Cytogenetic and molecular markers for detecting Aegilops uniaristata chromosomes in a wheat background

Genome ◽  
2014 ◽  
Vol 57 (9) ◽  
pp. 489-497 ◽  
Author(s):  
Wenping Gong ◽  
Guangrong Li ◽  
Jianping Zhou ◽  
Genying Li ◽  
Cheng Liu ◽  
...  
Keyword(s):  
Molecular Markers ◽  
Expressed Sequence Tag ◽  
Triticum Turgidum ◽  
Wheat Breeding ◽  
Addition Lines ◽  
Chromosome Engineering ◽  
Aegilops Uniaristata ◽  
Expressed Sequence ◽  
Simple Sequence

Aegilops uniaristata has many agronomically useful traits that can be used for wheat breeding. So far, a Triticum turgidum – Ae. uniaristata amphiploid and one set of Chinese Spring (CS) – Ae. uniaristata addition lines have been produced. To guide Ae. uniaristata chromatin transformation from these lines into cultivated wheat through chromosome engineering, reliable cytogenetic and molecular markers specific for Ae. uniaristata chromosomes need to be developed. Standard C-banding shows that C-bands mainly exist in the centromeric regions of Ae. uniaristata but rarely at the distal ends. Fluorescence in situ hybridization (FISH) using (GAA)8 as a probe showed that the hybridization signal of chromosomes 1N–7N are different, thus (GAA)8 can be used to identify all Ae. uniaristata chromosomes in wheat background simultaneously. Moreover, a total of 42 molecular markers specific for Ae. uniaristata chromosomes were developed by screening expressed sequence tag – sequence tagged site (EST–STS), expressed sequence tag – simple sequence repeat (EST–SSR), and PCR-based landmark unique gene (PLUG) primers. The markers were subsequently localized using the CS – Ae. uniaristata addition lines and different wheat cultivars as controls. The cytogenetic and molecular markers developed herein will be helpful for screening and identifying wheat – Ae. uniaristata progeny.

scholarly journals Molecular cytogenetic identification of a wheat – Thinopyrum ponticum substitution line with stripe rust resistance

Genome ◽  
2017 ◽  
Vol 60 (10) ◽  
pp. 860-867 ◽  
Author(s):  
Chen Zhu ◽  
Yanzhen Wang ◽  
Chunhuan Chen ◽  
Changyou Wang ◽  
Aicen Zhang ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Stripe Rust ◽  
Expressed Sequence Tag ◽  
Wheat Breeding ◽  
Substitution Line ◽  
Stripe Rust Resistance ◽  
Fish Analysis ◽  
Thinopyrum Ponticum ◽  
Expressed Sequence

Thinopyrum ponticum (Th. ponticum) (2n = 10x = 70) is an important breeding material with excellent resistance and stress tolerance. In this study, we characterized the derivative line CH1113-B13-1-1-2-1 (CH1113-B13) through cytological, morphological, genomic in situ hybridization (GISH), fluorescence in situ hybridization (FISH), expressed sequence tag (EST), and PCR-based landmark unique gene (PLUG) marker analysis. The GISH analysis revealed that CH1113-B13 contained 20 pairs of common wheat chromosomes and one pair of JSt genomic chromosomes. Linkage analysis of Th. ponticum using seven EST and seven PLUG markers indicated that the pair of alien chromosomes belonged to the seventh homeologous group. Nulli-tetrasomic and FISH analysis revealed that wheat 7B chromosomes were absent in CH1113-B13; thus, CH1113-B13 was identified as a 7JSt (7B) substitution line. Finally, adult-stage CH1113-B13 exhibited immunity to wheat stripe rust. This substitution line is therefore a promising germplasm resource for wheat breeding.

scholarly journals Characterization of Chromosomal Rearrangement in New Wheat—Thinopyrum intermedium Addition Lines Carrying Thinopyrum—Specific Grain Hardness Genes

Agronomy ◽  
2019 ◽  
Vol 9 (1) ◽  
pp. 18 ◽  
Author(s):  
Zhihui Yu ◽  
Hongjin Wang ◽  
Yunfang Xu ◽  
Yongshang Li ◽  
Tao Lang ◽  
...  
Keyword(s):  
Molecular Markers ◽  
Chromosome Rearrangement ◽  
Snp Array ◽  
5S Rdna ◽  
Wheat Breeding ◽  
Addition Lines ◽  
Thinopyrum Intermedium ◽  
Group 5 ◽  
Partial Amphiploids

