马铃薯病毒外壳蛋白融合基因转化马铃薯及其抗病性分析

将多种病毒的有效核酸片断拼接成融合基因转入马铃薯可获得多抗马铃薯材料。针对马铃薯生产中分布广泛、危害严重并经常混合感染的马铃薯X病毒(PVX)、马铃薯Y病毒(PVY)、马铃薯卷叶病毒(PLRV)和马铃薯S病毒(PVS),开展了利用基因工程方法获得兼抗4种马铃薯病毒转基因马铃薯材料的研究。试验在前期获得含4种马铃薯病毒外壳蛋白基因片段的质粒pART27-XSYV-rh的基础上,通过根癌农杆菌(Agrobacterium tumefaciens)介导转化马铃薯(Solanum tuberosum)品种‘陇薯3号’,PCR扩增和PCR-Southern杂交证明,4价融合基因已整合到马铃薯基因组中。qRT-PCR分析表明,该融合基因在转基因植株中能正常表达。3株转基因植株的抗病性鉴定结果表明,2株对4种病毒同时具有抗性;1株对PLRV侵染表现阳性,对另外3种病毒同时具有抗性。

Transformation of Potato Virus Coat Protein Fusion Gene into Potato and Analysis of Its Disease Resistance

The effective nucleic acid fragments of multiple viruses were spliced into fusion genes and transferred to potato to obtain multi-resistant potato materials.Potato diseases,such as potato virus X(PVX),potato virus Y(PVY),potato leafroll virus(PLRV)and potato virus S(PVS),are becoming more and more rampant,which greatly reduces potato yield,especially multi-virus complex infection.Genetic engineering methods were used to obtain transgenic potato materials with the ability to resist these four potato virus infections.On the basis of the previously obtained plasmid pART27-XSYV-rh containing four potato virus coat protein gene fragments,the potato(Solanum tuberosum)variety'Longshu 3'was transformed using Agrobacterium tumefaciens as a vector,and PCR and PCR-Southern analyses proved that the fusion gene has been integrated into the potato genome.The results of qRT-PCR indicated that the fusion gene could be normally expressed in the transgenic plants. The disease resistance identification results of threetransgenic plants showed that two of them were highly resistant to the PVX, PVY, PVS and PLRV infection;the otherone was positive for PLRV infection, but was resistant to other three viruses.