农杆菌介导的小麦遗传转化研究
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摘要
遗传转化技术可以将外源DNA片段导入到受体基因组中,是进行遗传改良和功能基因研究的重要技术手段。农杆菌介导法是目前应用较为广泛的转化方法之一,但在小麦中,只有包括Bobwhite和扬麦158在内的少数几个基因型被成功用于农杆菌介导的小麦遗传转化中,为了拓宽小麦农杆菌转化的受体基因型范围,本研究对10个不同基因型的材料进行了转化实验。B31706和B31575的转化率明显高于对照材料Bobwhite
其中,B31706转化后再生率及总抗性苗愈伤比例均为最高,在受到农杆菌侵染损伤后再生效率下降幅度最小。B31575在可再生愈伤组织中的抗性苗愈伤比例最高,该基因型的分化能力最强,但是受到农杆菌侵染后再生率会大幅下降。同时,研究了不同载体对转化效率的影响,结果显示,转化效率在不同载体之间不存在明显差异。
The genetic transformation technique is the most important tool for research in genetic improvements and functionanl genomics. The method of Agrobacterium-mediated transformation is the most widely used in genetic transformation. To this day, only several genotypes such as Bobwhite and Yangmai158 have been used in transformation successfully. To widen the recipient genotypes, ten different genotypes were transformated by Agrobacterium-mediated In our study, transformation efficiency of B31706 and B31575 were significantly higher than Bobwhite. Moreover, regeneration rate and resistant plantlet rate of Agrobacterium-mediated genetic transformation in B 31706 immature embryos callus was the highest one among the genotypes. At mean time, the decline of regeneration rate after Agrobacterium co-cultivation of B31706 was less than others. B31575, the resistant plantlet rate of regeneration plants was higher than others. Though the Regeneration rate without transformation of B31575 was the highest, it was dropped after co-cultivation. In addition, the difference of transformation efficiency among vectors was studied. From the results, there was no significant difference among different vectors in transformation efficiency.
其中,B31706转化后再生率及总抗性苗愈伤比例均为最高,在受到农杆菌侵染损伤后再生效率下降幅度最小。B31575在可再生愈伤组织中的抗性苗愈伤比例最高,该基因型的分化能力最强,但是受到农杆菌侵染后再生率会大幅下降。同时,研究了不同载体对转化效率的影响,结果显示,转化效率在不同载体之间不存在明显差异。
The genetic transformation technique is the most important tool for research in genetic improvements and functionanl genomics. The method of Agrobacterium-mediated transformation is the most widely used in genetic transformation. To this day, only several genotypes such as Bobwhite and Yangmai158 have been used in transformation successfully. To widen the recipient genotypes, ten different genotypes were transformated by Agrobacterium-mediated In our study, transformation efficiency of B31706 and B31575 were significantly higher than Bobwhite. Moreover, regeneration rate and resistant plantlet rate of Agrobacterium-mediated genetic transformation in B 31706 immature embryos callus was the highest one among the genotypes. At mean time, the decline of regeneration rate after Agrobacterium co-cultivation of B31706 was less than others. B31575, the resistant plantlet rate of regeneration plants was higher than others. Though the Regeneration rate without transformation of B31575 was the highest, it was dropped after co-cultivation. In addition, the difference of transformation efficiency among vectors was studied. From the results, there was no significant difference among different vectors in transformation efficiency.
引文
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