National Key Laboratory for Germplasm Innovation & Utilization of Horticultural Crops, Huazhong Agricultural University.
National Key Laboratory for Germplasm Innovation & Utilization of Horticultural Crops, Huazhong Agricultural University;
J Vis Exp. 2023 Jun 30(196). doi: 10.3791/65056.
Citrus uses S-RNase-based self-incompatibility to reject self-pollen and, therefore, requires nearby pollinizer trees for successful pollination and fertilization. However, identifying suitable varieties to serve as pollinizers is a time-consuming process. To solve this problem, we have developed a rapid method for identifying pollination-compatible citrus cultivars that utilizes agarose gel electrophoresis and aniline blue staining. Pollen compatibility is determined based on the identification of S genotypes by extracting total DNA and performing PCR-based genotyping assays with specific primers. Additionally, styles are collected 3-4 days after manual pollination, and aniline blue staining is performed. Finally, the growth status of the pollen tubes is observed with a fluorescence microscope. Pollen compatibility and incompatibility can be established by observing whether the pollen tube growth is normal or suppressed, respectively. Due to its simplicity and cost-effectiveness, this method is an effective tool for determining the pollen compatibility and incompatibility of different citrus varieties to establish incompatibility groups and incompatibility relationships between different cultivars. This method provides information essential for the successful selection of suitable pollinizer trees and, thus, facilitates the establishment of new orchards and the selection of appropriate parents for breeding programs.
柑橘类植物利用 S-RNase 基的自交不亲和性来排斥自花花粉，因此需要附近的传粉树种来实现成功的授粉和受精。然而，识别合适的品种作为传粉者是一个耗时的过程。为了解决这个问题，我们开发了一种利用琼脂糖凝胶电泳和苯胺蓝染色来快速识别亲和柑橘品种的方法。花粉亲和性是通过提取总 DNA 并使用特定引物进行基于 PCR 的基因分型检测来确定 S 基因型来确定的。此外，在人工授粉后 3-4 天收集花柱，并进行苯胺蓝染色。最后，用荧光显微镜观察花粉管的生长情况。通过观察花粉管生长是否正常或受到抑制，可以确定花粉的亲和性和不亲和性。由于其简单性和成本效益，这种方法是确定不同柑橘品种花粉亲和性和不亲和性的有效工具，有助于建立不亲和组和不同品种之间的不亲和关系。该方法提供了成功选择合适传粉树的必要信息，从而促进了新果园的建立和育种计划中合适亲本的选择。