The wild species, Thinopyrum intermedium. (Genome StStJSJSJJ), serves as a valuable germplasm resource providing novel genes for wheat improvement. In the current study, non-denaturing fluorescence in situ hybridization (ND-FISH) with multiple probes and comparative molecular markers were applied to characterize two wheat-Th. intermedium chromosome additions. Sequential ND-FISH with new labeled Th. intermedium specific oligo-probes were used to precisely determine the chromosomal constitution of Th. intermedium, wheat—Th. intermedium partial amphiploids and addition lines Hy36 and Hy37. The ND-FISH results showed that the added JS-St translocated chromosomes in Hy36 had minor Oligo-5S ribosomal DNA (rDNA) signals at the short arm, while a pair of J-St chromosomes in Hy37 had major Oligo-pTa71 and minor Oligo-5S rDNA signals. The 90K SNP array and PCR-based molecular markers that mapped on wheat linkage group 5 and 3 facilitated the identification of Thinopyrum chromosome introgressions in the addition lines, and confirmed that added chromosomes in Hy36 and Hy37 were 5JSS.3StS and 5JS.3StS, respectively. Complete coding sequences at the paralogous puroindoline-a (Pina) loci from Th. intermedium were cloned and localized on the short arm of chromosome 5JS of Hy36. Line Hy36 showed a reduction in the hardness index, which suggested that Th. intermedium-specific Pina gene sequences may be associated with the softness trait in wheat background. The molecular cytogenetic identification of novel wheat—Th. intermedium derivatives indicated that the frequent chromosome rearrangement occurred in the progenies of wheat-Thinopyrum hybridization. The new wheat-Thinopyrum derived lines may increase the genetic diversity for wheat breeding.

Characterization of expressed sequence tag-derived simple sequence repeat markers for 17 Linum species

Botany ◽  
2010 ◽  
Vol 88 (5) ◽  
pp. 537-543 ◽  
Author(s):  
Yong-Bi Fu ◽  
Gregory W. Peterson
Keyword(s):  
Simple Sequence Repeat ◽  
Expressed Sequence Tag ◽  
Sequence Repeat ◽  
Simple Sequence Repeat Markers ◽  
Linum Usitatissimum L ◽  
Evolutionary Studies ◽  
Expressed Sequence ◽  
Simple Sequence ◽  
Cultivated Flax

One major challenge in genetic and evolutionary studies of wild flax species is the lack of informative molecular markers. A set of 100 informative expressed sequence tag-derived simple sequence repeat (EST-SSR) primer pairs developed in cultivated flax ( Linum usitatissimum L.) were characterized on 35 Linum accessions representing 17 Linum species for their transferability to other Linum species. Ninety-nine primer pairs displayed scorable polymorphisms across 35 Linum samples and generated 627 bands likely from 121 loci. About 50% of the detected bands occurred only in three or fewer samples. A total of 393 bands, likely from 116 loci, were detected by 97 primer pairs in Linum bienne Mill. samples, but only up to 60 bands, likely from up to 39 loci, were revealed by 6 to 37 primer pairs in the samples of the other 15 Linum species. The L. bienne samples displayed 23.7% more EST-SSR variation than the L. usitatissimum samples. These characterized EST-SSR markers should be useful for future genetic diversity and evolutionary studies of Linum species, particularly for the progenitor of cultivated flax.

scholarly journals Development of EST-derived SSR Markers with Long-core Repeat in Olive and Their Use for Paternity Testing

2013 ◽  
Vol 138 (4) ◽  
pp. 290-296 ◽  
Author(s):  
Raúl De la Rosa ◽  
Angjelina Belaj ◽  
Antonio Muñoz-Mérida ◽  
Oswaldo Trelles ◽  
Inmaculada Ortíz-Martín ◽  
...  
Keyword(s):  
Ssr Markers ◽  
Simple Sequence Repeats ◽  
Expressed Sequence Tag ◽  
Paternity Testing ◽  
High Level ◽  
Repeated Motifs ◽  
Expressed Sequence ◽  
Simple Sequence ◽  
Olive Breeding ◽  
Genomic Project

In the present work, a set of eight new hexa-nucleotide simple sequence repeats (SSRs) is reported in olive (Olea europaea L). These SSRs loci were generated on the basis of expressed sequence tag (EST) sequences in the frame of an olive genomic project. The markers showed a high level of polymorphism when tested on a set of cultivars used as genitors in the olive breeding program of Córdoba, Spain. The long-core repeat motif of these markers allows a wider separation among alleles, thus permitting an accurate genotyping. Besides, these markers showed comparable levels of polymorphism to di-nucleotide SSRs, the only ones so far reported in olive. Selected on the basis of their discrimination capacity, four of the eight SSRs were used to test their ability for paternity testing in a total of 81 seedlings coming from 12 crosses. The paternity testing showed that seven crosses matched the alleged paternity and the remaining five were products of illicit pollinations. These results exactly matched with previous paternity testing performed with di-nucleotide SSR markers. These results demonstrate the usefulness of the developed hexa-nucleotide repeated motifs for checking the paternity of breeding progenies and suggest their use on variability studies.

