Linear-motion tattoo machine and prefabricated needle sets for the delivery of plant viruses by vascular puncture inoculation |
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Authors: | John J Weiland Michael C Edwards |
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Institution: | (1) USDA, Agricultural Research Service, Cereal Crops Research Unit, 1605 Albrecht Blvd. N., Fargo, ND 58102–2765, USA; |
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Abstract: | Vascular puncture inoculation (VPI) of plant viruses previously has been conducted either manually or by use of a commercial
engraving tool and laboratory-fabricated needle arrays. In an effort to improve this technique, a linear-motion tattoo machine
driving industry-standard needle arrays was tested as a means of delivering plant viruses into maize and small grain seed
embryos. The new method was applied in the successful transmission of maize rayado fino virus (MRFV), the type member of the
genus Marafivirus, from an archived sample to maize. Subsequent transfer of MRFV from the sap of an infected plant using the method produced
an average infection rate in maize of 70% (range 39–93%). Maize, oat, and triticale were successfully infected with oat blue
dwarf virus (OBDV) using the method; similar infection rates were observed between maize seeds inoculated with the tattoo
machine and those inoculated with the engraving machine when using prefabricated needle arrays. No infection was obtained
in repeated tests with barley yellow dwarf virus (BYDV-PAV) or cereal yellow dwarf virus (CYDV-RPV) using either sap or RNA
from infectious cloned cDNA. Replacement of the original engraving-tool with a linear-motion tattoo machine in VPI provides
greater flexibility and convenience in a quiet, readily-available instrument, while improving reproducibility through the
use of prefabricated needle arrays. |
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