Biosynthesis of Antibiotic Leucinostatins in Bio-control Fungus Purpureocillium lilacinum and Their Inhibition on Phytophthora Revealed by Genome Mining

Abstract

Purpureocillium lilacinum of Ophiocordycipitaceae is one of the most promising and commercialized agents for controlling plant parasitic nematodes, as well as other insects and plant pathogens. However, how the fungus functions at the molecular level remains unknown. Here, we sequenced two isolates (PLBJ-1 and PLFJ-1) of P. lilacinum from different places Beijing and Fujian. Genomic analysis showed high synteny of the two isolates, and the phylogenetic analysis indicated they were most related to the insect pathogen Tolypocladium inflatum. A comparison with other species revealed that this fungus was enriched in carbohydrate-active enzymes (CAZymes), proteases and pathogenesis related genes. Whole genome search revealed a rich repertoire of secondary metabolites (SMs) encoding genes. The non-ribosomal peptide synthetase LcsA, which is comprised of ten C-A-PCP modules, was identified as the core biosynthetic gene of lipopeptide leucinostatins, which was specific to P. lilacinum and T. ophioglossoides, as confirmed by phylogenetic analysis. Furthermore, gene expression level was analyzed when PLBJ-1 was grown in leucinostatin-inducing and non-inducing medium, and 20 genes involved in the biosynthesis of leucionostatins were identified. Disruption mutants allowed us to propose a putative biosynthetic pathway of leucinostatin A. Moreover, overexpression of the transcription factor lcsF increased the production (1.5-fold) of leucinostatins A and B compared to wild type. Bioassays explored a new bioactivity of leucinostatins and P. lilacinum: inhibiting the growth of Phytophthora infestans and P. capsici. These results contribute to our understanding of the biosynthetic mechanism of leucinostatins and may allow us to utilize P. lilacinum better as bio-control agent.

VelA and LaeA are key regulators of theEpichloë festucaetranscriptomic response during symbiosis with perennial ryegrass

“…EfM3.055320 was only expressed in the in planta comparisons and is a homologue of Cu/Zn superoxide dismutase from Claviceps purpurea [6] which are involved in reducing superoxide radicals generated by host plant defence mechanisms. EfM3.079420 is a homologue of a fungal hydrophobin domain-containing protein in Pochonia chlamydosporia [62].…”

Significant in vitro antagonism of the laurel wilt pathogen by endophytic fungi from the xylem of avocado does not predict their ability to control the disease

“…() reported that C. globosum produced a secondary metabolite that reduced in vitro growth of the northern corn leaf blight pathogen, Setosphaeria turcica , and inhibited development of the disease on detached corn leaves. Likewise, P. lilacinum (Wang et al ., ) and P. citrinum (Melouk & Akem, ; Park et al ., ) produced inhibitory secondary metabolites in vitro . Lan et al .…”

A genome scan of diversifying selection in Ophiocordyceps zombie‐ant fungi suggests a role for enterotoxins in co‐evolution and host specificity

“…Putative heat‐labile enterotoxin genes were retrieved for 14 entomopathogenic fungi (one strain per species) (Supporting Information Table ): Metarhizium anisopliae ARSEF23 (24 genes), M. acridum CQMa 102 (three genes) (Pattemore et al., ); M. album ARSEF1941 (12 genes), M. brunneum ARSEF3297 (32 genes), M. guizhouense ARSEF977 (32 genes), M. majus ARSEF297 (32 genes) (Hu et al., ); M. rileyi RCEF4871 (three genes), Isaria fumosorosea ARSEF2679 (five genes), Aschersonia aleyrodis RCEF2490 (14 genes), Cordyceps confragosa RCEF1005 (six genes), C. brongniartii RCEF3172 (30 genes) (Shang et al., ), Cordyceps militaris CM01 (one gene, Zheng et al., ), Beauveria bassiana ARSEF2860 (six genes, Xiao et al., ); and Ophiocordyceps sinensis Co18 (13 genes, Xia et al., ). We also included putative heat‐labile enterotoxin sequences from two nematode‐killing fungi: Purpureocillium lilacinum PLBJ‐1 (two genes, Wang et al., ) and Pochonia chlamydosporia 170 (four genes). Orthologs between these sequences and the putative enterotoxins of O. unilatealis species studied here were identified.…”