Application And Breeding For Seedlessness With Cold-resistance Grape Germplasms Using Embryo Rescue

Although conventional hybridization is still the main method for breeding seedless grapes,the breeding efficiency of this traditional mode is low.The application of embryo rescue technique in hybrid breeding can significantly improve the efficiency.At present,most seedless grapevine varieties in the world are European cultivars(Vitis vinifera L.),which are not resistant to low temperature and the current situation severely limits the development of the grape industry.In this study,8 seedless grapevine varieties(Vitis vinifera L.)were selected as female parents,and 4 cold-resistant Chinese wild grapevine varieties(Vitis amurensis Rupr.or V.vinifera×V.amurensis)were used as male parents.A batch of hybrid seedlings were obtained by embryo rescue.New materials of seedless,cold-resistant plants were also selected by marker-assisted selection.Main results are as follows:1.In this study,13 hybrid combinations were designed,10712 berrises were obtained,11489 ovules were cultured in vitro using embryo rescue technique,2304 developing embryos and 794 hybrid plantlets were obtained eventually.The rate of developed embryos was 20.05%,and the seedling rate reached 6.91%.2.The maternal genotype has a great influence on the result of embryo rescue in this study.The embryo development rate and seedling rate of Ruby Seedless × Beibinghong were the highest(41.09% and 17.00%)among five hybrid combinations whose male parent was Beibinghong,using Flameless Seedless、Heshi Seedless、Ruby Seedless、Crimson Seedless and Black Monukka as female parents.The embryo development rate and seedling rate of Kunxiang Seedless × Xuelanhong were the highest(29.97% and 8.96%)among three hybrid combinations whose male parent was Xuelanhong,using Kunxiang Seedless、Heshi Seedless and Sultanina Rose as female parents.Between two hybrid combinations using Beichun as the male parent,the embryo development rate and seedling rate of Kunxiang Seedless × Beichun(28.13% and 8.57%)were higher than Perlette ×Beichun(21.96%,5.93%).The hybrid combinations using Ruby Seedless and Kunxiang Seedless as female parents have the better embryo rescue results than other combinations.Therefore,it is considered that Ruby Seedless and Kunxiang seedless are suitable as male parents in embryo rescue breeding of seedless grapes.Paternal genotype also affects embryo rescue results.3.740 individual plantlets(530 lines)of the 11 hybrid combinations in the experiment were transplanted in the greenhouse.Finally,570 individual plantlets(453 lines)survived.The survival rate reached 77.03%.4.302 hybrid lines were detected by using three molecular markers,SCF27-2000,GSLP1-569 linked to the seedlessness gene and S241-717 linked to the hardiness gene derived from V.amurensis.Three seedless lines were preliminarily identified using the marker SCF27-2000,70 seedless lines were preliminarily identified using the marker GSLP1-569,and 151 cold-resistant lines were preliminarily identified using the marker S241-717.37 seedless and cold-resistant hybrid lines were preliminarily identified using both GSLP1-569 and S241-717.