Where did the northern peatland species survive the dry glacials: cloudberry (Rubus chamaemorus) as an example

Aim During the last ice age large parts of the north boreal and subarctic zones were covered by ice, while the climate in ice‐free regions of northern Asia was extremely cold and dry. The extensive peatlands of these zones with their characteristic vegetation developed at the beginning of the Holocene. We combine a phylogeographical approach with maps of pollen records to identify regions where Rubus chamaemorus, a plant of moist, peaty soils, was likely to grow during this period. Location Circumarctic/circumboreal. Methods Samples were collected from 45 locations throughout much of the range of R. chamaemorus and 398 plants were analysed with amplified fragment length polymorphism (AFLP). Estimates of diversity and differentiation, principal coordinates analysis and Bayesian clustering methods were used for the analysis of genetic data. Dated pollen records were retrieved from the European and the Global Pollen Databases. Results The plants from Sakhalin are highly divergent from the rest of the material and represent the previously described var. pseudochamaemorus. The main genetic division in R. chamaemorus sensu stricto is found in the Taymyr region in central Eurasia. Genetic diversity and the relative number of rare markers are highest in central Siberia and eastern Asia and decrease towards Europe and to a lesser extent eastwards through North America. Pollen dating back to the last ice age is found in central and eastern Siberia and Alaska. The maximum observed clone size is about 250 m, and more than one clone is found in nearly every local population. Main conclusions The genetic data are consistent with the pollen records and indicate that R. chamaemorus was growing in several areas of northern central Siberia and Beringia during the last glaciation. This finding suggests that sufficient humidity for this and other species of peaty soils was present locally in different parts of the generally dry ice‐free areas of northern Asia, as had been previously documented for Beringia. The AFLP data show that var. pseudochamaemorus, which is also morphologically quite divergent, clearly represents a distinct genetic entity.     

The “humanized” N-glycosylation pathway in CRISPR/Cas9-edited Nicotiana benthamiana significantly enhances the immunogenicity of a S/preS1 Hepatitis B Virus antigen and the virus-neutralizing antibody response in vaccinated mice

The recent SARS-CoV-2 pandemic has taught the world a costly lesson about the devastating consequences of viral disease outbreaks but also, the remarkable impact of vaccination in limiting life and economic losses. Vaccination against human Hepatitis B Virus (HBV), a major human pathogen affecting 290 million people worldwide, remains a key action towards viral hepatitis elimination by 2030. To meet this goal, the development of improved HBV antigens is critical to overcome non-responsiveness to standard vaccines based on the yeast-produced, small (S) envelope protein. We have recently shown that combining relevant immunogenic determinants of S and large (L) HBV proteins in chimeric antigens markedly enhances the anti-HBV immune response. However, the demand for cost-efficient, high-quality antigens remains challenging. This issue could be addressed by using plants as versatile and rapidly scalable protein production platforms. Moreover, the recent generation of plants lacking β-1,2-xylosyltransferase and α-1,3-fucosyltransferase activities (FX-KO), by CRISPR/Cas9 genome editing, enables production of proteins with “humanized” N-glycosylation. In this study, we investigated the impact of plant N-glycosylation on the immunogenic properties of a chimeric HBV S/L vaccine candidate produced in wild-type and FX-KO Nicotiana benthamiana. Prevention of β-1,2-xylose and α-1,3-fucose attachment to the HBV antigen significantly increased the immune response in mice, as compared with the wild-type plant-produced counterpart. Notably, the antibodies triggered by the FX-KO-made antigen neutralized more efficiently both wild-type HBV and a clinically relevant vaccine escape mutant. Our study validates in premiere the glyco-engineered Nicotiana benthamiana as a substantially improved host for plant production of glycoprotein vaccines.