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ARS Home » Southeast Area » Griffin, Georgia » Plant Genetic Resources Conservation Unit » Research » Publications at this Location » Publication #105420
Title: TESTING PEANUT SEED LOTS FOR PEANUT STRIPE AND PEANUT MOTTLE POTYVIRUS INFECTION BY IMMUNOCAPTURE-REVERSE TRANSCRIPTION-POLYMERASE CHAIN REACTION

Author
item Gillaspie, Athey - Graves
item Pittman, Roy
item Pinnow, David
item CASSIDY, B - NOBLE FOUNDATION

Submitted to: Plant Disease
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 11/1/1999
Publication Date: 1/24/2000
Citation: Gillaspie Jr, A.G., Pittman, R.N., Pinnow, D.L., Cassidy, B.G. 2000. Testing peanut seed lots for peanut stripe and peanut mottle potyvirus infection by immunocapture-reverse transcription-polymerase chain reaction. Plant Disease Series. Plant Disease 84: 559-561 (2000).

Interpretive Summary: Each year, numerous lines of peanut are introduced into the U.S. Each of these peanut seed lots must be tested for the presence of seedborne viruses to ensure that no new viruses are introduced that may jeopardize commercial peanut production in the U.S. We developed a new molecular method called immunocapture-reverse transcription-polymerase chain reaction (IC-RT-PCR) for testing large numbers of seed lots for infection with peanut stripe an peanut mottle viruses. The technique is very sensitive in that it can detect one infected seed in a seed lot of 100 seeds, and thus allows evaluation of more seed lots in a shorter period of time than do other techniques currently in use.

Technical Abstract: An immunocapture-reverse transcription-polymerase chain reaction (IC-RT-PCR) method was developed for testing peanut (Arachis hypogaea) seed lots for seedborne infection by peanut stripe (PStV) and peanut mottle (PeMV) viruses. A small slice was removed from each seed distal to the radicle of a random 100-seed sample, the slices were extracted in buffer, centrifuged, and a portion was incubated in a tube coated with antiserum. After washing, the RT-PCR mix (with primers designed from the capsid protein region of the viruses) was placed in the same tubes and the test completed. Results obtained on 15 seed lots from virus- infected plants indicated good correlation between virus detected by the IC-RT-PCR method and virus detected from the same seed lots by ELISA testing of five-seed samples. The IC-RT-PCR method yielded three lots with PeMV and none with PStV from 106 seed lots grown in Ecuador (results confirmed by ELISA). The IC-RT- PCR method is useful for testing large numbers of seed lots of peanut germ plasm.