【作者】 唐冬梅；

【导师】 王跃进； 张剑侠；

【作者基本信息】 西北农林科技大学 ， 果树学， 2005， 硕士

【摘要】 2002 年5 月～2003 年5 月,实施无核葡萄作母本、抗病的中国野生葡萄作父本的杂交组合7 个,无核×无核组合6 个,同时对影响胚挽救效果的取样时期、培养基、培养条件等因素进行了研究,并对胚挽救获得的幼苗进行了移栽定植,2004年以森田尼无核、无核白、克瑞森无核为材料,用不同浓度的植物生长调节物质矮壮素、BA、腐胺和乙烯利并添加矿物元素,在不同时期处理果穗,以促进无核葡萄胚珠的发育,提高胚挽救成苗率。主要取得的研究成果如下:　1.获得一批无核抗病葡萄杂交胚挽救幼苗　运用胚挽救技术,以无核葡萄作母本,中国野生葡萄和欧洲无核葡萄作父本,共进行了13 个组合,获得322 株杂交苗。其中无核葡萄×中国野生葡萄组合7 个,成苗257 株;无核葡萄×无核葡萄组合6 个,成苗65 株。无核抗病杂交后代包括:底来特×江西-3　　17 株、底来特×江西-2　　18 株、底来特×双优114 株、爱莫无核×泰山-2　　32 株、爱莫无核×甘肃-91　　26 株、火焰无核×平利-5　　15 株、火焰无核×黑龙江实生35 株。不同的杂交结合胚挽救成苗结果差异较大,以底来特、爱莫无核和赫什无核作母本的组合成苗率较高。其中底来特×双优的胚发育率最高,达24.48%,成苗率10.86%。共移栽杂交株系255 个1471 株,出圃211 个株系601 株,温室移栽成活率和苗圃成活率分别为67.0%和83.9%,出圃杂交苗已于2005年春定植大田。　2.优化了胚挽救技术体系　比较了不同杂交组合胚挽救取材时间对成苗的影响。底来特×江西-3　58 天取材的胚发育率高于50 天;赫什无核×火焰无核54 天和58 天取材差异不显著;无核紫×森田尼无核54 天取材的效果优于52 天,底来特×双优51 天取材的优于47天,火焰无核×黑龙江实生50 天取材的优于46 天。研究表明在一定范围内,较晚时期取材接种胚珠,能得到较高的成苗率。对胚挽救的培养基和培养环境条件进行对比,结果表明:在胚珠发育阶段,胚发育宜使用ER 液体培养基,用培养皿作培养容器,进行暗培养,添加2～5　m　mol/L 的腐胺有助于底来特胚的发育;在胚萌发阶段宜使用1/2MS 附加IBA 或IAA　0.1～0.2　mg/L 的固体培养基,使用试管作培养容器,光照培养。胚接种在上述萌发培养基中,萌发后能直接成苗,省去转接成苗培养基的步骤,简化培养程序。更多还原

