T/CI 026-2022 Method for detecting and evaluating the resistance of cassava white-leaf disease

Scope

Scope: This standard specifies the terms and definitions, abbreviations, symptoms, and epidemic characteristics of sugarcane white leaf disease pathogen detection and disease resistance identification, as well as the technical methods and standards for pathogen detection and disease resistance identification. This standard applies to the molecular detection of sugarcane white leaf disease and the artificial inoculation and evaluation of sugarcane germplasm resources for their resistance to sugarcane white leaf disease. Main technical content: 3.1 Nested PCR: Nested PCR is a method that uses two pairs of PCR primers to amplify complete fragments. The first pair of PCR primers amplifies fragments similar to ordinary PCR, and the second pair of primers binds within the first PCR product, making the sensitivity of the second PCR amplification higher than that of the first PCR amplification. 3.2 Sugarcane White Leaf Disease (SCWL): A plant-pathogenic disease caused by the Sugarcane White Leaf Phytoplasma, which systematically infects sugarcane and results in the appearance of one or several white stripes on the leaves, or even the entire leaf turning white. The leaves become soft, and the infected plants develop more tillers and stunted growth. 3.3 Susceptible Control: Sugarcane varieties with a disease incidence rate of more than 20.1% after infection by the Sugarcane White Leaf Phytoplasma. 3.4 Resistant Control: Sugarcane varieties with a disease incidence rate of less than 10% after infection by the Sugarcane White Leaf Phytoplasma. 4. Abbreviations: bp: Base pair CTAB: Hexadecyl Trimethyl Ammonium Bromide dNTPs: Deoxyribonucleoside Triphosphates mixture ddH2O: Double distilled water EDTA: Ethylene Diamine Tetraacetic Acid PCR: Polymerase Chain Reaction Tris-base: Tris (hydroxymethyl) aminomethane TAE: A buffer solution composed of Tris (hydroxymethyl) aminomethane, acetic acid, and ethylene diamine tetraacetic acid

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