The Transformation Of’Manicure Finger’ Grapevine With PGIP Gene

An effective adventitious bud regeneration system and somatic embrogenesis system of ’Manicure Finger’ grapevine was developed in this study. Agrobacterium-mediated transformation was adopted to introduce the PGIP gene into’Manicure Finger’grapevine. The results were listed as follows:1. The effects of different explant types (leaves, petioles and stems), hormones and their concentration combinations as well as dark-treated periods on the regeneration of the grape ’Manicure Finger’ were investigated. The results showed that adventitious shoots regenerated from the leaves, petioles and stems, and the regenerate rate of the petiole is maximum. TDZ enhanced regeneration of leaves better than BA. IBA enhanced regeneration of leaves better than NAA and2,4-D. Petiole explants displayed the highest rate of regeneration after3weeks dark treatment. The explants growing on followed media produced the most efficient regeneration:60.71%regeneration efficiency and5.65shoots per explant for leaves explants growing on MS+1.0mg-L-1TDZ+0.1mg-L-1IBA medium;93.10%generation efficiency and5.91shoots per explant for the petioles on the same medium; and47.37%generation rate and5.00shoots per explant for stems on the MS+1.0mg-L-1TDZ+0.2mg-L-1IBA medium. Adventitious shoots were rooted on3/4MS+0.35mg-L-1IBA medium, and the rooted plantlets were survived after acclimatization and then were transplanted to the greenhouse.2. Somatic embryogenesis and plant regeneration from anthers and gynoecia in ’Manicure Finger’ were performed. Anthers and gynoecia of’Manicure Finger’ at uninucleate stage were collected and cultured on MS (Murashige and Skoog,1962) basal medium supplemented with6-BA and2,4-D at various concentrations to induce embryogenic callus, at25±2℃, in the dark. After3months, embryogeneic callus was produced from primary callus, at a rate of3.51%in anthers, and4.55%in gynoecia. Somatic embryos were differentiated from embryogenic callus transferred onto MS basal medium supplemented with1.0mg·L-16-BA. The isolated somatic embryos at torpedo stage cultured on growth-regulator-free mediums displayed a germination rate of 83.89±8.39%. And, plantlets cultured on growth mediums presented a normal grown-up rate of72.33±12.21%. The embryogenic culture could be maintained for as long as one and a half years by repeatedly recallusing the embryos on the embryogenic callus induction medium, which could ensure sufficient materials for genetic transformation.3. Nine’Manicure Finger’grapevine plants lines with hyg resistance were obtained by optimizing Agrobacterium mediated transformation. There were7PCR positive plants and3RT-PCR positive plants. It proved that introduced PGIP gene could express in’Manicure Finger’ grapevine.