Fine Tuning of Real Time PCR as a First Tool for the Detection of G143A Substitution in <i>Venturia inaequalis</i>Samples  

作　　者：Ceren Turan Irene Maja Nanni Lucia Landi Alessandro Pirondi Marina Collina Ceren Turan;Irene Maja Nanni;Lucia Landi;Alessandro Pirondi;Marina Collina(Department of Agricultural and Food Sciences, University of Bologna, Bologna, Italy;Department of Agricultural, Food and Environmental Sciences, Polytechnic University of Marche, Ancona, Italy)

机构地区：[1]Department of Agricultural and Food Sciences, University of Bologna, Bologna, Italy [2]Department of Agricultural, Food and Environmental Sciences, Polytechnic University of Marche, Ancona, Italy

出　　处：《American Journal of Plant Sciences》2021年第6期960-974,共15页美国植物学期刊（英文）

摘　　要：Apple scab caused by </span><i><span style="font-family:Verdana;">Venturia inaequalis</span></i><span style="font-family:Verdana;"> (Cke.) Wint. is the most important disease of apple trees worldwide and requires a high number of fungicide applications. The G143A substitution in the inhibitor binding site of cytochrome </span><i><span style="font-family:Verdana;">b</span></i><span style="font-family:Verdana;"> of </span><i><span style="font-family:Verdana;">V. inaequalis</span></i><span style="font-family:Verdana;"> confers a high level of resistance to strobilurins targeting the </span><i><span style="font-family:Verdana;">bc</span></i>_{<span style="font-family:Verdana;">1</span>}<span style="font-family:Verdana;"> complex. The aim of this work was to substitute the labor intensive </span><i><span style="font-family:Verdana;">in vitro</span></i><span style="font-family:Verdana;"> assays, with the faster quantitative PCR. An allele-specific qPCR method with a newly designed primer set was successfully developed to quantitatively determine the frequency of QoI-resistant A143 allele in populations of </span><i><span style="font-family:Verdana;">V. inaequalis. </span></i><span style="font-family:Verdana;">To be able to suggest that the molecular method could be applied as unique and robust technique, we carried out </span><i><span style="font-family:Verdana;">in vitro</span></i><span style="font-family:Verdana;"> sensitivity test to trifloxystrobin;first testing the relative germination and subsequently confirmed with the quantification of mutated allele frequencies by qPCR on forty-nine Italian </span><i><span style="font-family:Verdana;">V. inaequalis</span></i><span style="font-family:Verdana;"> populations. qPCR gave a similar pattern to that obtained using </span><i><span style="font-family:Verdana;">in vitro</span></i><span style="font-family:Verdana;"> conidial germination test in predominantly sensitive and resistant populations, the variability between these two tests was observed in some heterogenApple scab caused by </span><i><span style="font-family:Verdana;">Venturia inaequalis</span></i><span style="font-family:Verdana;"> (Cke.) Wint. is the most important disease of apple trees worldwide and requires a high number of fungicide applications. The G143A substitution in the inhibitor binding site of cytochrome </span><i><span style="font-family:Verdana;">b</span></i><span style="font-family:Verdana;"> of </span><i><span style="font-family:Verdana;">V. inaequalis</span></i><span style="font-family:Verdana;"> confers a high level of resistance to strobilurins targeting the </span><i><span style="font-family:Verdana;">bc</span></i>_{<span style="font-family:Verdana;">1</span>}<span style="font-family:Verdana;"> complex. The aim of this work was to substitute the labor intensive </span><i><span style="font-family:Verdana;">in vitro</span></i><span style="font-family:Verdana;"> assays, with the faster quantitative PCR. An allele-specific qPCR method with a newly designed primer set was successfully developed to quantitatively determine the frequency of QoI-resistant A143 allele in populations of </span><i><span style="font-family:Verdana;">V. inaequalis. </span></i><span style="font-family:Verdana;">To be able to suggest that the molecular method could be applied as unique and robust technique, we carried out </span><i><span style="font-family:Verdana;">in vitro</span></i><span style="font-family:Verdana;"> sensitivity test to trifloxystrobin;first testing the relative germination and subsequently confirmed with the quantification of mutated allele frequencies by qPCR on forty-nine Italian </span><i><span style="font-family:Verdana;">V. inaequalis</span></i><span style="font-family:Verdana;"> populations. qPCR gave a similar pattern to that obtained using </span><i><span style="font-family:Verdana;">in vitro</span></i><span style="font-family:Verdana;"> conidial germination test in predominantly sensitive and resistant populations, the variability between these two tests was observed in some heterogen

关 键 词：Venturia inaequalis qPCR STROBILURIN Cytochrome b 

分 类 号：O17[理学—数学]