Kon Tatsuya, Yoshikawa Nubuyuki
Plant Pathology Laboratory, Faculty of Agriculture, Iwate University, 3-18-8 Ueda, Morioka, 020-8550, Japan.
Methods Mol Biol. 2015;1287:191-9. doi: 10.1007/978-1-4939-2453-0_14.
Virus infection leads to the synthesis of double-stranded RNA during virus replication, and then this infection produces small RNA molecules in the antiviral RNA silencing pathway. Here, we develop an Agrobacterium-mediated inoculation system for ALSV-based vectors. This system is more effective and convenient for inoculation of the ALSV vectors into plants compared to direct inoculation of ALSV-RNA2-based vectors in pUC plasmids reported previously. In addition, cointroduction of various plant viral RNA-silencing suppressors increased the efficiency of agroinoculation of the ALSV-based vector. An ALSV vector could be successfully used to silence an endogenous gene in plants. Therefore, the ALSV-based VIGS/agroinoculation system provides a valuable tool for functional genomics among a broad range of plant species.
病毒感染在病毒复制过程中导致双链RNA的合成，然后这种感染在抗病毒RNA沉默途径中产生小RNA分子。在此，我们开发了一种用于基于ALSV的载体的农杆菌介导接种系统。与先前报道的将基于ALSV-RNA2的载体直接接种到pUC质粒中相比，该系统在将ALSV载体接种到植物中时更有效且更方便。此外，共导入各种植物病毒RNA沉默抑制子提高了基于ALSV的载体的农杆菌接种效率。一种ALSV载体可成功用于沉默植物中的内源基因。因此，基于ALSV的VIGS/农杆菌接种系统为广泛植物物种的功能基因组学提供了一种有价值的工具。
