An efficient pipeline for ancient DNA mapping and recovery of endogenous ancient DNA from whole‐genome sequencing data

Ancient DNA research has developed rapidly over the past few decades due to improvements in PCR and next‐generation sequencing (NGS) technologies, but challenges still exist. One major challenge in relation to ancient DNA research is to recover genuine endogenous ancient DNA sequences from raw sequencing data. This is often difficult due to degradation of ancient DNA and high levels of contamination, especially homologous contamination that has extremely similar genetic background with that of the real ancient DNA. In this study, we collected whole‐genome sequencing (WGS) data from 6 ancient samples to compare different mapping algorithms. To further explore more effective methods to separate endogenous DNA from homologous contaminations, we attempted to recover reads based on ancient DNA specific characteristics of deamination, depurination, and DNA fragmentation with different parameters. We propose a quick and improved pipeline for separating endogenous ancient DNA while simultaneously decreasing homologous contaminations to very low proportions. Our goal in this research was to develop useful recommendations for ancient DNA mapping and for separation of endogenous DNA to facilitate future studies of ancient DNA.     

Desiccation limits recruitment in the pleometrotic desert seed‐harvester ant Veromessor pergandei

The desert harvester ant Veromessor pergandei displays geographic variation in colony founding with queens initiating nests singly (haplometrosis) or in groups (pleometrosis). The transition from haplo‐ to pleometrotic founding is associated with lower rainfall. Numerous hypotheses have been proposed to explain the evolution of cooperative founding in this species, but the ultimate explanation remains unanswered. In laboratory experiments, water level was positively associated with survival, condition, and brood production by single queens. Queen survival also was positively influenced by water level and queen number in a two‐factor experiment. Water level also was a significant effect for three measures of queen condition, but queen number was not significant for any measure. Foundress queens excavated after two weeks of desiccating conditions were dehydrated compared to alate queens captured from their natal colony, indicating that desiccation can be a source of queen mortality. Long‐term monitoring in central Arizona, USA, documented that recruitment only occurred in four of 20 years. A discriminant analysis using rainfall as a predictor of recruitment correctly predicted recruitment in 17 of 20 years for total rainfall from January to June (the period for mating flights and establishment) and in 19 of 20 years for early plus late rainfall (January–March and April–June, respectively), often with a posterior probability > 0.90. Moreover, recruitment occurred only in years in which both early and late rainfall exceeded the long‐term mean. This result also was supported by the discriminant analysis predicting no recruitment when long‐term mean early and late rainfall were included as ungrouped periods. These data suggest that pleometrosis in V. pergandei evolved to enhance colony survival in areas with harsh abiotic (desiccating) conditions, facilitating colonization of habitats in which solitary queens could not establish even in wet years. This favorable‐year hypothesis supports enhanced worker production as the primary advantage of pleometrosis.     

Geographical pattern of genetic diversity in Capsella bursa‐pastoris (Brassicaceae)—A global perspective

We analyzed the global genetic variation pattern of Capsella bursa‐pastoris (Brassicaceae) as expressed in allozymic (within‐locus) diversity and isozymic (between‐locus) diversity. Results are based on a global sampling of more than 20,000 C. bursa‐pastoris individuals randomly taken from 1,469 natural provenances in the native and introduced range, covering a broad spectrum of the species’ geographic distribution. We evaluated data for population genetic parameters and F‐statistics, and Mantel tests and AMOVA were performed. Geographical distribution patterns of alleles and multilocus genotypes are shown in maps and tables. Genetic diversity of introduced populations is only moderately reduced in comparison with native populations. Global population structure was analyzed with structure, and the obtained cluster affiliation was tested independently with classification approaches and macroclimatic data using species distribution modeling. Analyses revealed two main clusters: one distributed predominantly in warm arid to semiarid climate regions and the other predominantly in more temperate humid to semihumid climate regions. We observed admixture between the two lineages predominantly in regions with intermediate humidity in both the native and non‐native ranges. The genetically derived clusters are strongly supported in macroclimatic data space. The worldwide distribution patterns of genetic variation in the range of C. bursa‐pastoris can be explained by intensive intra‐ and intercontinental migration, but environmental filtering due to climate preadaption seems also involved. Multiple independent introductions of genotypes from different source regions are obvious. “Endemic” genotypes might be the outcome of admixture or of de novo mutation. We conclude that today's successfully established Capsella genotypes were preadapted and found matching niche conditions in the colonized range parts.     

