| Abstract Detail
Plant Protection Atallah, Zahi [1], Stevenson, Walter [2]. Detecting and quantifying pathogens causing potato tuber decay using real-time quantitative PCR, to predict tuber storage potential. WE present a methodology to detect and quantify the causal agents of the five diseases from whole potato tubers, using real-time quantitative-PCR (Q-PCR). We developed specific Q-PCR primer pairs for the detection and quantification of Phytophthora infestans, Phytophthora erythroseptica, Pythium ultimum, Fusarium sambucinum, and Erwinia carotovora subsp. carotovora and subsp. atroseptica. Amplification efficiencies ranged between 95 and 100 % over a five-log dilution series and were unaffected by the presence of large amounts of host DNA. The detection level of the primers reached 0.5 pg of target DNA. Pathogens were detected in 100 pg of total DNA extracted from 170-250 g tubers, four days after inoculation, regardless of the presence of symptoms. In 2005, we compared tuber sampling strategies from potato fields prior to harvest. We collected 2,000 tubers from four fields in Wisconsin, one field in North Dakota and one in Minnesota. In each field, tubers were collected following two sampling schemes: stratified and completely random. Sampling sites were pre-identified from aerial photographs using the ARC-GIS software, and tubers were collected from specific geo-referenced sites. Q-PCR amplifications were conducted on the collected tubers to detect and quantify the five aforementioned pathogens. This study provides a method for the molecular detection and quantification of minute numbers of pathogenic propagules, and suggests the most appropriate sampling scheme to identify the risk of tuber decay prior to harvest. Additionally, we provide the first step in a methodology to predict the potential for storability of potato tubers.
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1 - University of Wisconsin, Plant Pathology, 285A Russell Labs, 1630 Linden Dr., Madison, Wisconsin, 53706, USA
2 - University of Wisconsin, Plant Pathology, 285B Russell Labs, 1630 Linden Dr., Madison, Wisconsin, 53706, USA
Keywords:
post-harvest diseases
real-time PCR
disease
field sampling.
Session: PAA03-8
Location: Hall of Ideas Room E/Monona Terrace
Date: Thursday, July 27th, 2006
Time: 10:45 AM
Abstract ID:268 |
