Studies On The Optimization Of Regeneration In Peach And The Transformation Of Antisense PG Gene

Peach〔Prunuspersica (L.) Batsch.〕is an important fruit in the worldwide. China is a major producing country of peach, whose output ranks first in the world, and Gansu Province is the original and the most appropriate cultivation area of peach. Peach is a tipical respiratory climacteric fruit. Soon ripe fruits picked appeared the respiratory peak and flesh became soft rapid, which caused great inconvenience in transport and storage and it is a major problem of restraining the production in peach .While it is unsatisfactory that the common method such as cold storage usually leads to chilling injury of peach. Therefore, using plant genetic engineering technology can control fruit softening, and changing fundamentally storage and transport properties in peach has an important significance.In this study, Baifen peach often planted in Gansu Province was used as test materials, and then established a peach regeneration system. with Baifen peach stem segments as materials, based on the optimization genetic transformation system of peach, taking advantage of Agrobacterium mediated transformed into antisense PG gene and obtained transformed plants, and then tested by PCR. The main results obtained are as follows:1. With tender stem and matural fruit in different time of Baifen after flowering as explants, optimize the genetic transformation system. The best medium for induction was MS+1.0mg/L6-BA+0.5mg/LNAA, and induction frequency was up to 93.5%. After flowering 75d embryo can directly differentiate into seedlings, induction frequency was up to 91.5%. The shoots dipped its base with 100mg/L1IBA were shifted to 1/2MS medium without hormone to induce roots whose rate was up to 78%.2. Transform into the stem of Baifen peach by expressive vector pCAMBIA2300 carrying the antisense PG gene fragments and 35S promoter. Receptor materials precultivated for 3 days, were co-cultured for 60h by Agrobacterium bacteria (OD600 value of 0.4) after being disseminated, transferred into the differentiative medium MS +1.0 mg/L6-BA +0.5 mg / LNAA +50 mg / L km +250 mg / LCef to induce adventitious buds, and anti-km adventitious buds were placed in the 50mg/Lkm mediums of growth, proliferation and rooting. Ultimately the whole plants were obtained by The PCR test which showed that the antisense PG gene has been introduced into the plants.