| In this study,7 hybrid combinations were designed using 5 seedless grapevine cultivars?Vitis vinifera?as female parents and 3 cold-resistant grapevine varieties?V.amurensis Rupr.or V.amurensis×V.vinifera?as male parents.Hybrid ovules and embryos were cultured in vitro and the ovule development medium was modified via embryo rescue.Four molecular markers were used to identify seedlessness and cold-resistance among hybrid parents and offspring at early stage.The main results were as follows:1.4341 berries,5079 ovules,1607 embryos and 537 hybrid offspring were obtained from 7 combinations using the embryo rescue technique.Among these crosses,Kunxiang Seedless×Shuangyou got the highest embryo development rate and seeding rate.49.37%and18.21%,respectively.The maximum seedlings,350 offspring were obtained from Ruby Seedless×Beichun.2.The results indicated that different female parents affected seedling rate of embryo rescue.For‘Shuangyou'as male parent and‘Blush Seedless',‘Thompson Seedless'and‘Kunxiang Seedless'as female parents,the embryo development rates were 39.00%,19.26%and 45.39%,and the seedling rates were 9.50%,8.15%and 16.31%,respectively;for‘Beibinghong'as male parent and‘Blush Seedless',‘Beauty Seedless'and‘Thompson Seedless'as female materials,the embryo development rates were 13.33%,10.34%and7.64%,and seedlings were 6.59%,1.48%and 1.91%,respectively.Different male parents also influenced on seedlings formation.‘Blush Seedless'בBeibinghong'and‘Blush Seedless'בShuangyou'had the embryo development rates 13.33%and 39.00%,seedling rates were6.59%and 9.50%,respectively;Thompson Seedless×Beibinghong and Thompson Seedless×Shuangyou were 7.64%and 19.26%of the embryo development rate,and 1.91%and8.15%of the seedling rates,respectively.3.The proportion of components in different medium for embryo development had affected the embryo and seedling development.Compared with ER,MM3 showed higher embryo development rate than ER.4.The role of amino acids in embryo rescue were compared between two combinations.For Kunxiang Seedless×Shuangyou,the percentage of embryo development in MM3 with asparagine,glutamine and serine were 47.86%,59.69%and 53.33%,respectively,all higher than the control?45.39%?;moreover,the MM3+2.5mmol·L-1 glutamine showed the highest embryo development rate?59.69%?;the basal medium with cysteine,glutamine and asparagines for ovules culture,got the seedling rates 18.67%,20.93%and 21.43%,respectively,all higher than that of CK?16.31%?.For Ruby Seedless×Beichun,the ovules in medium with Cysteine and Asparagines got the percentage of embryo development 47.13%and 55.71%,respectively,higher than the basal medium.The seedling rate in the medium containing asparagine,glutamine and serine?21.43%,22.22%and 18.82%,respectively?was superior to CK.Therefore,MM3+2.5mmol·L-1 glutamine or asparagine could improve the seedling rate of the two cross combinations.5.In the 83 hybrid offsprings from 6 cross combinations,there were 49 strains with specific bands of the molecular markers GSLP-569 and SCC8-1018 linked to the seedlessness and 55 strains with specific bands of the markers S241-717 for cold-resistance.Importantly,35 strains with both seedlessness and cold-resistance were identified in the study. |
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