Inheritance And Molecular Markers Of Potato Virus Y Resistance In Tobacco

Tobacco is one of the important economic crops in China. Tobacco potato virus (PVY) has become a serious disease in tobacco. The yield of attacked plant reduced largely and even lead to no output when seriously. Besides the yield loss, the color, aroma and flavor of tobacco leaves were seriously poor after baking, which can reduce its commercial value significantly. Cultivating resistant varieties is considered to be the most economical and effective methods to control the disease. But the conventional breeding ways make the breeding process time-consuming and low efficiency. Molecular marker-assisted selection breeding can shorten the breeding process, avoid the multiplication of the vaccination evaluation process, save resources and improve the efficiency of breeding. In this paper, we study the inheritance of resistance genes of PVY in tobacco and analyse the linked molecular markers with resistance genes. The main content include three aspects as follows: (1) Preparing anti-sense combination of (VAM×Zhongyan100)and (VAM×G140),By manual inoculation identification of F1 and F2 at the seedling stage, investigating the inheritance of the resistance gene of N lines of the tobacco Potato virus Y (PVYN); (2) Using the orthogonal design to establish the reaction system of tobacco SRAP molecular markers; (3) Using bulked segregant analysis to screen the SRAP molecular markers of tobacco PVYN resistance gene, and determining molecular markers linked to the PVYN resistance gene. The main results are as follows:(1) VAM (burley tobacco varieties) was highly resistant to the PVYN, Both of the anti-sence of F1 were susceptible to the PVYN,that was recessive gene, F2 generation of anti-sence isolation than were 7:9,χ2 test showed that the resistance was probably controlled by two recessive genes which had the complementary effect.(2) In this paper, the orthogonal design was used to optimize SRAP-PCR amplification system on Tobacco. In four levels of five factors (Tap DNA polymerase, Mg2+, DNA template, d NTP, and primer) respectively. The result of PCR was analyzed by software Spss V 13.0, and the result showed that:①The affections of each factor in different levels were found and the quantity of Tap DNA polymerase was the most effective factor to the result of PCR;②A most suitable SRAP-PCR system for Tobacco was established, it's 25ul reaction system which contained 1.0U Tap DNA polymerase, 1.5mmol/L Mg2+, 30.00～120.00ngDNA template, 0.15mmol/L d NTP, and 0.40umol/L primer. Finally, the application of tobacco SRAP-PCR reaction system to the best of PCR procedures for the annealing temperature and cycle number of screening, drawn SRAP-PCR with annealing temperature of 52℃, 35 cycles for the best. This optimization of the system established for future use SRAP marker of tobacco conduct basic research provides a standardized procedure.(3) Obtained two SRAP markers closely linked to the resistance gene by using the BSA, which named as F3R13610 and F26R23450, Genetic distances were 6.51cM and 14.13cM, respectively.