New Germplasm Of Seedless And Disease Resistant Grape Was Creation By Embryo Rescue Technique

It is an important goal and task to breed seedless and disease-resistant grape over the world.Embryo rescue technique is a method to improve the breeding efficiency.In this paper,there were 16 combinations by using seedless varieties as female parent,hybrid with strong disease resistant Chinese wild grape varieties and their progeny as the male parent.The research in optimizing sampling time and medium to obtain hybrid progeny.To screen the seedless and disease-resistant trait preliminarily from the hybrid progeny to obtain new seedless and strong disease resistant germplasm resources.Results show that:1.European seedless grape was used as female parent to hybrid with‘Beichun’（Vitis vinifera L.×Vitis amurensis Rupr）,‘Shuangyou’（Vitis amurensis Rupr.）and high disease resistance cultivar‘Tangwei’（Vitis davidii）and‘Xuefeng’（Vitis davidii）as male parents.A total of 13341 hybrid fruits,8589 ovules inoculated and 1338 developed embryos were obtained.The average rate of embryo development was 15.59%,and 234 lines were grown into normal seedlings,the average rate of seedling formation was 2.73%.2.Among the 16 combinations,the embryo development rate,seedling formation rate and seedling formation of‘Ruby seedless’בTangwei’were significantly higher than those of other crosses,which were 40.17%,7.76%,respectively.Among the three combinations,when using‘shuangyou’as male parent to hybrid with,male parent‘Thompson Seedless’the seedling formation rate is highest with 3.08%.Among the five combinations using‘Beichun’as male parent hybrid with‘Flame seedless’as female parent the seedling formation rate is highestwith 1.98%.Among the three combinations,when using‘Tangwei’as male parent,hybrid with‘Ruby seedless’the seedling formation rate is highest with 7.76%.Compared with the reciprocal crosses,the seedling formation rate（2.01%）is higher of‘Flame seedless’בKunxiang seedless’.The highest seedling formation rate are 40 d and 38 d after pollination of‘Flame seedless’בBeichun’,‘Jingkejing’בBeichun’with 3.52%and 1.08%,respectively.3.To rescue embryos of‘Ruby seedless’בTangwei’combinations,the highest embryo development rate and seedling formation rate is 55.83%and 20%when MM3 medium adding3 mmol·L^-1 putroamine,followed embryo development rate and seedling formation rate was50.37%and 12.78%when adding 1 mmol·L^-1 spermidine.exogenous polyamine,the results show that adding results is best.4.Among four natural pollination varieties‘Ruby Seedless’,‘Beauty Seedless’and‘Heshi Seedless’when culturing embryos on E20A medium the embryo development rate which significant difference was 14.33%,4.50%and 10.61%,respectively.5.A total of 226 strains and 388 plants of five combinations‘Flames seedless’בKunxiang seedless’,‘Flames seedless’בBeichun’,‘Flames seedless’בTangwei’,‘Ruby seedless’בTangwei’,‘Ruby seedless’בXuefeng’are transplanted in the field.There are 200plants survived,the average survival rate was 63.72%.6.The seedless and disease-resistant traits of 226 hybrid lines of five combinations were detected by SCF27-2000 and GSLP1-569,S294-369 and S382-615,and OPY13-661.The result showed that 155 lines with seedless trait,79 lines with downy mildew resistant,43 lines with powdery mildew resistant,and 16 lines with seedless,downy mildew and powdery mildew resistant.In conclusion,among 16 combinations the highest embryo development rate and seedling formation rate is‘Ruby seedless’בTangwei’.By embryo rescue technique there are 16 lines with seedless,downy mildew and powdery mildew resistance screen from 226 hybrid progenies,which provide technical basis and new germplasm for seedless and disease-resistant breeding of grape.