Abstract
BackgroundPokkah boeng disease caused by the Fusarium species complex results in significant yield losses in sugarcane. Thus, the rapid and accurate detection and identification of the pathogen is urgently required to manage and prevent the spreading of sugarcane pokkah boeng.MethodsA total of 101 isolates were recovered from the pokkah boeng samples collected from five major sugarcane production areas in China throughout 2012 and 2013. The causal pathogen was identified by morphological observation, pathogenicity test, and phylogenetic analysis based on the fungus-conserved rDNA-ITS. Species-specific TaqMan real-time PCR and conventional PCR methods were developed for rapid and accurate detection of the causal agent of sugarcane pokkah boeng. The specificity and sensitivity of PCR assay were also evaluated on a total of 84 isolates of Fusarium from China and several isolates from other fungal pathogens of Sporisorium scitamineum and Phoma sp. and sugarcane endophyte of Acremonium sp.ResultTwo Fusarium species (F. verticillioides and F. proliferatum) that caused sugarcane pokahh boeng were identified by morphological observation, pathogenicity test, and phylogenetic analysis. Species-specific TaqMan PCR and conventional PCR were designed and optimized to target their rDNA-ITS regions. The sensitivity of the TaqMan PCR was approximately 10 pg of fungal DNA input, which was 1,000-fold over conventional PCR, and successfully detected pokkah boeng in the field-grown sugarcane.Conclusions/SignificanceThis study was the first to identify two species, F. verticillioides and F. proliferatum, that were causal pathogens of sugarcane pokkah boeng in China. It also described the development of a species-specific PCR assay to detect and confirm these pathogens in sugarcane plants from mainland China. This method will be very useful for a broad range of research endeavors as well as the regulatory response and management of sugarcane pokkah boeng.
Highlights
Sugarcane pokkah boeng is an economically important fungal disease worldwide [1], which was first described in Java by Walker and Went in 1896, and the name was a Javanese term denoting a malformed or distorted top
The sequences of the internal transcribed spacer from 103 isolates were amplified by PCR using fungus-conserved ITS1 and ITS4 primers, which resulted in a fragment in the size range of 485 bp to 603 bp
The phylogenetic tree was constructed using neighbor-joining (NJ) method whereas some known ITS sequences from Fusarium species complex were used as references for the out-group (Fig. 1)
Summary
Sugarcane pokkah boeng is an economically important fungal disease worldwide [1], which was first described in Java by Walker and Went in 1896, and the name was a Javanese term denoting a malformed or distorted top. Pokkah boeng has been recorded in almost all cane growing countries, but it only causes severe damage in areas where susceptible varieties are widely planted during a hot and dry season followed by a wet season [2]. A survey of different sugarcane areas in India found that the incidence of pokkah boeng increased from 2007 to 2013 and affected almost all of the sugarcane cultivars, which was recommended for general cultivation for different agricultural climatic regions. The incidence and severity of pokahh boeng has been reported from major sugarcane growing areas during all seasons, rather than only during the wet and hot summer seasons. Pokkah boeng disease caused by the Fusarium species complex results in significant yield losses in sugarcane. The rapid and accurate detection and identification of the pathogen is urgently required to manage and prevent the spreading of sugarcane pokkah boeng
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