Cloning And Expression Of ZmPP2C Gene In Maize Roots And Its Transformation

The maize variety of stress-resistance LD9002 was cultivated by solution in the mode of sitmulated treatments with signalling regulators, low-temperature and drought stress. A homologous gene of protein phosphatase 2C was cloned from Zea mays roots for the first time, the state of the gene in genome, the expression characteristics, the physiology functions were studied in this thesis by Southern blot, Northern blot and transforming technology and so on. Researching is to explore the molecular mechanisms of stress-resistant of maize and other crops, and to privide the basis for the screening and utilizing of stress-tolerance resource. The main results were as follows: 1. According to the conserved motifs of plant protein phosphatase 2C gene, degenerate oligonucleotides were designed. A full cDNA sequence of PP2C gene from Zea mays roots was cloned by RT-PCR. It was named ZmPP2C and had 936 bp . The accession number is AY621066 in GenBank. 2. Genomic DNA was isolated from normal growth maize, and completely digested with three restriction endonucleases: BamH I, EcoR V, Sac I. Using N-sequence of ZmPP2C gene as probe, southern blot showed that the ZmPP2C gene was a low copy in the Zea mays genome, and there was probably a small PP2C gene family. 3. Total RNA was isolated form root, stem and leaf in maize. Using N-sequence of ZmPP2C gene as probe, Northern blot showed that the...