Differential Gene Expression Analysis Of Copper Stress In Rice Roots By Cdna-Aflp

With the development of modern industry, the heavy metal pollution has become an serious social problem. Heavy metals such as Cu and Zn are essential for physiological process sincethey are constituents of many enzymes and other proteins. However, elevated concentrations of both essential and nonessential heavy metals can lead to toxicity. Plants possess several potential metal tolerance mechanisms including chelation, compartmentation and exclusion. Rice(Oryza sative L.)is considered as the main staple food in many countries of the world, it is also monocotyledon model plant. Rice Cu-tolerant variety (No.1139) and Cu-sensitive variety (No.1195), which was previously screened and identified by our research group, was used as the plant material throughout the experiments. We used the cDNA-amplified fragment length polymorphism to research the differences of gene expression in rice roots with Copper stress. It will Provide a theoretical basis for the heavy metal tolerance mechanism to the plant.cDNA-AFLP is a common analysis techniques for the differentially expressed genes. cDNA was digested by AseI and TaqI, ligated by two annealed anchors. And then, amplified by primers complementary to the anchors with two selective bases at3’end. The result of electrophoresis showed different primer combinations produced different banding pattern, while same banding patterns could be repeatedly obtained by a same primer combination Sequence of the specific between B1139and B1195showed the cloned fragments had the primer sequence and the two selective bases.These suggested cDNA-AFLP was of high reliability and repeatability.Totally32of combinations of8A-NN and4T-N primers were used.More than20bands were detected from each primers combination, and approximately700expression bands obtained in sum. Of which,22bands were singled out. They could be divided into seven categories:transcription factor, stress response gene, antioxidant genes, photosynthesis genes, cell wall synthesis genes, cell signal transduction and other. The genes of zinc finger protein、PR10、MT、Trxs、GTS、and CAM2maybe were the key genes in the Plant copper resistance mechanism.