Karyotyping Dasypyrum breviaristatum chromosomes with multiple oligonucleotide probes reveals the genomic divergence in Dasypyrum

Genome ◽  
2021 ◽  
Author(s):  
Zhihui Yu ◽  
Hongjin Wang ◽  
Wenxi Jiang ◽  
Chengzhi Jiang ◽  
Weiguang Yuan ◽  
...  
Keyword(s):  
Repetitive Sequences ◽  
Wheat Breeding ◽  
Evolutionary Divergence ◽  
Oligonucleotide Probes ◽  
Addition Lines ◽  
Partial Amphiploid ◽  
Standard Karyotype ◽  
Molecular Marker Analysis ◽  
Dasypyrum Breviaristatum

The perennial species <i>Dasypyrum breviaristatum</i> (genome V<sup>b</sup>) contains many potentially valuable genes for the improvement of common wheat. Construction of a detailed karyotype of <i>D. breviaristatum</i> chromosomes will be useful for the detection of <i>Dasypyrum</i> chromatin in wheat background. We established the standard karyotype of 1V<sup>b</sup>-7V<sup>b</sup> chromosomes through non-denaturing fluorescence <i>in situ</i> hybridization (ND-FISH) technique using 28 oligonucleotide probes from the wheat-<i>D. breviaristatum</i> partial amphiploid TDH-2 (AABBV<sup>b</sup>V<sup>b</sup>) and newly identified wheat-<i>D. breviaristatum</i> disomic translocation and addition lines D2138 (6V<sup>b</sup>S.2V<sup>b</sup>L), D2547 (4V<sup>b</sup>) and D2532 (3V<sup>b</sup>S.6V<sup>b</sup>L) by comparative molecular marker analysis. The ND-FISH with multiple oligo probes were conducted on the durum wheat-<i>D. villosum</i> amphiploid TDV-1 and large karyotype differences between <i>D. breviaristatum</i> and <i>D. villosum</i> was revealed. These ND-FISH probes will be valuable for screening the wheat-<i>Dasypyrum</i> derivative lines for chromosome identification, and newly developed wheat-<i>D. breviaristatum</i> addition lines may broaden the gene pool of wheat breeding. The differences between <i>D. villosum</i> and <i>D. breviaristatum</i> chromosomes revealed by ND-FISH will help us understand evolutionary divergence of repetitive sequences within the genus <i>Dasypyrum</i>.

scholarly journals Does the genetic diversity among pubescent white oaks in southern Italy, Sicily and Sardinia islands support the current taxonomic classification?

2020 ◽  
Author(s):  
Romeo Di Pietro ◽  
Antonio Luca Conte ◽  
Piera Di Marzio ◽  
Paola Fortini ◽  
Emmanuele Farris ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Expressed Sequence Tag ◽  
Taxonomic Classification ◽  
Genetic Assignment ◽  
Molecular Analyses ◽  
Genetic Groups ◽  
Ecological Features ◽  
Weak Separation ◽  
Expressed Sequence ◽  
Simple Sequence

AbstractMolecular diversity analysis of deciduous pubescent oaks was conducted for populations from Calabria, Sicily and Sardinia. The aims of this study were twofold. First, to provide data on the genetic diversity of pubescent oaks from an understudied area which currently exhibits one of the highest concentrations of pubescent oak species in Europe. Second, to verify if these groups of oaks are genetically distinct and if their identification is in accordance with the current taxonomic classification. Molecular analyses of leaf material of 480 trees from seventeen populations belonging to putatively different pubescent oak species (Quercus amplifolia, Q. congesta, Q. dalechampii, Q. ichnusae, Q. leptobalanos, Q. virgiliana) were performed. Twelve gene-based Expressed Sequence Tag-Simple Sequence Repeat markers were selected, and genetic diversity and differentiation were calculated. The results showed relatively high values of allelic richness, heterozygosity and number of private alleles for the populations investigated. A weak but positive correlation between geographical and genetic distance was detected. Genetic assignment (STRUCTURE) and principle coordinate analyses exhibited a weak separation into two genetic groups which, however, did not correspond to the taxonomic, chorological and ecological features of the populations investigated. Sardinian populations formed one group which was separated from the Calabrian and Sicilian populations. In light of the results obtained, the taxonomic classification for the pubescent white oaks currently reported in the major Italian floras and checklists for the study area was not confirmed by molecular analyses.

