Herrera Sara, Lora Jorge, Hormaza José I, Rodrigo Javier
Unidad de Hortofruticultura, Centro de Investigación y Tecnología Agroalimentaria de Aragón (CITA); Instituto Agroalimentario de Aragón - IA2 (CITA-Universidad de Zaragoza);
Instituto de Hortofruticultura Subtropical y Mediterránea La Mayora (IHSM La Mayora-UMA-CSIC).
J Vis Exp. 2020 Jun 16(160). doi: 10.3791/60241.
Self-incompatibility in Rosaceae is determined by a Gametophytic Self-Incompatibility System (GSI) that is mainly controlled by the multiallelic locus S. In apricot, the determination of self- and inter-(in)compatibility relationships is increasingly important, since the release of an important number of new cultivars has resulted in the increase of cultivars with unknown pollination requirements. Here, we describe a methodology that combines the determination of self-(in)compatibility by hand-pollinations and microscopy with the identification of the S-genotype by PCR analysis. For self-(in)compatibility determination, flowers at balloon stage from each cultivar were collected in the field, hand-pollinated in the laboratory, fixed, and stained with aniline blue for the observation of pollen tube behavior under the fluorescence microscopy. For the establishment of incompatibility relationships between cultivars, DNA from each cultivar was extracted from young leaves and S-alleles were identified by PCR. This approach allows establishing incompatibility groups and elucidate incompatibility relationships between cultivars, which provides a valuable information to choose suitable pollinizers in the design of new orchards and to select appropriate parents in breeding programs.
蔷薇科的自交不亲和性由配子体自交不亲和系统（GSI）决定，该系统主要由多等位基因座S控制。在杏树中，确定自交和杂交（不）亲和关系变得越来越重要，因为大量新品种的推出导致了授粉需求未知的品种数量增加。在此，我们描述了一种方法，该方法将通过人工授粉和显微镜观察来确定自交（不）亲和性与通过PCR分析鉴定S基因型相结合。为了确定自交（不）亲和性，在田间采集每个品种处于气球期的花朵，在实验室进行人工授粉，固定后用苯胺蓝染色，以便在荧光显微镜下观察花粉管行为。为了确定品种间的不亲和关系，从每个品种的幼叶中提取DNA，并通过PCR鉴定S等位基因。这种方法可以建立不亲和组并阐明品种间的不亲和关系，这为在新果园设计中选择合适的授粉树以及在育种计划中选择合适的亲本提供了有价值的信息。
