Abstract
Non-denaturing FISH (ND-FISH) technology has been widely used to study the chromosomes of Triticeae species because of its convenience. The oligo probes for ND-FISH analysis of wheat (Triticum aestivum L.) chromosomes are still limited. In this study, the whole genome shotgun assembly sequences (IWGSC WGA v0.4) and the first version of the reference sequences (IWGSC RefSeq v1.0) of Chinese Spring (T. aestivum L.) were used to find new tandem repeats. One hundred and twenty oligo probes were designed according to the new tandem repeats and used for ND-FISH analysis of chromosomes of wheat Chinese Spring. Twenty nine of the 120 oligo probes produce clear or strong signals on wheat chromosomes. Two of the 29 oligo probes can be used to conveniently distinguish wheat A-, B-, and D-genome chromosomes. Sixteen of the 29 oligo probes only produce clear or strong signals on the subtelomeric regions of 1AS, 5AS, 7AL, 4BS, 5BS, and 3DS arms, on the telomeric regions of 1AL, 5AL, 2BS, 3BL, 6DS, and 7DL arms, on the intercalary regions of 4AL and 2DL arms, and on the pericentromeric regions of 3DL and 6DS arms. Eleven of the 29 oligo probes generate distinct signal bands on several chromosomes and they are different from those previously reported. In addition, the short and long arms of 6D chromosome have been confirmed. The new oligo probes developed in this study are useful and convenient for distinguishing wheat chromosomes or specific segments of wheat chromosomes.
Highlights
Genomic in situ hybridization (GISH) and Fluorescent in situ hybridization (FISH) techniques are often used to identify plant chromosomes (Kato et al, 2005)
One hundred and fourteen new tandem repeats were detected from whole genome shotgun assembly sequences (IWGSC WGA v0.4) and the first version of the reference sequences (IWGSC RefSeq v1.0) of wheat Chinese Spring
Non-denaturing FISH (ND-FISH) technology has provided a convenient and efficient way to identify the chromosomes of Triticeae species (Cuadrado and Jouve, 2010; Carmona et al, 2013; Carvalho et al, 2013; Cuadrado et al, 2013; Cabo et al, 2014; Tang et al, 2014; Fu et al, 2015; Delgado et al, 2016, 2017; Li et al, 2016; Li J. et al, 2017; Li H. et al, 2017; Du et al, 2017; Duan et al, 2017; Jiang et al, 2017; Xiao et al, 2017)
Summary
Genomic in situ hybridization (GISH) and Fluorescent in situ hybridization (FISH) techniques are often used to identify plant chromosomes (Kato et al, 2005). Multicolor GISH has been successfully used to identify the A, B, and D genomes of common wheat (Mukai et al, 1993; Han et al, 2003, 2004; Zhao et al, 2011). FISH technology using genes and repetitive DNA sequences as probes is often used to establish chromosomal landmarks of the Triticeae tribe (Cuadrado and Schwarzacher, 1998; Yuan and Tomita, 2009; Wang et al, 2010; Linc et al, 2012; Cuadrado et al, 2013; Molnár et al, 2014; Danilova et al, 2017). The multicolor GISH using total genomic DNA as probes and the FISH using the repetitive DNA sequences as probes can unambiguously identify the different genome chromosomes or individual chromosome of wheat, their procedures are time-consuming because of the preparation of probe sequences, the labeling of DNA sequences, and the denaturing of chromosomes and probes
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have