Neuropeptide Bursicon and its receptor-mediated the transition from summer-form to winter-form of Cacopsylla chinensis
- Department of Entomology and MOA Key Lab of Pest Monitoring and Green Management, College of Plant Protection, China Agricultural University, China
- Sanya Institute of China Agricultural University, China
Figures
Figure 1 with 4 supplements
Molecular characteristic of CcBurs-α and CcBurs-β in C. chinensis.
(A) Multiple alignments of the amino acid sequences of CcBurs-α with homologs from five other insect species. Black represents 100% identity, red represents 75% identity, and blue represents <75% …
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Figure 1—source data 1
Labelled file for the western blot analysis in Figure 1D (reduced gel).
- https://cdn.elifesciences.org/articles/97298/elife-97298-fig1-data1-v1.tif
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Figure 1—source data 2
Original, uncropped file for the western blot analysis in Figure 1D (reduced gel).
- https://cdn.elifesciences.org/articles/97298/elife-97298-fig1-data2-v1.tif
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Figure 1—source data 3
Labelled file for the western blot analysis in Figure 1D (non-reduced gel).
- https://cdn.elifesciences.org/articles/97298/elife-97298-fig1-data3-v1.tif
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Figure 1—source data 4
Original, uncropped file for the western blot analysis in Figure 1D (non-reduced gel).
- https://cdn.elifesciences.org/articles/97298/elife-97298-fig1-data4-v1.tif
Figure 1—figure supplement 1
Investigation of the relationship between nymph phenotype, cuticle pigment absorbance, and cuticle thickness during the transition from summer-form to winter-form in C. chinensis.
(A) The nymph morphology during the transition from summer form to winter form of C. chinensis at varying time intervals (3, 6, 9, 12, 15 days). (B–C) The absorbance of total cuticle pigment and …
Figure 1—figure supplement 2
Phylogenetic tree analysis of CcBurs-α and CcBurs-β with its homologs in other insect species.
HvBurs-α (Homalodisca vitripennis, XP_046670477.1), PsBurs-α (Phenacoccus solenopsis, QEY08365.1), DvBurs-α (Daktulosphaira vitifoliae, XP_050533746.1), RmBurs-α (Rhopalosiphum maidis, …
Figure 1—figure supplement 3
Spatio-temporal expression patterns of CcBurs-α and CcBurs-β.
(A–B) Melting curve for qRT-PCR primers of CcBurs-α and CcBurs-β. (C-D) The relative mRNA expression of CcBurs-α and CcBurs-β at different development ages of summer-form and winter-form by qRT-PCR …
Figure 1—figure supplement 4
RNAi efficiency of CcTRPM after dsRNA treatment at 3, 6, and 10 days by qRT-PCR under 10 °C condition (n=3).
Figure 2
Neuropeptide Bursicon was essential for the transition from summer-form to winter-form in C. chinensis.
(A-B) RNAi efficiency of CcBurs-α and CcBurs-β after dsRNA treatment at 3, 6, and 10 days by qRT-PCR under 10 °C condition (n=3). (C-I) Effect of RNAi-mediated knockdown of CcBurs-α and CcBurs-β on …
Figure 3 with 2 supplements
CcBurs-R was identified as the Bursicon receptor in C. chinensis.
(A) Multiple alignments of the amino acid sequences of the CcBurs-R transmembrane domain with homologs from four other insect species. The transmembrane domain from TM1 to TM6 is indicated by blue …
Figure 3—figure supplement 1
Spatio-temporal expression patterns of CcBurs-R in both summer-form and winter-form by qRT-PCR (n=3).
Figure 3—figure supplement 2
Concentration-response relationships for peptides tested on Ccburs-R.
Figure 4
CcBurs-R directly mediated the transition from summer-form to winter-form in C. chinensis. (A) RNAi efficiency of CcBurs-R after dsRNA treatment at 3, 6, and 10 days by qRT-PCR under 10 °C condition (n=3). (B-H) Effect of RNAi-mediated knockdown of CcBurs-R on the absorbance of total cuticle pigment, relative cuticle chitin content, cuticle thickness of the thorax, transition percent, and phenotypic changes of first instar nymphs compared to dsEGFP treatments under 10 °C condition (n=9). (I-J) Relative mRNA expression of CcTre1 and CcCHS1 afterknockdown of CcBurs-α,CcBurs-β, and CcBurs-R at 10 d, separately (n=3). Data in 4 A, 4I, and 4 J are shown as the mean ± SE with three independent biological replications, with at least 50 nymphs for each replication. Data in 4B, 4 C, and 4E are presented as mean ± SE with three biological replications, with three technical replications for each biological replication. Data in 4 G are presented as mean ± SE with nine biological replications. Statistically significant differences were determined using pair-wise Student’s t-test, and significance levels were denoted by **p<0.01 and ***p<0.001. Different letters above the bars indicate statistically significant differences (p<0.05), as determined by ANOVA followed by a Turkey’s HSD multiple comparison test in SPSS 26.0 software.
Figure 5 with 1 supplement
miR-6012 directly targeted CcBurs-R to inhibit its expression.
(A) Predicted binding sites of four miRNAs in the 3’UTR of CcBurs-R. (B) In vitro confirmation of the target relationship between miR-6012 and CcBurs-R using dual luciferase reporter assays. (C) In …
Figure 5—figure supplement 1
Enrichment of miR-6012 by antibody against Ago1 in agomir-6012 treated group compared with agomir-NC group.
Figure 6
miR-6012 targeted CcBurs-R to mediate the seasonal polyphenism in C. chinensis.
(A) Expression of miR-6012 after agomir-6012 treatment at 3, 6, and 10 days by qRT-PCR under 10 °C condition (n=3). (B-H) Effect of agomir-6012 treatment on absorbance of total cuticle pigment, …
Figure 7
Schematic model of the novel functions of Bursicon signaling in the seasonal polyphenism of C. chinensis in response to low temperature.
Under 10 °C conditions, low temperature significantly upregulated the expression of the Bursicon signaling pathway. CcBurs-α and CcBurs-β then formed a heterodimeric neuropeptide to activate their …
Additional files
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Supplementary file 1
The primers used in current study and comparison of pigmentation and cuticle thickness after genes knockdown.
(a) List of primers used in this study. (b) Comparison of pigmentation and cuticle thickness after CcTRPM, CcBurs-a, CcBurs-β, and CcBurs-R knockdown.
- https://cdn.elifesciences.org/articles/97298/elife-97298-supp1-v1.docx
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MDAR checklist
- https://cdn.elifesciences.org/articles/97298/elife-97298-mdarchecklist1-v1.pdf
