Progress towards a Definitive Diagnostic Test for Sugarcane Mosaic Virus Infection
نویسندگان
چکیده
A new, rapid and sensitive protocol is reported for diagnosis of sugarcane mosaic virus (SCMV) using small (2 cm/) samples of leaf tissue as starting material. The procedure involves total RNA isolation followed by reverse transcription and amplification by the polymerase chain reaction (RT-PCR) of a substantial portion (884bp) of the coat protein gene of SCMV. This can be followed by corroborative amplification of a smaller region (360bp) within the first amplified fragment (nested PCR) to reduce the potential for false positives. Results have shown that both the 884bp and 360bp fragments are reproducibly amplified from infected leaf but not from control (visually symptom free) material of the same variety. Comparison of the sequence of the larger fragment to the international database has confirmed the identity of the source material as SCMV. So far the method has successfully diagnosed SCMV in infected sugarcane from two widely separated regions of KwaZulu-Natal: from the midlands (Eston) and from the extreme north coast (Pongola). This augurs well for its eventual application as a reliable and definitive test for SCMV regardless of strain. In addition, there is the potential to use sequence information from the larger fragment for specific strain identification. This could, in turn, lead to the design of tests for specific strain diagnosis.
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