小油桐外植体的KN1基因遗传转化及其超量表达的转基因植株

Genetic Transformation of Explant with KN1 Gene and Its Overexpression on Transgenic Plant in Jatropha Curcas L.

本研究采用农杆菌介导法将KN1基因遗传转化小油桐,并获得了转基因植株。在研究中分析了农杆菌菌液菌液的浓度、侵染时间和外植体的大小对遗传转化效率的影响以及KN1基因超量表达对转基因植株再生的影响。研究结果表明:以苗龄15d左右的小油桐无菌苗子叶为外植体,农杆菌菌液浓度OD600为0.6～0.8时,侵染8min,外植体大小为(0.8×0.8)～(1.0×1.0)mm时,遗传转化效果最好;对抗性芽及再生植株进行GUS及PCR检测结果表明,KN1基因已经整合到小油桐植物基因组中。KN1基因的超量表达可提高小油桐再生芽分化,影响转化芽及植株的外观形态及叶片的表型,包括芽及植株矮小,茎杆粗壮;叶片缩小,边缘分裂,对称性丧失,无子叶柄等。

In this research,KN1 gene was introduced into Jatropha curcas via Agrobacterium tumefaciens mediated transformation and transgenic plants were achieved.A series of parameter tests such as Agrobacterium concentration,infection time,size of explant and overexpression of KN1 gene were used to evaluate for their effect on transformation efficiency.The results showed that the genetic transformation efficiency was more better when the 15-day-old asepsis cotyledon leaf explants（0.8×0.8）～（1.0×1.0） mm were infected 8 minutes with Agrobacterium concentration（OD600=0.6～0.8）.The results of GUS staining analysis and PCR of transformants indicated that exogenes KN1 gene have been inserted into Jatropha curcas genomes.Overexpression of KN1 gene improved shoot differentiation and affected the appearance and plant morphological and leaf phenotype of Jatropha curcas,including dwarf shoots and plants,stem stout,leaves grow downwards,lobed,split,out of symmetry,no such loss petiole phenotype.