title: Development of a loop-mediated isothermal amplification method for the rapid diagnosis of Ascochyta rabiei L. in chickpeas.
abstract: Ascochyta blight (AB) is a devastating fungal disease of chickpeas that has spread to nearly all of the chickpea cultivating regions of the world. The rapid diagnosis of Ascochyta rabiei L. (A. rabiei), the pathogen that causes AB, plays an important role in A. rabiei epidemic tracking and AB management. In this study, a group of loop-mediated isothermal amplification (LAMP) primers was designed to detect A. rabiei in chickpea plants and seeds via a LAMP method and a conventional PCR method based on an internal transcribed spacer (ITS) sequence analysis of A. rabiei. Compared with the conventional PCR method, the LAMP method not only exhibited greater sensitivity and specificity in the detection of A. rabiei but also used simpler equipment and required less operational time. The minimum detectable concentration of the A. rabiei genomic DNA solution with the LAMP method was 6.01 × 10(-6 )ng/μl, which was 100 times lower than that of the conventional PCR method with the same outer primers. The greatest advantage of the LAMP method is that results can be observed via the visualization of color changes in SYBR Green I dye with the naked eye, and it does not require expensive instruments, also with less time consumption.
authors.affiliation: College of Agronomy, Xinjiang Agricultural University, Key Laboratory of the Pest Monitoring and Safety Control of Crops and Forests of the Universities of the Xinjiang Uygur Autonomous Region, Xinjiang Agricultural University, No. 311 Nongda East Road, 830052, Urumqi, Xinjiang, People's Republic of China.
authors.forename: Xiaolu
authors.initials: X
authors.lastname: Chen
authors.affiliation: College of Agronomy, Xinjiang Agricultural University, Key Laboratory of the Pest Monitoring and Safety Control of Crops and Forests of the Universities of the Xinjiang Uygur Autonomous Region, Xinjiang Agricultural University, No. 311 Nongda East Road, 830052, Urumqi, Xinjiang, People's Republic of China.
authors.forename: Lijuan
authors.initials: L
authors.lastname: Ma
authors.affiliation: College of Agronomy, Xinjiang Agricultural University, Key Laboratory of the Pest Monitoring and Safety Control of Crops and Forests of the Universities of the Xinjiang Uygur Autonomous Region, Xinjiang Agricultural University, No. 311 Nongda East Road, 830052, Urumqi, Xinjiang, People's Republic of China.
authors.forename: Song
authors.initials: S
authors.lastname: Qiang
authors.affiliation: College of Agronomy, Xinjiang Agricultural University, Key Laboratory of the Pest Monitoring and Safety Control of Crops and Forests of the Universities of the Xinjiang Uygur Autonomous Region, Xinjiang Agricultural University, No. 311 Nongda East Road, 830052, Urumqi, Xinjiang, People's Republic of China.
authors.forename: Deying
authors.initials: D
authors.lastname: Ma
mesh_terms: D001203:Ascomycota / Q000235:genetics*; D029863:Cicer / Q000382:microbiology*; D017931:DNA Primers / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000235:genetics; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation / Q000379:methods*; D010935:Plant Diseases / Q000382:microbiology*; D015203:Reproducibility of Results; D012639:Seeds / Q000382:microbiology
chemical_list: D017931:DNA Primers; D004271:DNA, Fungal; D021903:DNA, Ribosomal Spacer