MRNA Differential Expression From Interaction Of Puccinia Recondita With Lr38 Based On DDRT-PCR

Wheat leaf rust disease, caused by Puccinia recondita f. sp. tritici, is one of the most important diseases on wheat production throughout all the world. Now we haven't found the cloning about wheat leaf rust resistance gene in cloned, and no wheat leaf rust races have been found to overcome resistance of Lr38. So Lr38 and two leaf rust races 99-5-30-2 which express type is 2 and 99-8-9-2-1 which express type is 0 as the materials of this research, isolated the total RNA of wheat leaf which inoculated by two leaf rust races and control, reverse tanscription the mRNA to complement DNA, then compare the bands of two test and control by DDRT-PCR methods. At last we make a break through in this research.Two races of leaf rust inoculated the Lr38 and no inoculated as the control. Combining three kinds of anchored primer T10N(N is A, G or C), which is containing a HindIII recongnition site, and with 24 kinds of 10 mer arbitrary primer, difference on mRNA level were analyzed among between the two tests and control by DDRT-PCR. We found the difference between differential primers comebination is big, and the The difference between differential primer combination is big, and the amplification products polymorphism is also difference.The differential time 12, 24, 36, 48, 60 and 72 hours of wheat leaf samples which is inoculated by two leaf rust races and control. We use the extracted total RNA of wheat leaves to replace the mRNA analysis the difference of resistance gene induced expression. We start to find the differential cDNA bands at 24 hours after inoculated, but the quantity is small. The high level resistance gene expression after 36 hours. Restrive the differential cDNA fragment from the PAG and reamplified it at the same parasites which can make us do the others research works, for example Northernblotting, molecular hybrid, cloning, sequence checking and so on.