title: Development of a SCAR marker for molecular detection and diagnosis of Tilletia controversa Kühn, the causal fungus of wheat dwarf bunt.
abstract: Tilletia controversa Kühn (TCK) is an important quarantine pathogen that causes wheat dwarf bunt and results in devastating damage to wheat production. The fungus is difficult to be distinguished from T. caries and T. laevis, which cause wheat common bunt, based on morphological, physiological and symptomatological characteristics of the pathogens. The traditional detection of the fungus can be a long and tedious process with poor accuracy. The inter-simple sequence repeat (ISSR) technique has been used for identifying molecular markers for detection of TCK. Of 28 ISSR primers screened, ISSR-859 amplified a specific 678 bp DNA fragment from all TCK isolates but not from any isolates of the common bunt fungi or other pathogenic fungi tested. Based on the fragment sequence, a pair of sequence characterized amplified region (SCAR) primers was designed, which amplified a 372 bp DNA fragment specifically in TCK. The SCAR marker was detected using as low as 1 ng template DNA of TCK, and was also detected using broken teliospores and DNA from asymptomatic wheat samples. We developed the SYBR Green I and TaqMan Green I and TaqMan real-time polymorphism chain reaction methods to detect TCK with the detection limit of 0.1 fg with asymptomatic wheat samples. Further work is needed to develop a rapid test kit for this pathogenic fungus using the designed specific primers.
authors.affiliation: State Key Laboratory for Biology of Plant Disease and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, 100193, China, xiaogaosx@hotmail.com.
authors.forename: Li
authors.initials: L
authors.lastname: Gao
authors.forename: Huixin
authors.initials: H
authors.lastname: Yu
authors.forename: Wensu
authors.initials: W
authors.lastname: Han
authors.forename: Fei
authors.initials: F
authors.lastname: Gao
authors.forename: Taiguo
authors.initials: T
authors.lastname: Liu
authors.forename: Bo
authors.initials: B
authors.lastname: Liu
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Kang
authors.forename: Jiguo
authors.initials: J
authors.lastname: Gao
authors.forename: Wanquan
authors.initials: W
authors.lastname: Chen
mesh_terms: D001487:Basidiomycota / Q000235:genetics* / Q000302:isolation & purification*; D052160:Benzothiazoles; D017931:DNA Primers / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics; D003959:Diamines; D005819:Genetic Markers; D009172:Mycology / Q000379:methods*; D009930:Organic Chemicals / Q000378:metabolism; D010935:Plant Diseases / Q000382:microbiology*; D011804:Quinolines; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods; D014908:Triticum / Q000382:microbiology*
chemical_list: D052160:Benzothiazoles; D017931:DNA Primers; D004271:DNA, Fungal; D003959:Diamines; D005819:Genetic Markers; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I