Verticillium wilt, a soil-borne disease caused by Verticillium spp., is one of the most important diseases in eggplant production. It seriously affects the yield and quality of eggplant. At present, the control methods for eggplant verticillium wilt are limited to grafting and chemical controls, but neither of them can prevent the occurrence of verticillium wilt. The best way to control Verticillium wilt is to explore the verticillium wilt resistance genes in eggplant and to breed some resistant eggplant varieties with genetic engineering technology. Solanum symbriifolium Lam. is a wild eggplant in Yunnan, China with a high Verticillium wilt resistance. PR genes encode pathogenesis-related proteins, which are closely related to plant disease defense. Among them, PR10 protein is a nuclease-like protein, and the defense mechanism of PR10 protein in S. sisymbriifolium is still unclear. In this study, a wild eggplant S. sisymbriifolium was used as the experimental material. The pathogenesis related protein 10 (PR10) homologous gene was isolated and cloned by reverse transcription PCR (RT-PCR) technology from the previously established transcriptome database of S. sisymbriifolium and named as SsPR-10. The cloned sequence length of SsPR-10 was 645 bp, and its coding region was 480 bp long, encoding a protein of 159 amino acids residues in total. And the bioinformatics analysis showed that SsPR-10 protein was an acidic hydrophilic protein with a molecular mass of 17.71 kDa and a theoretical isoelectric point (pI) of 5.54. The prediction of transmembrane region and signal peptide showed that SsPR-10 protein had no transmembrane region and signal peptide. The prediction for subcellular localization showed that SsPR-10 was localized in the cytoplasm of plant cell. The results of conserved domain prediction showed that SsPR-10 contained a "P-LOOP" circle structure (sequence positions at 47-52 bp) and conserved sequence PATHOGENESIS_BETVI (sequence positions at 88-120 bp). The sequence alignment and phylogenetic tree analysis showed that SsPR-10 protein had the highest homology with PR10 protein from S. virginianum, followed by STH-2 protein from S. tuberosum. The expression of SsPR-10 gene was analyzed by real-time fluorescence quantitative PCR (RT-qPCR) experiments, and the expression level of SsPR-10 was the highest in the root than in other organs of S. sisymbriifolium, showing a tissue specificity. The expression level of SsPR-10 was multiplied to 8.16 times at 24 h after inoculation with V. dahliae, indicating that verticillium wilt might induce the expression of SsPR-10. In this study, SsPR-10 gene was cloned and its expression pattern was analyzed in the wild eggplant S. sisymbriifolium for the first time, which will provide a theoretical basis for further studying the mechanism of SsPR-10 responding to Verticillium wilt and other biological stresses.
| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |