【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， 2002， 硕士

【摘要】 2000年5月～2001年5月通过田间人工杂交，共获得以无核葡萄作母本中国野生葡萄山葡萄(黑龙江实生、左山74-1-326)、蘡薁葡萄(泰山-2)、燕山葡萄、秦岭葡萄(平利-5)和北醇作父本的种间组合17个和无核葡萄品种间组合6个及自交组合2个。然后离体培养上述25个组合的胚珠，对影响无核抗病葡萄胚挽救的取样时期、基本培养基、培养方式、珠被处理方式、幼胚萌发蔗糖浓度等因素进行了研究。主要取得了以下研究结果： 1．获得了一批杂交后代的胚挽救幼苗 通过胚挽救技术，以无核葡萄为母本的23个杂交组合共获得了764株幼苗，其中无核×抗病组合17个，成苗377株，4个无核×无核组合和2个自交组合，成苗387株。获得的无核抗病杂交后代包括底来特×北醇117株，爱莫无核×北醇107株，底来特×燕山葡萄32株，底来特×黑龙江实生29株，底来特×平利-5 28株，火焰无核×北醇27株，爱莫无核×泰山-215株，森田尼无核×北醇、爱莫无核×左山74-1-326各3株，森田尼无核×左山74-1-326 2株，京可晶×左山74-1-326、火焰无核×燕山葡萄各1株。不同的杂交组合胚挽救成苗的结果差异较大。炼苗移栽102株，成功定植于大田86株，成活率达84.31％。 2．筛选出了最适宜无核抗病葡萄胚挽救的基本培养基 筛选出的适宜胚挽救的培养基是改良ER培养基。胚发育阶段采用附加6％蔗糖的改良ER液体培养基，胚萌发及成苗阶段采用附加1.0～1.5％蔗糖的Wood Plant固体培养基，并附加0.2mg／LBA。 3．确定了各杂交组合的最佳取材时期 不同杂交组合胚挽救的最佳取材时期不同，本研究的不同组合是在授粉后40～52天之间。具体表现为：底来特×北醇、底来特×黑龙江实生、底来特×红地球、底来特×爱莫无核、底来特自交、爱莫无核×北醇及爱莫无核×底来特是授粉后48天，底来特×平利-5是授粉后52天，底来特×燕山葡萄是授粉后49天，火焰无核×北醇是授粉后40天。 4．建立了适宜无核抗病葡萄胚挽救的应用操作程序 本研究获得的无核抗病葡萄胚挽救的操作方法可分为三个培养阶段。首先将杂交胚珠取出接种在胚发育培养基上进行胚珠内幼胚的培养8周，挽救其中的幼胚以防败育，使幼胚在离体条件下进一步得到发育；然后解剖镜下无菌剥取裸胚转移到胚萌发培养基上诱导胚萌发；最后将萌发10～15天的胚转移到成苗培养基上使之生根直接成苗。 5．温室炼苗的关键环节是无菌条件下将胚挽救苗移栽到以珍珠岩为基质的营养钵中，杀菌剂的浇灌及湿度的调控。更多还原

【Abstract】 17 pollinations between seedless grapes andChinese wild grapes (K amurensis (Heilongjiang seedling,Zuoshan741-326) V.thunbergii(raKhanr2),V.yeshanensis,V. qinlingensis (Pingli-5) and Beichun (a specific hybrid of V.amurensisJ), 6 pollinations between seedless grapes, 2 crosses of self-pollinated seedless grapes were carried out in May of 2000 and May of 2001.Fertilized ovules of 25 crosses above were cultured in vitro to study effect of harvest time, basic medium, embryo culture styles, ovule excision on embryo rescue and sugar concentrations on embryo germination.The results were as follows:1. 764 embryo rescue plants were obtained from 23 crosses, including 377 plants from 17 hybrids of seedless grapes x Chinese wild grapes, 387 plants fr om 4 hybrids of seedless grapes x seedless grapes and 2 crosses of self-pollination. Progenies of hybrids of seedless grapesxChinese wild grapes included 117 new plants from DelightxBeichun,107 plants from Olmo SeedlessxBeichun, 32 plants from Delightx V.yeshanensis, 29 Plants from DelightxHeilongjiang seedling, 28 plants from DelightxPingli-5, 27 Plants from Flame SeedlessxBeichun, 15 Plants from Olmo SeedlessxTaishan-2, 3 plants from Centennial SeedlessxBeichun or Olmo SeedlessxZuoshan 74-1-326, 2 plants from Centennial SeedlessxZuoshan 74-1-326, 1 plant from JingkejingxZuoshan74-l-326 or Flame SeedlessxV.yeshanensis. The percentage of recovered embryos differed significantly among crosses. 102 recovered plants were transfered and 86 plants were established in the vineyard and survival percentage was 84.31%.2. The suitable basic media were selected for embryo rescue. The best medium for embryo development was modified liquid ER medium plus 6% sucrose; the best medium for embryo germination and plant establi shment was solid Wood Plant (WP) medium plus 1.0-1.5% sucrose and 0.2mg/L BA.3. Different harvest time was investigated to determine the best for embryo rescue. It was found that the best time was different among crosses. Harvest time of 48 days after pollination was the best for DelightxBeichun, DelightxHeilongjiang seedling, DelightxRed Globe, DelightxOlmo Seedless, self-pollinated Delight, Olmo SeedlessxBeichun and OlmoSeedlessxDelight; 52 days after pollination for DelightxPingli-5; 49 days after pollination for Delightx V.yeshanensis and 40 days after pollination for Flame SeedlessxBeichun.4. Simple procedure of embryo rescue was established, which was classified into three stages. First, ovules were excised from seedless fruits to recover embryos and in-vitro ovules were cultured on medium for 8 weeks till embryo became mature. After that, the developed embryos were excised in sterile under anatomic microscope and were transferred into another medium for embryo germination. 10~15days after germination, the recovered embryos were transferred into WP medium to establish seedlings.5. The key way to establish seedlings was that the domesticated plants should be transferred in sterile from Erenmeyer flasks to pots as matrix of pearlite, with irrigation of fungicide solution and with moisture control.更多还原