【作者】 刘可慧；

【导师】 易干军； 张秋明；

【作者基本信息】 湖南农业大学 ， 果树学， 2004， 硕士

【摘要】 沙田柚（Citrus grandis cv.shatianyou.）属芸香科（Rutaceac）柑桔属，（Aurantioodeac）果实风味甜美，颜色润泽。但种子数目较多，一定程度影响了果实的可食性和商品性。本试验以华中农业大学邓秀新教授提供的异源四倍体体细胞杂种为父本（异源四倍体粗柠檬（Citrus jambhiri Lush）与哈姆林甜橙（C.sinesis Osbeck）体细胞杂种，简称HR；酸柚（Sour orange）与粗柠檬（Rough lemon）体细胞杂种，简称SR：墨西哥来檬（C.aurantifolia Swingle）与伏令夏甜橙（C.sinensis Osbeck）体细胞杂种，简称KV），以二倍体沙田柚为母本进行有性杂交，获得瘪籽沙田柚。同时利用胚挽救技术，对杂种合子胚（在遗传上高度不育）进行早期离体培养，再经微芽嫁接等技术获得完整的三倍体杂种后代植株。 主要研究结果如下： （1）瘪籽沙田柚果实与对照果实比较发现：①瘪籽沙田柚种子败育十分明显（授以HR、SR和KV体细胞杂种花粉的果实，其瘪籽率分别为96.0％、89.3％和41.4％而对照瘪籽率仅为2.0％）；②瘪籽沙田柚单果重、果皮厚度、全糖、可溶性固形物含量等比对照略低，但无显著性差异；③瘪籽沙田柚的可食率、可滴定酸和维生素C含量略高于对照，但无明显差异。④比较适合于瘪籽沙田柚生产的授粉体细胞杂种是HR和SR。 （2）将正交试验设计运用到沙田柚的组织培养中，初步摸索出适合于沙田柚胚挽救的培养基配方。较适合于沙田柚幼胚直接成苗的培养基是：MT+GA₂1.0mg/L+蜂皇桨400mg/L+LH250mg/L（试材来自幼果）和MT+麦芽提取物400mg/L（试材来自成熟果）：不定芽增殖的培养基是：MT+BA0.5mg/L+NAA0.1mg/L+GA₃0.2mg/L；试管嫁接的培养基是：MT+BA0.5mg/L；不定根诱导的培养基是：MT+IBA3.0mg/L+NAA1.0mg/L+活性碳0.1％。 （3）经胚挽救试验共获得了杂种后代植株46株，其中沙田柚×HR杂交组合12株、沙田柚×SR杂交组合23株、沙田柚×KV杂交组合11株。随机选取3株杂种后代植株叶片经流式细胞仪进行倍性检测，有2株为三倍体植株。 （4）成熟果实中未完全败育的种子，在适宜的条件下进行离体培养，比能再生出三倍体植株。更多还原

【Abstract】 Shatian pomelo(Cilrus grandis var. shatianyou Hort)is a subtropical evengreen fruit known for its delicious flavor. While too many seeds in the fruit effects its edible character and commercial character in some extent. Alloteraploid somatic hybrid (provided by Pro,D.X.X) were cross-pollinated with pomelo for seed abortion. The hybrid embryo coming from the sexual hybridization (crossed between tetraploid and diploid) was highly sterile. While new triploid germplasm in Citrus were obtained, with the technique of embryo rescue. The main study results as follows:1 Comparing the abortive seed fruits with Con, I Seed of cross-pollinated fruits were significantly found to be abortive; II The weight, size, total sugar content, and solubility of solid substance of fruit were less than that of Con, but no significant difference ; III The percentage of edible, sour and Vc were higher than that of Con, but no significant difference as well; IV The suitable allotetraploid somatic hybrids for seed abortion were HR and SR2 It was a useful way to apply the orthogonal design in the pomelo tissue culture. The results showed that the optimal medium for embryo rescue was solid MT medium supplemented with GA31mgL-1, LH40mgL-1 and queen bee oar 500mgL-1or solid MT medium supplemented with ME400mgL-1; The optimal medium for adventitious shoots multiplication was solid MT medium supplemented with BA0.5mgL-1, NAA0.1mgL-1 and GA30.2mgL-1; The optimal medium for tube-grafting was solid MT medium supplemented with BA0.5mgL-1; And The optimal medium for adventitious roots induction was solid MT medium supplemented with IBA3.0mgL-1, NAA1.0mgL-1 and AC0.1%.3 A total of 46 plants from 3 different combinations were obtained.Randomly selected 3 of them and examined by the apparatus of PA-1, the results showed that 2 of them were triploid plants.4 Triploid plants could also be obtained from the seed abortive mature fruit under proper conditions.更多还原