Monosomic lines of Nicotiana tabacum are helpful to confirm the location of genes on specific chromosomes. In the cross N. nudicaulis and N. tabacum , hybrid seedlings express lethal symptoms, which are controlled by the S subgenome of N. tabacum . To identify the responsible chromosome, we needed to produce chromosome lacking lines (CLLs) of N. tabacum L. “ Red Russian ” and use them to cross with N. nudicaulis . From a cross of ( N. tabacum × N. tomentosiformis ) × N. tabacum , 380 BC1 individuals were obtained. Using a Haplo-Q line (a monosomic line lacking the single linkage group 11 ) and N. tabacum , we found that qPCR is a simple and reliable screening method for CLLs of N. tabacum . The marker PT30342 is located on linkage group 11 , and the -Ct value (Ct Actin - Ct PT30342) was 2.0 for a disomic line and was 1.097 for a Haplo-Q line. By the use of flow cytometry, qPCR and chromosome counting together as a screening method, we identified 6 CLLs lacking 2 to 6 chromosomes. Compared with conventional methods, our method is a rapid technique for making and screening CLLs ofthe S or S/T subgenome of N. tabacum . Further, these CLLs will be useful to identify the location of two or more factors on chromosomes controlling a variety of genetic problems affecting breeding. Here, we only made CLLs of the S or S/T subgenome of N. tabacum . W e will use the method established in this study to produce CLLs of the T subgenome of N. tabacum , and gather a full set of CLLs of N. tabacum . qPCR could also be applied to the identification of chromosome aberrations in other plants.
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