【作者】 赵维峰；

【导师】 孙光明；

【作者基本信息】 华南热带农业大学 ， 果树学， 2005， 硕士

【摘要】 毛叶枣(Zizyphus mauritiana Lam.)又名台湾青枣,具有高产、优质等特点,被誉为果中佳品。但是,毛叶枣也有一些缺陷如果实甜度较低,果肉中含糖仅为12%-14%,抗寒性不强等等。而北方枣恰好可以弥补这些缺陷,北方枣的含糖量一般为30%以上,抗寒性要比毛叶枣强很多,因此将二者的优势结合育成的新品种,不仅种植面积可以扩大而且在品质上将会有更大的竞争优势。目前,枣树育种工作多采用选优的方法进行,未形成种质创新。本研究采用原生质体融合技术,以期通过电融合诱导枣树不同种间产生融合杂种,增加杂交后代遗传组成的多样性,通过培育再生植株,结合常规育种方法,选出优良的新品种。 本研究探讨了毛叶枣及冬枣的原生质体分离与纯化条件,毛叶枣与冬枣融合的最适电融合参数,为以后应用体细胞融合这一技术创造优异的毛叶枣体细胞杂种材料提供可用的技术路线。获得的主要结果如下: 1.取组培苗叶片和悬浮培养物作为分离原生质体研究对象,比较了二者的分离效果,结果反映出:叶片分离出的原生质体活性显著高于悬浮培养物,但是产量上又低于悬浮培养物分离出的原生质体。 2.获得了分离枣原生质体酶液的最佳浓度:毛叶枣酶解所需酶液最佳浓度为纤维素酶10g/L+离析酶4g/L+甘露醇0.7mol/L;冬枣酶解所需酶液最佳浓度为纤维素酶15g/L+离析酶4g/L+甘露醇0.7mol/L。 3.对毛叶枣和冬枣分离原生质体所需的离析酶浓度进行了研究。当离析酶浓度为5g/L时,毛叶枣分离的原生质体产量最高,而冬枣为4g/L;当离析酶浓度为4g/L时,毛叶枣和冬枣分离的原生质体活性都达到最高。综合考虑产量和活性两个方面,离析酶浓度都可以取4g/L。 4.毛叶枣和冬枣两个参试品种,都在14-16小时达到最佳酶解效果。 5.本研究的结果表明交变电场强度为100 V/cm,交变电场作用时间50 s,直流脉冲强度1000 V/cm,直流脉冲时间50μs,枣树原生质体融合效果较好。 6.冬枣与毛叶枣的融合比例为1:2,融合密度为0.5×10~5个/ml,可以使融合率达到18%。更多还原

【Abstract】 Ziziyphus mauritiana Lam. is known for high yield and good quality. But its content of soluble sugar and resistance to coldness are worse than Ziziphus jujuba Mill.. The hybrid induced by the combination of their advantages can improve the quality of fruit, and increase the cultivable area. This is the first time to establish Ziziphus jujuba protoplast electrofusion procedure in order to create new germplasm and breed new cultivars.In this study protoplast isolation and purification methods were developed and optimum parameters of protoplast fusion for Ziziyphus mauritiana Lam. and Ziziphus jujuba Mill.were determined. The main results as follows:1. Leaves of in vitro plants and suspension culture was studied. The results showed viability of protoplasts isolated from leaves was significantly higher than that of suspension culture. But for yield of protoplasts the result was just the opposition.2. The optimal enzyme solution used for protoplasts isolation for Ziziyphus mauritiana Lam was Cellulase10g/L+Macerozyme 4g/L+Mannital 0.7mol/L. and The optimal enzyme solution used for protoplasts isolation for Ziziphus jujuba Mill was Cellulase15g/L+Macerozyme 4g/L+Mannital 0.7mol/L.3. Yield of Ziziyphus mauritiana Lam. protoplast was highest when 5g/L Macyorozyme was used. The highest yield of Ziziphus jujuba Mill protoplast was obtained when 4g/L Macyorozyme was used. Viabilty of Ziziyphus mauritiana Lam. and Ziziphus jujuba Mill.protopalsts were highest when 4g/L Macyorozyme was used. So 4g/L Macyorozyme should be the optimal.4. Yield and vigor of Ziziyphus mauritiana Lam. and Ziziphus jujuba Mill.protoplasts were highest when enzyme was used for 14—16hours.5. The optimum electric parameters were alternating current (AC) 100V/cm,duration 50sdirect current (DC ) intensity 1000 V/cm,50μs per hours for electrofusion in Ziziphus jujube.6. When the fusion proportion of Ziziyphus mauritiana Lam. and Ziziphus jujuba Mill. Protoplast wasl to 2 and fusion density was 5×104/ml, 18 percent of pair fusion was obtained.更多还原