Molecular Cytogenetic Analysis of the Introgression between Agropyroncristatum P Genome and Wheat Genome

Agropyroncristatum (2n = 4x = 28, PPPP) is an important wild relative of common wheat (Triticum aestivum L., 2n = 6x = 42). A previous report showed that the wheat-A. cristatum 6P translocation line WAT655 carrying A. cristatum 6PS (0.81–1.00) exhibited high resistance to prevalent physiological races of stripe rust (CYR32 and CYR33). In this study, three disease resistance-related transcripts, which were mapped to A. cristatum 6PS (0.81–1.00) through the analysis of specific molecular markers, were acquired from among A. cristatum full-length transcripts. The BC5F2 and BC5F2:3 genetic populations of the translocation line WAT655 were analyzed by using three disease resistance-related gene markers, A. cristatum P genome-specific markers, and fluorescence in situ hybridization (FISH). The results revealed that the introgression between A. cristatum P genome and wheat genome was observed in progenies of the genetic populations of the translocation line WAT655 and the physical positions of the three genes were considerably adjacent on A. cristatum 6PS (0.81–1.00) according to the FISH results. Additionally, kompetitive allele-specific PCR (KASP) markers of the three genes were developed to detect and acquire 24 breeding lines selected from the progenies of the distant hybridization of wheat and A. cristatum, which showed resistance to physiological races of stripe rust (CYR32 and CYR33) and other desirable agronomic traits according to the field investigation. In conclusion, this study not only provides new insights into the introgression between A. cristatum P genome and wheat genome but also provides the desirable germplasms for breeding practice.

Cytogenetic identification and molecular marker development for the novel stripe rust-resistant wheat–Thinopyrum intermedium translocation line WTT11

Stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is one of the most destructive diseases of wheat (Triticum aestivum L.) worldwide. Xiaoyan 78829, a partial amphidiploid developed by crossing common wheat with Thinopyrum intermedium, is immune to wheat stripe rust. To transfer the resistance gene of this excellent germplasm resource to wheat, the translocation line WTT11 was produced by pollen irradiation and assessed for immunity to stripe rust races CYR32, CYR33 and CYR34. A novel stripe rust-resistance locus derived from Th. intermedium was confirmed by linkage and diagnostic marker analyses. Molecular cytogenetic analyses revealed that WTT11 carries a TTh·2DL translocation. The breakpoint of 1B was located at 95.5 MB, and the alien segments were found to be homoeologous to wheat-group chromosomes 6 and 7 according to a wheat660K single-nucleotide polymorphism (SNP) array analysis. Ten previously developed PCR-based markers were confirmed to rapidly trace the alien segments of WTT11, and 20 kompetitive allele-specific PCR (KASP) markers were developed to enable genotyping of Th. intermedium and common wheat. Evaluation of agronomic traits in two consecutive crop seasons uncovered some favorable agronomic traits in WTT11, such as lower plant height and longer main panicles, that may be applicable to wheat improvement. As a novel genetic resource, the new resistance locus may be useful for wheat disease-resistance breeding.

Targeted Sequencing of the Short Arm of Chromosome 6V of a Wheat Relative Haynaldia villosa for Marker Development and Gene Mining

The short arm of chromosome 6V (6VS) of Haynaldia villosa has been used in wheat breeding programs to introduce Pm21 resistance gene against powdery mildew (Pm) and some other genes. In this this study, 6VS was flow-sorted from wheat-H. villosa ditelosomic addition line Dt6VS and sequenced by Illumina technology. An assembly of 230.39 Mb was built with contig N50 of 9.788 bp. In total, 3.276 high-confidence genes were annotated and supported by RNA sequencing data. Repetitive elements represented 74.91% of the 6VS assembly. The 6VS homologous genes were identified on homologous group 6 in six Triticeae species confirming their synteny relationships. Out of 45 NB-ARC domain proteins identified on 6VS, 15 were upregulated and might also be involved in the innate immunity of H. villosa to Pm. High thousand grain weight (TGW) for 6VS/6AL translocation line was not attributable to GW2-6V gene. Based on the intron size differences, 119 intron-target (IT) markers were developed to trace the 6VS chromatins introduced into wheat background. The assembled 6VS genome sequence and the developed 6VS specific IT markers in this work will facilitate the gene mining and utilization of agronomic important genes on 6VS.

