【作者】 潘学军；

【导师】 王跃进；

【作者基本信息】 西北农林科技大学 ， 果树学， 2005， 博士

【摘要】 无核葡萄是当今世界葡萄消费和育种的重要方向之一。无核葡萄的合子胚在发育过程中败育而不能发育成正常的种子,因而常规育种只能以无核品种作父本杂交选育无核葡萄品种,但后代中无核几率低(0～15.9%),育种效率低下。胚挽救技术可用于无核葡萄作母本的品种间杂交,后代无核性状比率高(45%～82%),育种效率高,但选育的欧洲葡萄无核品种的突出缺点是缺乏抗病性。国外已将该技术应用于无核葡萄与圆叶葡萄育种中,但又存在杂交障碍,成苗率极低的问题。中国葡萄属野生种抗病性强,与欧亚种无核品种杂交亲合性强。因此,利用胚挽救技术获得欧洲葡萄无核品种与中国葡萄属野生种杂交后代,以培育无核抗病葡萄新品种(系)是完全可行的。　本研究田间人工杂交38 个葡萄种间杂交组合和13 个种内杂交组合。在前期胚挽救初步研究的基础上,本试验用19 个以无核葡萄为母本的杂交组合为试材,进行了无核抗病葡萄胚挽救的进一步研究,主要取得了如下研究结果: 1.　通过田间杂交和胚挽救获得了51 个杂交组合的葡萄新种质2998 份。利用胚挽救技术获得19 个杂交组合幼苗814 株,其中13 个无核品种×中国葡萄属野生种(杂种)组合成苗257 株;常规田间杂交组合32 个,成苗2184 株,其中20 个中国葡萄属野生种(杂种)×无核品种组合成苗1606 株;　　2.　获得了适宜无核抗病葡萄即将败育的幼胚继续发育的最佳培养基:MM3+10mmol/L　甘氨酸;建立了其最优培养方式:固液双层培养;筛选到了适宜作胚挽救母本的欧洲葡萄无核品种是爱莫无核(Olmo　Seedless)、底来特(Delight)和火焰无核(Flame　Seedless)。适宜培养基、培养方式和胚挽救母本的最佳搭配使无核抗病葡萄胚发育率高达66.67%,成苗率达64.58%。获得了一种适宜诱导合子胚胚状体再生的培养基配方:ER 固体+1.0umol/L　BA。利用该培养基对底来特×北醇的合子胚诱导率达82.61%。　　3.　建立了葡萄胚挽救苗移栽技术体系:春季选择壮苗→强光锻炼1 周→无菌条件更多还原

【Abstract】 Seedless grapes are one of the principle objectives of grape breeding in the world. Zygotic embryos of seedless grapes can’t develop normal seeds because of abortion, so seedless grape cultivars are only taken as paternal parents in breeding new seedless grapes by conventional crossing, but the seedless progeny is only 0 to 15.9%. By embryo rescue technique, seedless×seedless crosses have produced 45% to 82% seedless offspring, but most of them are susceptible to diseases. Hybridization of seedless grapes and V. rotundifolia are performed by embryo rescue, but having a crossing barrier. The wild grapes whose origin of species in China are not only resistant to diseases but also easy to cross with V. vinifera. With the embryo rescue technique coming, it is a feasible way to cross between the seedless grapes (V. vinifera) and Chinese Wild Vitis in order to breed new seedless grape cultivars which are resistant to diseases. 38 interspecific crosses and 13 crosses of V. vinifera have been obtained by artificial pollination in the vineyard in this study. The embryo rescue technique was studied by using 19 crosses as materials that were taking the seedless grapes as maternal parents. The main results were as follows: 1. 2998 new grape seedlings of 51 crosses were obtained, including 2184 seedlings of 32 crosses by normal cross breeding and 814 grapes by embryo rescue breeding. In all of them, the seedlings from crosses between Chinese Wild Vitis (or hybrids) and seedless grapes were respectively 1606 of 20 crosses by pollinations and 257 grapes of 13 crosses through embryo rescue. 2. The optimum medium and the best suitable cultural way were obtained for abortive embryos development of diseases resistance seedless grapes. It was MM3+10mmol/L Gly under the solid and liquid dual layers medium. And the best seedless maternal parents of V. vinifera for embryo rescue were ‘Olmo seedless’, ‘Delight’, and ‘Flame seedless’. It could improve the embryo development ratio to 66.67%, the ratio of seedling emergence to 64.58%. The optimum medium for somatic embryogenesis was obtained. It was ER (solid medium)+1.0umol/L BA. The somatic embryos coming from zygotic embryos of ‘Delight’בBeichun ’were induced to 82.61% cultured on this medium. 3. Transplanting technique for grape seedlings by embryo rescue has been found in this research. Strong seedlings were selected in spring, then the seedlings were domesticated for 1 week in Erenmeyer flasks under intense light. The next stage was to transplant into pots filled pearlite to domesticate in culture house, in this stage the seedlings must be held moisture, sterilized and sprinkled nutrient solution. And then domesticated into nutrient soil in glasshouse. Finally the plants were transplanted into fields. 4. RAPD analysis was performed using 25 10-mer primers in 6 parent pairs, in order to find specific primers, which could amplify polymorphic fragments in paternal parent. Finally, 7 specific primers(S76、S111、S117、S183、S238、S264、S449) were obtained which could be used to differentiate the parents. In all of them, S183 and S238 were able to select true hybrids. 105 true hybrids coming from seedless cultivars (V. vinifera) ×Chinese wild Vitis(or its hybrid) by embryo rescue were selected by using both morphological characterization and RAPD analysis. The system of marker-assisted breeding for grapes was established. 202 embryo rescue seedlings were chose by both morphological characterization and marker-assisted breeding. 23 seedless grapes resistance to anthracnose, 16 seedless grapes resistance to dowdy mildew and 10 seedless grapes resistance to both diseases were selected. 5. The breeding techniques for seedless grape resistant to disease including pollination technique in fields, embryo rescue breeding technique and marker-assisted breeding technique were selected. A new seedless cultivar 00-3-1 with resistance to disease and a new seedless cultivar 00-2-7 were bred by using these techniques.更多还原