Holstein Friesian dairy cattle edited for diluted coat color as a potential adaptation to climate change

Background High-producing Holstein Friesian dairy cattle have a characteristic black and white coat, often with large proportions of black. Compared to a light coat color, black absorbs more solar radiation which is a contributing factor to heat stress in cattle. To better adapt dairy cattle to rapidly warming climates, we aimed to lighten their coat color by genome editing. Results Using gRNA/Cas9-mediated editing, we introduced a three bp deletion in the pre-melanosomal protein 17 gene (PMEL) proposed as causative variant for the semi-dominant color dilution phenotype observed in Galloway and Highland cattle. Calves generated from cells with homozygous edits revealed a strong color dilution effect. Instead of the characteristic black and white markings of control calves generated from unedited cells, the edited calves displayed a novel grey and white coat pattern. Conclusion This, for the first time, verified the causative nature of the PMEL mutation for diluting the black coat color in cattle. Although only one of the calves was healthy at birth and later succumbed to a naval infection, the study showed the feasibility of generating such edited animals with the possibility to dissect the effects of the introgressed edit and other interfering allelic variants that might exist in individual cattle and accurately determine the impact of only the three bp change.

Coat Polymorphism in Eurasian Lynx: Adaptation to Environment or Phylogeographic Legacy?

We studied the relationship between the variability and contemporary distribution of pelage phenotypes in one of most widely distributed felid species and an array of environmental and demographic conditions. We collected 672 photographic georeferenced records of the Eurasian lynx throughout Eurasia. We assigned each lynx coat to one of five phenotypes. Then we fitted the coat patterns to different environmental and anthropogenic variables, as well as the effective geographic distances from inferred glacial refugia. A majority of lynx were either of the large spotted (41.5%) or unspotted (uniform, 36.2%) phenotype. The remaining patterns (rosettes, small spots and pseudo-rosettes) were represented in 11.0%, 7.4%, and 3.9% of samples, respectively. Although various environmental variables greatly affected lynx distribution and habitat suitability, it was the effect of least-cost distances from locations of the inferred refugia during the Last Glacial Maximum that explained the distribution of lynx coat patterns the best. Whereas the occurrence of lynx phenotypes with large spots was explained by the proximity to refugia located in the Caucasus/Middle East, the uniform phenotype was associated with refugia in the Far East and Central Asia. Despite the widely accepted hypothesis of adaptive functionality of coat patterns in mammals and exceptionally high phenotypic polymorphism in Eurasian lynx, we did not find well-defined signs of habitat matching in the coat pattern of this species. Instead, we showed how the global patterns of morphological variability in this large mammal and its environmental adaptations may have been shaped by past climatic change.

Towards Forensic DNA Phenotyping for Predicting Visible Traits in Dogs

The popularity of dogs as human companions explains why these pets regularly come into focus in forensic cases such as bite attacks or accidents. Canine evidence, e.g., dog hairs, can also act as a link between the victim and suspect in a crime case due to the close contact between dogs and their owners. In line with human DNA identification, dog individualization from crime scene evidence is mainly based on the analysis of short tandem repeat (STR) markers. However, when the DNA profile does not match a reference, additional information regarding the appearance of the dog may provide substantial intelligence value. Key features of the dog’s appearance, such as the body size and coat colour are well-recognizable and easy to describe even to non-dog experts, including most investigating officers and eyewitnesses. Therefore, it is reasonable to complement eyewitnesses’ testimonies with externally visible traits predicted from associated canine DNA samples. Here, the feasibility and suitability of canine DNA phenotyping is explored from scratch in the form of a proof of concept study. To predict the overall appearance of an unknown dog from its DNA as accurately as possible, the following six traits were chosen: (1) coat colour, (2) coat pattern, (3) coat structure, (4) body size, (5) ear shape, and (6) tail length. A total of 21 genetic markers known for high predicting values for these traits were selected from previously published datasets, comprising 15 SNPs and six INDELS. Three of them belonged to SINE insertions. The experiments were designed in three phases. In the first two stages, the performance of the markers was tested on DNA samples from dogs with well-documented physical characteristics from different breeds. The final blind test, including dogs with initially withheld appearance information, showed that the majority of the selected markers allowed to develop composite sketches, providing a realistic impression of the tested dogs. We regard this study as the first attempt to evaluate the possibilities and limitations of forensic canine DNA phenotyping.

