Semester of Graduation
Fall 2025
Degree
Master of Science (MS)
Department
Plant Pathology and Crop Physiology
Document Type
Thesis
Abstract
Leaf scald (LS) and ratoon stunt disease (RSD) are important sugarcane diseases that can cause yield losses. LS and RSD are caused by the vascular system-inhabiting bacteria Xanthomonas albilineans and Leifsonia xyli subsp. xyli, respectively. As using pathogen-free sugarcane propagative material is an important management strategy against LS and RSD, accurate and efficient pathogen detection is crucial. Our study aimed to increase pathogen detection sensitivity and efficiency through a cost-effective real-time PCR protocol and high-throughput DNA extraction methods. A SYBR-based qPCR assay detecting the RSD and LS causal agents was 100 times more sensitive than traditional PCR. Additionally, multiplex reactions accurately identified the RSD and LS pathogens without primer interference. Furthermore, two high-throughput nucleic acid extraction methods, a crude nucleic acid extraction using a general buffer and the internode biopsy disc method reduced time and costs, albeit sensitivity was slightly reduced. Our results demonstrate that alternative nucleic acid extraction methods can facilitate a statewide RSD and LS survey in Louisiana utilizing a multiplex SYBR-based qPCR assay that is both sensitive and cost effective.
Date
11-23-2025
Recommended Citation
Pelizzaro Morales, Joao Vitor, "STREAMLINED REAL-TIME qPCR DETECTION ASSAY AND NUCLEIC ACID EXTRACTION PROTOCOLS FOR LEAF SCALD AND RATOON STUNT DISEASE IN LOUISIANA" (2025). LSU Master's Theses. 6261.
https://repository.lsu.edu/gradschool_theses/6261
Committee Chair
Gama, Andre B.