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A rapid real-time PCR assay for detecting columnar growth in apples (Malus x domestica)

机译:用于检测苹果柱状生长的快速实时PCR测定(Malus x Domestica)

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摘要

Columnar growth in apples can have considerable positive consequences for orchard maintenance, but the lack of columnar cultivars producing fruits with desired market quality prevents their broader utilization in orchards. New crossings focused on their breeding are therefore conducted throughout the world every year. Unfortunately, the reliable discrimination between columnar growth and normal phenotype is difficult until seedlings are 2-3 years of age. However, molecular marker-assisted selection (MAS) can be used in seedlings several weeks old providing immense help in breeding programmes focused on columnar apple production. Here, we describe a rapid and reliable method for the detection of both wild-type (WT) and columnar (Co) alleles originating from 'McIntosh Wijcik' in a single reaction using a real-time PCR allelic discrimination approach. Our assay was tested in 130 genotypes (80 normal and 50 columnar habit) from various crossings and their parents, and in all cases, genotype corresponded to phenotype. The assay is thus suitable for laboratories interested in high-throughput MAS for columnar tree habit breeding.
机译:苹果的柱状生长对果园的维护有相当大的积极影响,但由于缺乏能产生理想市场质量果实的柱状品种,阻碍了它们在果园中的广泛应用。因此,世界各地每年都会进行新的杂交,重点是繁殖它们。不幸的是,柱状生长和正常表型之间的可靠区分在幼苗2-3岁之前是困难的。然而,分子标记辅助选择(MAS)可用于数周龄的幼苗,为以柱状苹果生产为重点的育种计划提供了巨大帮助。在这里,我们描述了一种快速可靠的方法,使用实时PCR等位基因鉴别方法,在单一反应中检测源自“McIntosh Wijcik”的野生型(WT)和柱状(Co)等位基因。我们对来自不同杂交组合及其亲本的130个基因型(80个正常基因型和50个柱状习惯基因型)进行了检测,在所有情况下,基因型都与表型相对应。因此,该检测方法适用于对柱状树习性育种的高通量MAS感兴趣的实验室。

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