Characterization of non-specific lipid transfer protein (nsLtp) gene families in the Brassica napus pangenome reveals abundance variation

Background Brassica napus is an important agricultural species, improving stress resistance was one of the main breeding goals at present. Non-specific lipid transfer proteins (nsLTPs) are small, basic proteins which are involved in some biotic or abiotic stress responses. B. napus is susceptible to a variety of fungal diseases, so identify the BnLTPs and their expression in disease responses is very important. The common reference genome of B. napus does not contain all B. napus genes because of gene presence/absence variations between individuals. Therefore, it was necessary to search for candidate BnLTP genes in the B. napus pangenome. Results In the present study, the BnLTP genes were identified throughout the pangenome, and different BnLTP genes were presented among varieties. Totally, 246 BnLTP genes were identified and could be divided into five types (1, 2, C, D, and G). The classification, phylogenetic reconstruction, chromosome distribution, functional annotation, and gene expression were analyzed. We also identified potential cis-elements that respond to biotic and abiotic stresses in the 2 kb upstream regions of all BnLTP genes. RNA sequencing analysis showed that the BnLTP genes were involved in the response to Sclerotinia sclerotiorum infection. We identified 32 BnLTPs linked to blackleg resistance quantitative trait locus (QTL). Conclusion The identification and analysis of LTP genes in the B. napus pangenome could help to elucidate the function of BnLTP family members and provide new information for future molecular breeding in B. napus.

Associations of Natural Variation in the CD163 and Other Candidate Genes on Host Response of Nursery Pigs to PRRSV Infection

Pigs with complete resistance to porcine reproductive and respiratory syndrome (PRRS) virus (PRRSV) have been produced by genetically knocking out the CD163 gene, which encodes a receptor of the PRRSV for entry into macrophages. The objectives of this study were to evaluate associations of naturally occurring SNPs in the CD163 gene and in three other candidate genes (CD169, RGS16, and TRAF1) with host response to PRRSV-only infection and to PRRS vaccination and PRRSV/porcine circovirus 2b (PCV2b) co-infection. SNPs in the CD163 gene were not included on SNP genotyping panels that were used for previous genome-wide association analyses of these data. An additional objective was to identify the potential genetic interaction of variants at these four candidate genes with a mutation in the GBP5 gene that was previously identified to be associated with host response to PRRSV infection. Finally, the association of SNPs with expression level of the nearby gene was tested. Several SNPs in the CD163, CD169, and RGS16 genes were significantly associated with host response under PRRSV-only and/or PRRSV/PCV2b co-infection. The effect of all SNPs that were significant in the PRRSV-only infection trials depended on genetic background. The effects of some SNPs in the CD163, CD169, and RGS16 genes depended on genotype at the putative causative mutation in the GBP5 gene, which indicates a potential biological interaction of these genes with GBP5. In addition, genome-wide association results for the PRRSV-only infection trials revealed that SNPs located in the CDK5RAP2 or MEGF9 genes, near the TRAF1 gene, had suggestive effects on PRRS viral load, which indicates that these SNPs might contribute to PRRSV neuropathogenesis. In conclusion, natural genetic variants in the CD163, CD169, and RGS16 genes are associated with resistance to PRRSV and/or PCV2b infection and appear to interact with the resistance quantitative trait locus in the GBP5 gene. The identified SNPs can be used to select for increased natural resistance to PRRSV and/or PRRSV-PCV2b co-infection.

Pyramiding of Fusarium Head Blight Resistance Quantitative Trait Loci, Fhb1, Fhb4, and Fhb5, in Modern Chinese Wheat Cultivars

Wheat production is increasingly threatened by the fungal disease, Fusarium head blight (FHB), caused by Fusarium spp. The introduction of resistant varieties is considered to be an effective measure for containment of this disease. Mapping of FHB-resistance quantitative trait locus (QTL) has promoted marker-assisted breeding for FHB resistance, which has been difficult through traditional breeding due to paucity of resistance genes and quantitative nature of the resistance. The lab of Ma previously cloned Fhb1, which inhibits FHB spread within spikes, and fine mapped Fhb4 and Fhb5, which condition resistance to initial infection of Fusarium spp., from FHB-resistant indigenous line Wangshuibai (WSB). In this study, these three QTLs were simultaneously introduced into five modern Chinese wheat cultivars or lines with different ecological adaptations through marker-assisted backcross in early generations. A total of 14 introgression lines were obtained. All these lines showed significantly improved resistance to the fungal infection and disease spread in 2-year field trials after artificial inoculation. In comparison with the respective recipient lines, the Fhb1, Fhb4, and Fhb5 pyramiding could reduce the disease severity by 95% and did not systematically affect plant height, productive tiller number, kernel number per spike, thousand grain weight, flowering time, and unit yield (without Fusarium inoculation). These results indicated the great value of FHB-resistance QTLs Fhb1, Fhb4, and Fhb5 derived from WSB, and the feasibility and effectiveness of early generation selection for FHB resistance solely based on linked molecular markers.

