Development of new cytogenetic markers for Thinopyrum ponticum (Podp.) Z.-W. Liu & R.-C. Wang

Abstract

Thinopyrum ponticum (Podpěra, 1902) Z.-W. Liu & R.-C.Wang, 1993 is an important polyploid wild perennial Triticeae species that is widely used as a source of valuable genes for wheat but its genomic constitution has long been debated. For its chromosome identification, only a limited set of FISH probes has been used. The development of new cytogenetic markers for Th. ponticum chromosomes is of great importance both for cytogenetic characterization of wheat-wheatgrass hybrids and for fundamental comparative studies of phylogenetic relationships between species. Here, we report on the development of five cytogenetic markers for Th. ponticum based on repetitive satellite DNA of which sequences were selected from the whole genome sequence of Aegilops tauschii Cosson, 1849. Using real-time quantitative PCR we estimated the abundance of the found repeats: P720 and P427 had the highest abundance and P132, P332 and P170 had lower quantity in Th. ponticum genome. Using fluorescence in situ hybridization (FISH) we localized five repeats to different regions of the chromosomes of Th. ponticum. Using reprobing multicolor FISH we colocalized the probes between each other. The distribution of these found repeats in the Triticeae genomes and its usability as cytogenetic markers for chromosomes of Th. ponticum are discussed.

Aegilops crassa Boiss. repeatome characterized using low-coverage NGS as a source of new FISH markers: Application in phylogenetic studies of the Triticeae

“…Real-time quantitative PCR qPCR using primers developed for repeat monomers (Table 2) was performed in triple technical replication with water as negative control and VRN1 as a reference gene (Yaakov et al, 2013) according to the protocol described in Kroupin et al (2019b). The amplification was performed on a CFX Real-Time PCR Detection System (Bio-Rad) and in Real-Time PCR Mix reaction mixture with Eva Green (Syntol Ltd., Moscow, Russia) according to the manufacturer's protocol.…”

Developing Oligo Probes for Chromosomes Identification in Hemp (Cannabis sativa L.)

“…Cytogenetic markers help to distinguish similar plant chromosomes, including sex chromosomes, which is important for the breeding process [ 19 , 30 ]. Cytogenetic markers also help us to associate genomic assemblies with physical chromosomes [ 31 , 32 ].…”

Cytogenetic Analysis and Molecular Marker Development for a New Wheat–Thinopyrum ponticum 1Js (1D) Disomic Substitution Line With Resistance to Stripe Rust and Powdery Mildew

“…Thinopyrum ponticum (2 n = 10 x = 70) often produces large amount of biomass and is tolerant of biotic and abiotic stresses such as drought and salt stress. It is also highly resistant to wheat leaf rust, stem rust, stripe rust, powdery mildew, barley yellow dwarf virus (BYDV), and Fusarium head blight (FHB) ( Ceoloni et al, 2014 ; Kroupin et al, 2019 ), and recently, Fhb7 was successfully cloned by Wang et al (2020a) from Th. elongatum .…”