fungi species richness index decreased in M. incognita -infected MiMIF-2 transgenic A. thaliana plants. The relative abundance of genera such as Clitopilus, Komagataeibacter, Lactobacillus, Prevotella, Moritella, Vibrio, Escherichia-Shigella , and Pseudomonas was reduced in MiMIF-2 transgenic A. thaliana plants compared to wild type, but was significantly increased after inoculation with M. incognita . The Cluster of Orthologous Genes (COG) function classification analysis revealed a decrease in the relative abundance of defense mechanisms, secondary metabolite biosynthesis, transport, and nematode infection catabolism-related functions in MiMIF-2 lines compared to the wild type. These differences may be the reason for the increased susceptibility of MiMIF-2 transgenic A. thaliana to nematode infection. Our results provide a new insight into RKN effector proteins and their association with the microbial community, host, and plant pathogens, which will lead to the exploration of new innovative ideas for future biological control of RKNs.
Background MicroRNAs (miRNAs) are considered to be very important in regulating the growth, development, behavior and stress response in animals and plants in post-transcriptional gene regulation. Pinewood nematode, Bursaphelenchus xylophilus, is an important invasive plant parasitic nematode in Asia. To have a comprehensive knowledge about miRNAs of the nematode is necessary for further in-depth study on roles of miRNAs in the ecological adaptation of the invasive species. Methods and Findings Five small RNA libraries were constructed and sequenced by Illumina/Solexa deep-sequencing technology. A total of 810 miRNA candidates (49 conserved and 761 novel) were predicted by a computational pipeline, of which 57 miRNAs (20 conserved and 37 novel) encoded by 53 miRNA precursors were identified by experimental methods. Ten novel miRNAs were considered to be species-specific miRNAs of B. xylophilus. Comparison of expression profiles of miRNAs in the five small RNA libraries showed that many miRNAs exhibited obviously different expression levels in the third-stage dispersal juvenile and at a cold-stressed status. Most of the miRNAs exhibited obviously down-regulated expression in the dispersal stage. But differences among the three geographic libraries were not prominent. A total of 979 genes were predicted to be targets of these authentic miRNAs. Among them, seven heat shock protein genes were targeted by 14 miRNAs, and six FMRFamide-like neuropeptides genes were targeted by 17 miRNAs. A real-time quantitative polymerase chain reaction was used to quantify the mRNA expression levels of target genes. Conclusions Basing on the fact that a negative correlation existed between the expression profiles of miRNAs and the mRNA expression profiles of their target genes (hsp, flp) by comparing those of the nematodes at a cold stressed status and a normal status, we suggested that miRNAs might participate in ecological adaptation and behavior regulation of the nematode. This is the first description of miRNAs in plant parasitic nematodes. The results provide a useful resource for further in-depth study on molecular regulation and evolution of miRNAs in plant parasitic nematodes.
AbstractN-methyltransferase (NMT)-catalyzed methylations are rarely reported at nonribosomal peptides (NRPs) terminuses. Here, we discovered a fungal NMT LcsG for the iterative terminal N-methyl formation of a family of NRPs, leucinostatins. Gene deletion suggested LcsG is essential to the methylation of leucinostatins. In vitro assay and HRESI-MS-MS analysis proved the methylation sites were the NH2, NHCH3 and N(CH3)2 in the C-terminal unit of various leucinostatins. Based on the protein structure predicted by artificial intelligence (AI), molecular docking, and site-directed mutagenesis, we proposed the catalytic mechanism of the LcsG-catalyzed reaction was an N atom coordinated by two negatively charged residues (Asp368, Asp395 for LcsG) towards the subsequent SN2 methylation. These findings not only provide an approach for enriching the variety of natural bioactivity of NPRs but also deepen the insight into the catalytic mechanism of N-methylation of NRPs.
Specific endophytes with biocontrol potential might occur in diseased plant tissues. We isolated an endophytic fungus from tomato root galls infected with Meloidogyne incognita and identified it as Acremonium implicatum based on morphology and internal transcribed spacer sequences. Its biocontrol potential was tested in vitro and in pot and field experiments. In the in vitro test, 96.0% of second-stage juveniles of M. incognita were killed by a culture filtrate of A. implicatum after 48 h. The fungus also suppressed egg hatching, with only 36.3% of treated eggs hatching compared with 87.3% of control eggs. Pot experiments showed that A. implicatum inhibited the formation of root galls, with 40.6 galls per treated plant compared with 121.6 on control plants. A. implicatum reduced the nematode population in soil, with 151.1 nematodes per 100 g treated soil and 375.1 in control soil. Field experiments demonstrated that the root gall index of treated plants (25) was markedly lower than that of control plants (96). In conclusion, A. implicatum has excellent potential for the biocontrol of M. incognita.
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