You Had Me at “MAGIC”!: Four Barley MAGIC Populations Reveal Novel Resistance QTL for Powdery Mildew

Blumeria graminis f. sp. hordei (Bgh), the causal agent of barley powdery mildew (PM), is one of the most important barley leaf diseases and is prevalent in most barley growing regions. Infection decreases grain quality and yields on average by 30%. Multi-parent advanced generation inter-cross (MAGIC) populations combine the advantages of bi-parental and association panels and offer the opportunity to incorporate exotic alleles into adapted material. Here, four barley MAGIC populations consisting of six to eight founders were tested for PM resistance in field trials in Denmark. Principle component and STRUCTURE analysis showed the populations were unstructured and genome-wide linkage disequilibrium (LD) decay varied between 14 and 38 Mbp. Genome-wide association studies (GWAS) identified 11 regions associated with PM resistance located on chromosomes 1H, 2H, 3H, 4H, 5H and 7H, of which three regions are putatively novel resistance quantitative trait locus/loci (QTL). For all regions high-confidence candidate genes were identified that are predicted to be involved in pathogen defense. Haplotype analysis of the significant SNPs revealed new allele combinations not present in the founders and associated with high resistance levels.

High-density genetic linkage map construction and white rot resistance QTL mapping for Vitis based on restriction site-associated DNA sequencing

Grape white rot (Coniothyrium diplodiella) is a major fungal disease affecting grape yield and quality. The present study aimed to provide a foundation to improve future grape white rot resistance breeding. To this end, interspecific hybridization was conducted between a white rot resistance cultivar ‘Zhuosexiang’ (Vitus vinifera L. × Vitus labrusca L.), and a susceptible cultivar ‘Victoria’ (Vitus vinifera L.), using 177 hybrid offspring and two parents for restriction site-associated DNA sequencing (RAD-seq) and high-density genetic linkage mapping. Female, male, and integrated map marker numbers were 2501, 4110, and 6249, respectively. The average genetic distance of adjacent markers was 1.25 cM, 0.77 cM, and 0.50 cM. White rot resistance identification of the two parents and 177 individuals was conducted in July and August of 2017 and 2018. Additionally, white rot resistance quantitative trait locus (QTL) mapping was conducted. In total, nine QTLs were detected and located on linkage groups LG1, LG4, LG7, LG12, LG14, and LG15, with overlapping QTLs on LG7 and LG15. Further, three candidate genes that may be responsible for grape white rot resistance were screened. The results will provide an important theoretical reference for future grape white rot resistance breeding.

A Recessive Resistance to Rice yellow mottle virus Is Associated with a Rice Homolog of the CPR5 Gene, a Regulator of Active Defense Mechanisms

RYMV2 is a major recessive resistance gene identified in cultivated African rice (Oryza glaberrima) which confers high resistance to the Rice yellow mottle virus (RYMV). We mapped RYMV2 in an approximately 30-kb interval in which four genes have been annotated. Sequencing of the candidate region in the resistant Tog7291 accession revealed a single mutation affecting a predicted gene, as compared with the RYMV-susceptible O. glaberrima CG14 reference sequence. This mutation was found to be a one-base deletion leading to a truncated and probably nonfunctional protein. It affected a gene homologous to the Arabidopsis thaliana CPR5 gene, known to be a defense mechanism regulator. Only seven O. glaberrima accessions showing this deletion were identified in a collection consisting of 417 accessions from three rice species. All seven accessions were resistant to RYMV, which is an additional argument in favor of the involvement of the deletion in resistance. In addition, fine mapping of a resistance quantitative trait locus in O. sativa advanced backcrossed lines pinpointed a 151-kb interval containing RYMV2, suggesting that allelic variants of the same gene may control both high and partial resistance.

Generation and Screening of a BAC Library from a Diploid Potato Clone to Unravel Durable Late Blight Resistance on Linkage Group IV

We describe the construction and screening of a large insert genomic library from the diploid potato clone HB171(13) that has been shown to express durable quantitative field resistance to Phytophthora infestans, the causal agent of potato late blight disease. Integrated genetic mapping of the field resistance quantitative trait locus with markers developed from populations segregating for Rpi-blb3, Rpi-abpt, R2, and R2-like resistance, all located on linkage group IV, has positioned the field resistance QTL within the proximity of this R gene cluster. The library has been successfully screened with resistance gene analogues (RGA) potentially linked to the R gene cluster. Over 30 positive BAC clones were identified and confirmed by PCR and Southern hybridisations to harbour RGA-like sequences. In addition, BAC end sequencing of positive clones has corroborated two BAC clones with a very high level of nucleotide similarity to the RGA probes utilised.