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

文献类型： 外文期刊

作者： Du, Pei 1 ; Zhuang, Lifang 1 ; Wang, Yanzhi 1 ; Yuan, Li 1 ; Wang, Qing 1 ; Wang, Danrui 1 ; Dawadondup 1 ; Tan, Lijun 1 ; S 1 ;

作者机构： 1.Nanjing Agr Univ, JCIC MCP, State Key Lab Crop Genet & Germplasm Enhancement, Nanjing 210095, Jiangsu, Peoples R China

2.Nanjing Agr Univ, JCIC MCP, State Key Lab Crop Genet & Germplasm

关键词： chromosome engineering;chromosome painting;genetic diversity;ND-FISH;oligonucleotide multiplexes

期刊名称：GENOME （ 影响因子：2.166； 五年影响因子：2.474 ）

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收录情况： SCI

摘要： 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.

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