Kim Jae Yoon, Moon Jun-Cheol, Kim Hyo Chul, Shin Seungho, Song Kitae, Kim Kyung-Hee, Lee Byung-Moo
Department of Plant Resources, College of Industrial Science, Kongju National University, Yesan 32439, Republic of Korea.
Agriculture and Life Science Research Institute, Kangwon National University, Chuncheon 24341, Republic of Korea.
Appl Plant Sci. 2017 Feb 14;5(2). doi: 10.3732/apps.1600132. eCollection 2017 Feb.
Positional cloning in combination with phenotyping is a general approach to identify disease-resistance gene candidates in plants; however, it requires several time-consuming steps including population or fine mapping. Therefore, in the present study, we suggest a new combined strategy to improve the identification of disease-resistance gene candidates.
Downy mildew (DM)-resistant maize was selected from five cultivars using a spreader row technique. Positional cloning and bioinformatics tools were used to identify the DM-resistance quantitative trait locus marker (bnlg1702) and 47 protein-coding gene annotations. Eventually, five DM-resistance gene candidates, including , , and , were identified by quantitative reverse-transcription PCR (RT-PCR) without fine mapping of the bnlg1702 locus.
The combined protocol with the spreader row technique, quantitative trait locus positional cloning, and quantitative RT-PCR was effective for identifying DM-resistance candidate genes. This cloning approach may be applied to other whole-genome-sequenced crops or resistance to other diseases.
定位克隆与表型分析相结合是鉴定植物抗病基因候选者的常用方法；然而，它需要几个耗时的步骤，包括群体或精细定位。因此，在本研究中，我们提出了一种新的组合策略来改进抗病基因候选者的鉴定。
利用传播行技术从五个玉米品种中筛选出抗霜霉病（DM）的玉米。使用定位克隆和生物信息学工具来鉴定DM抗性数量性状位点标记（bnlg1702）和47个蛋白质编码基因注释。最终，通过定量逆转录PCR（RT-PCR）鉴定出五个DM抗性基因候选者，包括 、 和 ，而无需对bnlg1702位点进行精细定位。
传播行技术、数量性状位点定位克隆和定量RT-PCR相结合的方案对于鉴定DM抗性候选基因是有效的。这种克隆方法可能适用于其他全基因组测序作物或对其他疾病的抗性研究。
