Tritordeum: Creating a New Crop Species—The Successful Use of Plant Genetic Resources

Hexaploid tritordeum is the amphiploid derived from the cross between the wild barley Hordeum chilense and durum wheat. This paper reviews the main advances and achievements in the last two decades that led to the successful development of tritordeum as a new crop. In particular, we summarize the progress in breeding for agronomic performance, including the potential of tritordeum as a genetic bridge for wheat breeding; the impact of molecular markers in genetic studies and breeding; and the progress in quality and development of innovative food products. The success of tritordeum as a crop shows the importance of the effective utilization of plant genetic resources for the development of new innovative products for agriculture and industry. Considering that wild plant genetic resources have made possible the development of this new crop, the huge potential of more accessible resources, such as landraces conserved in gene banks, goes beyond being sources of resistance to biotic and abiotic stresses. In addition, the positive result of tritordeum also shows the importance of adequate commercialization strategies and demonstrative experiences aimed to integrate the whole food chain, from producers to end-point sellers, in order to develop new products for consumers.

Durum Wheat (Triticum durum L.) Landraces Reveal Potential for the Improvement of Grain Carotenoid Esterification in Breeding Programs

Carotenoids are essential in the human diet for their important functions in health. Besides, they are responsible for the yellow pigments desirable for industrial quality in durum wheat. The remarkable carotenoid content of durum wheat endosperm is mostly due to lutein. However, lutein esters have not been previously detected in durum wheat as in other cereals such as common wheat, tritordeum or Hordeum chilense. Esterification increases carotenoid stability and allows greater retention and accumulation through the food chain. Therefore, carotenoid esterification is revealed as a new key target in breeding. We characterized the carotenoid profile of 156 accessions of the Spanish durum wheat collection, searching for landraces with esterification ability. Interestingly, four accessions produced lutein monoesters and diesters. Also, traces of lutein monoesters were detected in eleven accessions. The identification of the first durum wheat accessions with esterification ability reported herein is a remarkable advance for carotenoid biofortification. Furthermore, variation for the relative content of zeaxanthin, α-carotene and β-carotene was also observed. This diversity for the β,ε and β,β branches of the carotenogenic pathway also represents a new opportunity for breeding for specific carotenoids in biofortification programs.

Wild and Cultivated Homoeologous Barley Chromosomes Can Associate and Recombine in Wheat in the Absence of the Ph1 Locus

Bread wheat is an allohexaploid that behaves as a diploid during meiosis, the cell division process to produce the gametes occurring in organisms with sexual reproduction. Knowledge of the mechanisms implicated in meiosis can contribute to facilitating the transfer of desirable traits from related species into a crop like wheat in the framework of breeding. It is particularly interesting to shed light on the mechanisms controlling correct pairing between homologous (equivalent) chromosomes and recombination, even more in polyploid species. The Ph1 (Pairing homoeologous 1) locus is implicated in recombination. In this work, we aimed to study whether homoeologous (equivalent chromosomes from different genomes) Hordeum chilense (wild barley) and H. vulgare (cultivated barley) chromosomes can associate and recombine during meiosis in the wheat background in the absence of the Ph1 locus. For this, we have developed H. chilense and H. vulgare double monosomic addition lines for the same and for different homoeology group in wheat in the ph1b mutant background. Using genomic in situ hybridization, we visualized the two (wild and cultivated) barley chromosomes during meiosis and we studied the processes of recognition, association, and recombination between homoeologous chromosomes in the absence of the Ph1 locus. Our results showed that the Ph1 locus does not prevent homoeologous chromosome pairing but it can regulate recombination.

