Rapid and sensitive detection of Phytophthora colocasiae responsible for the taro leaf blight using conventional and real‐time PCR assay. Issue 2 (24th February 2014)

• Record Type: Journal Article
• Authors: Nath, Vishnu S.; Hegde, Vinayaka M.; Jeeva, Muthulekshmi L.; Misra, Raj S.; Veena, Syamala S.; Raj, Mithun; Unnikrishnan, Suresh K.; Darveekaran, Sree S.
• Is Part Of: FEMS microbiology letters. Volume 352:Issue 2(2014:Mar.)
• Page Start: 174
• Page End: 183
• Publication Date: 2014-02-24
• Subjects: Microbiology -- Periodicals; 579
• DOI: 10.1111/1574-6968.12395
• Languages: English
• ISSNs: 0378-1097

Abstract

<abstract abstract-type="main" id="fml12395-abs-0001"> <title>Abstract</title> <p>Conventional and real‐time PCR assays were developed for sensitive and specific detection of <italic>Phytophthora colocasiae</italic>, an oomycete pathogen that causes leaf blight and corm rot of taro. A set of three primer pairs was designed from regions of the RAS‐related protein (<italic>Ypt</italic>1), G protein alpha‐subunit (<italic>GPA</italic>1) and phospho‐ribosylanthranilate isomerase (<italic>TRP</italic>1) genes. In conventional PCR, the lower limit of detection was 50 pg DNA, whereas in real‐time PCR, the detection limit was 12.5 fg for the primer based on <italic>Ypt</italic>1 gene. The cycle threshold values were linearly correlated with the concentration of the target DNA (range of <italic>R</italic>² = 0.911–0.999). All the primer sets were successful in detecting <italic>P. colocasie</italic> from naturally infected leaves and tubers of taro. <italic>Phytophthora colocasiae</italic> was detected from artificially infested samples after 18 and 15 h of postinoculation in conventional and real‐time PCR assay, respectively. The developed PCR assay proved to be a robust and reliable technique to detect <italic>P. colocasiae</italic> in taro planting material and for assessing the distribution of pathogen within fields, thus aid in mitigating taro leaf blight.</p> </abstract>