Garcia Melissa, Mather Diane E
Australian Centre for Plant Functional Genomics, School of Agriculture, Food and Wine, Waite Research Institute, University of Adelaide, Hartley Grove, Urrbrae, Glen Osmond, SA, Australia,
Methods Mol Biol. 2014;1145:21-36. doi: 10.1007/978-1-4939-0446-4_2.
Once the sequence is known for a gene of interest, it is usually possible to design markers to detect polymorphisms within the gene. Such markers can be particularly useful in plant breeding, especially if they detect the causal polymorphism within the gene and are diagnostic of the phenotype. In this chapter, we (1) discuss how gene sequences are obtained and aligned and how polymorphic sites can be identified or predicted; (2) explain the principles of PCR primer design and PCR amplification and provide guidelines for their application in the design and testing of markers; (3) discuss detection methods for presence/absence (dominant) polymorphisms, length polymorphisms and single nucleotide polymorphisms (SNPs); and (4) outline some of the factors that affect the utility of markers in plant breeding and explain how markers can be evaluated (validated) for use in plant breeding.
一旦已知目标基因的序列，通常就有可能设计标记来检测该基因内的多态性。这类标记在植物育种中可能特别有用，尤其是当它们能检测到基因内的因果多态性并可用于诊断表型时。在本章中，我们：（1）讨论如何获取和比对基因序列，以及如何识别或预测多态性位点；（2）解释PCR引物设计和PCR扩增的原理，并为其在标记设计和测试中的应用提供指导；（3）讨论存在/缺失（显性）多态性、长度多态性和单核苷酸多态性（SNP）的检测方法；（4）概述一些影响标记在植物育种中效用的因素，并解释如何对标记进行评估（验证）以用于植物育种。