R-locus for roaned coat is associated with a tandem duplication in an intronic region of USH2A in dogs and also contributes to Dalmatian spotting

Structural variations (SVs) represent a large fraction of all genetic diversity, but how this genetic diversity is translated into phenotypic and organismal diversity is unclear. Explosive diversification of dog coat color and patterns after domestication can provide a unique opportunity to explore this question; however, the major obstacle is to efficiently collect a sufficient number of individuals with known phenotypes and genotypes of hundreds of thousands of markers. Using customer-provided information about coat color and patterns of dogs tested on a commercial canine genotyping platform, we identified a genomic region on chromosome 38 that is strongly associated with a mottled coat pattern (roaning) by genome-wide association study. We identified a putative causal variant in this region, an 11-kb tandem duplication (11,131,835–11,143,237) characterized by sequence read coverage and discordant reads of whole-genome sequence data, microarray probe intensity data, and a duplication-specific PCR assay. The tandem duplication is in an intronic region of usherin gene (USH2A), which was perfectly associated with roaning but absent in non-roaned dogs. We detected strong selection signals in this region characterized by reduced nucleotide diversity (π), increased runs of homozygosity, and extended haplotype homozygosity in Wirehaired Pointing Griffons and Australian Cattle Dogs (typically roaned breeds), as well as elevated genetic difference (FST) between Wirehaired Pointing Griffon (roaned) and Labrador Retriever (non-roaned). Surprisingly, all Dalmatians (N = 262) carried the duplication embedded in identical or similar haplotypes with roaned dogs, indicating this region as a shared target of selection during the breed’s formation. We propose that the Dalmatian’s unique spots were a derived coat pattern by establishing a novel epistatic interaction between roaning “R-locus” on chromosome 38 and an uncharacterized modifier locus. These results highlight the utility of consumer-oriented genotype and phenotype data in the discovery of genomic regions contributing to phenotypic diversity in dogs.

How unique is unique? Quantifying geometric differences in stripe patterns of Cape mountain zebra, Equus zebra zebra (Perissodactyla: Equidae)

Quantified coat pattern dissimilarity provides a visible surface for individual animal traceability to populations. We determined the feasibility in quantifying uniqueness of stripe patterns of Cape mountain zebra (CMZ; Equus zebra zebra) using geometric morphometrics. We photogrammetrically created dense surface models of CMZ (N = 56). Stripe edges were landmarked, superimposed and compared for shape variation across replicates and the population. Significant allometry in stripe patterns prompted allometric correction to remove increased curvature of stripes at the rump, belly and back with larger adult individuals, to facilitate equilibrated comparison between individuals. Re-landmarked replicates showed lower dissimilarity (Di) than non-replicates (Dp), representing minimal landmarking error. Individuals were 78.07 ± 1.79% unique (U=1−DiDp×100%) relative to the study population. Size, the number of torso stripes and degree of branching in four rear torso stripes described the most shape variation (36.79%) but a significant portion could only be distinguished with geometric morphometrics (41.82%). This is the first known use of geometric morphometrics to quantify coat pattern uniqueness, using a model species to provide baseline individual morphological variation. Measures of coat pattern similarity have a place in phenotypic monitoring and identification.

Macro- and micro-morphometrical seed characters in seven species of Egyptian Datura and Nicotiana (Solanaceae)

This work describes and compares the external macro- and micro-morphometrical features of the seed of seven species of Datura and Nicotiana belonging to two tribes of the Solanaceae. Fourteen unique characters were measured and described, the results of which reveal marked differences, both among and within genera. The species of Datura exhibited higher values than those of Nicotiana in all morphometrically measured parameters, such as seed weight, size, perimeter, and surface area, with no or little variation in roundness and eccentricity. For qualitative features, the notched outline, rough texture, pitted ornamentation, circular hilum shape, and stellate coat pattern characterized all examined species of Nicotiana. The yellow colour of the seeds can be used to distinguish the seeds of Nicotiana glauca from the brown seeds of other Nicotiana species examined in this study. The circular shape of the hilum distinguishes the seeds of Datura innoxia from those of Datura metel, which have oral-shaped hilum. Considerable differences in seed coat morphology were found both among and within species. However, the size and shape of the seed can be useful and stable characters, and the position of the hilum had some taxonomic significance.

Seed Coat Pattern QTL and Development in Cowpea (Vigna unguiculata [L.] Walp.)

The appearance of the seed is an important aspect of consumer preference for cowpea (Vigna unguiculata [L.] Walp.). Seed coat pattern in cowpea has been a subject of study for over a century. This study makes use of newly available resources, including mapping populations, a reference genome and additional genome assemblies, and a high-density single nucleotide polymorphism genotyping platform, to map various seed coat pattern traits to three loci, concurrent with the Color Factor (C), Watson (W), and Holstein (H) factors identified previously. Several gene models encoding proteins involved in regulating the later stages of the flavonoid biosynthesis pathway have been identified as candidate genes, including a basic helix-loop-helix gene (Vigun07g110700) for the C locus, a WD-repeat gene (Vigun09g139900) for the W locus and an E3 ubiquitin ligase gene (Vigun10g163900) for the H locus. A model of seed coat development, consisting of six distinct stages, is described to explain some of the observed pattern phenotypes.