【Abstract】 7 crosses between seedless grapes and Chinese wild Vitis ,6 crosses between seedless grapes were carried out from May of 2002 to May of 2003 in order to obtain new grape germplasms of seedless and disease resistance.The effects of sampling dates,basic medium,culture environment on embryo rescue were studied.The plantlets converted from embryo rescue were transplanted and established on the vineyard .In 2004,plant growth regulators:chlormequat,BAP,putrescine,ethephon,appending nourish-ment ,were evaluated at different concentrations and times by spraying berries for promotion of ovules development and plants convertion of three stenospermocarpic grape cultivars(Vitis vinifera L.):“Centennial Seedless”,“Thompson Seedless”and “Crimson Seedless”.The results were as follows: 1. 322 embryo rescue seedlings were obtained from 13 crosses,including 257 seedlings from 7 crosses of seedless grapes×Chinese wild Vitis,65 seedlings from 6 crosses of seedless grapes×seedless grapes.Progenies of hybrids of seedless grapes×Chinese wild vitis including 17 new seedlings from Delight(V.vinifera L.) ×Jiangxi-3(V.hancockii Hance),18 seedlings from Delight ×Jiangxi-2 (V.romanetii Roman.),114 seedlings from Delight ×Shuangyou(V.amurensis Rupr.),32 seedlings from Olmo Seedless(V.vinifera L.) ×Taishan-2(V.adstricta Hance), 26 seedlings from Olmo Seedless ×Gansu-91 (V.piasezkii Maxim.), 15 seedlings from Flame Seedless(V.vinifera L.) ×Pinli-5 (V.qinlingensis P.C.He), 35 seedlings from Flame Seedless ×Heilongjiang Seedling (V.amurensis Rupr.).The percentage of seedlings convertion of embryo rescue differed significantly among crosses. The percentages of seedlings convertion in the crosses of regarding Delight,Olmo Seedless and K????? X????y as female parent seperately were greater.Of Delight ×Shuangyou, the percentage(24.48%) of embryo development was greatest, the percenage of seedling obtainment was 10.86%. 1471 plants of 255 strains were transplanted,601 plants of 211 strains survived from nursery,the percentage surviving from greenhouse and nursery were 67.0% and 83.9% respectively.The survived plants from nursery were established on the vineyard in spring of 2005. 2. The system of embryo rescue was optimized.The effect of different sampling dates to embryos development in a few crosses was studied.Of Delight×Jiangxi-3,the greater percentage of embryo development was obtained by cultured in 58 day than in 50 day after pollination.Of K????? X????y ×Flame Seedless,the effect on embryos development is similar to cultured in 58 and 54 day after pollination.Of Black Monukka ×Centennial Seedless ,the result of embryos development in 54 day is better than in 52 day.Of Flame Seedless ×Heilongjiang Seedling,the result in 50 day is better than 46 day.It was showed that the greater percentage of embryo development was obtained by cultured in the later date during the limited period.The effect of medium and culture environment to embryo rescue was studied.As a result,The greater percentage of embryo development was obtained on ER liquid medium in polycarbonate petri dishes cultured in dark during the stage of ovules development.The addition of 2～5 m mol/L putrescine to ER medium promoted the ovules development of “Delight”.While during the stage of embryos germination,1/2MS solid medium supplemented IBA or IAA 0.1～0.2 mg/L with test tubes using as culture vessels and in light environment was preferable.Embryos transferred in germination medium could be converted into plants directly.This was an advanced system because it eliminated the step of transferring convertion medium and simplifed the embryo rescue procedure. 3.The preliminary method was investigated to promote stenospermic grape ovules development with plant growth regulator.The studies were conducted on “Centennial Seedless”,“Thompson Seedless”and “Crimson Seedless”.Treatments consisted of four plant growth regulators(chlormequat, BAP, putrescine, ethephon) appending nourish-ment applied at two different concentrations each,once and twice applied each and three controls,for a total of 19 treatments.The plant growth regulators had no effect on ovules number of per berry.No evidence indicated that the nourishment could stimulate ovule development. The ovule weight and percentage of ovules upper grade 2 of“Centennial Seedless”,“Thompson Seedless”and “Crimson Seedless”were increased by sigle applied at two weeks pre-bloom and double applied at pre-bloom and post-bloom of 100～1000 mg/L chlormequat.With the treatment of 1000 mg/L chlormequat double applied,the weight and big size ovules percentage increased signifinantly compared to controls in maturation,moreover,the increasing was observed in 50 and 60 day postbloom.The increasing was also significant in maturation and 50 day postbloom of“Crimson Seedless”compared to control.The effect of 100 mg/L BA and 20 mg/L putrescine showed that ovules development of“Centennial Seedless”,“Crimson Seedless”were promoted.Ethephon had no effect on development of ovules.Treatment above increased the percentage of developed embryos by means of promoting the development of ovules,as the result,raised the percentage of plant convertion.The ovules were biger,the percentage of embryo development was greater.Moreover,the embryos were biger,the percentage of embryo germination was greater,the embryos of primarier than globular stage didn’t germinate.更多还原