New St-chromosome-specific molecular markers for identifying wheat–Thinopyrum intermedium derivative lines

Lang

et al. 2016

Planta

Self Cite

New molecular markers were developed for targeting Thinopyrum intermedium 1St#2 chromosome, and novel FISH probe representing the terminal repeats was produced for identification of Thinopyrum chromosomes. Thinopyrum intermedium has been used as a valuable resource for improving the disease resistance and yield potential of wheat. A wheat-Th. intermedium ssp. trichophorum chromosome 1St#2 substitution and translocation has displayed superior grain protein and wet gluten content. With the aim to develop a number of chromosome 1St#2 specific molecular and cytogenetic markers, a high throughput, low-cost specific-locus amplified fragment sequencing (SLAF-seq) technology was used to compare the sequences between a wheat-Thinopyrum 1St#2 (1D) substitution and the related species Pseudoroegneria spicata (St genome, 2n = 14). A total of 5142 polymorphic fragments were analyzed and 359 different SLAF markers for 1St#2 were predicted. Thirty-seven specific molecular markers were validated by PCR from 50 randomly selected SLAFs. Meanwhile, the distribution of transposable elements (TEs) at the family level between wheat and St genomes was compared using the SLAFs. A new oligo-nucleotide probe named Oligo-pSt122 from high SLAF reads was produced for fluorescence in situ hybridization (FISH), and was observed to hybridize to the terminal region of 1St#L and also onto the terminal heterochromatic region of Th. intermedium genomes. The genome-wide markers and repetitive based probe Oligo-pSt122 will be valuable for identifying Thinopyrum chromosome segments in wheat backgrounds.

Section: Discussionmentioning

confidence: 99%

“…The cycling parameters were 94°C for 3 min for denaturation; followed by 35 cycles at 94°C for 1 min, 55°C for 1 min, 72°C for 2 min; and a final extension at 72°C for 10 min. The amplified products were separated by 1.5 % agarose gel as described by Hu et al (2012).…”

Section: Transposable Element (Te) Comparisonmentioning

confidence: 99%

New molecular markers and cytogenetic probes enable chromosome identification of wheat-Thinopyrum intermedium introgression lines for improving protein and gluten contents

Guang-rong

Lang

et al. 2016

Planta

Self Cite

“…, Hu et al. ), collinear gene relationships among Triticeae species and sequence polymorphism among different species, but also determine homoeologous group relationships between wheat and alien chromosomes. Hu et al.…”

Section: Discussionmentioning

confidence: 99%

“…Hu et al. () revealed genomic rearrangement between wheat and Th . elongatum using mapped functional molecular markers (EST–SST and PLUG markers).…”

Section: Discussionmentioning

confidence: 99%

Development and molecular cytogenetic identification of a new wheat–Leymus mollis Lm#6Ns disomic addition line

Yang

et al. 2016

Plant Breeding

We developed a new disomic addition line M11028‐1‐1‐5 (2n = 44 = 21” + 1”) from a cross between wheat cv. ‘7182’ and octoploid Tritileymus M47 (2n = 8x = 56, AABBDDNsNs). Cytological observations demonstrated that M11028‐1‐1‐5 contained 44 chromosomes and formed 22 bivalents during meiotic metaphase I. The genomic in situ hybridization (GISH) investigations showed this line contained 42 wheat chromosomes and a pair of L. mollis chromosomes. SSR, EST and PCR‐based landmark unique gene (PLUG) markers were screened to determine the homoeologous relationships of the introduced L. mollis chromosomes in wheat background. Nine markers, i.e. Xwmc256, Xgpw312, Swes123, CD452568, BF483643, BQ169205, TNAC1748, TNAC1751 and TNAC1752, all of which were located on the homoeologous group 6 chromosomes of common wheat, amplified bands unique to L. mollis in M11028‐1‐1‐5. Gliadin analysis also confirmed that the added chromosomes in M11028‐1‐1‐5 were correlated with the sixth group chromosome. This indicated that M11028‐1‐1‐5 contained a pair of introduced L. mollis chromosome belonging to homoeologous group 6, which we designated it as Lm#6Ns disomic addition line. This is the first report of a common wheat–L. mollis disomic addition line.

“…, Thinopyrum elongatum (Host) D.R. Dewey (Xu et al, 2012), and Thinopyrum intermedium (Host) Barkworth and Dewey (Hu et al, 2012). Thus, SCAR markers are the most practical method for screening large number of samples in a time-efficient and labor-saving manner because they are accurate, easy to use, and cost efficient (Kasai et al, 2000).…”

Section: Introductionmentioning

confidence: 99%

A novel PCR-based marker for identifying Ns chromosomes in wheat-Psathyrostachys huashanica Keng derivative lines

Lü

et al. 2013

Span. j. agric. res.

The genome of Psathyrostachys huashanica Keng (2n = 2x = 14, NsNs) contains many desirable characteristics, such as resistance to biotic and abiotic stresses, which make it suitable for wheat improvement (Kang et al., 2009). Several P. huashanica genes have already been introgressed successfully into the wheat (Triticum aestivum L.) genome (Zhao et al., 2010; Du et al., 2013a,b). In the 1990s, we generated (Chen et al., 1991(Chen et al., , 1996 the F 1 hybrid H881 (2n = 28, ABDNs) and a derived heptaploid H8911 (2n = 49, AABBDDNs) of common wheat cv. 7182 and P. huashanica (GenBank Acc. No. 0503383) via embryo culture, two backcrosses, and selfing. As a result, we generated a large and complex range of wheat-P. huashanica offspring that carried different chromosome number or structure variants, i.e., chromosome additions, deletions, and rearrangements. However, a major problem was the selection of recombinant plants derived from the large interbred population. After several years of screening and identification, a complete set of wheat-P. huashanica disomic addition lines (1Ns -7Ns, 2n = 44 = 22II) was developed. This was a repetitive and difficult work. Although cytogenetic techniques proved to be very useful tools for alien identification, they had limitations because of the small population number, the complexity of the experimental techniques, and envi- AbstractPsathyrostachys huashanica Keng is an endangered species that is endemic to China, which provides an important gene pool for wheat improvement. We developed a quick and reliable PCR-based diagnostic assay to accurately and efficiently detect P. huashanica DNA sequences from introgression lines, which was based on a species-specific marker derived from genomic DNA. The 900-bp PCR-amplified band used as a P. huashanica-specific RAPD marker was tested with 21 different plant species and was converted into a sequence-characterized amplified region (SCAR) marker by cloning and sequencing the selected fragments (pHs11). This SCAR marker, which was designated as RHS-23, could clearly distinguish the presence of P. huashanica DNA repetitive sequences in wheat-P. huashanica derivative lines. The specificity of the marker was validated using 21 different plant species and a complete set of wheat-P. huashanica disomic addition lines (1Ns -7Ns, 2n = 44 = 22II). This specific sequence targeted the Ns genome of P. huashanica and it was present in all the seven P. huashanica chromosomes. Therefore, this SCAR marker is specific for P. huashanica chromosomes and may be used in the identification of alien repetitive sequences in large gene pools. This diagnostic PCR assay for screening the target genetic material may play a key role in marker-assisted selective breeding programs.Additional key words: addition lines; marker-assisted selection; repetitive sequences; RAPD; SCAR.* Corresponding author: cxh2089@126.com; wj2105@163.com Received: 24-03-13. Accepted: 15-10-13.This work has one Supplementary Figure that does not appear in the printed article but that accompan...