Introgression of the Powdery Mildew Resistance Genes Pm60 and Pm60b from Triticum urartu to Common Wheat Using Durum as a ‘Bridge’

Powdery mildew, caused by the fungus Blumeria graminis f. sp. tritici (Bgt), has limited wheat yields in many major wheat-production areas across the world. Introducing resistance genes from wild relatives into cultivated wheat can enrich the genetic resources for disease resistance breeding. The powdery mildew resistance gene Pm60 was first identified in diploid wild wheat Triticum urartu (T. urartu). In this study, we used durum as a ‘bridge’ approach to transfer Pm60 and Pm60b into hexaploid common wheat. Synthetic hexaploid wheat (SHW, AABBAuAu), developed by crossing T. urartu (AuAu) with durum (AABB), was used for crossing and backcrossing with common wheat. The Pm60 alleles were tracked by molecular markers and the resistance to powdery mildew. From BC1F1 backcross populations, eight recombinant types were identified based on five Pm60-flanking markers, which indicated different sizes of the introgressed chromosome segments from T. urartu. Moreover, we have selected two resistance-harboring introgression lines with high self-fertility, which could be easily used in wheat breeding system. Our results showed that the durum was an excellent ‘bridge’ for introducing the target gene from diploid T. urartu into the hexaploid cultivated wheat. Moreover, these introgression lines could be deployed in wheat resistance breeding programs, together with the assistance of the molecular markers for Pm60 alleles.

Frequent numerical and structural chromosome changes in early generations of synthetic hexaploid wheat

Modern hexaploid wheat (Triticum aestivum L.; AABBDD) evolved from a hybrid of tetraploid wheat (closely related to Triticum turgidum L. ssp. durum (Desf.) Husn., AABB) and goatgrass (Aegilops tauschii Coss., DD). Variations in chromosome structure and ploidy played important roles in wheat evolution. How these variations occurred and their role in expanding the genetic diversity in modern wheat is mostly unknown. Synthetic hexaploid wheat (SHW) can be used to investigate chromosome variation that occurs during the early generations of existence. SHW lines derived by crossing durum wheat ‘Langdon’ with twelve Ae. tauschii accessions were analyzed using oligonucelotide probe multiplex fluorescence in situ hybridization (FISH) to metaphase chromosomes and SNP markers. Cluster analysis based on SNP markers categorized them into three groups. Among 702 plants from the S8 and S9 generations, 415 (59.12%) carried chromosome variations involving all 21 chromosomes but with different frequencies for each chromosome and sub-genome. Total chromosome variation frequencies varied between lines, but there was no significant difference among the three groups. The non-random chromosome variations in SHW lines detected in this research may be an indication that similar variations occurred in the early stages of wheat polyploidization and played important roles in wheat evolution.

Resistance of synthetic hexaploid wheat to the leaf rust pathogen

Background. One of the promising sources for enrichment of the common wheat (Triticum aestivum L.) gene pool with new alleles is synthetic hexaploid wheat (SHW), or allopolyploids from crossing tetraploid wheats (2n = 4x = 28, BBAA) with accessions of Aegilops tauschii Coss. (2n = 2x = 14, DD), and subsequent doubling of the chromosome number in the hybrids. Objectives of the study were to evaluate the SHW accessions from the VIR collection for resistance to Puccinia triticina Erikss. populations collected in Russia; genotype the accessions; and summarize information from the published sources concerning the resistance of the studied accessions to other harmful diseases and pests.Materials and methods. Resistance of 36 SHW accessions from the VIR collection to the populations of P. triticina was assessed in the laboratory and in the field, under artificial infection pressure, using the techniques developed by the Institute of Plant Protection. A phytopathological test and PCR markers were used to identify the Lr genes.Results and conclusion. The SHW accessions were characterized according to their resistance to the Russian populations of the wheat leaf rust pathogen. The sources of resistance in the phase of emergence and in adult plants were identified. The phytopathological test isolated three accessions with Lr23; the PCR marker of Lr21=Lr40 was found in 11 accessions, Lr39=Lr41 in 19, and Lr22a in 3. At the same time, k-65496, k-65515 and k-65517 had si multaneously Lr21=Lr40 and Lr39=Lr41, while k-65497, k-65503 and k-65508 had Lr22a and Lr39=Lr41. The analysis of published data showed that many of the studied SHW accessions were also resistant to other harmful diseases and insect pests, so they are of interest for further studying and possible use in domestic breeding.

Genome-wide Analysis of NPF, NRT2, CLC and SLAC1/SLAH Nitrate Transporters in Hexaploid Wheat (Triticum Aestivum)

Nitrogen transport is one of the most important processes in plants mediated by specialized transmembrane proteins. Plants have two main systems for nitrogen uptake from soil and its transport within the system - a low-affinity transport system and a high-affinity transport system. Nitrate transporters are of special interest in cereal crops because large amount of money is spent on N fertilizers every year to enhance the crop productivity. Till date four gene families of nitrate transporter proteins; NPF (nitrate transporter 1/peptide transporter family), NRT2 (nitrate transporter 2 family), the CLC (chloride channel family), and the SLAC/SLAH (slow anion channel-associated homologues) have been reported in plants. In our study, in silico mining of nitrate transporter genes along with their detailed structure, phylogenetic and expression analysis was carried out. A total of 412 nitrate transporter genes were identified in hexaploid wheat genome using HMMER based homology searches in IWGSC Refseq v2.0. Out of those twenty genes were root specific, 11 leaf/shoot specific and 17 genes were grain/spike specific. The identification of nitrate transporter genes in the close proximity to the previously identified 67 marker-traits associations associated with the nitrogen use efficiency related traits in nested synthetic hexaploid wheat introgression library indicated the robustness of the reported transporter genes. The detailed crosstalk between the genome and proteome and the validation of identified putative candidate genes through expression and gene editing studies may lay down the foundation to improve nitrogen use efficiency of cereal crops.

