Using FOX Hunting System To Screen Genes Which Related With Salt Stress In Puccinellia Tenuifolra

Puccinellia tenuifolra can widely survive in highly saline soils of northeast plain of China, it can naturally grow at foul saline-alkali soil, even around alkaline earth. This research uses a novel functional genes screening technology-FOX hunting system to mine potential genes resources which related with salt stress tolerance in yeast InVscI firstly. We constructed Puccinellia tenuifolra partial full-length pAUR101-cDNA gene library, then transformed to yeast InVscI and analyzed. We wanted to screen some functional genes which related with abiotic stress of Puccinellia tenuifolra full-length pAUR101-cDNA yeast library. Puccinellia tenuifolra full-length pAUR101-cDNA yeast library was initially screened 56 transgenic yeasts under NaCl salt stress, then confirmed 15 ones to proceed to study. The 15 transgenic yeasts were analyzed with salt-stress of NaCl、NaHC03 and drought-stress of Mannitol. We found 15 transgenic yeasts exhibit differently tolerance of stress, but they grown better than contrast clearly.Puccinellia tenuifolra cDNAs were identified by PCR amplification from 10 transgenic yeasts in 15 ones, then analyzed sequence homology after DNA sequencing. Expect transgenic yeast Y011, novel small protein encoding genes from 9 transgenic yeasts were discovered, and their functions remain unknown.After sequence homology analyzed the gene from transgenic yeasts Y011, we detected the gene full length is 1162bp (including full length of Open Reading Frame is 756), it encoded 252 amino acid. The protein that gene encoded was gamma type tonoplast intrinsic proteins (TIP), and exhibited high sequence homology of tonoplast intrinsic proteins from Triticum aestivum, Hordeum vulgare, Phyllostachys edulis and Zea mays.