Wheat, Rye, and Barley Genomes Can Associate during Meiosis in Newly Synthesized Trigeneric Hybrids

Polyploidization, or whole genome duplication (WGD), has an important role in evolution and speciation. One of the biggest challenges faced by a new polyploid is meiosis, in particular, discriminating between multiple related chromosomes so that only homologs recombine to ensure regular chromosome segregation and fertility. Here, we report the production of two new hybrids formed by the genomes of species from three different genera: a hybrid between Aegilops tauschii (DD), Hordeum chilense (HchHch), and Secale cereale (RR) with the haploid genomic constitution HchDR (n = 7× = 21); and a hybrid between Triticum turgidum spp. durum (AABB), H. chilense, and S. cereale with the constitution ABHchR (n = 7× = 28). We used genomic in situ hybridization and immunolocalization of key meiotic proteins to establish the chromosome composition of the new hybrids and to study their meiotic behavior. Interestingly, there were multiple chromosome associations at metaphase I in both hybrids. A high level of crossover (CO) formation was observed in HchDR, which shows the possibility of meiotic recombination between the different genomes. We succeeded in the duplication of the ABHchR genome, and several amphiploids, AABBHchHchRR, were obtained and characterized. These results indicate that recombination between the genera of three economically important crops is possible.

Mediation of a GDSL Esterase/Lipase in Carotenoid Esterification in Tritordeum Suggests a Common Mechanism of Carotenoid Esterification in Triticeae Species

Carotenoids are essential in human diet, so that the development of programs toward carotenoid enhancement has been promoted in several crops. The cereal tritordeum, the amphiploid derived from the cross between Hordeum chilense Roem. et Schulz. and durum wheat has a remarkable carotenoid content in the endosperm. Besides, a high proportion of these carotenoids are esterified with fatty acids. The identification of the gene(s) responsible for xanthophyll esterification would be useful for breeding as esterified carotenoids show an increased ability to accumulate within plant cells and have a higher stability during post-harvest storage. In this work, we analyzed five genes identified as candidates for coding the xanthophyll acyltransferase (XAT) enzyme responsible for lutein esterification in H. chilense genome. All these genes were expressed during grain development in tritordeum, but only HORCH7HG021460 was highly upregulated. Sequence analysis of HORCH7HG021460 revealed a G-to-T transversion, causing a Glycine to Cysteine substitution in the protein of H290 (the only accession not producing quantifiable amounts of lutein esters, hereinafter referred as zero-ester) of H. chilense compared to the esterifying genotypes. An allele-specific marker was designed for the SNP detection in the H. chilense diversity panel. From the 93 accessions, only H290 showed the T allele and the zero-ester phenotype. Furthermore, HORCH7HG021460 is the orthologue of XAT-7D, which encodes a XAT enzyme responsible for carotenoid esterification in wheat. Thus, HORCH7HG021460 (XAT-7Hch) is a strong candidate for lutein esterification in H. chilense and tritordeum, suggesting a common mechanism of carotenoid esterification in Triticeae species. The transference of XAT-7Hch to wheat may be useful for the enhancement of lutein esters in biofortification programs.

A Comparative Study of Organic and Conventional Management on the Rhizosphere Microbiome, Growth and Grain Quality Traits of Tritordeum

Tritordeum is a novel hexaploid cereal derived from the cross between a wild Chilean barley species (Hordeum chilense Roem. et Schultz) and durum wheat (Triticum turgidum ssp. durum Desf.) that is potentially of great interest for human nutrition. In this study, a commercial and an experimental Tritordeum cultivar were analyzed in comparison with a reference durum wheat under conventional and organic management. We demonstrate that Tritordeum is better adapted to organic farming through an increase in the below-ground rhizosphere community of the Bacteroidetes phylum, which includes many bacteria species known to exert beneficial effects on plants, particularly for root growth. Despite a considerably lower grain yield, Tritordeum had better quality traits than durum wheat, particularly under organic farming vs. conventional management, with respect to total protein contents, high molecular weight glutenin subunits, antioxidant free phenols and nutrients (i.e., calcium, potassium, sulphur, iron, and zinc), depending on the cultivar. We conclude that Tritordeum is a promising cereal in light of its quality traits and adaptability to sustainable crop management practices, such as organic farming, although further improvement in yield potential should be pursued by breeding and by optimising the cultivation method.

