Shunzong Ning

Evaluation of Aegilops tauschii for heading date and its gene location in a re-synthesized hexaploid wheat.

The successful worldwide cultivation of hexaploid wheat in a diverse range of environments is because of, in part, breeding and selection for appropriate heading date. To adjust and fine-tune the heading time of hexaploid wheat to particular geographical regions and specific environment within these, there is an urgent need to evaluate and use alternative alleles for heading time. Aegilops tauschii, the donor species of D-genome of hexaploid wheat, has a wide geographic distribution. The present study revealed a wide variation for heading time among 56 Ae. tauschii accessions. All the accessions with short heading dates belonged to the ssp. tauschii, whereas most of ssp. strangulata accessions showed very long heading date. The heading date was also related to distribution of this species. The monotelosomic and monosomic analysis of a synthetic hexaploid wheat showed that chromosome 2D derived from ssp. tauschii accession AS60 had a major effect on promoting heading time with a reduction of more than 5 days. It is postulated that this Ae. tauschii genotype possess the allele Ppd-Dt1 responsible for the insensitivity to photoperiod. This allele is probably different from Ppd-D1 existing in hexaploid wheat. The new allele Ppd-Dt1 derived from Ae. tauschii might be used as a source for hexaploid wheat breeding on photoperiod response.G156S with part drooping leaf,dark-green leaf color and normal flower organs was respectively crossed with restoring lines Minghui 63,Minghui 86,Qianhui 085,Shuhui 527 with straight leaf to study heredity and gene loci of drooping leaf character.The results showed that the drooping leaf character was controlled by a pair of recessive genes according to the phenotypes of F1,F2,BC1 populations and χ2 test,and the gene controlling drooping leaf character was between RM5628 and RM6849 of the short arm of chromosome 3 and its genetic distance was 1.0 cM and 0.9 cM.

QTug.sau-3B is a major quantitative trait locus for wheat hexaploidization

Meiotic nonreduction resulting in unreduced gametes is thought to be the predominant mechanism underlying allopolyploid formation in plants. Until now, however, its genetic base was largely unknown. The allohexaploid crop common wheat (Triticum aestivum L.), which originated from hybrids of T. turgidum L. with Aegilops tauschii Cosson, provides a model to address this issue. Our observations of meiosis in pollen mother cells from T. turgidum×Ae. tauschii hybrids indicated that first division restitution, which exhibited prolonged cell division during meiosis I, was responsible for unreduced gamete formation. A major quantitative trait locus (QTL) for this trait, named QTug.sau-3B, was detected on chromosome 3B in two T. turgidum×Ae. tauschii haploid populations. This QTL is situated between markers Xgwm285 and Xcfp1012 and covered a genetic distance of 1 cM in one population. QTug.sau-3B is a haploid-dependent QTL because it was not detected in doubled haploid populations. Comparative genome analysis indicated that this QTL was close to Ttam-3B, a collinear homolog of tam in wheat. Although the relationship between QTug.sau-3B and Ttam requires further study, high frequencies of unreduced gametes may be related to reduced expression of Ttam in wheat.

Cytological identification of an Aegilops variabilis chromosome carrying stripe rust resistance in wheat

Aegilops variabilis (UUSvSv), an important sources for wheat improvement, originated from chromosome doubling of a natural hybrid between Ae. umbellulata (UU) with Ae. longissima (SlSl). The Ae. variabilis karyotype was poorly characterized by fluorescent in situ hybridization (FISH). The FISH probe combination of pSc119.2, pTa71 and pTa-713 identified each of the 14 pairs of Ae. variabilis chromosomes. Our FISH ideogram was further used to detect an Ae. variabilis chromosome carrying stripe rust resistance in the background of wheat lines developed from crosses of the stripe rust susceptible bread wheat cultivar Yiyuan 2 with a resistant Ae. variabilis accession. Among the 15 resistant BC1F7 lines, three were 2Sv + 4Sv addition lines (2n = 46) and 12 were 2Sv(2B) or 2Sv(2D) substitution lines that were confirmed with SSR markers. SSR marker gwm148 can be used to trace 2Sv in common wheat background. Chromosome 2Sv probably carries gametocidal(Gc) gene(s) since cytological instability and chromosome structural variations, including non-homologous translocations, were observed in some lines with this chromosome. Due to the effects of photoperiod genes, substitution lines 2Sv(2D) and 2Sv(2B) exhibited late heading with 2Sv(2D) lines being later than 2Sv(2B) lines. 2Sv(2D) substitution lines were also taller and exhibited higher spikelet numbers and longer spikes.

