Molecular Cytogenetic Identification of Wheat-Aegilops Biuncialis 5Mb Disomic Addition Line with Tenacious and Black Glumes

Abstract

Production of wheat-alien disomic addition lines is of great value to the exploitation and utilization of elite genes originated from related species to wheat. In this study, a novel wheat-Aegilops biuncialis 5Mb disomic addition line WA317 was characterized by in situ hybridization (ISH) and specific-locus amplified fragment sequencing (SLAF-seq) markers. Compared to its parent Chinese Spring (CS), the glumes of WA317 had black color and were difficult to remove after harvesting, suggesting chromosome 5Mb carried gene(s) related to glume development and Triticeae domestication process. A total of 242 Ae. biuncialis SLAF-based markers (298 amplified patterns) were developed and further divided into four categories by Ae. biuncialis Y17, Ae. umbellulata Y139 and Ae. comosa Y258, including 172 markers amplifying the same bands of U and M genome, six and 102 markers amplifying U-specific and M-specific bands, respectively and eighteen markers amplifying specific bands in Y17. Among them, 45 markers had the specific amplifications in WA317 and were 5Mb specific markers. Taken together, line WA317 with tenacious and black glumes should serve as the foundation for understanding of the Triticeae domestication process and further exploitation of primitive alleles for wheat improvement. Ae. biuncialis SLAF-based markers can be used for studying syntenic relationships between U and M genomes as well as rapid tracking of U and M chromosomal segments in wheat background.

Identification of a Wheat-Psathyrostachys huashanica 7Ns Ditelosomic Addition Line Conferring Early Maturation by Cytological Analysis and Newly Developed Molecular and FISH Markers

“…ponticum ( Yang et al, 2021 ), S. cereale ( Du et al, 2018 ), and Ae. biuncialis ( Song et al, 2020 ). To date, no P. huashanica -specific SLAF-based markers have been reported.…”

Genetic analysis of resistance to powdery mildew on 7Mg chromosome of Wheat–Aegilops geniculata,development and utilization of specific molecular markers

“…designed the 242 markers showed Ae. biuncialis speci c bands with a success rate up to 40.33% [49]. Sixtyone pairs of molecular markers were screened about Ae.…”

“…In recent years, in the eld of molecular markers, the development of speci c heterogeneous chromosome markers is of great signi cance in breeding [48]. Then, SLAF-seq of NA0973-5-4-1-2-9-1 and W16998 were performed with some modi cation by the Beijing Biomarker Technologies Corporation [49]. To improve the speci city and e ciency of the marker, rstly, the SLAF-seq of NA0973-5-4-1-2-9-1 and W16998 was compared with the CS sequences (http://www.wheatgenome.org/News/Latest-news/IWGSC-Reference-Sequencev1.0-browser-now-available-at-URGI), and the sequences with similarity less than 50% were reserved.…”

Molecular Cytogenetics and SLAF-seq-based Specific Molecular Markers for a Wheat–Aegilops geniculata 3M g Alien Addition Line with Resistance to Stripe Rust and Powdery Mildew

“…elongatum with e ciency of 65.9%. Markers speci c to Aegilops biuncialis were developed with a success rate of up to 40.33% [51]; The success rate of 1JS-speci c marker development from an alien chromosome of Th. ponticum was 52.98% [52].…”

“…The SLAF-seq method is a simpli ed genome sequencing technology independently developed by Biomarker [63] and shows three distinguishing characteristics: (1) deep sequencing to ensure genotyping accuracy, (2) a reduced representation strategy to reduce sequencing costs, and (3) a predesigned reduced representation scheme to optimize marker e ciency [31]. To increase marker speci city and e ciency, sequences that showed 50% similarity to CS (http://www.wheatgenome.org/News/Latest-news/IWGSC-Reference-Sequence-v1.0-browser-now-available-at-URGI) were selected using the BWA tool [51], From these markers, sequences that showed at least 90% similarity to SY159 were selected. After PCR ampli cation, speci c bands were detected in 1% agarose gel [63].…”