In plant breeding, in new breeding and in the routine production of seeds or the production of young plants, there are a number of scenarios in which natural or induced variations in the number of chromosomes, i.e. aneuploidy, can occur.
These are often undesirable, sometimes also desirable changes (induced processes). Aneuploidies usually arise due to errors in cell division, in particular due to non-disjunction – that is, chromosome pairs do not separate correctly.
Aneuploidy can be both useful and problematic in plant breeding: While it is used specifically in experimental breeding and the development of new plant varieties, it can lead to undesirable effects, especially in seed production and plant propagation.
For example, aneuploid individuals are used to assign certain genes to certain chromosomes, as a source of genetic variation or to transfer useful genes from wild species to cultivated lines (introgression).
On the other hand, unwanted, i.e. spontaneous aneuploidy, leads to unstable inheritance with lower fertility or sterility. It leads to growth and developmental disorders or to increased sensitivity to stress factors. In production, such aneuploid individuals are not desirable, as they negatively affect the homogeneity and quality of the seed or young plants. If seed with a high proportion of aneuploid seeds is sown, a significant reduction in yield is to be expected. This harms the farmer directly and indirectly the seed producer. In plant production, on the other hand, aneuploid individuals are often phenotypically conspicuous and thus produce a higher level of waste for the producer, thus directly reducing their profit.
Various technologies are available for determining aneuploidy. These are very complex and slow (chromosome counting), very specific and expensive (FISH via gene probes, PCR/qPCR via specific markers) or a combination of both (genome-wide sequencing) and are therefore unsuitable for the search for aneuploidy on an industrial level.
The rapid, cost-effective and reliable quantification of DNA using flow cytometry represents a methodology adapted to industrial standards for the search for aneuploid individuals. The cell nuclei are extracted from the tissue and the DNA of the cell nuclei is labeled with a fluorescent dye.
The fluorescence intensity of the cell nuclei is measured via flow cytometry and correlated with the regular ploidy level of the species under investigation. Aueuploid plants differ from plants with a normal chromosome number and regular DNA content and can thus be clearly identified. This process can be largely automated, so that a large number of individual plant samples can be measured in a short time with few resources. This makes the methodology perfectly suited for screening for aneuploidy in large populations, in production, breeding and research. Leaves, seeds and callus material can be used as samples.
The CyStain™ reagents for ploidy determination, together with the specially matched CyFlow™ RP Ploidy Analyzers from Sysmex Partec GmbH, represent a complete package of precisely fitting reagents and analysis device. Thanks to further accessories, such as the fast CyFlow™ AL-30 Autoloader and the competent advice from our product specialists based on 50 years of experience in the market, Sysmex is the only provider worldwide to offer a coordinated complete package for flow cytometric determination of ploidy and aneuploidy.
CyStain™ reagents, such as CyStain™ UV Precise P, CyStain™ UV Ploidy and CyStain™ UV OxProtect, are available in various versions and packaging units and can thus be precisely tailored to the user profile and the plants to be examined. In addition, like all Sysmex Partec products, they are produced in accordance with the strict quality requirements of EN ISO 13485:2016 and ISO 9001:2015, so you can always be on the safe side with our CyStain™ reagents.
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