|CUI, YU - Shandong Agricultural University|
|ZHANG, YANPING - Shandong Agricultural University|
|QI, JUAN - Shandong Agricultural University|
|WANG, HONGGANG - Shandong Agricultural University|
|BAO, YINGUANG - Shandong Agricultural University|
|LI, XINGFENG - Shangdong Agricultural University|
Submitted to: Genome
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 3/31/2018
Publication Date: 5/8/2018
Citation: Cui, Y., Zhang, Y., Qi, J., Wang, H., Wang, R., Bao, Y., Li, X. 2018. Identification of chromosomes in Thinopyrum intermedium and wheat-Th. intermedium amphiploids based on multiplex oligonucleotides probes. Genome. 61(7):515-521. https://doi.org/10.1139/gen-2018-0019.
Interpretive Summary: Chromosomes of intermediate wheatgrass had been studied by genomic in situ hybridiation (GISH) and single probe fluorescence in situ hybridization (FISH). Although chromosomes could be distinguished into three different subgenomes, homologous pairs within a subgenome could not be identified with certainty. In this study, multiple synthesized oligonucleotides were used in FISH to characterize chromosomes of wheat, intermediate wheatgrass, and wheat-intermediate wheatgrass ampliploid. Chromosome identity could be reliably ascertained regarding to not only the subgenome but also the homologous pair within subgenome. Changes in chromosomal structures in wheat and grass chromosomes could also be detected. This multiplex FISH techique will be very useful in general chromosome identification and alien chromosome transfer.
Technical Abstract: Synthesized obligonucleotide (oligo) can be used as effective probes as plasmid clones for chromosomes identification in fluorescence in situ hybridization (FISH) analysis, making oligoFISH procedure a simpler and more efficient molecular cytogenetic technique in studying of plants. In this study, multiplex oligonucleotide probes including pSc119.2-1, pAS1-4, (GAA)10, (AAC)6 and 45S rDNA were combined and used in FISH to identify chromosomes in common wheat, Thinopyrum intermedium and a wheat-Th, intermedium amphiploids TE256-1. In comparison with general FISH probes, signals generated by the multiplex probes were more abundant, colorful and characteristic. While combining the results of genomic in situ hybridization (GISH) with FISH, Th. intermedium chromosomes and alien chromosomes in TE256-1 could be classified and identified more precisely, especially the J- and Js-genome chromosomes. Moreover, based on the FISH results of multiplex probes, more structural variations in wheat chromosomes of TE256-1 were detected. Results indicated that multiplex oligo probes would have a wide range of application prospects in creation and identification of wheat-Th. intermedium germplasms.