Microdissection and Chromosome Painting of the Alien Chromosome in an Addition Line of Wheat - Thinopyrum intermedium

Abstract

In this study, chromosome painting was developed and used to identify alien chromosomes in TAi-27, a wheat - Thinopyrum intermedium addition line, and the chromosomes of the three different genomes of Th. Intermedium. The smallest alien chromosome of TAi-27 was microdissected and its DNA amplified by DOP-PCR was used as a probe to hybridize with metaphase chromosomes of TAi-27 and Th . intermedium . Results showed that hybridization signals were observed in all regions of a pair of the smallest alien chromosomes and the pericentromeric area of another pair of alien chromosomes in TAi-27, indicating that the probe from microdissected chromosome is species specific. In Th . intermedium , 14 chromosomes had wide and strong hybridization signals distributed mainly on the pericentromere area and 9 chromosomes with narrow and weak signals on the pericentromere area. The remaining chromosomes displayed a very weak or no signal. Sequential FISH/GISH on Th . intermedium chromosomes using the DNAs of microdissected chromosome, Pseudoroegneria spicata (St genome) and pDbH12 (a Js genome specific probe) as the probes indicated that the microdissected chromosome belonged to the St genome, three genomes (Js, J and St) in Th . intermedium could be distinguished, in which there is no hybridization signal on J genome that is similar to the genome of Th . bessarabicum . Our results showed that the smallest alien chromosomes may represent a truncated chromosome and the repetitive sequence distribution might be similar in different chromosomes within the St genome. However, the repetitive sequence distributions are different within the Js genome, within a single chromosome, and among different genomes in Th . intermedium . Our results suggested that chromosome painting could be feasible in some plants and useful in detecting chromosome variation and repetitive sequence distribution in different genomes of polyploidy plants, which is helpful for understanding the evolution of different genomes in polyploid plants.

Simultaneous visualization of different genomes (J, JSt and St) in a Thinopyrum intermedium × Thinopyrum ponticum synthetic hybrid (Poaceae) and in its parental species by multicolour genomic in situ hybridization (mcGISH)

“…They proposed the J vs J r St genome designation, where J vs and J r represented ancestral genomes of the present-day J b of Thinopyrum bessarabicum and J e of Thinopyrum elongatum , J vs being the more ancient. The change of J s to J vs is based on the study of Mahelka et al (2011) and Deng et al (2013), as all 14 chromosomes of J s showed GISH/FISH hybridization signals from V-genome probes [ Dasypyrum villosum (Linnaeus, 1753) P. Candargy, 1901], but only 8 to 11 of the 14 chromosomes have the centromeric region being hybridized by the St genome probe (Chen et al 1998, Zhang et al 1996, Kishii et al 2005, Tang et al 2011, Deng et al 2013). FISH analysis using pMD232-500 as probe (originating from Secale cereale Linnaeus, 1753 cv.…”

Degenerate Oligonucleotide Primed–Polymerase Chain Reaction‐Based Chromosome Painting of P Genome Chromosomes in Agropyron cristatum and Wheat–A. cristatum Addition Lines

“…From the distribution patterns of DOP–PCR products from microdissected 6PS and 6PL, we found that the repetitive sequences were all over the P genome chromosomes, an observation different from that observed on the St genome chromosome. The DOP–PCR products of the smallest St chromosome from Tai‐27 only distributed in the pericentromeric region of St genome chromosomes (Deng et al, 2013a). In addition, the DOP–PCR products of 1R and 1RS yielded weaker signals in the terminals of chromosomes than in the other regions (Deng et al, 2014).…”

“…A DNA library of Gossypium arboreum chromosome 5 containing approximately 173,000 clones was created by chromosome microdissection and microcloning (Peng et al, 2012). In addition, there were chromosomes microdissected from wheat, Thinopyrum intermedium , and spinach ( Spinacia oleracea L.) (Deng et al, 2013a, 2013b; Vega et al, 1994). The microdissected DNA would facilitate specific probe screening, molecular mapping, and gene cloning.…”

“…In mammalian species, chromosome painting can identify individual chromosome, so it is a powerful tool to study synteny and chromosomal evolution (Wienberg and Stanyon, 1997). In plant species, due to the presence of many repetitive DNA sequences in all chromosomes, this technique can only be used for identification of genome, and alien chromosomes in addition lines or translocation lines (Deng et al, 2014; Deng et al, 2013a). If the repetitive sequences were excluded from the painting probes, chromosome painting may identify individual chromosome of plant genome chromosomes (Jiang and Gill, 2006).…”

Development and molecular cytogenetic identification of a new wheat–Leymus mollis Lm#6Ns disomic addition line

“…, Deng et al. ). In addition, molecular marker techniques are powerful tools for determining the efficiency of the introgression of useful genes from related species into common wheat (Niu et al.…”