Genome-Wide Identification of GDSL-Type Esterase/Lipase Gene Family in Dasypyrum villosum L. Reveals That DvGELP53 Is Related to BSMV Infection

GDSL-type esterase/lipase proteins (GELPs) characterized by a conserved GDSL motif at their N-terminus belong to the lipid hydrolysis enzyme superfamily. In plants, GELPs play an important role in plant growth, development and stress response. The studies of the identification and characterization of the GELP gene family in Triticeae have not been reported. In this study, 193 DvGELPs were identified in Dasypyrum villosum and classified into 11 groups (clade A–K) by means of phylogenetic analysis. Most DvGELPs contain only one GDSL domain, only four DvGELPs contain other domains besides the GDSL domain. Gene structure analysis indicated 35.2% DvGELP genes have four introns and five exons. In the promoter regions of the identified DvGELPs, we detected 4502 putative cis-elements, which were associated with plant hormones, plant growth, environmental stress and light responsiveness. Expression profiling revealed 36, 44 and 17 DvGELPs were highly expressed in the spike, the root and the grain, respectively. Further investigation of a root-specific expressing GELP, DvGELP53, indicated it was induced by a variety of biotic and abiotic stresses. The knockdown of DvGELP53 inhibited long-distance movement of BSMV in the tissue of D. villosum. This research provides a genome-wide glimpse of the D. villosum GELP genes and hints at the participation of DvGELP53 in the interaction between virus and plants.

Morphological, Genetic and Biochemical Evaluation of Dasypyrum villosum (L.) P. Candargy in the Gene Bank Collection

The Dasypyrum villosum gene bank collection, comprising 32 accessions, was characterized morphologically and genetically for resistance to leaf diseases and for quality parameters of seeds with specific accent to protein polymorphism and protein and starch composition. The collected material represented nearly the whole distribution area in the Mediterranean. For SSR analysis, a set of 40 SSR markers for wheat was selected. A matrix of distances between genotypes was calculated using Simple Matching dissimilarity coefficient in the DARwin software. The collection was scored for resistance to powdery mildew, brown, stripe and stem rusts. A modified SDS-PAGE method with clear interpretation of high and low molecular glutenin subunits (HMW, LMW) was used for characterization of accessions. Morphological phenotyping revealed considerable diversity allowing the distinguishing of clusters tracing the geographical origin of accessions. Genetic diversity showed three groups but without significant bootstrap support. All tested accessions were resistant to the applied races of powdery mildew and leaf rust. Three accessions were moderately susceptible to currently available races of yellow rust. Biochemical analyses of seeds in selected populations showed a high content of crude proteins with a significant proportion of prolamins and Ʃglutelins. The SDS-PAGE of HMW and LMW glutelins confirmed both the high population polymorphism and the intra-population differences. Apart from the recent research in CWR breeding, Dasypyrum villosum is still an underrepresented species in germplasm collections and an underutilized species in breeding.

Genotypic and Phenotypic Characterization of Two Triticum aestivum L.—Dasypyrum villosum Translocations Lines in the Same Wheat Genetic Background

A wheat 660K chip was used to genotype two wheat-Dasypyrum villosum 6V#4S.6DL and 6V#2S.6AL translocation lines (A303 and B303) and their common wheat recurrent parent Wan7107. The results showed that these three lines have similar characteristics of base composition except for the translocation chromosomes. The alien translocation chromosomes have fewer homozygous and more heterozygous genotypes with more invalid probes. Distributions of SNPs between the translocation lines and Wan7107 were mainly dense on the regions of 6AS or 6DS as expected, but unexpectedly also on near the telomere of 2BS, and some regions of other wheat chromosomes. Meanwhile, the translocation lines A303 and B303 have 99.44% and 98.81% identical genotypes to Wan7107, respectively. Under the same genetic background, A303 and B303 showed different reactions to Blumeria graminis f. sp. tritici (Bgt) strains of powdery mildew. Both translocation lines have higher grain weight and plant height, and B303 has fewer spikelets compared to Wan7107. These results provide us a new insight into the genomic variation between the backcross generation plant and the recurrent parent, which is valuable information for understanding the relationship between wheat and the 6VS chromosome of D. villosum as well as the application potential of the alien chromosome arms.

Molecular analysis of gibberellin receptor gene GID1 in Dasypyrum villosum and development of DNA marker for its identification

Dasypyrum villosum is an annual cereal used as a donor of agronomic traits for wheat. Productivity is one of the most important traits that breeding is aimed at. It is a very complex trait, the formation of which is influenced by many different factors, both internal (the genotype of the plant) and external. The genes responsible for the gibberellin sensitivity played a large role in multiplying yields of cereal crops. Another such gene is the Gid1, which encodes a receptor for gibberellins. This article compares the DNA sequences of the Gid1 gene obtained from six Dasypyrum villosum samples. Using a sequence of wheat and rye taken from the GenBank database (NCBI), we selected primers for regions of different genomes (A, B, and D subgenomes of wheat and the R genome of rye), and carried out a polymerase chain reaction on D. villosum accessions of diverse geographical origin. The resulting PCR product was sequenced by an NGS method. Based on the assembled sequences, DNA markers have been created that make it possible to differentiate these genes of the V genome and homologous genes of wheat origin. Using monosomic addition, substitution, and translocation wheat lines, the localization of the Gid1 gene of D. villosum was established on the long arm of the first V chromosome. A phenotypic assessment of common wheat lines carrying substituted, translocated, or added D. villosum chromosomes in their karyotype was performed. Tendency of disappearance of the first chromosome of D. villosum in the lines with added chromosomes was revealed.

Molecular Characterization of the Dwarf53 Gene Homolog in Dasypyrum Villosum

The Dwarf53 (D53) gene, first studied in rice, encodes a protein that acts as a repressor of the physiological response of plants to strigolactones—substances that regulate the activity of axillary buds, stem growth, branching of roots and other physiological processes. In this work, we isolated and sequenced the homolog of the D53 gene in several accessions of the wild grass Dasypyrum villosum of different geographical origins, resulting in the discovery of large allelic variety. A molecular marker was also created that allows us to differentiate the D. villosum D53 gene from common wheat genes. Using this marker and monosomic addition, substitution and translocation wheat lines carrying the known D. villosum chromosomes, the D53 gene was localized on the long arm of the 5V chromosome.