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Chromosome Dosage Analysis in Plants Using Whole Genome Sequencing   

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Abstract

Relative chromosome dosage, i.e., increases or decreases in the number of copies of specific chromosome regions in one sample versus another, can be determined using aligned read-counts from Illumina sequencing (Henry et al., 2010). The following protocol was used to identify the different classes of aneuploids that result from uniparental genome elimination in Arabidopsis thaliana, including chromosomes that have undergone chromothripsis (Tan et al., 2015). Uniparental genome elimination results in the production of haploid progeny from crosses to specific strains called “haploid inducers” (Ravi et al., 2014). On the other hand, chromothripsis, which was first discovered in cancer genomes, is a phenomenon that results in clustered, highly rearranged chromosomes. In plants, chromothripsis has been observed as a result of genome elimination (Tan et al., 2015). Detecting variation in chromosome dosage has multiple applications beside those linked to genome elimination. For example, a dosage variant population of poplar hybrids was created by gamma-irradiation of pollen grains. Hundreds of dosage lesions, insertions and deletions, were identified using this technique and provide a way to associate loci with the phenotypic consequences observed in this population (Henry et al., 2015).

This method has been successfully used to detect changes in chromosome dosage in many different species, including Arabidopsis thaliana (Tan et al., 2015), Arabidopsis suecica (Ravi et al., 2014), rice (Henry et al., 2010) and poplar (Henry et al., 2015). It is important to note that dosage plots always indicate dosage variation relative to the control sample used (Note 1). Therefore, this approach is not suitable to detect ploidy variants (diploid vs triploid, for example). Similarly, this technique does not allow the detection of balanced chromosomal rearrangements such as reciprocal translocations.

Keywords: Dosage Analysis, Whole Genome Sequencing, Chromothripsis, Genome Elimination, Aneuploidy

Copyright Tan et al. This article is distributed under the terms of the Creative Commons Attribution License (CC BY 4.0).
How to cite:  Readers should cite both the Bio-protocol article and the original research article where this protocol was used:
  1. Tan, E. H., Comai, L. and Henry, I. M. (2016). Chromosome Dosage Analysis in Plants Using Whole Genome Sequencing. Bio-protocol 6(13): e1854. DOI: 10.21769/BioProtoc.1854.
  2. Tan, E. H., Henry, I. M., Ravi, M., Bradnam, K. R., Mandakova, T., Marimuthu, M. P., Korf, I., Lysak, M. A., Comai, L. and Chan, S. W. (2015). Catastrophic chromosomal restructuring during genome elimination in plants. Elife 4.06516.
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