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. 2022 Mar 30;13(4):338.
doi: 10.3390/insects13040338.

Full-Length Transcriptome Sequencing-Based Analysis of Pinus sylvestris var. mongolica in Response to Sirex noctilio Venom

Chenglong Gao  1   2 , Lili Ren  1   2 , Ming Wang  1   2 , Zhengtong Wang  1   2 , Ningning Fu  1   2 , Huiying Wang  3 , Juan Shi  1   2
Affiliations

Full-Length Transcriptome Sequencing-Based Analysis of Pinus sylvestris var. mongolica in Response to Sirex noctilio Venom

Chenglong Gao et al. Insects. .

Abstract

Sirex noctilio is a major international quarantine pest that recently emerged in northeast China to specifically invade conifers. During female oviposition, venom is injected into the host together with its symbiotic fungus to alter the normal Pinus physiology and weaken or even kill the tree. In China, the Mongolian pine (Pinus sylvestris var. mongolica), an important wind-proof and sand-fixing species, is the unique host of S. noctilio. To explore the interplay between S. noctilio venom and Mongolian pine, we performed a transcriptome comparative analysis of a 10-year-old Mongolian pine after wounding and inoculation with S. noctilio venom. The analysis was performed at 12 h, 24 h and 72 h. PacBio ISO-seq was used and integrated with RNA-seq to construct an accurate full-length transcriptomic database. We obtained 52,963 high-precision unigenes, consisting of 48,654 (91.86%) unigenes that were BLASTed to known sequences in the public database and 4309 unigenes without any annotation information, which were presumed to be new genes. The number of differentially expressed genes (DEGs) increased with the treatment time, and the DEGs were most abundant at 72 h. A total of 706 inoculation-specific DEGs (475 upregulated and 231 downregulated) and 387 wounding-specific DEGs (183 upregulated and 204 downregulated) were identified compared with the control. Under venom stress, we identified 6 DEGs associated with reactive oxygen species (ROS) and 20 resistance genes in Mongolian pine. Overall, 52 transcription factors (TFs) were found under venom stress, 45 of which belonged to the AP2/ERF TF family and were upregulated. A total of 13 genes related to the photosystem, 3 genes related photo-regulation, and 9 TFs were identified under wounding stress. In conclusion, several novel putative genes were found in Mongolian pine by PacBio ISO seq. Meanwhile, we also identified various genes that were resistant to S. noctilio venom, such as GAPDH, GPX, CAT, FL2, CERK1, and HSP83A, etc.

Keywords: Pinus sylvestris var. mongolica; Sirex noctilio; full-length transcriptome; venom; wounding.

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Conflict of interest statement

The authors declare no potential conflicts of interest.

Figures

Figure 1
Figure 1
Unigene annotation of PacBio sequencing. (A) Overlap between the number of all unigenes according to five databases. (B) Distribution of unigene annotations based on the NR database for the species-distribution statistics.
Figure 2
Figure 2
GO and KEGG classification of unigenes. (A) GO functional classification of all unigenes. (B) KEGG classification of all unigenes.
Figure 3
Figure 3
Number of DEGs induced by wounding and inoculation at different times.
Figure 4
Figure 4
GO (A) and KEGG (B) analyses of DEGs induced by inoculation specificity.
Figure 5
Figure 5
GO (A) and KEGG (B) analysis of DEGs induced by wounding specificity.
Figure 6
Figure 6
GO (A) and KEGG (B) analysis of DEGs co-induced by inoculation and wounding.
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