Abstract: In order to rapidly detect the Plum pox virus (PPV) from the introduced apricot resources, and screening PPV-resistant materials from domestic resources, we employed RT-PCR and quantitative Real-time PCR, and the rapid detection technology system of PPV was established in apricot, by comparing and verifying these two methods. The PCR specific parameters of the PCR amplification system and reaction conditions were determined.RT-PCR amplifications were performed in 20 μL reaction mixture containing 10×PCR Buffer 2 μL, 25 mmol/L Mg²⁺ 1.6 μL, 10 mmol/L dNTP 0.4 μL, 0.5 U/μL Taq DNA polymerase 0.2 μL, both forward and reverse primer 0.4 μL, cDNA 1 μL, and ddH₂O 14 μL. Reaction parameters for RT-PCR were as follows:94℃ for 5 min; 35 cycles of 94℃ for 45 s, 55℃ for 30 s, and 72℃ for 45 s; with a final extension step at 72℃ for 10 min. In addition, F₁ population crossed from two different PPV-resistant apricot parents had been constructed by Faculty of Horticulture, Mendel University. Based on these F₁ individual materials, we used SSR method coupled with BSA method to screen the markers linked to PPV-resistant genes. Firstly, the polymorphic markers were screened between the PPV-resistant gene pool and susceptible one, and six SSR loci (PGS1.23, PGS1.24, PaCITA5, UDAP-414, Pchcms4 and Pchgms4) had been screened out. The polymorphism between two gene pools and between two parents was consistent. Then these six locus were further verified among the F₁ progenies. As the final results, one SSR locus PGS1.23 with a high degree of linkage to PPV resistance was identified. There was a high coincidence of 78.3% between expected marker genotype and phenotype for the PGS1.23. Furthermore, this linkaged marker was used to evaluate the PPV resistance for 37 Chinese apricot germplasms, and five cultivars with PPV-resistance including Caizihuang, Yangjiyuan, Chuizhishanxing, Dazaoshu and Xifuxing were screened out. These results would provide better parents materials for PPV-resistant apricot breeding, and also lay the foundation for the further study on the candidate PPV-resistant gene.

Key words: Plum pox virus, Apricot, Molecular detection, Simple sequence repeat, Bulked segregant analysis

CLC Number: 

• Q78

• S662.03