红籽鸢尾瞬时表达体系的建立

A System Establishment of Transient Expression on Iris foetidissima L.

本试验通过瞬时表达解决红籽鸢尾遗传转化效率低和转化周期长的问题。以红籽鸢尾叶片为材料,用转入植物表达载体pBI121的根癌农杆菌EHA105进行遗传转化。通过研究菌液不同浓度(OD600)、乙酰丁香酮浓度(AS)、侵染时间及共培养时间等因素对红籽鸢尾叶片中GUS基因表达效果的影响。结果表明:在农杆菌浓度OD600为0.6,添加1×10^-4mol/L乙酰丁香酮的侵染液中真空抽吸30 min后,黑暗培养4 d,红籽鸢尾叶片的GUS基因表达效果最好。本试验建立的瞬时表达方法操作简单、转化效率高,为后期红籽鸢尾的基因功能研究及分子育种提供了依据。

We transformed the Iris foetidissima through the transient transformation,to resolve its poor transformation efficiency and long transformation periodin this study.An experiment was conducted to investigate the transient expression mediated by Agrobacterium using the leaves of Iris foetidissima as explants and the Agrobacterium tumefaciens EHA105 containing the plant expression vector pBI121 for genetic transformation.We investigated the effects of Agrobacterium tumefaciens concentration(OD600),acetosyringone concentration(AS),infection time and co-culture time on transient GUS gene expression in Iris foetidissima leaves.A higher transient expression level of GUS gene could be obtained as OD600 value of A.tumefaciens 0.6,1×10^-4 mol/L acetosyringone,infiltrating for 30 min,and co-culture for 4 days in darkness.An efficient and simple protocol of transient transformation was developed,providing technical support for the study of gene function and molecular breeding of Iris foetidissima.