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Research on Crops

The potential for physical gene localization on compact chromosomes of Citrus plants using Tyramide-FISH


DOI: 10.31830/2348-7542.2026.ROC-1285    | Article Id: ROC-1285 | Page : 99-104
Citation :- The potential for physical gene localization on compact chromosomes of Citrus plants using Tyramide-FISH. Res. Crop. 27: 99-104
MIKHAIL MONAKHOS, YULIYA SMOTROVA, OLGA RAZUMOVA, VARVARA KOROBKOVA AND DMITRY ROMANOV mihaluch1864@gmail.com
Address : All-Russian Research Institute of Agricultural Biotechnology, 127550 Moscow, Russia
Submitted Date : 13-01-2026
Accepted Date : 27-01-2026
Online Published:

Abstract

Citrus species have very small, compact chromosomes that hinder physical gene localization by conventional FISH technique. This study evaluates whether Tyramide-FISH can reliably localize short, gene-derived cDNA probes on metaphase chromosomes and provides a cDNA resource for mapping. Total RNA from 21 Rutaceae accessions (Citrus, Fortunella, Poncirus) was used to construct random-fragment cDNA libraries (216 clones, 86 Sanger-sequenced). Selected large-insert cDNAs were DIG-labeled and hybridized to root-tip metaphase spreads. Signals were detected with anti-DIG-POD and tyramide-Cy3 amplification. Images were acquired by epifluorescence microscopy and analyzed with DRAWID to place probes on ideograms. Tyramide-FISH produced reproducible, locus-specific signals for four cDNA probes on homologous chromosome pairs in different Rutaceae taxa, demonstrating successful localization on distal chromosome arms. Some accessions yielded too few high-quality spreads for mapping. Overall, Tyramid-FISH is shown to be a practical approach for physical gene mapping on compact Citrus chromosomes, and the generated cDNA collection constitutes a useful resource for further cytogenetic and genomic work.

Keywords

Citrus cDNA library mRNA probes Rutaceae Tyramide-FISH

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