Table 1.
List of primers used in RT-PCR, RACE-PCR, and real time PCR.
Table 2.
Characteristics of the four Hsps in O. chinensis.
Fig 1.
Phylogenetic analysis of O. chinensis Hsps and Hsps from other insects.
Multiple alignment of the sequences used in the analysis was conducted using CLUSTAL W. A neighbor-joining tree construction program (MEGA 5.0) was used for this phylogenetic analysis. Information on the sequences used in this analysis is shown in Table 3.
Table 3.
A complete list of species used in the phylogenetic analysis and accession numbers for Hsp sequences.
Fig 2.
The tissue expression pattern of O. chinensis Hsps in 5th instar nymphs.
The bars represent the mean ± SE (n = 3) of target gene mRNA expression in different tissues. The small letters on the bars indicate significant differences between tissues (Duncan’s multiple comparison, P< 0.05). BR: brain; EP: epidermis; FG: foregut; GC: gastric caecum; MG: midgut; HG: hindgut; MT: Malpighian tubule; FB: fat body. The mRNA level in the brain was arbitrarily taken as 1.0.
Fig 3.
The developmental expression pattern of O. chinensis Hsps in eggs, nymphs and adults.
The bars represent the mean ± SE (n = 3) of target gene mRNA expression at different developmental stages. The different small letters on the bars indicate significant differences between developmental stages (Duncan’s multiple comparison, P< 0.05). EG: eggs; 1st: first instar nymphs; 2nd: second instar nymphs; 3rd: third instar nymphs; 4th: fourth instar nymphs: 5th: fifth instar nymphs; AD: adults. The mRNA level in the eggs was arbitrarily taken as 1.0.
Fig 4.
mRNA expression of O. chinensis Hsps at different times after acute Cd exposure.
The bars represent the mean ± SE (n = 3) of target gene mRNA expression in 5th instar nymphs 0 h, 2 h, 6 h, 12 h, 24 h and 48 h after the injection of 2.7 mg·kg-1 Cd. The small letters on the bars indicate significant differences between the different treatment times (Duncan’s multiple comparison, P< 0.05). The mRNA level at 0 h was arbitrarily taken as 1.0.
Fig 5.
mRNA expression of O. chinensis Hsps 6 h after exposure to different Cd concentrations.
The bars represent the mean ± SE (n = 3) of target gene mRNA expression in 5th instar nymphs 6 h after the injection of different Cd concentrations. The small letters on the bars represent significant differences between different Cd concentrations (Duncan’s multiple comparison, P< 0.05). Distilled water was used as a control. The value of the control was arbitrarily taken as 1.0.
Fig 6.
mRNA expression of O. chinensis Hsps after chronic Cd exposure.
Wheat seedlings cultured in distilled water were used as a control. During this experiment, significant numbers of insect deaths did not occur. The bars represent the mean ± SE (n = 3) of target gene mRNA expression after chronic Cd exposure. The small letters on the bars represent significant differences between different Cd concentrations after the insects fed on dietary Cd (Duncan’s multiple comparison, P< 0.05). The value of the control was arbitrarily taken as 1.0.