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基于 I 类 S 单倍型鉴定的分子标记检测自交不亲和油菜(甘蓝型油菜)植株。

Detection of self-incompatible oilseed rape plants (Brassica napus L.) based on molecular markers for identification of the class I S haplotype.

机构信息

Biotechnological Centre, Faculty of Agriculture , University of South Bohemia , Ceské Budejovice , Czech Republic .

Crop Research Institute , Praha , Czech Republic .

出版信息

Genet Mol Biol. 2014 Sep;37(3):556-9. doi: 10.1590/s1415-47572014000400012.

Abstract

The selection of desirable genotypes with recessive characteristics, such as self-incompatible plants, is often difficult or even impossible and represents a crucial barrier in accelerating the breeding process. Molecular approaches and selection based on molecular markers can allow breeders to overcome this limitation. The use of self-incompatibility is an alternative in hybrid breeding of oilseed rape. Unfortunately, stable self-incompatibility is recessive and phenotype-based selection is very difficult and time-consuming. The development of reliable molecular markers for detecting desirable plants with functional self-incompatible genes is of great importance for breeders and allows selection at early stages of plant growth. Because most of these reliable molecular markers are based on discrimination of class I S-locus genes that are present in self-compatible plants, there is a need to use an internal control in order to detect possible PCR inhibition that gives false results during genotyping. In this study, 269 double haploid F2 oilseed rape plants obtained by microspore embryogenesis were used to verify the applicability of an improved PCR assay based on the detection of the class I SLG gene along with an internal control. Comparative analysis of the PCR genotyping results vs. S phenotype analysis confirmed the applicability of this molecular approach in hybrid breeding programs. This approach allows accurate detection of self-incompatible plants via a different amplification profile.

摘要

具有隐性特征的理想基因型的选择,如自交不亲和植物,通常是困难的,甚至是不可能的,这是加速育种过程的一个关键障碍。基于分子标记的分子方法和选择可以使育种者克服这一限制。自交不亲和性的利用是油菜杂种育种的一种替代方法。不幸的是,稳定的自交不亲和性是隐性的,基于表型的选择非常困难和耗时。开发可靠的分子标记来检测具有功能性自交不亲和基因的理想植物对育种者来说非常重要,并且可以在植物生长的早期进行选择。因为这些可靠的分子标记大多基于存在于自交亲和植物中的 I 类 S 座位基因的区分,所以需要使用内部对照来检测在基因型检测过程中可能出现的导致假阳性结果的 PCR 抑制。在这项研究中,使用了 269 株通过小孢子胚胎发生获得的双单倍体 F2 油菜植物,以验证基于 I 类 SLG 基因检测和内部对照的改良 PCR 检测的适用性。PCR 基因分型结果与 S 表型分析的比较分析证实了这种分子方法在杂种育种计划中的适用性。这种方法可以通过不同的扩增谱准确检测自交不亲和植物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5531/4171774/2dc936655a39/gmb-37-556-g001.jpg

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