Application of RNA Interference in the Pinewood Nematode, <em>Bursaphelenchus xylophilus</em>

The pinewood nematode, Bursaphelenchus xylophilus, is one of the most destructive invasive species worldwide, causing the wilting and eventual death of ...

Zhejiang agricultural & forestry University

Pandora neoaphidis overwintering had been investigated by monitoring its prevalence in Myzus persicae populations in open fields. Cabbage plants in field plots were weekly taken after mycosis initiation, to count and examine the living and dead aphids infected by P. neoaphidis . Based on the field data, infection levels ( I) varied with field temperature (T), relative humidity (RH) and aphid count (numbers of living aphids per plant, N) over days ( D), fitting well to the modified logistic equation I =0.91/[1+exp(8.5+(2.0 H T H RH-20.2 NI 0) D)] ( r (2)=0.897), where H T indicated daily hours of low temperature (<4°C), H RH daily hours of high air humidity (>90% RH) and I 0 primary infection level. The model demonstrated the abiotic and biotic factors influencing P. neoaphidis mycosis development in winter, and also verifies the fungal overwintering by infecting available host aphids without a resting stage. Ultimately, P. neoaphidis mycosis reduced 81.4% of aphid populations, presenting great potential for biocontrol. 

Winter prevalence of obligate aphid pathogen Pandora neoaphidis mycosis in the host Myzus persicae populations in southern China: modeling description and biocontrol implication.

The brown planthopper (BPH), Nilaparvata lugens (Staparvata luera: Delphacidae), is a serious rice pest that easily develops resistance to chemical insecticides and resistant rice varieties. This study evaluated the infectivity of the BPH fungal pathogen, Pandora delphacis, and developed a novel formulation as an alternative means of BPH control.

A floatable formulation and laboratory bioassay of Pandora delphacis (Entomophthoromycota: Entomophthorales) for the control of rice pest Nilaparvata lugens Stål (Hemiptera: Delphacidae).

Two aerobic, rod-shaped, non-motile, non-sporulating and Gram-staining-negative bacterial strains, namely NH6-24T and Za3-11, were isolated from the surface seawater of the South China Sea and the estuary of the Yangtze River, respectively. The two isolates grew at 14–44 °C (optimum 37–40 °C) and pH 6.0–8.5 (optimum pH 7.0–7.5). The sea salt ranges for growth were 0.5–10 % (w/v) (optimum 1–2.5 %) for strain NH6-24T and 0–12 % (w/v) (optimum 0.5–4.5 %) for strain Za3-11.Both strains could grow in the absence of NaCl. Results of phylogenetic analysis based on 16S rRNA gene sequences indicated that the two isolates showed closest affinity to the genera Fontimonas (96.0 %) and Solimonas (94.1–95.1 %) and formed a single lineage in the cluster of the family Solimonadaceae. The predominant isoprenoid quinone was ubiquinone-8.The major fatty acids were C18 : 1ω7c, iso-C16 : 0 and C16 : 0.The dominant polar lipids were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. The DNA G+C content was 65 mol%. Based on the polyphasic taxonomic characterization, strains NH6-24T and Za3-11 are considered to represent a novel species of a novel genus, for which the name Sinimarinibacterium flocculans gen. nov., sp. nov. is proposed. The type strain of the type species is NH6-24T ( = CGMCC 1.10815T = JCM 17607T) and an additional strain is Za3-11 ( = CGMCC 1.10816 = JCM 17606).

Sinimarinibacterium flocculans gen. nov., sp. nov., a gammaproteobacterium from offshore surface seawater.

An aerobic, coccoid to short rod, yellow-pigmented, non-sporulating and Gram-staining-negative bacterium, designated NH6-79T, was isolated from surface seawater of the South China Sea. The isolate was motile with a polar flagellum. Growth was observed at 4-42 °C (optimum 37 °C), at pH 6.0-8.5 (optimum pH 7.0), and with 0.5-11 % (w/v) NaCl (optimum 4.5 %) and 1.5-17 % (w/v) sea salt (optimum 3.5-5 %). Strain NH6-79T could decompose peptone to produce H2S, but could not hydrolyse skimmed milk. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain NH6-79T had the closest affinity to the genus Parvularcula, sharing the highest 16S rRNA gene sequence similarity with 'Parvularcula oceanus' JLT2013 (94.1 %), Parvularcula lutaonensis CC-MMS-1T (93.4 %), Parvularcula dongshanensis SH25T (92.9 %) and Parvularcula bermudensis HTCC2503T (92.7 %), and lower sequence similarities (<90 %) with all other genera. The dominant fatty acids were C18 : 1ω7c and C16 : 0. The polar lipid profile was mainly composed of three unidentified glycolipids. The predominant isoprenoid quinone was ubiquinone-10. The DNA G+C content was 60.7 mol%. Based on the polyphasic taxonomic characterization, strain NH6-79T is considered to represent a novel species of the genus Parvularcula, for which the name Parvularcula flava sp. nov. is proposed. The type strain is NH6-79T (=CGMCC 1.14984T=JCM 30557T=MCCC 1K00277T).

