Table 1.
Primers used in the experiments.
Table 2.
Overview of the sequencing reads.
Table 3.
Summary statistics for assemblies.
Table 4.
Summary of annotations of the R. wuchangensis unigenes against major public databases.
Fig 1.
The x-axis indicates the subcategories, the y-axis indicates the number of unigenes.
Table 5.
Infection rate and sex ratio of R. wuchangensis in different situations.
Fig 2.
Photomicrographs of pathological mosquito larvae infected by R. wuchangensis and normal larvae (A). Photomicrographs of many nematodes were obtained from infected group (B). Arrow: the nematode existed in the thorax of mosquito C. quinquefasciatus.
Fig 3.
Multiple sequence alignment by DNAMAN of Rwucmab-3 with that of other nematodes.
Identical and similar amino acid residues are shaded in black and gray, respectively. Sequences from the following nematode were used in this analysis: RwucMAB-3 (KU201268); CeleMAB-3 (CE14902); CbreMAB-3 (CN05170); CjapMAB-3 (JA51043); BmalMAB-3 (BM23119); OvolMAB-3 (OVP11339).
Fig 4.
Phylogenetic trees based on the deduced amino acid sequences of sex differentiation gene mab-3.
Amino acid sequences of Rwucmab-3 were analyzed using the Mega 6.0 program by the neighbor-joining method, respectively. The numbers at each node represent the bootstrap value with 1000 replicates. Sequences from the following nematode were used in this analysis: RwucMAB-3 (KU201268); CbriMAB-3(CBP05898), CremMAB-3 (RP15394), CeleMAB-3 (CE14902), CjapMAB-3 (JA51043), CbreMAB-3 (CN05170), BmalMAB-3 (BM23119), OvolMAB-3 (OVP11339), PpacMAB-3 (PP31573), DrerMAB-3 (Q71MM5).
Fig 5.
Expression pattern of sex differentiation genes Rwucmab-3 (A), Rwuclaf-1 (B) and Rwuctra-2 (C) in various developmental stages of R. wuchangensis.
P3-P5: the 3rd day of parasitic stage to the 5th day of parasitic stage; PP1-PP7: the 1st day of post-parasitic stage to the 7th day of post-parasitic stage; A: adult stage; G: Gravid adult nematode. *P < 0.05, **P < 0.01.
Fig 6.
FITC fluorescence of R. wuchangensis incubated in RNase-free water.
Images in Figures A-J show FITC fluorescence of R. wuchangensis incubated for 16 h with different concentrations of FITC: A = 20 μg/mL; B = 40 μg/mL; C = 60 μg/mL; D = 80 μg/mL; E = 100 μg/mL; F = 200 μg/mL; G = 400 μg/mL; H = 600 μg/mL; I = 800 μg/mL; J = 1000 μg/mL. Scale bar represents 50 μm.
Fig 7.
Sex ratio displayed by R. wuchangensis following soaking in dsRNA of Rwucmab-3 (A), Rwuclaf-1 (B) and Rwuctra-2 (C) for 16h.
*P < 0.05, **P < 0.01.