Serological and genetic analysis of a rare CisAB01/O01 blood group

The paper aims to analyze a rare blood sample in Ganzhou City Hospital with CisAB subtype and explore a feasible pattern for blood typing of rare blood type patients, so as to ensure clinical transfusion safety. The routine serological methods were used for ABO forward and reverse blood typing and the fluorescence real-time PCR technique was used for sample genotyping. A human ABO blood group 6-7 exon sequencing kit was used for sequence analysis. The nucleic acid sequence of the sample was compared with reference sequences. The forward typing results demonstrated that the sample was ABw, RhD positive. The sample exhibited 4+ agglutination with anti-H and anti-AB antibodies. Reverse typing by microcolumn gel method showed an AB result, but the serum sample demonstrated weak agglutination with B cell under room temperature, 4 °C and 37 °C in saline when tested with tube method respectively. The serological results matched with the A₂B₃ serotype. The fluorescent real-time PCR genotyping results displayed A/O01. The sequence analysis demonstrated deletion of guanine in 261-position 467C>T (heterozygote) and 803G>T (heterozygote) mutation respectively. The mutation caused the A glycosyltransferase peptide chain to change from proline to leucine (P156L) at 156 and from glutamate to alanine (G268A) at 268. The result demonstrated that the sample''s genotype was CisAB01/O01. The mutation of glycosyltransferase coding gene leads to an abnormal serological reaction pattern. Only by combining the results of genetic analysis can we get the true sample blood type and better ensure the safety of clinical blood transfusion.     

Next Generation Semiconductor Based-Sequencing of a Nutrigenetics Target Gene (GPR120) and Association with Growth Rate in Italian Large White Pigs

The GPR120 gene (also known as FFAR4 or O3FAR1) encodes for a functional omega-3 fatty acid receptor/sensor that mediates potent insulin sensitizing effects by repressing macrophage-induced tissue inflammation. For its functional role, GPR120 could be considered a potential target gene in animal nutrigenetics. In this work we resequenced the porcine GPR120 gene by high throughput Ion Torrent semiconductor sequencing of amplified fragments obtained from 8 DNA pools derived, on the whole, from 153 pigs of different breeds/populations (two Italian Large White pools, Italian Duroc, Italian Landrace, Casertana, Pietrain, Meishan, and wild boars). Three single nucleotide polymorphisms (SNPs), two synonymous substitutions and one in the putative 3′-untranslated region (g.114765469C > T), were identified and their allele frequencies were estimated by sequencing reads count. The g.114765469C > T SNP was also genotyped by PCR-RFLP confirming estimated frequency in Italian Large White pools. Then, this SNP was analyzed in two Italian Large White cohorts using a selective genotyping approach based on extreme and divergent pigs for back fat thickness (BFT) estimated breeding value (EBV) and average daily gain (ADG) EBV. Significant differences of allele and genotype frequencies distribution was observed between the extreme ADG-EBV groups (P < 0.001) whereas this marker was not associated with BFT-EBV.     

Restoration and Science: A Practitioner/Scientist's View from Rare Habitat Restoration at a Southern California Preserve

Researchers reexamining the relationship between restoration science and practice report a continuing scientist‐practitioner gap. As a land manager with scientific training, I offer my perspective of the chasm and describe a restoration practice infused with as much science as the realities of limited budget and time allow. The coastal sage scrub (CSS) restoration project at Starr Ranch, a 1,585 ha Audubon preserve in southern California, combines non‐chemical invasive species control, restoration, and applied research. Our practices evolve from modified scientific approaches and the scientific literature. Results from experiments with non‐optimum replication (on effects of seed rates, soil tamping, and timing of planting) nonetheless had value for management decisions. A critical practice came from academic research that encouraged cost‐effective passive restoration. Our passive restoration monitoring data showed 28–100% total native cover after 3–5 years. Another published study found that restoration success in semiarid regions is dependent on rainfall, a finding vital for understanding active restoration monitoring results that showed a range of 0–88% total native cover at the end of the first season. Work progresses through a combination of applied research, a watchful eye on the scientific literature, and “ecological intuition” informed by the scientific literature and our own findings. I suggest that it is less critical for academic scientists to address the basic questions on technique that are helpful to land managers but rather advocate practitioner training in methods to test alternative strategies and long‐term monitoring.     

Spontaneous Return of Biodiversity in Restored Subtropical Thicket: Portulacaria afra as an Ecosystem Engineer

An accepted criterion for measuring the success of ecosystem restoration is the return of biodiversity relative to intact reference ecosystems. The emerging global carbon economy has made landscape‐scale restoration of severely degraded Portulacaria afra (spekboom)‐dominated subtropical thicket, by planting multiple rows of spekboom truncheons, a viable land‐use option. Although large amounts of carbon are sequestered when planting a monoculture of spekboom, it is unknown whether this is associated with the return of other thicket biodiversity components. We used available carbon stock data from degraded, restored, and intact stands at one site, and sampled carbon stocks at restored stands at another site in the same plant community. We also sampled plant community composition at both sites. The total carbon stock of the oldest (50 years) post‐restoration stand (250.8 ± 14 t C ha^?1) approximated that of intact stands (245 t C ha^?1) and we observed a general increase in carbon content with restoration age (71.4 ± 24 t C ha^?1 after 35 and 167.9 ± 20 t C ha^?1 after 50 years). A multiple correspondence analysis separated degraded stands from stands under restoration based on ground cover, floristic composition, and total carbon stock. Older post‐restoration and intact stands were clustered according to woody canopy recruit abundance. Our results suggest that spekboom is an ecosystem engineer that promotes spontaneous return of canopy species and other components of thicket biodiversity. The spekboom canopy creates a cooler micro‐climate and a dense litter layer, both likely to favor the recruitment of other canopy species.     

