Not AvailableChaetomium globosum is a potential biocontrol agent
against various seed and soil-borne pathogens. To ensure
proper use of C. globosum in agriculture, accurate data is
essential for population monitoring. A PCR-based marker
has been developed for detection of this biocontrol agent,
which will help to detect the fungus at the place of its
application. Out of twelve URP primers tested against 15
isolates of C. globosum and other Chaetomium species,
URP 2R amplified a monomorphic band of 1,900 bp only
in C. globosum isolates. This amplicon was cloned and sequenced, and based on the sequence obtained, four primer
sets were designed, one of which in PCR assays amplified
a region (SCAR; SCCgRA1900) of the expected size (1.9kb)
in C. globosum isolates. The specific marker also detected
the presence of C. globosum in soil, roots and leaves. The
detection limit of marker in conventional PCR assay was
75 pg. The sensitivity and usefulness of SCAR marker was
further enhanced by developing qPCR using the primer
set SCCgQF/SCCgQR designed from SCCgRA1900, which
detected as much as 1 pg of DNA (4.83×105 copy number
of target DNA). The initial population of C. globosum in
terms of target DNA in C. globosum-amended soil was
equivalent to 2.5×108 copy number/g soil (0.51ng target
DNA/g soil) which increased approximately 10 times after 15 days of application i.e., 2×109copy number/g soil
(3.1ng/g soil). However, with, Bipolaris sorokiniana the
quantity of C. globosum target DNA increased slowly
reaching 4.32×108 copy number/g soil after 15 days. Conventional PCR-based detection using SCAR marker and
subsequent qPCR provided a rapid and reliable tool for
efficient detection and monitoring of C. globosum at the
site of its application.Not Availabl
