Cloning and physical localization of male-biased repetitive DNA sequences in Spinacia oleracea (Amaranthaceae)

By

zhou, jian
wang, Shaojing
Yu, Li'ang
Li, Ning
Li, Shufen
Zhang, Yulan
Qin, Ruiyun
Gao, Wujun
Deng, Chuanliang

Type

Article

Date of Publication

2021-4-23

Original Publication

Comparative cytogenetics : international journal of plant & animal cytogenetics, karyosystematics and molecular systematics

Volume

15

Series / Issue

Issue: 2

Pages

101--118

Subjects

Fish , Genomic subtraction hybridization , retrotransposon , Sex chromosome evolution , Spinach

Contributed by

Pensoft Publishers

Language

English

Published by

St. Petersburg, Russia [Published] with technical support of Zoological Institute RAS [2007]-

Identifiers

DOI: https://doi.org/10.3897/CompCytogen.v15.i2.63061

 

Abstract

Spinach (Spinacia oleracea Linnaeus, 1753) is an ideal material for studying molecular mechanisms of early-stage sex chromosome evolution in dioecious plants. Degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR) technique facilitates the retrotransposon-relevant studies by enriching specific repetitive DNA sequences from a micro-dissected single chromosome. We conducted genomic subtractive hybridization to screen sex-biased DNA sequences by using the DOP-PCR amplification products of micro-dissected spinach Y chromosome. The screening yielded 55 male-biased DNA sequences with 30 576 bp in length, of which, 32 DNA sequences (12 049 bp) contained repeat DNA sequences, including LTR/Copia, LTR/Gypsy, simple repeats, and DNA/CMC-EnSpm. Among these repetitive DNA sequences, four DNA sequences that contained a fragment of Ty3-gypsy retrotransposons (SP73, SP75, SP76, and SP77) were selected as fluorescence probes to hybridization on male and female spinach karyotypes. Fluorescence in situ hybridization (FISH) signals of SP73 and SP75 were captured mostly on the centromeres and their surrounding area for each homolog. Hybridization signals primarily appeared near the putative centromeres for each homologous chromosome pair by using SP76 and SP77 probes for FISH, and sporadic signals existed on the long arms. Results can be served as a basis to study the function of repetitive DNA sequences in sex chromosome evolution in spinach.