Development and Identification of Four New Synthetic Hexaploid Wheat Lines with Solid Stems

Stem solidness is an important agronomic trait for increasing the ability of wheat to resist lodging. In this study, four new synthetic hexaploid wheat with solid stems were developed from natural chromosome doubling of F1 hybrids between a solid-stemmed durum wheat (Triticum turgidum ssp. durum, 2n = 4x = 28, AABB) and four Aegilops tauschii (2n = 2x = 14, DD) accessions. The solid expression of the second internode at the base of the stem was stable for two synthetic hexalpoid wheat Syn-SAU-117 and Syn-SAU-119 grown in both the greenhouse and field. The lodging resistance of four synthetic solid-stem wheats is stronger than that of CS, and Syn-SAU-116 has the strongest lodging resistance, followed by Syn-SAU-119. The paraffin sections of the second internode showed that four synthetic wheat lines had large outer diameters, well-developed mechanical tissues, large number of vascular bundles, and similar anatomical characteristics with solid-stemmed durum wheat. The chromosomal composition of four synthetic hexaploid wheat was identified by FISH (fluorescence in situ hybridization) using Oligo-pSc119.2-1 and Oligo-pTa535-1. At adult stage, all four synthetic hexaploid wheat showed high resistance to mixed physiological races of stripe rust pathogen (CYR31, CYR32, CYR33, CYR34). These synthetic hexaploid wheat lines provide new materials for the improvement of common wheat.

Characterization of Synthetic Wheat Line Largo for Resistance to Stem Rust

Resistance breeding is an effective approach against wheat stem rust caused by Puccinia graminis f. sp. tritici (Pgt). The synthetic hexaploid wheat line Largo (pedigree: durum wheat ‘Langdon’ × Aegilops tauschii PI 268210) was found to have resistance to a broad spectrum of Pgt races including the Ug99 race group. To identify the stem rust resistance (Sr) genes, we genotyped a population of 188 recombinant inbred lines developed from a cross between the susceptible wheat line ND495 and Largo using the wheat Infinium 90 K SNP iSelect array and evaluated the population for seedling resistance to the Pgt races TTKSK, TRTTF, and TTTTF in the greenhouse conditions. Based on genetic linkage analysis using the marker and rust data, we identified six quantitative trait loci (QTL) with effectiveness against different races. Three QTL on chromosome arms 6AL, 2BL, and 2BS corresponded to Sr genes Sr13c, Sr9e, and a likely new gene from Langdon, respectively. Two other QTL from PI 268210 on 2DS and 1DS were associated with a potentially new allele of Sr46 and a likely new Sr gene, respectively. Additionally, Sr7a was identified as the underlying gene for the 4AL QTL from ND495. Knowledge of the Sr genes in Largo will help to design breeding experiments aimed to develop new stem rust-resistant wheat varieties. Largo and its derived lines are particularly useful for introducing two Ug99-effective genes Sr13c and Sr46 into modern bread wheat varieties. The 90 K SNP-based high-density map will be useful for identifying the other important genes in Largo.

Chromosome Karyotype and Stability Identification of New Synthetic Hexaploid Wheat

Synthetic hexaploid wheats offer breeder ready access to potentially novel genetic variations in wild ancestral species. In present study, we used MY3478 (2n = 4x = 28, AABB) as female crossing with SY41 (2n = 2x = 14, DD) of stripe rust resistant as male through natural chromosome doubling to constructed the new hexaploid wheat line NA0928. Agronomic traits and cytological analysis were characterized in NA0928 of S8-S9 generations. The major study and results were described as follows, agronomic character variation coefficient in NA0928 of S8 generation showed that the effective tiller number (55.3%) > spike length (15.3%) > number of spikelets (13.9%) > plant height (8.7). It is suggested that the effective tiller number and spike length have great utilization value in breeding. Cytological observation and fluorescence in situ hybridization (FISH) results showed that the chromosome number and configuration have rich variations in S9 of NA0928. Chromosome number variation range of 36–44. Numerous chromosome karyotype variations were almost detected in A and B subgenomes. In addition, more diverse types of chromosomal structure variations were observed in the stripe rust resistant strains with more excellent performance than susceptible strains in agronomic traits. Especially, the tillering number of the resistant strains were much higher. Here, Meiosis stage of pollen mother cells and multicolor-GISH (Mc-GISH) results showed that two new synthetic hexaploid wheat lines were obtained, which showed genetic stability, one line was resistance to stripe rust, and the other one line was susceptible stripe rust, at the same time, there had two excellent characteristics with high 1000-grain weight and multiple tillers. They will be valuable germplasm materials in wheat breeding utilization.