Parvularcula flava sp. nov., an alphaproteobacterium isolated from surface seawater of the South China Sea.

Conidiobolus obscurus is a widespread fungal entomopathogen with aphid biocontrol potential. This study focused on a de novo transcriptomic analysis of C. obscurus.

Transcriptomic insight into pathogenicity-associated factors of Conidiobolus obscurus, an obligate aphid-pathogenic fungus belonging to Entomopthoromycota.

Dickeya dadantii is a plant pathogen that causes soft rot disease on vegetable and potato crops. To successfully cause infection, this pathogen needs to coordinately modulate the expression of genes encoding several virulence determinants, including plant cell wall degrading enzymes (PCWDEs), type III secretion system (T3SS) and flagellar motility. Here, we uncover a novel feed-forward signalling circuit for controlling virulence. Global RNA chaperone Hfq interacts with an Hfq-dependent sRNA ArcZ and represses the translation of pecT, encoding a LysR-type transcriptional regulator. We demonstrate that the ability of ArcZ to be processed to a 50 nt 3'- end fragment is essential for its regulation of pecT. PecT down-regulates PCWDE and the T3SS by repressing the expression of a global post-transcriptional regulator- (RsmA-) associated sRNA encoding gene rsmB. In addition, we show that the protein levels of two cyclic di-GMP (c-di-GMP) diguanylate cyclases (DGCs), GcpA and GcpL, are repressed by Hfq. Further studies show that both DGCs are essential for the Hfq-mediated post-transcriptional regulation on RsmB. Overall, our report provides new insights into the interplays between ubiquitous signalling transduction systems that were most studied independently and sheds light on multitiered regulatory mechanisms for a precise disease regulation in bacteria.

A feed-forward signalling circuit controls bacterial virulence through linking cyclic di-GMP and two mechanistically distinct sRNAs, ArcZ and RsmB.

 is a plant-pathogenic bacterium that causes soft-rot in a wide range of plants. Although we have previously demonstrated that cyclic bis-(3'-5')-cyclic dimeric guanosine monophosphate (c-di-GMP), a bacterial secondary messenger, plays a central role in virulence regulation in , the upstream signals that modulate c-di-GMP remain enigmatic. Using a genome-wide transposon mutagenesis approach of a Δ mutant strain that has high c-di-GMP and reduced motility, we uncovered transposon mutants that recovered the c-di-GMP-mediated repression on swimming motility. A number of these mutants harbored transposon insertions in genes encoding tricarboxylic acid (TCA) cycle enzymes. Two of these TCA transposon mutants were studied further by generating chromosomal deletions of the  gene (encoding fumarase) and the  operon (encoding succinate dehydrogenase). Disruption of the TCA cycle in these deletion mutants resulted in reduced intracellular c-di-GMP and enhanced production of pectate lyases (Pels), a major plant cell wall-degrading enzyme (PCWDE) known to be transcriptionally repressed by c-di-GMP. Consistent with this result, addition of TCA cycle intermediates such as citrate also resulted in increased c-di-GMP levels and decreased production of Pels. Additionally, we found that a diguanylate cyclase GcpA was solely responsible for the observed citrate-mediated modulation of c-di-GMP. Finally, we demonstrated that addition of citrate induced not only an overproduction of GcpA protein but also a concomitant repression of the c-di-GMP-degrading phosphodiesterase EGcpB which, together, resulted in an increase in the intracellular concentration of c-di-GMP. In summary, our report demonstrates that bacterial respiration and respiration metabolites serve as signals for the regulation of c-di-GMP signaling.

Tricarboxylic Acid (TCA) Cycle Enzymes and Intermediates Modulate Intracellular Cyclic di-GMP Levels and the Production of Plant Cell Wall-Degrading Enzymes in Soft Rot Pathogen .

Cytolytic (Cyt)-like genes are known by omics analyses to exist widely in bacterial and fungal pathogens, but their insecticidal activities in fungi remains unknown. A full-length coding sequence of a Cyt-like gene was first amplified from Conidiobolus obscurus (an obligate aphid-pathogenic fungus) through RACE (rapid-amplification of cDNA ends). The deduced protein structure was structurally characterized by a single Cyt-typical α/β domain. The expression level of the Cyt-like gene in conidia correlated well with the fungal virulence against aphids (r = 0.97). The results demonstrate the Cyt-like gene acts as an important virulence factor of C. obscurus against aphids, and has potential for exploitation in aphid control.

Xiang Zhou

College of Forestry and Biotechnology

Zhejiang agricultural & forestry University

Application of RNA Interference in the Pinewood Nematode, Bursaphelenchus xylophilus

Xiang Zhou

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Zhejiang agricultural & forestry University

Application of RNA Interference in the Pinewood Nematode, Bursaphelenchus xylophilus

College of Forestry and Biotechnology