【作者】 崔妍；

【导师】 朱宝成；

【作者基本信息】 河北大学 ， 植物学， 2004， 硕士

【摘要】 水稻是世界上最重要的禾谷类作物之一，但是在生产中由于病虫害和不良气候与逆境的影响危害，水稻的高产、稳产受到了严重的制约。水稻的抗性转基因研究在过去的十几年中取得了重大的进展。 以水稻“中作8593”的成熟胚作为材料，在LS+2,4-D2.0培养基上诱导和继代培养愈伤组织。选择生长状态良好的胚性愈伤组织作为受体组织，利用基因枪，将含有来自大麦的胚胎发生晚期丰富蛋白LEA基因导入水稻愈伤组织细胞。将转化后的愈伤组织接种到CARS+Bialaphos8.0和KPRS+Bialaphos8.0的选择培养基中进行选择培养，连续选择2次，将抗性愈伤组织转接到附加KT1.0+ZT1.0+BA1.0+NAA0.1的MS培养基上进行再生培养，两个月后得到了转化再生苗。 提取转化植株的总DNA进行PCR扩增，检测到转化植株中含有目的基因片段。电泳结果显示转化植株的电泳条带大小与质粒的电泳条带大小相同，未转化植株在相应的位置没有条带出现。 在苗期对转化植株进行了抗性生理检测。在盐胁迫实验中，使用0.1％、0.3％、0.5％、0.7％、1％五个浓度的NaCl溶液持续胁迫处理转化植株和对照植株。转化植株出现萎蔫死亡的时间较对照推迟了10天以上；在持续胁迫处理的情况下，转化植株的全部萎蔫死亡的时间较对照推迟了15天以上。由此明显地观察到转化植株对不同浓度的盐胁迫都较对照表现出良好的抗性性状。 在水杨酸诱导胁迫实验中，使用转化植株依然表现出较对照具有较强的抗性。但是对照的POD活性高峰点仅次于No.2。 综合实验结果表明，LEA3基因导入并整合到水稻基因组；与对照相比转化植株抗旱耐盐性明显增强，并且在受到盐胁迫时，转化株萎蔫症状出现晚，受害程度轻，到最终死亡的间隔时间也较长。在水杨酸诱导的渗透胁迫下转化植株表现也出良好的抗性。更多还原

【Abstract】 Rice is one of the most important graminaceous crops. Every year virus and pests cause serious damage to rice production. Research on resistance gene engineering of rice has great improvement in past ten years. Recently reports for resistance-transgenic rice are mainly about virus and pests resistance.Chosing mature embryo of "Zhongzuo8593" as material to inducte callus on LS+2,4-D 2.0. Calli of rice were transformed by particle bombardment with plasmid pBY520 containing late embryogenesis abundant protein gene, LEA3, from barley (Hordeum vulgare L.). The bombarded calli were transformed to media CARS+Bialaphos8.0 and KPRS+Bialaphos8.0 for selection. After 2 times selected culture by Bialaphos, three lines resistant calli were obtained. The resistant calli were transformed to LS media with KT1.0mg/l and ZT1.0mg/l and BA1.0mg/l and NAA0.lmg/1 and got three regeneration lines planets two months later.Positive plants were selected quickly by PCR analysis, and the electrophoresis result showed that foreign LEA3 gene integrated into rice genome.Under 0.!% 0.3%0.5% 0.7% 1%NaCl salt stress conditions, positive plants began wilting were late than contradistinguish plants for l0days. All positive plants died later than all contradistinguish plants for 15days. The results showed that the salt tolerance of transgenic rice was increased, the development of damage symptoms (wilting) caused by salt stress delayed.Under 0.002mol/1 0.004mol/1 0.008mol/1 0.0 lmol/1 salicylic acid solution stress conditions, positive plants showed strong OA ability .Though the POD activity of the control is taller than those of No.3 and No.4, but No.2 has the strongest ability to OA. The result of physiological analysis confirmed the positive plants have strong resistance to salt and water deficit than the controls.更多还原