Multiplex PCR For Detection Of Multitraits In Wheat

In this paper,in order to check the PCR condition of the primer pairs,51 pairs of marker of different traits in wheat were used to amplified the 54 materials respectively,used the optimized markers and established the MPCR to distinguish the different traits in wheat,and the results are as the follows:Six pairs of PCR marker linked to Pm4,2 pairs of PCR marker linked to Pm6,3 pairs of PCR marker linked to Pm13,1 pairs of PCR marker linked to Pm16,3 pairs of PCR markers linked to Pm21,2 pairs of PCR markers linked to Pm24,1 pairs of PCR markers linked to Pam and Pox respectively were used to amplified the materials containing different resistance genes in wheat by different annealing temperature in different PCR program.The results showed that using the Mark of 2ab and 4G+4I can detect the powdery mildew resistance gene Pm4.detect the powdery mildew resistance genes Pm13 by using the Mark of 9ab and 10ab and the Mark of 12ab is closely linked to the powdery mildew resistance gene Pm21.1 pairs of PCR marker linked to Yr2,3 pairs of PCR markers linked to Yr5,1 pairs of PCR marker linked to Yrc591,Yr8 and Yr10 respectively,1 pairs of PCR marker linked to Lr9,Lr19 and Lr24 respectively,3 pairs of PCR marker linked to Lr35,1 pairs of PCR marker linked to Lr28 were used to amplified the materials containing different resistance genes in wheat by different annealing temperature in different PCR program. The results showed that using the Mark of 20ab and 21ab can detect stripe rust resistance gene Yr5.detect t stripe rust resistance gene Yr10 by using the Mark of 25ab. The materials "bainong3217" was detected to have Yr10 gene and "jingshuang 16","danmail","fengkang8"and "luzhenl"catained the Yr5,"jingshang16" also had the Lr28 gene.1 pairs of PCR markers linked to AX1,Ay,BX,By,BX7,BX14 and BX17 respectively,2 pairs of PCR markers linked to Dx2,6 pairs of PCR markers linked to Dx5,pairs of PCR markers linked to Dy10,were used to amplified the materials containing different HWM-GS subunits in wheat by different annealing temperature in different PCR program.The results showed that using the Mark of 43ab and 47 can detect HWM-GS subunit gene Dx5.and the Mark of 33ab is closely linked to the HWM-GS subunit gene Ax1.1 pairs of PCR markers linked to Rht8,Rht-B1b and Rht8-D1b respectively,were used to amplified the materials containing different dwarfs genes in wheat by different annealing temperature in different PCR program.The results showed that using the Mark of 49ab can detect the dwarf gene Rht8.and the Mark of 5lab is closely linked to the dwarf gene Rht8-D1b.By optimizing the PCR condition,a target DNA fragment was amplified to distinguished Pm4,Pm13 and Pm21,Yr5 and Yr10,Ax1 and DX5,Rht8 and Rht8-D1b. A multiplexed PCR system was established to distinguish HMW-GS subunits between plant height,powdery mildew resistant genes between HMW-GS subunits,wheat stripe rust resistance genes between wheat leaf rust resistance genes in a single PCR reaction and PAGE.Molecular markers have many advantages over other kinds of marker.Using molecular markers,it is quickly and properly to identify the resistances genes against wheat and HMW-GS composition of the hybridization progeny.This method shows excellentapplied property in marker-assisted selection for wheat quality breeding and polymerization breeding.