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Genome. 2017 Feb;60(2):93-103. doi: 10.1139/gen-2016-0095. Epub 2016 Sep 14.

Development of oligonucleotides and multiplex probes for quick and accurate identification of wheat and Thinopyrum bessarabicum chromosomes.

Author information

1
a State Key Laboratory of Crop Genetics and Germplasm Enhancement, JCIC-MCP, Nanjing Agricultural University, Nanjing 210095, China.
2
b Faculty of Biology and Environmental Science, Nanjing Forestry University, Nanjing 210037, China.
3
c Provincial Key Laboratory of Agrobiology, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China.
4
d Monsanto Company, 21120 Hwy 30, Filer, ID 83328, USA.

Abstract

In comparison with general FISH for preparing probes in terms of time and cost, synthesized oligonucleotide (oligo hereafter) probes for FISH have many advantages such as ease of design, synthesis, and labeling. Low cost and high sensitivity and resolution of oligo probes greatly simplify the FISH procedure as a simple, fast, and efficient method of chromosome identification. In this study, we developed new oligo and oligo multiplex probes to accurately and efficiently distinguish wheat (Triticum aestivum, 2n = 6x, AABBDD) and Thinopyrum bessarabicum (2n = 2x = 14, JJ) chromosomes. The oligo probes contained more nucleotides or more repeat units that produced stronger signals for more efficient chromosome painting. Four Th. bessarabicum-specific oligo probes were developed based on genomic DNA sequences of Th. bessarabicum chromosome arm 4JL, and one of them (oligo DP4J27982) was pooled with the oligo multiplex #1 to simultaneously detect wheat and Th. bessarabicum chromosomes for quick and accurate identification of Chinese Spring (CS) - Th. bessarabicum alien chromosome introgression lines. Oligo multiplex #4 revealed chromosome variations among CS and eight wheat cultivars by a single round of FISH analysis. This research demonstrated the high efficiency of using oligos and oligo multiplexes in chromosome identification and manipulation.

KEYWORDS:

ND-FISH; chromosome engineering; chromosome painting; coloration des chromosomes; diversité génétique; genetic diversity; ingénierie chromosomique; multiplexes d’oligonucléotides; oligonucleotide multiplexes

PMID:
27936984
DOI:
10.1139/gen-2016-0095
[Indexed for MEDLINE]

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