Reducing Biotic and Abiotic Land‐Use Legacies to Restore Invaded,Abandoned Citrus Groves

Land‐use legacies associated with agriculture, such as increased soil fertility and elevated soil pH, promote invasions by non‐native plant species on former agricultural lands. Restoring natural soil conditions (i.e. low fertility and low pH) may be an effective, long‐term method to control and reduce the abundance of non‐native and ruderal species that invade abandoned agricultural lands. In this study, we examined how soil manipulation treatments of lowering soil fertility with carbon additions and lowering soil pH by applying sulfur affect non‐native and ruderal native plant species abundance in two former citrus groves in central Florida. Non‐native plant biomass was removed by one of two methods (tilling or topsoil removal), and was combined with a soil amendment of sulfur, carbon, sulfur + carbon, or none. The biomass removal treatments significantly decreased non‐native abundance, with topsoil removal as the most effective. Carbon additions did not affect soil fertility or vegetation. Sulfur and sulfur + carbon additions significantly decreased soil pH in both groves for at least 1 year post‐treatment; however, we did not see a significant vegetation response. Overall, our results suggest that removing vegetation by tilling and topsoil removal is an effective method for reducing non‐target species cover. Although we did not see a response of vegetation to our treatments, we were able to restore the initial soil characteristics, which can be a first step toward complete restoration.     

Integration of Biological Control and Native Seeding to Restore Invaded Plant Communities

Disturbed natural areas frequently experience invasion by introduced plant species that can reduce native biodiversity. Biological control can suppress these introduced species, but without restoration another introduced species can invade. Integration of biological control with concurrent revegetation can both aid in weed reduction via interspecific plant competition and establish a restored native plant community. This 3‐year study investigated an integrated approach to controlling the introduced annual Mile‐a‐minute weed (Persicaria perfoliata [L.] H. Gross [Polygonaceae]) using the biocontrol weevil Rhinoncomimus latipes Korotyaev (Coleoptera: Curculionidae) and restoration planting using a native seed mix. A fully factorial design tested weevils and seeding, separately and together, using insecticide to eliminate weevils. The weevils together with the native seed mix reduced P. perfoliata percent cover in 2009 and 2010, and peak seed cluster production in 2010, compared to the insecticide ? no seed control treatment. Persicaria perfoliata final dry biomass was reduced by 75% in 2010 and by 57% in 2011 in the weevils plus seed treatment compared to the control, with weevils having the greatest effect in 2010 and the seed treatment having the greatest impact in 2011. Results suggest an additive effect of biocontrol and seeding in suppressing P. perfoliata. Seeded treatments also developed the highest native plant species richness and diversity, comprised of spontaneous recolonization in addition to species from the seed mix. Results support the use of integrated management of this invasive weed, with suppression through biological control and native revegetation together helping prevent reinvasion while restoring native plant biodiversity.     

Ecosystem Services from Keystone Species: Diversionary Seeding and Seed‐Caching Desert Rodents Can Enhance Indian Ricegrass Seedling Establishment

Seeds of Indian ricegrass (Achnatherum hymenoides), a native bunchgrass common to sandy soils on arid western rangelands, are naturally dispersed by seed‐caching rodent species, particularly Dipodomys spp. (kangaroo rats). These animals cache large quantities of seeds when mature seeds are available on or beneath plants and recover most of their caches for consumption during the remainder of the year. Unrecovered seeds in caches account for the vast majority of Indian ricegrass seedling recruitment. We applied three different densities of white millet (Panicum miliaceum) seeds as “diversionary foods” to plots at three Great Basin study sites in an attempt to reduce rodents' over‐winter cache recovery so that more Indian ricegrass seeds would remain in soil seedbanks and potentially establish new seedlings. One year after diversionary seed application, a moderate level of Indian ricegrass seedling recruitment occurred at two of our study sites in western Nevada, although there was no recruitment at the third site in eastern California. At both Nevada sites, the number of Indian ricegrass seedlings sampled along transects was significantly greater on all plots treated with diversionary seeds than on non‐seeded control plots. However, the density of diversionary seeds applied to plots had a marginally non‐significant effect on seedling recruitment, and it was not correlated with recruitment patterns among plots. Results suggest that application of a diversionary seed type that is preferred by seed‐caching rodents provides a promising passive restoration strategy for target plant species that are dispersed by these rodents.