Cloning And Analysis Of Cold Induced Gene From Volvariella Volvacea

Volvariella volvacea is an important edible fungus of the tropics and subtropics. With its delicious taste and nutritious qualities, V. volvacea has a long cultivation history in china. Like some tropical and subtropical plants, V. volvacea can not be stored in low temperature. When it is stored at 4℃, the mycelia of V. volvacea will autolyze quickly and the fruit bodies will liquefy and perish more easily than other mushrooms, which seriously restricts the production, currency, storage and export of V. volvacea , even the development in china and the world. Now, we do not know the detailed hypostasis and mechanism why the mushroom autolyzes in low temperature yet.In this paper, the change of gene expression of V. volvacea under cold stress condition was studied with molecular biology methods. At first, the gpd gene was cloned. In order to studying the relationship between gpd gene and V. volvacea cold shock response, the expression pattern of gpd was performed in low temperature. Meanwhile, the sequence of the gpd upstream promoter was amplified, which provided an important element for establishing the genetic transformation system of V. volvacea. Then, a cold full-length cDNA library of V. volvacea was established using SMART cDNA Library Construction kit. The cold induced gene fragments labeled with DIG, isolated by DDRT-PCR, were used to screen the cDNA library. Cold induced full-length genes were obtained. The 5'-flanking sequence was also isolated. The role of cold induced gene in cold shock response of V. volvacea was investigated through the sequence analysis. The study not only helps us find out the straw mushroom's autolyzing molecular hypostasis under cold condition, but also provides the foundation for breeding new variety and improving storage skills with genetic engineering of V. volvacea.1 Cloning and analysis of gpd from V. volvaceaThe degenerate oligonucleotide primers reported in article was synthesized for PCR amplification of genomic DNA. Sequencing of the amplification product revealed a fragment of 685 bp encoding an amino acid sequence corresponding to GPD. Two specific primers were designed on the basis of the 685 bp of V. volvacea gpd gene. Collaborated...