Study On Embryo Rescue Breeding Technology Of Seedless Cold-resistant Grape

Grape is one of the most widely planted fruit tree in the world.With the continuous development of social demand,seedless grapes occupy an increasingly important position in the market,and their development is of great value in terms of fresh food and Production of preserved fruit.This research starts from exploring the characteristics of the formation and development of seedless grapes,and uses seedless European grapes as pistillate parent,with certain cold-resistant ’Beibinghong’,’Beichun’,’Shuangyou’,and’Xinyu’,’Shine-Muscat’ and other varieties were used as paternal plant,and 14 hybrid combinations were prepared according to the characteristics of the paternal plant.The ovules were cultured in vitro using embryo rescue technology to obtain hybrid embryos,and the sampling period,days of embryo development and different parent plant genotypes were explored.The influence of factors such as seedless grape embryo development rate and Percentage of seedling emergency.And using molecular labeling technology,preliminary identification and testing of the seedless traits of its hybrid offspring,the results are as follows:1.The genotype of the stock plant has a certain impact on the efficiency of embryo rescue.The comparison of different maternal plant found that when ’Beibinghong’ is selected as the paternal plant and’Ruby Seedless’ is selected as the maternal plant material,the embryo development rate(39.9%)and seedling growth rate(21.5%)was the highest in the combination.The embryo development rate(10%)and seedling formation rate(5%)were compared with ’Cuibao without seed’ as the female parent and ’Yangguang Rose’ as the male parent.2.The determination of sampling time plays a certain role in the efficiency of embryo rescue.By comparing the efficiency of embryo rescue with that of sampling by stages,The best sampling time for ’Yuehong Seedless’ × ’SP740’,’Molesia Seedless’ × ’Xinyu’,’Ruby Seedless’ × ’Shine-Muscat’,’Flame Seedless’ × ’Munage’ and ’Cuibao Seedless’ × ’Xinyu’were 37 d,52 d,55 d,38 d and 56 d after pollination,respectively.3.Comparison of medium with different sucrose concentrations: by adding sucrose of different gradient into embryo germination medium,it was found that when sucrose concentration was 1%-1.5%,embryo rescue efficiency was the highest.4.IAA concentrations of different exogenous additives were added to compare the seedling formation rate,and the influences of the three combinations on embryo rescue breeding efficiency during embryo embryo culture were compared,and it was found that different hybrid combinations had different influences.Among them,the addition of 1.75 mg/L IAA in ’Flame Seedless’ × ’Shuangyou’ and ’Ruby Seedless’ × ’Beibinghong’ promoted the growth of the embryos,while IAA inhibited the development of the embryos in ’Huozhou hongyu’ × ’Beibinghong’.5.Detection of hybrid progeny by non-nucleus labeling: GLSP1-569 and SCF27-2000 probes are selected for preliminary detection of the parent’s non-nucleus traits,and then the progeny of the hybrid combination are tested,and the seedless plants are preliminarily determined through GLSP1-569 There were 90 strains,and SCF27-2000 detected the seedless traits of 15 strains.6.Detection of cold-resistant markers on hybrid progeny: S241-717 cold-resistant genes were selected to detect cold-resistant traits of the parents,and 18 cold-resistant lines were initially identified.Based on the above test results,this study configured 14 hybrid combinations,obtained 5807 hybrid fruits,obtained 427 hybrid lines,and detected 105 seedless embryos saved grape strains and cold resistant embryos saved through molecular marker detection.There are 18 of them.