基于荧光的抗性基因类似物分析在植物遗传资源基因分型中的应用。

Application of fluorescence-based resistance gene analog analysis for genotyping plant genetic resources.

作者信息

Cheng Jianping, Yan Jun, Krugman Tamar, Fahima Tzion

机构信息

Institute of Evolution, University of Haifa, Mount Carmel, Haifa, 31905, Israel.

出版信息

Biotechnol Lett. 2005 Jan;27(2):83-9. doi: 10.1007/s10529-004-6933-2.

DOI:10.1007/s10529-004-6933-2

PMID:15703869

Abstract

Conventional approaches for detecting disease resistance gene analogs (RGAs) in plants are based on agarose gels or on polyacrylamide gel electrophoresis (PAGE) in combination with silver staining or radioactive labeling. A modified method for RGA analysis has been developed by using fluorescence-labeled primers for PCR amplifications. The amplified fragments are detected by denaturing PAGE using an automated laser fluorescence DNA sequencer and analyzed by fragment analysis software. This technique is not limited to specific plant species and is suitable for high-throughput genotyping plant genetic resources. We demonstrate here the efficiency of this method for comparison of RGA patterns in diverse plant species and for genotyping of natural populations of the wheat progenitor, Triticum dicoccoides.

摘要

检测植物中抗病基因类似物（RGAs）的传统方法是基于琼脂糖凝胶或聚丙烯酰胺凝胶电泳（PAGE），并结合银染或放射性标记。通过使用荧光标记引物进行PCR扩增，开发了一种改良的RGA分析方法。扩增片段通过变性PAGE使用自动激光荧光DNA测序仪进行检测，并通过片段分析软件进行分析。该技术不限于特定植物物种，适用于对植物遗传资源进行高通量基因分型。我们在此展示了该方法在比较不同植物物种的RGA模式以及对小麦祖先野生二粒小麦自然群体进行基因分型方面的效率。