Analysis of expressed sequence tags from the ectoparasitic nematode Xiphinema index

Nematology ◽  
2005 ◽  
Vol 7 (1) ◽  
pp. 95-104 ◽  
Author(s):  
Cleber Furlanetto ◽  
Linda Cardle ◽  
Derek J.F. Brown ◽  
John T. Jones
Keyword(s):  
Expressed Sequence Tag ◽  
In Situ Hybridisation ◽  
Cdna Libraries ◽  
Small Scale ◽  
Xiphinema Index ◽  
Genes Encoding ◽  
Predicted Signal Peptide ◽  
Basal Bulb ◽  
Expressed Sequence

AbstractWe report the results of a small-scale expressed sequence tag project performed on the ectoparasitic nematode Xiphinema index. Approximately 1400 genes were sequenced, 70% from a cDNA library generated from dissected basal bulbs (containing the pharyngeal gland cells) and 30% from cDNA libraries generated from whole mixed stage nematodes. A large portion of the bulb library (48%) was composed of proteins with no matches in the database. Further analysis of these genes revealed that a total of 51 contigs were present, half of which encoded novel secreted proteins. By contrast, the whole nematode library contained more housekeeping and nematode-specific genes. Only one of the novel genes from the whole nematode library had a predicted signal peptide at the N-terminus. Genes encoding transthyretin-like proteins were abundant in the bulb library and in situ hybridisation confirmed that one of these is expressed in the basal bulb. Genes encoding a variety of proteases, which were shown using in situ hybridisation to be expressed in the gut, were also identified.

Development and characterization of microsatellite markers from tropical forage Stylosanthes species and analysis of genetic variability and cross-species transferability

Genome ◽  
2011 ◽  
Vol 54 (12) ◽  
pp. 1016-1028 ◽  
Author(s):  
Amaresh Chandra ◽  
K.K. Tiwari ◽  
D. Nagaich ◽  
N. Dubey ◽  
S. Kumar ◽  
...  
Keyword(s):  
Genetic Improvement ◽  
Expressed Sequence Tag ◽  
Intraspecific Diversity ◽  
Genomic Libraries ◽  
Tropical Forage ◽  
Genomic Ssr ◽  
Functional Molecular Markers ◽  
Expressed Sequence ◽  
Simple Sequence

A limited number of functional molecular markers has slowed the desired genetic improvement of Stylosanthes species. Hence, in an attempt to develop simple sequence repeat (SSR) markers, genomic libraries from Stylosanthes seabrana B.L. Maass & ’t Mannetje (2n = 2x = 20) using 5′ anchored degenerate microsatellite primers were constructed. Of the 76 new microsatellites, 21 functional primer pairs were designed. Because of the small number of primer pairs designed, 428 expressed sequence tag (EST) sequences from seven Stylosanthes species were also examined for SSR detection. Approximately 10% of sequences delivered functional primer pairs, and after redundancy elimination, 57 microsatellite repeats were selected. Tetranucleotides followed by trinucleotides were the major repeated sequences in Stylosanthes ESTs. In total, a robust set of 21 genomic–SSR (gSSR) and 20 EST–SSR (eSSR) markers were developed. These markers were analyzed for intraspecific diversity within 20 S. seabrana accessions and for their cross-species transferability. Mean expected (He) and observed (Ho) heterozygosity values with gSSR markers were 0.64 and 0.372, respectively, whereas with eSSR markers these were 0.297 and 0.214, respectively. Dendrograms having moderate bootstrap value (23%–94%) were able to distinguish all accessions of S. seabrana with gSSR markers, whereas eSSR markers showed 100% similarities between few accessions. The set of 21 gSSRs, from S. seabrana, and 20 eSSRs, from selected Stylosanthes species, with their high cross-species transferability (45% with gSSRs, 86% with eSSRs) will facilitate genetic improvement of Stylosanthes species globally.

Sign in / Sign up

Export Citation Format

Share Document