Abstract

Viral cross protection in plants is known as an acquired immunity phenomenon, where a mild virus isolate/strain can protect plants against economic damage caused by a severe challenge strain/isolate of the same virus. Mild strain cross protection (MSCP) has been used extensively to control losses caused by a few major virus diseases in some parts of the world. So far, none of the many proposed mechanisms can fully explain the intact process of MSCP. In fact, it may be that different mechanisms are involved in MSCP against different viruses, even when different research approaches are used for the same virus, different mechanisms could be proposed. The molecular detail of MSCP still remains unclear, although several lines of evidence imply that the resistance is protein and/or RNA mediated. Some data to date have shown that a minimum time (a few days to less than a month) is required for the mild virus strain to establish MSCP. To investigate interference among virus strains and the plant host at an early stage of MSCP at a subcellular level, we developed a rapid micro-extraction method for the preparation of total nucleic acid (TNA), combined with other molecular methods, to monitor the interaction of virus strains at short time intervals in young plants. This method was initially developed to further study the mechanism of MSCP against Citrus tristeza virus, but has potentially widespread application to other viruses after having been efficiently used to extract over 50,000 TNA samples of citrus viruses, viroids, and bacteria.

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