Hypoxia-induced tracheal elasticity in vector beetle facilitates the loading of pinewood nematode
- State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences, China
- University of Chinese Academy of Sciences, China
- College of Life Science/Hebei Basic Science Center for Biotic Interactions, Institute of Life Science and Green Development, Hebei University, China
Figures
Figure 1
Negative correlation between oxygen level and pine wood nematode (PWN) loading in tracheal system of M. alternatus.
(A) Schematic representation of experimental setup used for microelectrode (top right) measurements inside a major trachea tube of the thorax (bottom left) and the atrium cavity of the largest …
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Figure 1—source data 1
Raw data for partial pressure of oxygen and corresponding PWN number.
- https://cdn.elifesciences.org/articles/84621/elife-84621-fig1-data1-v1.xlsx
Figure 2 with 4 supplements
Effects of pine wood nematode (PWN) loading and hypoxia on mechanical properties and apical extracellular matrix (aECM) of the trachea tubes of M. alternatus.
(A) Calculated Young’s modulus in different experimental groups according to values in tensile test in the longitudinal direction on the longest tubes linking two homolateral spiracles. Groups …
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Figure 2—source data 1
Raw data for Young’s modulus and aECM thinkness in different experimental groups.
- https://cdn.elifesciences.org/articles/84621/elife-84621-fig2-data1-v1.xlsx
Figure 2—figure supplement 1
Morphological changes of trachea of M. alternatus after pine wood nematode (PWN) loading.
(A) Morphology of thoracic tracheal tubes without and with heavy PWN under stereoscopic microscope. Images in the upper (scale bar = 500 μm) or lower (scale bar = 50 μm) panel show trachea in vivo …
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Figure 2—figure supplement 1—source data 1
Raw data for the relationship between PWN number and rubberization degree.
- https://cdn.elifesciences.org/articles/84621/elife-84621-fig2-figsupp1-data1-v1.xlsx
Figure 2—figure supplement 2
Pulling test of trachea of M. alternatus after pine wood nematode (PWN) loading.
Snapshots from videos imaged under stereotype microscope showed the pulling result of thoracic tracheal tubes with or without PWN. Red circles show one representative tubes in two groups before and …
Figure 2—video 1
Pulling test of trachea of M. alternatus without pine wood nematode (PWN).
Figure 2—video 2
Pulling test of trachea of M. alternatus with pine wood nematode (PWN).
Figure 3 with 1 supplement
Transcriptome analysis showed changed expression pattern of apical extracellular matrix (aECMs)-related genes in trachea treat with pine wood nematode (PWN).
(A) Volcano plot of RNA-seq data from trachea of beetles with and without PWN. Gray dots indicate all differentially expressed genes (DEGs), and blue dots indicate aECM-related DEGs. The red dots …
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Figure 3—source data 1
Raw data for FPKM values of 45 selected aECM genes in trachea with and without PWN.
- https://cdn.elifesciences.org/articles/84621/elife-84621-fig3-data1-v1.xlsx
Figure 3—figure supplement 1
Heat map of differential expressed genes related to mechanical property of tracheal tubes of beetles after 1 day, 3 days, 5 days, and 7 days post eclosion (PAE 1, PAE 3, PAE 5 and PAE 7, respectively).
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Figure 3—figure supplement 1—source data 1
Raw data for FPKM values of 45 selected aECM genes during tracheal maturation post adult eclosion.
- https://cdn.elifesciences.org/articles/84621/elife-84621-fig3-figsupp1-data1-v1.xlsx
Figure 4
Heavy pine wood nematode (PWN) loading and hypoxia enhance Muc91C expression at the mRNA and protein level.
(A) Relative RNA abundance of three genes belonging to mucin family in trachea of beetles with null, light, (<10,000 nematodes) or heavy (>10,000 nematodes) PWN loading. Data are represented as mean …
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Figure 4—source data 1
Raw data for mRNA level of Muc91C in different experimental groups and tissues.
- https://cdn.elifesciences.org/articles/84621/elife-84621-fig4-data1-v1.xlsx
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Figure 4—source data 2
Raw data for protein level of Muc91C in tracheal tubes with PWN.
- https://cdn.elifesciences.org/articles/84621/elife-84621-fig4-data2-v1.zip
Figure 5
The localization of Muc91C at the apical extracellular matrix (aECM) layer in tracheal tubes.
Immunostaining images of tracheal tube with null (left panels) and heavy pine wood nematode (PWN) (right panels) loading. Boxed regions (pink) in the top-left fluorescence images (bars, 50 μm) are …
Figure 6 with 1 supplement
Phylogenetic relationship among resilin-related proteins in insects.
The phylogenetic analysis was performed using the full-length amino acid sequences of representative resilin-related proteins. Am, Apis melifera; Mp, Myzus persicae; Nv, Nasonia vitripennis; Aga, Ano…
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Figure 6—source data 1
Raw data for sequences for MaMucins.
- https://cdn.elifesciences.org/articles/84621/elife-84621-fig6-data1-v1.docx
Figure 6—figure supplement 1
A phylogenetic tree constructed using M. alternatus mucin sequences (orange dots) and other species' mucin sequences.
Muc91C proteins were highlighted in red font. The full-length amino acid sequences of representative proteins were retrieved from the GenBank database. The phylogenetic tree was constructed with the …
Figure 7 with 1 supplement
The role of Muc91C in tracheal elasticity and pine wood nematode (PWN) loading.
(A) Electron microscopy images showed the thickened tracheal apical extracellular matrix (aECM) (red arrow) under 1% O2 for 12 hr after dsGFP or dsMuc91C injection. Bars, 1 μm. The right panel shows …
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Figure 7—source data 1
Raw data for aECM thickness, Young’s Module values and PWN numbers of dsMuc91C-treated beetles.
- https://cdn.elifesciences.org/articles/84621/elife-84621-fig7-data1-v1.xlsx
Figure 7—figure supplement 1
RNAi efficiency of the Muc91C gene and survival rates in dsRNA-treated adult beetles.
(A). Relative abundance of Muc91C in GFPi and Muc91Ci beetles. Data are presented as mean ± SEM. N = 5. p-Value is determined by Mann–Whitney U test. (B). Accumulated survival rate of adults in RNAi …
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Figure 7—figure supplement 1—source data 1
Raw data for mRNA level of Muc91C and survival rate of dsMuc91C-injected beetle.
- https://cdn.elifesciences.org/articles/84621/elife-84621-fig7-figsupp1-data1-v1.xlsx
Figure 8
Schematic diagram of the feedback regulation of pine wood nematode (PWN) loading through O2 and CO2 level in the tracheal system of M. alternatus.
When LIV dispersal nematodes continuously enter the vector beetle through spiracles, the consequent hypoxia enhances the elasticity of tracheal tubes by upregulating Muc91C. The more elastic tube …
Additional files
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Supplementary file 1
Species and GenBank accession no. of pro-resilins and mucins.
- https://cdn.elifesciences.org/articles/84621/elife-84621-supp1-v1.docx
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Supplementary file 2
Primers for real-time qPCR and RNAi experiments.
- https://cdn.elifesciences.org/articles/84621/elife-84621-supp2-v1.docx
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MDAR checklist
- https://cdn.elifesciences.org/articles/84621/elife-84621-mdarchecklist1-v1.docx