Characterization of WAP2 gene in Aegilops tauschii and comparison with homoeologous loci in wheat

Abstract  The Q/q gene, also known as WAP2, is an important gene for wheat domestication and is a member of the AP2 (APETALA2) class of transcription factors. In the present study, we first isolated the WtAP2 allele (where the superscript “t” refers to the speciese source, in this case “tauschii”) on chromosome 5D from Aegilops tauschii Coss., the D‐genome donor species of common wheat. We found that WtAP2 and the AP2 gene from Arabidopsis share a central core of the AP2 polypeptide, a highly basic 10‐amino acid domain, and an AASSGF box, although there are many differences in the 37‐amino acid serine‐rich acidic domain and the remaining regions. In addition, WtAP2 was highly homologous to the homoeologous loci on 5A and 5B of wheat at both the nucleotide and amino acid level. However, there were some variations that are probably related to gene function. In the first AP2 domain, the amino acids VYL on the 5D and 5A loci were replaced with LLR on 5B. In the 37‐amino acid serine‐rich acidic domain, WtAP2 on 5D had an extra amino acid insertion. There was also a variation at the 329 amino acid position, which is thought to be related to the appearance of free‐threshing wheat. At this position, the amino acid is isoleucine on 5A for the Q allele and valine for the q allele, whereas the amino acid is leucine on 5D and 5B. Furthermore, a Stowaway miniature terminal inverted repeat element (MITE) insertion was present in the ninth intron of WAP2 on 5B of all common wheats and partial tetraploid Triticum turgidum wheats. These results provide new clues for studies into the evolutionary biology of WAP2 and the origin of common wheat.

High Transferability of Homoeolog-Specific Markers between Bread Wheat and Newly Synthesized Hexaploid Wheat Lines

Bread wheat (Triticum aestivum, 2n = 6x = 42, AABBDD) has a complex allohexaploid genome, which makes it difficult to differentiate between the homoeologous sequences and assign them to the chromosome A, B, or D subgenomes. The chromosome-based draft genome sequence of the ‘Chinese Spring’ common wheat cultivar enables the large-scale development of polymerase chain reaction (PCR)-based markers specific for homoeologs. Based on high-confidence ‘Chinese Spring’ genes with known functions, we developed 183 putative homoeolog-specific markers for chromosomes 4B and 7B. These markers were used in PCR assays for the 4B and 7B nullisomes and their euploid synthetic hexaploid wheat (SHW) line that was newly generated from a hybridization between Triticum turgidum (AABB) and the wild diploid species Aegilops tauschii (DD). Up to 64% of the markers for chromosomes 4B or 7B in the SHW background were confirmed to be homoeolog-specific. Thus, these markers were highly transferable between the ‘Chinese Spring’ bread wheat and SHW lines. Homoeolog-specific markers designed using genes with known functions may be useful for genetic investigations involving homoeologous chromosome tracking and homoeolog expression and interaction analyses.

Amphitelic orientation of centromeres at metaphase I is an important feature for univalent-dependent meiotic nonreduction.

Univalent-dependent meiotic nonreduction, which leads to the production of unreduced gametes, is thought to be the predominant mechanism underlying allopolyploid plant formation. However, little is known about the underlying cytological mechanism. In the present study, we observed male sporogenesis in F1 amphihaploid hybrids of wheat–rye by FISH with the help of diagnostic PrCEN-1 specific for the rye centromere. Our observations indicated that at meiotic metaphase I, the chromosomes were accumulated on the equatorial plate. At this stage, the elongated centromeres were amphitelically oriented perpendicular to the equatorial plate, indicating tension from opposite poles. At late metaphase, the centromeres and sister chromatids started separating. Subsequently, the sister chromatids and centromeres split finally resulting in dyads. Our observations indicate that bipolar orientation of the sister kinetochores of univalents at the equatorial plate in metaphase I is important for the subsequent bipolar separation of sister chromatids in the first meiotic division. Allopolyploids are common in plants. Wide hybridization, the first step for the origination of allopolyploids, brings divergent genomes from different species together into an amphihaploid hybrid. Because only one set of homologous chromosomes is present, amphihaploids (analogous to haploid plants) are usually sterile due to reduced meiosis. Meiotic nonreduction (meiotic restitution), however, can lead to production of functionally unreduced gametes, and their union immediately generates an amphidiploid (allopolyploid). Meiotic nonreduction is thought to be the

The abundance of homoeologue transcripts is disrupted by hybridization and is partially restored by genome doubling in synthetic hexaploid wheat

