Detection of Ratoon Stunting Disease in Virus-free Seedcane via Real-time Fluorescence Quantitative PCR

This study was to develop the real-time fluorescence quantitative PCR technique for detecting the ratoon stunting disease （RSD） in virus-free seedcane seedlings. Healthy tissue culture seedlings were obtained from six plants of sugarcane ROC22, which had been confirmed RSD-positive by detecting the sugarcane juice, by employing the sugarcane seedlings production protocol. Real-time fluorescence quantitative PCR was used to detect RSD pathogens in tissue culture sam- pies. The results showed that target fragment of RSD pathogens was not found in all 10 samples in real-time fluorescence quantitative PCR, with the Ct values of 37 - 39. The healthy tissue culture sugarcane seedlings do not carry RSD pathogens, indicating that adopting healthy seedcane seedlings production technique could thoroughly get rid of RSD pathogens.

Detection of Ratoon Stunting Disease in Virus-free Seedcane of Saccharum officinarum by PCR

[Objective]The study was carried out for providing good method to detect ratoon stunting disease（RSD）in virus-free seedcane of Saccharum officinarum.[Method]PCR method was used for the detection of RSD in virus-free seedcane of sugarcane.[Result]During the different growth stages of healthy virus-free seedling including proliferation stage,rooting stage,sand culture stage and temporary planting stage,the RSD detection results were negative.PCR detection sensitivity in the present study showed that PCR could detect RSD pathogen in the solution with bacterium concentration 10-3.[Conclusion]PCR method showed a good specificity and higher sensitivity,so it was suitable for the batch detection in the production of healthy virus-free seedlings of sugarcane.