【作者】 萨如拉；

【导师】 马庆；

【作者基本信息】 内蒙古农业大学 ， 作物遗传育种， 2006， 硕士

【摘要】 紫花苜蓿是重要的豆科牧草之一。由于它改良土壤,保持水土而引起人们的广泛关注。我国苜蓿品种资源非常丰富,但是可消化率较低。如何提高苜蓿的可消化率,改良它的品质。是目前育种工作者的首要任务。本文介绍了在紫花苜蓿木质素合成代谢过程的主要酶4CL(4-coumarate:CoA ligase, 4一香豆酸辅酶A连接酶)基因的克隆,反向插入植物表达载体,并利用基因枪转化法将反义4CL基因转入紫花苜蓿中的全过程,结果表明:基因枪转化法简单易行,但是对设备有特殊的要求,且转化率较低,适于瞬时表达和整合表达的研究,Bio-Rad基因枪转化苜蓿的瞬时表达率为50%。利用基因枪将目的基因导入紫花苜蓿子叶中,在卡那霉素浓度为70mg/L选择培养基上,经过35天的继代和分化2次选择及45天连续3次选择后分化再生成植株。PCR结果表明,目的基因已导入并整合到紫花苜蓿基因组中,基因枪2次轰击的转化频率显著高于1次轰击和三次轰击。并且建立了紫花苜蓿组织培养体系。更多还原

【Abstract】 Medicago.sativa is one of the most important forage species, due to its modified soil and conserve water and top soil..How to improve its utilization efficiency remains a problem for breeder.This study focuses on reducing its lignin content by manipulation of lignin biosynthesis enzyme gene.The aim gene was translated into the cotyledon of the alfalfa by the way of gene gun translation. As a result: the measure was easy and convenient, but there were many special requirements for the facility, and the race of the translation was low, it was fit to expressing instantaneously and conformal. The alfalfa of the race of instantaneous expression was 50 percent, the race of conformity expression is lower than 0.1 percent. The aim gene was translated into the cotyledon of the alfalfa by the way of gene gun, the concentration of Kanamycin Sulfate was 50 mg/l in the culture medium, the continued culture needed 35 days, polarization needed two choices, and needed continue three choices during the 45 days, then polarizated the new plant. The results of the PCR showed: the aim gene has been translated into the gene groups of the alfalfa, two bombardments is largely higher than one or three bombardments. Meanwhile, the system of tissue culture has been set up.更多还原