A Rapid and Specific Nest-PCR Method for Detection of Alternaria alternata in Wheat
QIN Zhao
LI Qiao-yun
DUAN Zong-biao
JIANG Yu-mei
NIU Ji-shan
Abstract：The aim of this study was to establish a method for detection of Alternaria alternata in wheat.Seven dominating isolates of A.alternata from wheat black point grains in Henan province were collected,and then their internal transcribed spacer(ITS)regions in rDNA were sequenced, and the sequence analysis showed that the ITS sequences were highly similar among isolates of A.alternata isolated in this study and in GenBank by 99%-100%.According to the comparative result of ITS sequences of Alternaria spp.(A.alternate,A.tenuis,A.tenuissima)and Bipolaris sorokiniana,a pair of primers,namely Aa1F and Aa1R,were designed.Specificity of the primers was tested with several major fungal pathogens of wheat in Henan province,and a 443 bp specific DNA fragment could be amplified only from A.alternata.A rapid and specific nest-PCR method for detection of A.alternata in wheat was established with fungal universal primers ITS1/ITS4 as the outer primers and Aa1F/Aa1R as the inner primers.The nest-PCR method was used to detect the samples with gradated concentrations of A.alternata DNA,and the minimum DNA concentration detected was 50 pg/μL.
Keywords：wheatAlternaria alternatadetectionnest-PCR
Publication Date：2014-10-01
Online Publishing Date：2026-09-12(First online date of this platform, not the publication date of the document)
Pages：7( 67-73 )