Development of wheat—Hordeum chilense Chromosome 2Hch Introgression Lines Potentially Useful for Improving Grain Quality Traits

The chromosome 2Hch of Hordeum chilense. has the potential to improve seed carotenoid content in wheat as it carries a set of endosperm carotenoid-related genes. We have obtained structural changes in chromosome 2Hch in a common wheat (Triticum aestivum L. “Chinese Spring”) background by crossing a wheat double disomic substitution 2Hch(2D) and 7Hch(7D) line with a disomic addition line carrying chromosome 2Cc from Aegilops cylindrica Host.. Seven introgressions of chromosome 2Hch into wheat were characterized by fluorescence in situ hybridization (FISH) and DNA markers. Chromosome-specific simple sequence repeats (SSRs) were used for identifying wheat chromosomes. In addition, we tested 82 conserved orthologous set (COS) markers for homoeologous group 2, of which 65 amplified targets in H. chilense and 26 showed polymorphism between H. chilense and wheat. A total of 24 markers were assigned to chromosome 2Hch with eight allocated to 2HchS and 16 to 2HchL. Among the seven introgressions there was a disomic substitution line 2Hch(2D), a ditelosomic addition line for the 2HchL arm, an isochromosome for the 2HchL arm, a homozygous centromeric 2HchS·2DL translocation, a double monosomic 2HchS·2DL plus 7HchS·D translocation, a homozygous centromeric 7HchS·2HchL translocation and, finally, a 2HchL·7HchL translocation. Wheat—H. chilense macrosyntenic comparisons using COS markers revealed that H. chilense chromosome 2Hch exhibits synteny to wheat homoeologous group 2 chromosomes, and the COS markers assigned to this chromosome will facilitate alien gene introgression into wheat. The genetic stocks developed here include new wheat—H. chilense recombinations which are useful for studying the effect of chromosome 2Hch on grain quality traits.

Wx Gene in Hordeum chilense: Chromosomal Location and Characterisation of the Allelic Variation in the Two Main Ecotypes of the Species

Starch, as the main grain component, has great importance in wheat quality, with the ratio between the two formed polymers, amylose and amylopectin, determining the starch properties. Granule-bound starch synthase I (GBSSI), or waxy protein, encoded by the Wx gene is the sole enzyme responsible for amylose synthesis. The current study evaluated the variability in Wx genes in two representative lines of Hordeum chilense Roem. et Schult., a wild barley species that was used in the development of tritordeum (×Tritordeum Ascherson et Graebner). Two novel alleles, Wx-Hch1a and Wx-Hch1b, were detected in this material. Molecular characterizations of these alleles revealed that the gene is more similar to the Wx gene of barley than that of wheat, which was confirmed by phylogenetic studies. However, the enzymatic function should be similar in all species, and, consequently, the variation present in H. chilense could be utilized in wheat breeding by using tritordeum as a bridge species.

Identification of candidate genes for lutein esterification in common wheat (Triticum aestivum) using physical mapping and genomics tools

A high carotenoid content is important for the production of pasta from durum wheat (Triticum durum Desf.) and yellow alkaline noodle from common wheat (T. aestivum L.). Carotenoid esters are more stable than free carotenoid during storage and processing, and thus they allow a higher retention through the food chain. Chromosome 7D carries gene(s) for lutein esterification. The aim of this study was the physical mapping of the gene(s) for lutein esterification on chromosome 7D and the identification of candidate genes for this trait. We developed crosses between a set of deletion lines for chromosome 7D in Chinese Spring (CS) background and the CS–Hordeum chilense substitution line CS(7D)7Hch. The F2 progeny derived from the deletion line 7DS4 produced a lower amount of lutein esters, which indicates that the main gene for lutein esterification is in the region of chromosome 7D lacking in 7DS4. Other gene(s) are contributing to lutein esterification because small amounts of lutein esters are produced in 7DS4. Genotyping by DArTSeq revealed that 7DS4 lacks a 127.7 Mb region of 7DS. A set of 10 candidate genes for lutein esterification was identified by using the wheat reference genome sequence along with the Wheat Expression Browser. This region contains the Lute locus previously identified in a different genetic background. Four genes with acyltransferase or GDSL esterase/lipase activity were identified in the vicinity of Lute. Our results indicate that the gene TraesCS7D01G094000 is a likely candidate for Lute but the gene TraesCS7D01G093200 cannot be ruled out. The candidate genes reported in this work are worthy for further investigation.