THE UNITED STATES OF AMERICA AS REPRESENTED BY THE SECRETARY OF AGRICULTURE
发明人:
FREDERICK REID D.,TOOLEY PAUL W.,BONDE MORRIS R.,KNORR DAVID A.,PETERSON GARY L.,SCHAAD NORMAN W.
申请号：
US19990471016
公开号：
US6316195(B1)
申请日:
1999.12.23
申请国别(地区):
美国
年份:
2001
代理人:
摘要：
Karnal bunt of wheat is caused by Tilletia indica Mitra. Recently, teliospores morphologically resembling T. indica were isolated from bunted ryegrass seeds and wheat seed washes. Previously developed PCR assays failed to differentiate T. indica from the newly discovered ryegrass pathogen, T. walkeri. The nucleotide sequence of a 2.3-kb region of mitochondrial DNA, previously amplified by PCR only from T. indica, was determined for three isolates of T. indica and for three isolates of T. walkeri. There was greater than 99% identity within either the T. indica group or the T. walkeri group of isolates, whereas there was approximately 3% divergence between isolates of these two Tilletia species. Five sets of PCR primers were made specific to T. indica, and three sets were designed specifically for T. walkeri based upon nucleotide differences within the mtDNA region. In addition, a 212 bp amplicon was developed as a target sequence in a fluorogenic 5' nuclease PCR assay using the TaqMan system for the detection and discrimination of T. indica and T. walkeri.
