Molecular Marker Detection And Resistance Identification Of Important Disease Resistance Genes In Common Tobacco

Tobacco is an important economic crop in China.Tobacco production and sales play an important role in China's national economy.Tobacco mosaic virus disease and black shank disease are extremely serious worldwide diseases that endanger the yield and quality of tobacco.In recent years,the incidence of two diseases in major tobacco producing areas in China has shown an increasing trend year by year,which seriously affects the yield and quality of tobacco in China.Studies have shown that the most economical and effective way to defend and control tobacco diseases in production is to promote the cultivation of tobacco resistant varieties.Excellent varieties are an important basis for crop production,and selection is an important part of cultivating good varieties.In recent years,with the development and application of molecular biology technology,and the continuous development of molecular markers of tobacco disease resistance and the continuous enhancement of the stability and accuracy of markers,the selection of parents and the resistance genotypes of offspring in tobacco resistance breeding Screening provides new technologies and methods.At present,the common tobacco has established a relatively stable specific molecular marker including a variety of important disease resistance characteristics including the anti-TMV N gene and the anti-black spot PH gene.To this end,based on a review of a large number of tobacco anti-TMV and anti-black spot molecular markers related research,using the specific molecular markers of screening and selection of tobacco TMV and black shank resistance characteristics,60 ordinary tobacco Breeding parental germplasm resources,65 sterile line hybrid materials,and 35 high-generation materials were tested for specific molecular markers of anti-TMV and black shank resistance genes,and the greenhouse disease resistance characteristics and tobacco of the above materials were also carried out.The correlation analysis between specific molecular markers of disease resistance genes and disease resistance characteristics in greenhouses yielded the following findings:1.Four specific molecular markers of 160 tobacco material tobacco mosaic resistance genes showed that 77,137,34,and 37 materials contained the markers NM2,F10,E1+E2,and N2,respectively.The proportion was 48.1%,85.6%,21.3% and 23.1%;9 materials did not contain the resistance mark of tobacco mosaic disease.Among the test materials,the F10 mark has the widest distribution rate.2.Detection of five specific molecular markers of 160 pieces of tobacco material black shank resistance gene showed that 123,143,140,138 and 140 materials contained the markers PT20383,INDEL10617,ST141,Scf₃0K and TM62,respectively The proportion was 76.9%,89.4%,87.5%,86.3% and 87.5%.6 materials did not contain the resistance marker of black shank.Among the tested materials,the distribution rate of five linkage markers of the black shank resistance gene was higher,both of which were more than 76%.3.Identification of resistance to tobacco mosaic virus disease in 160 tobacco materials indicated that 20 materials showed resistance to TMV?HR,MR and R?,and four linked markers of resistance genes.Among them,NC71,9J22,CV87,Shuangkang 70,anti-88 and Coker176 showing high resistance;high-generation materials 16F5-32,breeding parent material 89-07-1-2,Jiyan 5,Coker86,Taiyan 8 and CV91 showed resistance characteristics;sterile materials 9D45,9C37,9G45,9G07 and 19J06,high-generation materials 6F5-18,16F5-16 and 16F5-4,a total of 8 materials performance Medium resistance characteristics.The 20 materials exhibit both resistance to TMV and linkage markers of resistance genes,and are good germplasm resources for TMV resistance breeding.4.The identification of resistance to black shank in 160 greenhouse tobacco seedlings showed that there were 42 materials containing 5 linkage markers of black shank resistance gene and resistance to black shank?HR,MR and R?,among which,Gexin 3,NC55-1,73,9J25,16F5-15,16F5-16,16F5-39,16F5-40 and 16F5-44,a total of 9 materials exhibit high resistance;breeding parent material Qinyan96 and sterile line material 9E07 showed resistance characteristics;parent material wrinkle red,Shuangkang70,Yunyan110,FLorida301 and Ti245,sterile material 9A15,9A45,9A11,9B37,9J02,9J05,9J06,9J11,9J20,9J21,9J38,QY201,9D37,9C15,9G45,9G38,high-generation materials 5FB10-3,16F5-18,16F5-29,16F5-32,16F5-46,16F5-52,6FA10-3,NC89,16F5-11,16F5-26,a total of 31 materials showed medium resistance.The 42 materials exhibit both resistance to black shank and linkage markers of resistance genes,and are good germplasm resources for tobacco black shank resistance breeding.5.The haplotype analysis of the disease index corresponding to the four linkage markers of tobacco mosaic resistance gene showed that the tobacco mosaic disease index of tobacco varieties was significantly correlated with the presence or absence of four linkage markers of mosaic resistance genes?p<0.01?.And the disease mosaic index of the material containing the resistance gene was significantly lower than that of the disease-free genetic material?p<0.01?.It indicated that the resistance gene of tobacco mosaic virus plays an important role in regulating the disease resistance of the tested materials.6.A haplotype analysis of the disease index corresponding to the five markers related closely in the black shank resistance gene showed that: the disease index of tobacco varieties was significantly correlated with the presence or absence of five closely linked markers of resistance genes?p<0.01?.And the disease index of the material containing the resistance gene was significantly lower than that of the material without the resistance gene?p<0.01?.It indicated that the resistance gene of black shank in the tested materials played an important regulatory role in the resistance to black shank disease.7.To detect the resistance gene specific molecules of tobacco mosaic disease and black shank disease for 160 tobacco varieties,and to carry out the disease resistance identification of TMV and black scorpion patients simultaneously with the above resources.The results show that a total of 31 materials showed resistance characteristics?HR,R and MR?to both tobacco mosaic disease and black shank disease.A total of 21 test materials contained 4 special markers of TMV resistance gene and 5 markers of black shank resistance gene,Among them,parental NC71,shuangkang70 and sterile lines 9J06,9D45,9G07,a total of 5 materials contain all the linkage markers of the resistance gene,and also show the pair of tobacco mosaic disease and black shank in resistance identification.Anti-characteristics are excellent breeding materials.