PMID- 36361999 OWN - NLM STAT- MEDLINE DCOM- 20221114 LR - 20230328 IS - 1422-0067 (Electronic) IS - 1422-0067 (Linking) VI - 23 IP - 21 DP - 2022 Oct 30 TI - Accurate Chromosome Identification in the Prunus Subgenus Cerasus (Prunus pseudocerasus) and its Relatives by Oligo-FISH. LID - 10.3390/ijms232113213 [doi] LID - 13213 AB - A precise, rapid and straightforward approach to chromosome identification is fundamental for cytogenetics studies. However, the identification of individual chromosomes was not previously possible for Chinese cherry or other Prunus species due to the small size and similar morphology of their chromosomes. To address this issue, we designed a pool of oligonucleotides distributed across specific pseudochromosome regions of Chinese cherry. This oligonucleotide pool was amplified through multiplex PCR with specific internal primers to produce probes that could recognize specific chromosomes. External primers modified with red and green fluorescence tags could produce unique signal barcoding patterns to identify each chromosome concomitantly. The same oligonucleotide pool could also discriminate all chromosomes in other Prunus species. Additionally, the 5S/45S rDNA probes and the oligo pool were applied in two sequential rounds of fluorescence in situ hybridization (FISH) localized to chromosomes and showed different distribution patterns among Prunus species. At the same time, comparative karyotype analysis revealed high conservation among P. pseudocerasus, P. avium, and P. persica. Together, these findings establish this oligonucleotide pool as the most effective tool for chromosome identification and the analysis of genome organization and evolution in the genus Prunus. FAU - Wang, Lei AU - Wang L AD - College of Horticulture, Sichuan Agricultural University, Chengdu 611130, China. AD - Institute of Pomology and Olericulture, Sichuan Agricultural University, Chengdu 611130, China. FAU - Feng, Yan AU - Feng Y AD - College of Horticulture, Sichuan Agricultural University, Chengdu 611130, China. AD - Institute of Pomology and Olericulture, Sichuan Agricultural University, Chengdu 611130, China. FAU - Wang, Yan AU - Wang Y AD - College of Horticulture, Sichuan Agricultural University, Chengdu 611130, China. AD - Institute of Pomology and Olericulture, Sichuan Agricultural University, Chengdu 611130, China. FAU - Zhang, Jing AU - Zhang J AD - College of Horticulture, Sichuan Agricultural University, Chengdu 611130, China. AD - Institute of Pomology and Olericulture, Sichuan Agricultural University, Chengdu 611130, China. FAU - Chen, Qing AU - Chen Q AUID- ORCID: 0000-0002-1276-1064 AD - College of Horticulture, Sichuan Agricultural University, Chengdu 611130, China. FAU - Liu, Zhenshan AU - Liu Z AD - College of Horticulture, Sichuan Agricultural University, Chengdu 611130, China. AD - Institute of Pomology and Olericulture, Sichuan Agricultural University, Chengdu 611130, China. FAU - Liu, Congli AU - Liu C AD - Zhengzhou Fruit Research Institute, Chinese Academy of Agricultural Sciences, Zhengzhou 410100, China. FAU - He, Wen AU - He W AD - College of Horticulture, Sichuan Agricultural University, Chengdu 611130, China. AD - Institute of Pomology and Olericulture, Sichuan Agricultural University, Chengdu 611130, China. FAU - Wang, Hao AU - Wang H AD - College of Horticulture, Sichuan Agricultural University, Chengdu 611130, China. AD - Institute of Pomology and Olericulture, Sichuan Agricultural University, Chengdu 611130, China. FAU - Yang, Shaofeng AU - Yang S AD - College of Horticulture, Sichuan Agricultural University, Chengdu 611130, China. AD - Institute of Pomology and Olericulture, Sichuan Agricultural University, Chengdu 611130, China. FAU - Zhang, Yong AU - Zhang Y AUID- ORCID: 0000-0002-2281-4481 AD - College of Horticulture, Sichuan Agricultural University, Chengdu 611130, China. FAU - Luo, Ya AU - Luo Y AD - College of Horticulture, Sichuan Agricultural University, Chengdu 611130, China. FAU - Tang, Haoru AU - Tang H AUID- ORCID: 0000-0001-6008-2747 AD - College of Horticulture, Sichuan Agricultural University, Chengdu 611130, China. AD - Institute of Pomology and Olericulture, Sichuan Agricultural University, Chengdu 611130, China. FAU - Wang, Xiaorong AU - Wang X AUID- ORCID: 0000-0002-9332-9571 AD - College of Horticulture, Sichuan Agricultural University, Chengdu 611130, China. AD - Institute of Pomology and Olericulture, Sichuan Agricultural University, Chengdu 611130, China. LA - eng GR - 31672114/National Natural Science Foundation of China/ GR - 31801826/National Natural Science Foundation of China/ GR - 2019JDTD0010/Sichuan Science and Technology Program/ GR - 202010626010/Cherry Resources Sharing and Service Platform of Sichuan Province, National Undergraduate Innovation Training Program/ GR - 2021065/Sichuan Science and Technology Innovation Project/ GR - P202107/Shuangzhi Project Innovation Team of Sichuan Agricultural University/ PT - Journal Article DEP - 20221030 PL - Switzerland TA - Int J Mol Sci JT - International journal of molecular sciences JID - 101092791 RN - 0 (Oligonucleotides) SB - IM MH - In Situ Hybridization, Fluorescence MH - *Prunus/genetics MH - *Prunus avium/genetics MH - Karyotype MH - Oligonucleotides PMC - PMC9653872 OTO - NOTNLM OT - FISH OT - Prunus (cerasus) OT - chromosome identification OT - oligonucleotide OT - rDNA COIS- The authors declare no conflict of interest. EDAT- 2022/11/12 06:00 MHDA- 2022/11/15 06:00 PMCR- 2022/10/30 CRDT- 2022/11/11 01:27 PHST- 2022/09/14 00:00 [received] PHST- 2022/10/26 00:00 [revised] PHST- 2022/10/27 00:00 [accepted] PHST- 2022/11/11 01:27 [entrez] PHST- 2022/11/12 06:00 [pubmed] PHST- 2022/11/15 06:00 [medline] PHST- 2022/10/30 00:00 [pmc-release] AID - ijms232113213 [pii] AID - ijms-23-13213 [pii] AID - 10.3390/ijms232113213 [doi] PST - epublish SO - Int J Mol Sci. 2022 Oct 30;23(21):13213. doi: 10.3390/ijms232113213.