Development and Molecular Cytogenetic Identification of a New Wheat–Psathyrostachys huashanica Keng Translocation Line Resistant to Powdery Mildew

Psathyrostachys huashanica Keng, a wild relative of common wheat with many desirable traits, is an invaluable source of genetic material for wheat improvement. Few wheat–P. huashanica translocation lines resistant to powdery mildew have been reported. In this study, a wheat–P. huashanica line, E24-3-1-6-2-1, was generated via distant hybridization, ethyl methanesulfonate (EMS) mutagenesis, and backcross breeding. A chromosome karyotype of 2n = 44 was observed at the mitotic stage in E24-3-1-6-2-1. Genomic in situ hybridization (GISH) analysis revealed four translocated chromosomes in E24-3-1-6-2-1, and P. huashanica chromosome-specific marker analysis showed that the alien chromosome fragment was from the P. huashanica 4Ns chromosome. Moreover, fluorescence in situ hybridization (FISH) analysis demonstrated that reciprocal translocation had occurred between the P. huashanica 4Ns chromosome and the wheat 3D chromosome; thus, E24-3-1-6-2-1 carried two translocations: T3DS·3DL-4NsL and T3DL-4NsS. Translocation also occurred between wheat chromosomes 2A and 4A. At the adult stage, E24-3-1-6-2-1 was highly resistant to powdery mildew, caused by prevalent pathotypes in China. Further, the spike length, numbers of fertile spikelets, kernels per spike, thousand-kernel weight, and grain yield of E24-3-1-6-2-1 were significantly higher than those of its wheat parent 7182 and addition line 24-6-3-1. Thus, this translocation line that is highly resistant to powdery mildew and has excellent agronomic traits can be used as a novel promising germplasm for breeding resistant and high-yielding cultivars.

New Insights Into the Introgression Between Agropyron Cristatum P Genome and Wheat Genome

Agropyron cristatum (2n = 4x = 28, PPPP) is an important wild relative of common wheat and confers desirable agronomic traits to common wheat. A previous report showed that the wheat-A. cristatum 6P translocation line WAT655 carrying A. cristatum 6PS (0.81–1.00) exhibited high resistance to prevalent physiological races (CYR32 and CYR33). In this study, three disease resistance-related transcriptomes, which were mapped to A. cristatum 6PS (0.81–1.00) through the analysis of specific molecular markers, were searched from among A. cristatum full-length transcriptomes. Then, three disease resistance-related gene markers, A. cristatum P genome-specific markers, and fluorescence in situ hybridization (FISH)/genomic in situ hybridization (GISH) probes made from the DNA of three bacterial artificial chromosome (BAC) clones, three genes, and A. cristatum “Z559” were used to analyze the BC5F2 and BC5F2:3 genetic populations of the translocation line WAT655. The results revealed the introgression can spontaneously occur between A. cristatum P genome and wheat genome, and indicated the three genes could constitute a gene cluster according to the positions of their FISH signals. Additionally, kompetitive allele-specific PCR (KASP) markers of the three genes were developed to detect and acquire 24 wheat-A. cristatum breeding materials, which showed resistance to physiological races (CYR32 and CYR33) and other desirable agronomic traits according to the field investigation. In conclusion, our study not only provides new insights into the introgression between A. cristatum P genome and wheat genome, but also provides the desirable breeding materials for breeding practice.

Identification of an Elite Wheat-Rye T1RS·1BL Translocation Line Conferring High Resistance to Powdery Mildew and Stripe Rust

Wheat-rye T1RS·1BL translocations have been widely used worldwide in wheat production for multiple disease resistance and superior yield traits. However, many T1RS·1BL translocations have successively lost their resistance to pathogens due to the coevolution of pathogen virulence with host resistance. Because of the extensive variation in rye (Secale cereale L.) as a naturally cross-pollinating relative of wheat, it still has promise to widen the variation of 1RS and to fully realize its application value in wheat improvement. In the present study, the wheat-rye breeding line R2207 was characterized by comprehensive analyses using genomic in situ hybridization (GISH), multicolor fluorescence in situ hybridization with multiple probes, multicolor GISH, and molecular marker analysis, and then was proven to be a cytogenetically stable wheat-rye T1RS·1BL translocation line. Based on the disease responses to different isolates of powdery mildew and genetic analysis, R2207 appears to possess a novel variation for resistance, which was confirmed to be located on the rye chromosome arm 1RS. Line R2207 also exhibited high levels of resistance to stripe rust at both seedling and adult stages, as well as enhanced agronomic performance, so it has been transferred into a large number of commercial cultivars using an efficient 1RS-specific kompetitive allele specific PCR marker for marker-assisted selection.

Molecular and Cytogenetic Characterization of a Wheat-Rye 7BS.7RL Translocation Line with Resistance to Stripe Rust, Powdery Mildew, and Fusarium Head Blight

Secale cereale is used as a source of genes for disease resistance in wheat cultivation. In this study, a homozygous translocation line (RT14-245) that originated from a cross between a commercial wheat cultivar (Mianyang 11) and a local Chinese variety of rye (Baili) was developed. Multicolor fluorescence in situ hybridization and PCR analysis demonstrated that the translocation chromosome was 7BS.7RL. Resistance analysis showed that RT14-245 was resistant to prevalent pathotypes of stripe rust and powdery mildew. RT14-245 also exhibited high resistance to Fusarium head blight, which was similar to the resistance exhibited by the wheat cultivar Sumai 3. The results indicated that RT14-245 simultaneously exhibited high levels of resistance against stripe rust, powdery mildew, and Fusarium head blight. These results indicate that chromosome arm 7RL in the translocation line RT14-245 is an excellent new resource for wheat breeding programs.