Original Research ARTICLE
Development and applications of chromosome-specific cytogenetic BAC-FISH probes in S. spontaneum
- 1Center for Genomics and Biotechnology, Haixia Institute of Science and Technology, Fujian Provincial Key Laboratory of Haixia Applied Plant Systems Biology, College of life science, Fujian Agriculture and Forestry University, China
- 2Key Laboratory of Sugarcane Biology and Genetic Breeding, Ministry of Agriculture, Fujian Agriculture and Forestry University, China
- 3College of Life Sciences, Fujian Normal University, China
- 4Agronomy Department, University of Florida, United States
- 5Department of Plant Pathology and Microbiology, Texas A&M University System, United States
- 6Department of Plant Biology, University of Illinois at Urbana–Champaign, United States
Saccharum spontaneum is a major Saccahrum species that contributed to the origin of modern sugarcane cultivars, and is considered to be a plant species with one of the most complex genetics due to a high degree of polyploidy. Fluorescence in situ hybridization is a powerful and widely used tool in the genome studies. Here we demonstrated that fluorescence in situ hybridization (FISH) based on bacterial artificial chromosome (BAC) clones can be used as a specific cytological marker to identify S.spontaneum individual chromosomes and study the relationship between S.spontaneum and other related species. We screened low-copy BACs as probes from the sequences of a high coverage of S. spontaneum BAC library based on BLAST search of the sorghum genome. In total, we isolated 49 positive BAC clones, and identified 27 BAC clones that can give specific signals on the S.spontaneum chromosomes. Of the 27 BAC probes, 18 were confirmed to be able to discriminate the eight basic chromosomes of S.spontaneum. Moreover, BAC-24, BAC-66, BAC-78, BAC-69; BAC-71, BAC-73 and BAC-77 probes were used to construct chromosome 1 and chromosome 2 physical mapping of S. spontaneum, indicating synteny in Sb01 between S.spontaneum and sorghum. Furthermore, we found that BAC-14 and BAC-19 probes, corresponding to the sorghum chromosomes 2 and 8, respectively, localized to different arms of the same S.spontaneum chromosome, suggesting that there must have been an inter-chromosomal rearrangement event between S. spontaneum and sorghum. Our study provided the first set of chromosome-specific cytogenetic makers in Saccharum, and is critical for future advances in cytogenetics and genome sequencing studies in Saccharum.
Keywords: Saccharum spontaneum, Sorghum, Polyploidy, Bacterial artificial chromosome (BAC), fluorescence in situ hybridization (FISH).
Received: 29 Nov 2017;
Accepted: 05 Feb 2018.
Edited by:Jun Yang, Shanghai Chenshan Plant Science Research Center (CAS), China
Reviewed by:Gabriel Rodrigues Alves Margarido, University of São Paulo, Brazil
Jianying Sun, Jiangsu Normal University, China
Copyright: © 2018 Dong, Shen, Zhang, Wang, Yu, Ming, Wang and Zhang. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Prof. Jisen Zhang, Fujian Agriculture and Forestry University, Center for Genomics and Biotechnology, Haixia Institute of Science and Technology, Fujian Provincial Key Laboratory of Haixia Applied Plant Systems Biology, College of life science, Fuzhou, China, email@example.com