【摘要】 【目的】提高无核葡萄胚挽救育种效率。【方法】以12个无核葡萄品种或株系的自然授粉胚珠和6个杂交组合的胚珠为试材,研究不同配方的培养基、不同培养基相态、低温处理、光照和暗培养对无核葡萄的胚发育率的影响。同时,研究花后不同时间取样,摘取果粒取出胚珠培养的适宜时间。【结果】在供试的8种培养基中,Nitsch+GA3 0.5mg·L-1+IAA 1.5mg·L-1和MM3培养基最适宜于无核葡萄离体幼胚的发育,其次是ER培养基。在供试的固相、液相和固液双相的3种相态培养基中,以固液双相培养基进行无核葡萄品种底来特离体胚珠培养为最好。在胚珠离体培养阶段进行低温培养,降低了胚发育率。暗培养有利于幼胚的发育,可提高胚发育率。不同无核葡萄品种和组合在花后摘取果粒,取出胚珠进行离体胚挽救的适宜时间不同,无核白为35d、森田尼无核为40d、火焰无核为40d、底来特为60d、黎明无核为55d、无核紫为70d、优无核为50d、皇家秋天为70d、奇妙无核为60d、奥迪亚无核为40d、红宝石无核×贵妃玫瑰为70d。【结论】本研究中不同无核葡萄品种进行胚挽救宜选择Nitsch+GA3 0.5mg·L-1+IAA 1.5mg·L-1、MM3或ER培养基,在(25±2)℃常温下进行固液双相暗培养,有助于提高胚挽救效率。更多还原

【Abstract】 【Objective】The objective of this study was to improve breeding efficiency of seedless grape through embryo rescue in vitro, to investigate the effects of various correlative factors on embryo development in vitro. 【Method】 The open pollination ovules from 12 seedless grape varieties (lines), including Thompson Seedless, Black Monukka, Centennial Seedless, Delight, Flame Seedless, Superior Seedless, Autumn Royal, Fantasy Seedless, Dawn Seedless, Otilia Seedless, 00-3-1, Blush Seedless and hybrid ovules of 6 cross combinations Ruby Seedless×Guifeimeigui, Ruby Seedless×Xinyu, Delight×Kyoho, Xinyu×Kyoho, Emerald Seedless×Taishan-2, Delight ×Jiangxi-2 were used to investigate the effect of different ovule development media, different medium phases, low temperature, and light/dark condition on the proportion of embryo development in vitro, and evaluate the optimal sampling dates after bloom for picking the young berries, culturing the excised ovules in vitro in different cultivars and crosses. 【Result】 The results indicated that among the 8 media, Nitsch medium supplemented with 0.5 mg·L-1 GA3 and 1.5 mg·L-1 IAA, MM3 medium were found to be most suitable for embryo development in vitro of seedless grapes, and the next was ER medium. Among the solid, liquid and solid-liquid double phase media, solid-liquid double phase medium had the most positive effect on embryo development of middling ovules of seedless grape cv. Delight, yet three phase media showed no significant different effect on embryo development of bigger ovules of grape line 00-3-1. The results also showed that low temperature at the stage of ovule culture in vitro had a negative effect on embryos development. Dark culture had a positive effect on embryo development in ovule cultures in vitro. The optimal sampling dates after bloom of culturing ovules in vitro dissected from berries varied among different cultivars and crosses, the optimal sampling dates was 35 day after bloom (DAB) in Thompson Seedless, 40 DAB in Centennial Seedless, 40 DAB in Flame Seedless, 60 DAB in Delight, 55 DAB in Dawn Seedless, 70 DAB in Black Monukka, 50 DAB in Superior Seedless, 70 DAB in Autumn Royal, 60 DAB in Fantasy Seedless, 40 DAB in Otilia Seedless, 70 DAB in Ruby Seedless×Guifeimeigui, respectively. 【Conclusion】 The results indicated that Nitsch medium supplemented with 0.5 mg·L-1 GA3 and 1.5 mg·L-1 IAA, MM3 and ER media were suitable for embryo rescue in vitro of seedless grapes. The culture environment of room temperature (25±2)℃, solid-liquid double phase medium and dark culture had a positive effect on improvement of embryo rescue efficiency.更多还原