Potrokhov Andrii, Sosnovska Daria, Ovcharenko Olga, Budzanivska Irena, Rudas Volodymyr, Kuchuk Mykola
Institute of Cell Biology and Genetic Engineering NAS Ukraine, Kyiv Ukraine.
Virology Department, Educational and Scientific Center, Institute of Biology and Medicine of Taras Shevchenko National University of Kyiv, Kyiv Ukraine.
Turk J Biol. 2021 Feb 9;45(1):79-87. doi: 10.3906/biy-2007-87. eCollection 2021.
Viral pathogens seriously decrease the yield of potato ( L.) -an important agricultural crop. Therefore, there is a demand for potato cultivars resistant to multiple viruses. Ribonucleases (RNases) are supposed to be engaged to antiviral response in plants. Heterologous RNase gene expression provides a tool for production of cultivars with multiple resistance to viruses and viroids. Transgenic potato cultivars Luhivs'ka and Lasynak with heterologous genes and of apoplastic RNases from and respectively were obtained via -mediated transformation. The presence of and transgenes was confirmed by PCR analysis. RNase activity was examined by modified Oleshko method. Plants with heterologous ribonuclease genes had higher level of RNase activity compared to nontransgenic ones. Transgenic plants inoculated with , PVY (genus , family Potyviridae) demonstrated delayed and less severe symptoms of viral infection. DAS-ELISA confirmed the presence of viral antigens both in transformed and control plants. Visual manifestations of viral infection in transgenic potatoes were milder than in control plants and their development was delayed, but complete elimination of the virus did not occur.
病毒病原体严重降低了重要农作物马铃薯(L.)的产量。因此,需要培育对多种病毒具有抗性的马铃薯品种。核糖核酸酶(RNases)被认为参与植物的抗病毒反应。异源核糖核酸酶基因表达为培育对病毒和类病毒具有多重抗性的品种提供了一种手段。通过农杆菌介导的转化分别获得了含有来自烟草和菠菜质外体核糖核酸酶的异源基因和的转基因马铃薯品种Luhivs'ka和Lasynak。通过PCR分析证实了和转基因的存在。采用改良的奥列什科方法检测核糖核酸酶活性。与非转基因植物相比,含有异源核糖核酸酶基因的植物具有更高水平的核糖核酸酶活性。用马铃薯Y病毒(马铃薯Y病毒属,马铃薯Y病毒科)接种的转基因植物表现出病毒感染症状延迟且较轻。双抗体夹心酶联免疫吸附测定(DAS-ELISA)证实转基因植物和对照植物中均存在病毒抗原。转基因马铃薯中病毒感染的视觉表现比对照植物轻,且其发展延迟,但并未完全消除病毒。