BackgroundThe formation of an allopolyploid is a two step process, comprising an initial wide hybridization event, which is later followed by a whole genome doubling. Both processes can affect the transcription of homoeologues. Here, RNA-Seq was used to obtain the genome-wide leaf transcriptome of two independent Triticum turgidum × Aegilops tauschii allotriploids (F1), along with their spontaneous allohexaploids (S1) and their parental lines. The resulting sequence data were then used to characterize variation in homoeologue transcript abundance.ResultsThe hybridization event strongly down-regulated D-subgenome homoeologues, but this effect was in many cases reversed by whole genome doubling. The suppression of D-subgenome homoeologue transcription resulted in a marked frequency of parental transcription level dominance, especially with respect to genes encoding proteins involved in photosynthesis. Singletons (genes where no homoeologues were present) were frequently transcribed at both the allotriploid and allohexaploid plants.ConclusionsThe implication is that whole genome doubling helps to overcome the phenotypic weakness of the allotriploid, restoring a more favourable gene dosage in genes experiencing transcription level dominance in hexaploid wheat.

Characterization of a novel y-type HMW-GS with eight cysteine residues from Triticum monococcum ssp. monococcum

The composition and number of high-molecular-weight glutenin subunits (HMW-GSs) play important roles in determining the grain-processing quality of common wheat. The Glu-1Ay allele is silent in common wheat. In this study, an active y-type HMW-GS allele termed 1Ay8.2 (GenBank No. KP137569) was identified from Triticum monococcum L. ssp. monococcum (AmAm, 2n=2x=14), a species with a genome related to the A-genome of common wheat. Compared with previously reported active 1Ay subunits, this novel subunit contained an extra cysteine residue at position 103 of the amino acid sequence in the N-terminal region, in addition to the six cysteines in the N- and C-terminal regions found in most active 1Ay subunits and the one in the repetitive region that appears in only a few 1Ay alleles. This subunit was expressed in an amphiploid (AAAmAmBB, 2n=6x=42) between Triticum turgidum L. ssp. dicoccon and T. monococcum ssp. monococcum. This amphiploid could be used as a bridge to transfer 1Ay8.2 into common wheat cultivars. Replacing the silenced 1Ay in common wheat with the active 1Ay8.2 allele harboring an extra cysteine residue is expected to improve the quality by increasing the number of HMW-GSs and promoting the formation of covalent interactions through disulfide bonds with the extra cysteine residue.

Fluorescence in situ hybridization karyotyping reveals the presence of two distinct genomes in the taxon Aegilops tauschii

BackgroundAegilops tauschii is the donor of the bread wheat D genome. Based on spike morphology, the taxon has conventionally been subdivided into ssp. tauschii and ssp. strangulata. The present study was intended to address the poor match between this whole plant morphology-based subdivision and genetic relationships inferred from genotyping by fluorescence in situ hybridization karyotyping a set of 31 Ae. tauschii accessions.ResultsThe distribution of sites hybridizing to the two probes oligo-pTa-535 and (CTT)10 split the Ae. tauschii accessions into two clades, designated Dt and Ds, which corresponded perfectly with a previously assembled phylogeny based on marker genotype. The Dt cluster was populated exclusively by ssp. tauschii accessions, while the Ds cluster harbored both ssp. strangulata and morphologically intermediate accessions. As a result, it is proposed that Ae. tauschii ssp. tauschii is restricted to carriers of the Dt karyotype: their spikelets are regularly spaced along the rachis, at least in the central portion of their spike. Accessions classified as Ae. tauschii ssp. strangulata carry the Ds karyotype; their spikelets are irregularly spaced. Based on this criterion, forms formerly classified as ssp. tauschii var. meyeri have been re-designated ssp. strangulata var. meyeri.ConclusionsAccording to the reworking of the taxon, the bread wheat D genome was most probably donated by ssp. strangulata var. meyeri. Chromosomal differentiation reveals intra-species taxon of Ae. tauschii. Ae. tauschii ssp. tauschii has more distant relationship with breed wheat than ssp. strangulata and can be used for breeding improving effectively.

Introgression of Powdery Mildew Resistance Gene Pm56 on Rye Chromosome Arm 6RS Into Wheat

Powdery mildew, caused by the fungus Blumeria graminis f. sp. tritici, represents a yield constraint in many parts of the world. Here, the introduction of a resistance gene carried by the cereal rye cv. Qinling chromosome 6R was transferred into wheat in the form of spontaneous balanced translocation induced in plants doubly monosomic for chromosomes 6R and 6A. The translocation, along with other structural variants, was detected using in situ hybridization and genetic markers. The differential disease response of plants harboring various fragments of 6R indicated that a powdery mildew resistance gene(s) was present on both arms of rye chromosome 6R. Based on karyotyping, the short arm gene, designated Pm56, was mapped to the subtelomere region of the arm. The Robertsonian translocation 6AL⋅6RS can be exploited by wheat breeders as a novel resistance resource.