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Development of oligonucleotides and multiplex probes for quick and accurate identification of wheat and Thinopyrum bessarabicum chromosomes.
Genome. 2017 Feb; 60(2):93-103.G

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.

Authors+Show Affiliations

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

Pub Type(s)

Journal Article

Language

eng

PubMed ID

27936984

Citation

Du, Pei, et al. "Development of Oligonucleotides and Multiplex Probes for Quick and Accurate Identification of Wheat and Thinopyrum Bessarabicum Chromosomes." Genome, vol. 60, no. 2, 2017, pp. 93-103.
Du P, Zhuang L, Wang Y, et al. Development of oligonucleotides and multiplex probes for quick and accurate identification of wheat and Thinopyrum bessarabicum chromosomes. Genome. 2017;60(2):93-103.
Du, P., Zhuang, L., Wang, Y., Yuan, L., Wang, Q., Wang, D., Dawadondup, ., Tan, L., Shen, J., Xu, H., Zhao, H., Chu, C., & Qi, Z. (2017). Development of oligonucleotides and multiplex probes for quick and accurate identification of wheat and Thinopyrum bessarabicum chromosomes. Genome, 60(2), 93-103. https://doi.org/10.1139/gen-2016-0095
Du P, et al. Development of Oligonucleotides and Multiplex Probes for Quick and Accurate Identification of Wheat and Thinopyrum Bessarabicum Chromosomes. Genome. 2017;60(2):93-103. PubMed PMID: 27936984.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Development of oligonucleotides and multiplex probes for quick and accurate identification of wheat and Thinopyrum bessarabicum chromosomes. AU - Du,Pei, AU - Zhuang,Lifang, AU - Wang,Yanzhi, AU - Yuan,Li, AU - Wang,Qing, AU - Wang,Danrui, AU - Dawadondup,, AU - Tan,Lijun, AU - Shen,Jian, AU - Xu,Haibin, AU - Zhao,Han, AU - Chu,Chenggen, AU - Qi,Zengjun, Y1 - 2016/09/14/ PY - 2016/12/13/pubmed PY - 2017/2/24/medline PY - 2016/12/13/entrez KW - ND-FISH KW - chromosome engineering KW - chromosome painting KW - coloration des chromosomes KW - diversité génétique KW - genetic diversity KW - ingénierie chromosomique KW - multiplexes d’oligonucléotides KW - oligonucleotide multiplexes SP - 93 EP - 103 JF - Genome JO - Genome VL - 60 IS - 2 N2 - 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. SN - 1480-3321 UR - https://www.unboundmedicine.com/medline/citation/27936984/Development_of_oligonucleotides_and_multiplex_probes_for_quick_and_accurate_identification_of_wheat_and_Thinopyrum_bessarabicum_chromosomes_ L2 - https://cdnsciencepub.com/doi/10.1139/gen-2016-0095?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -