pubmed.ncbi.nlm.nih.gov

Transcriptome Analyses of Diaphorina citri Midgut Responses to Candidatus Liberibacter Asiaticus Infection - PubMed

  • ️Wed Jan 01 2020

Transcriptome Analyses of Diaphorina citri Midgut Responses to Candidatus Liberibacter Asiaticus Infection

Hai-Zhong Yu et al. Insects. 2020.

Abstract

The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama (Hemiptera: Liviidae), is an important transmission vector of the citrus greening disease Candidatus Liberibacter asiaticus (CLas). The D. citri midgut exhibits an important tissue barrier against CLas infection. However, the molecular mechanism of the midgut response to CLas infection has not been comprehensively elucidated. In this study, we identified 778 differentially expressed genes (DEGs) in the midgut upon CLas infection, by comparative transcriptome analyses, including 499 upregulated DEGs and 279 downregulated DEGs. Functional annotation analysis showed that these DEGs were associated with ubiquitination, the immune response, the ribosome, endocytosis, the cytoskeleton and insecticide resistance. KEGG enrichment analysis revealed that most of the DEGs were primarily involved in endocytosis and the ribosome. A total of fourteen DEG functions were further validated by reverse transcription quantitative PCR (RT-qPCR). This study will contribute to our understanding of the molecular interaction between CLas and D. citri.

Keywords: CLas infection; Diaphorina citri; transcriptome sequencing.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1

The structure of the D. citri midgut, and the PCR detection of Candidatus Liberibacter asiaticus (CLas). (A) The structure of the whole D. citri midgut was observed under a Lecia S8AP0 stereomicroscope. (B) Agarose gel electrophoresis analysis in CLas-free groups and CLas-infected groups. M, 2000 DNA marker; PC, positive control; NC, negative control.

Figure 2
Figure 2

The identification and hierarchical cluster analysis of differentially expressed genes. (A) A scatter diagram for each gene. The blue, red and green points represent no difference in expression, upregulated genes and downregulated genes, respectively. (B) Hierarchical clustering of differentially expressed genes (DEGs) between the CLas-free groups and CLas-infected groups. Columns indicate different samples. Rows represent different DEGs. Blue bands indicate a low expression level, and red bands indicate a high gene expression level.

Figure 3
Figure 3

Gene Ontology (GO) enrichment analysis of DEGs. A scatter diagram of GO categories. The X-axis indicates the gene ratio. The Y-axis indicates different categories. (A) Upregulated DEGs. (B) Downregulated DEGs.

Figure 4
Figure 4

KEGG (Kyoto Encyclopedia of Genes and Genomes) enrichment analysis of DEGs. A scatter diagram of KEGG pathways. The X-axis indicates the gene ratio. The Y-axis indicates different pathways. (A) Upregulated DEGs. (B) Downregulated DEGs.

Figure 5
Figure 5

The correlation between the gene expression ratios obtained from the transcriptome data and RT-qPCR data. (A) The differential expression levels of 14 differentially expressed genes in the CLas-free and CLas-infected D. citri midgut. The relative expression levels were calculated using the 2−∆∆Ct method. Statistical analysis was performed using the SPSS software. The significant differences are indicated by * (p < 0.05) or ** (p < 0.01); (B) Lineage analysis between the transcriptome and RT-qPCR data. The ratios obtained by RT-qPCR (Y-axis) were plotted against the ratios obtained by the transcriptome (X-axis).

Figure 6
Figure 6

Hierarchical analysis for DEGs related to ubiquitination, the immune response and ribosomes between CLas-free and CLas-infected D. citri midgut. DEG expression is shown with a pseudocolor scale (from −3 to 3), with a red color indicating high expression levels and a green color indicating low expression. Each group represents three biological replicates.

Figure 7
Figure 7

Hierarchical analysis for DEGs associated with endocytosis, the cytoskeleton and insecticide resistance between CLas-free and CLas-infected D. citri midgut. A hierarchical cluster analysis was performed using the Genesis software. DEG expression is shown with a pseudocolour scale (from −3 to 3), with the red colour indicating high expression levels and the green color indicating low expression. Each group represents three biological replicates.

Similar articles

Cited by

References

    1. Bove J.M. Huanglongbing: A destructive, newly-emerging, century-old disease of citrus. J. Plant Pathol. 2006;88:7–37.
    1. Wang N., Pierson E.A., Setubal J.C., Xu J., Levy J.G., Zhang Y., Li J., Rangel L.T., Martins J.J. The Candidatus Liberibacter-host interface: Insights into pathogenesis mechanisms and disease control. Annu. Rev. Phytopathol. 2017;55:451–482. doi: 10.1146/annurev-phyto-080516-035513. - DOI - PubMed
    1. Slisz A.M., Breksa A.P., Mishchuk D.O., McCollum G., Slupsky C.M. Metabolomic analysis of citrus infection by ‘Candidatus Liberibacter’ reveals insights into pathogenicity. J. Proteome Res. 2012;22:4223–4230. doi: 10.1021/pr300350x. - DOI - PubMed
    1. Hu J.H., Jiang J.X., Wang N. Control of citrus Huanglongbing (HLB) via trunk injection of plant activators and antibiotics. Phytopathology. 2018;108:186–195. doi: 10.1094/PHYTO-05-17-0175-R. - DOI - PubMed
    1. Qureshi J.A., Kostyk B.C., Stansly P.A. Insecticidal suppression of Asian citrus psyllid Diaphorina citri (Hemiptera: Liviidae) vector of huanglongbing pathogens. PLoS ONE. 2014;9:e112331. doi: 10.1371/journal.pone.0112331. - DOI - PMC - PubMed

LinkOut - more resources