Fluorescence In Situ Hybridization (FISH) Analysis of the Locations of the Oligonucleotides 5S rDNA, (AGGGTTT)3, and (TTG)6 in Three Genera of Oleaceae and Their Phylogenetic Framework.

Xiaomei Luo,Juncheng Liu

doi:10.3390/genes10050375

Abstract

We report the cytogenetic map for a collection of species in the Oleaceae, and test similarities among the karyotypes relative to their known species phylogeny. The oligonucleotides 5S ribosomal DNA (rDNA), (AGGGTTT)3, and (TTG)6 were used as fluorescence in situ hybridization (FISH) probes to locate the corresponding chromosomes in three Oleaceae genera: Fraxinus pennsylvanica, Syringa oblata, Ligustrum lucidum, and Ligustrum × vicaryi. Forty-six small chromosomes were identified in four species. (AGGGTTT)3 signals were observed on almost all chromosome ends of four species, but (AGGGTTT)3 played no role in distinguishing the chromosomes but displayed intact chromosomes and could thus be used as a guide for finding chromosome counts. (TTG)6 and 5S rDNA signals discerned several chromosomes located at subterminal or central regions. Based on the similarity of the signal pattern (mainly in number and location and less in intensity) of the four species, the variations in the 5S rDNA and (TTG)6 distribution can be ordered as L. lucidum < L. × vicaryi < F. pennsylvanica < S. oblata. Variations have observed in the three genera. The molecular cytogenetic data presented here might serve as a starting point for further larger-scale elucidation of the structure of the Oleaceae genome, and comparison with the known phylogeny of Oleaceae family.

Highlights

The Oleaceae family comprises approximately 30 genera and over 600 species in tropical, subtropical and temperate regions of the world, mainly in Asia [1]
fluorescence in situ hybridization (FISH) images of mitotic metaphases of Fraxinus pennsylvanica, Syringa oblata, Ligustrum lucidum, and Ligustrum × vicaryi are illustrated in Figures 1 and 2
Forty-six chromosomes were counted in all four species

Summary

Introduction

The Oleaceae family comprises approximately 30 genera and over 600 species in tropical, subtropical and temperate regions of the world, mainly in Asia [1]. Ten genera and more than 160 species (95 endemic) whose chromosome complement is arranged from 2n = 22 to 2n = 52. 2n = 28/42, Chionanthus 2n = 46, Osmanthus 2n = 46, Olea 2n = 46, Fraxinus 2n = 46, Ligustrum. Determination of the location of a chromosome through fluorescence in situ hybridization (FISH) has only been reported in one species, Olea europaea ssp. Sativa in Oleaceae; in this species, Katsiotis et al [6] identified two tandemly repeated DNA sequences in the chromosomes of this species: the 81-bp family and pOS218 co-localization. Only approximately five genetic maps of Oleaceae, including several integrated maps with a higher marker density [8,9,10,11,12], have been published, these constitute a foundational tool and a resource for marker-assisted selection and genomic studies

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