Effects of nitrogen addition and mowing on nitrogen- and water-use efficiency of Artemisia frigida in a grassland restored from an abandoned cropland

氮添加和刈割对内蒙古弃耕草地冷蒿氮和水分利用效率的影响在氮和水分限制的区域,植物氮利用效率(NUE)和水分利用效率(WUE)决定了它们在群落中的竞争优势。冷蒿(Artemisia frigida)是半干旱草地重度退化的先锋物种,在不同退化程度的草地中具有不 同的优势度,经常被认为是退化草地群落演替的指示物种。退化草地恢复过程中,氮添加和割草如何影响冷蒿的NUE和WUE尚不清晰。以内蒙古多伦县弃耕草地为研究对象,选取两个不同群落斑块(禾草和冷蒿为优势物种的斑块),经过长期(2006–2013)氮添加和刈割(对照、氮添加、刈割、氮添加+刈割)处理后,研究冷蒿的NUE (叶片碳氮比)和WUE (叶片碳同位素,δ¹³C)对氮添加、刈割及其交互作用的响应; 结合植物和土壤的碳、氮同位素(δ¹³C和δ¹⁵N)及碳、氮库探究退化草地恢复过程中植物的资源利用策略及其机制。研究结果表明：(1)氮添加对冷蒿的WUE没有显著影响(P > 0.05),但NUE 在禾草和冷蒿斑块 中分别显著降低了42.9%和26.6% (P < 0.05);(2)植物对不同氮源(NH₄₊或NO_3-)的利用会引起植物和土壤δ¹⁵N的分馏,研究表明叶片和土壤的δ¹⁵N与NUE呈现相反的变化趋势,因此冷蒿的NUE对氮添加的响应与不同氮源的利用有关;(3)刈割不影响冷蒿的NUE (P > 0.05),但在禾草斑块,冷蒿的WUE在刈割处理下显著提高了2.3% (P < 0.05);(4)在禾草斑块,氮添加减缓了割草对冷蒿WUE的促进作用;(5)结构方程模型显示,土壤含水量直接或间接的调控着冷蒿的WUE和NUE。综上所述,在禾草斑块,氮添加+刈割处理维持较低的NUE和WUE,不利于冷蒿对资源的竞争,进一步降低其优势度,这也预示着氮添加+ 刈割处理会促进退化草地的恢复。     

Toolboxes for cyanobacteria: Recent advances and future direction

Photosynthetic cyanobacteria are important primary producers and model organisms for studying photosynthesis and elements cycling on earth. Due to the ability to absorb sunlight and utilize carbon dioxide, cyanobacteria have also been proposed as renewable chassis for carbon-neutral “microbial cell factories”. Recent progresses on cyanobacterial synthetic biology have led to the successful production of more than two dozen of fuels and fine chemicals directly from CO₂, demonstrating their potential for scale-up application in the future. However, compared with popular heterotrophic chassis like Escherichia coli and Saccharomyces cerevisiae, where abundant genetic tools are available for manipulations at levels from single gene, pathway to whole genome, limited genetic tools are accessible to cyanobacteria. Consequently, this significant technical hurdle restricts both the basic biological researches and further development and application of these renewable systems. Though still lagging the heterotrophic chassis, the vital roles of genetic tools in tuning of gene expression, carbon flux re-direction as well as genome-wide manipulations have been increasingly recognized in cyanobacteria. In recent years, significant progresses on developing and introducing new and efficient genetic tools have been made for cyanobacteria, including promoters, riboswitches, ribosome binding site engineering, clustered regularly interspaced short palindromic repeats/CRISPR-associated nuclease (CRISPR/Cas) systems, small RNA regulatory tools and genome-scale modeling strategies. In this review, we critically summarize recent advances on development and applications as well as technical limitations and future directions of the genetic tools in cyanobacteria. In addition, toolboxes feasible for using in large-scale cultivation are also briefly discussed.     

The comparison of genetic variation in the envelope protein between various immunodeficiency viruses and equine infectious anemia virus

The envelope protein (Env) of lentiviruses such as HIV, SIV, FIV and EIAV is larger than that of other retroviruses. The Chinese EIAV attenuated vaccine is based on Env and has helped to successfully control this virus, demonstrating that envelope is crucial for vaccine. We compared Env variation of the four kinds of lentiviruses. Phylogenetic analysis showed that the evolutionary relationship of Env between HIV and SIV was the closest and they appeared to descend from a common ancestor, and the relationship of HIV and EIAV was the furthest. EIAV had the shortest Env length and the least number of potential N-linked glycosylation sites (PNGS) as well as glycosylation density compared to various immunodeficiency viruses. However, HIV had the longest Env length and the most PNGS. Moreover, the alignment of HIV and SIV showed that PNGS were primarily distributed within extracellular membrane protein gp120 rather than transmembrane gp41. It implies that the size difference among these viruses is associated with a lentivirus specific function and also the diversity of env. There are low levels of modification of glycosylation sites of Env and selection of optimal protective epitopes might be useful for development of an effective vaccine against HIV/AIDS.     

Whole-genome synthesis and characterization of viable S13-like bacteriophages

Background

Unprecedented progresses in high-throughput DNA sequencing and de novo gene synthesis technologies have allowed us to create living organisms in the absence of natural template.

Methodology/Principal Findings

The sequence of wild-type S13 phage genome was downloaded from GenBank. Two synonymous mutations were introduced into wt-S13 genome to generate m1-S13 genome. Another mutant, m2-S13 genome, was obtained by engineering two nonsynonymous mutations in the capsid protein coding region of wt-S13 genome. A chimeric phage genome was designed by replacing the F capsid protein open reading frame (ORF) from phage S13 with the F capsid protein ORF from phage G4. The whole genomes of all four phages were assembled from a series of chemically synthesized short overlapping oligonucleotides. The linear synthesized genomes were circularized and electroporated into E.coli C, the standard laboratory host of S13 phage. All four phages were recovered and plaques were visualized. The results of sequencing showed the accuracy of these synthetic genomes. The synthetic phages were capable of lysing their bacterial host and tolerating general environmental conditions. While no phenotypic differences among the variant strains were observed when grown in LB medium with CaCl₂, the S13/G4 chimera was found to be much more sensitive to the absence of calcium and to have a lower adsorption rate under calcium free condition.

Conclusions/Significance

The bacteriophage S13 and its variants can be chemically synthesized. The major capsid gene of phage G4 is functional in the phage S13 life cycle. These results support an evolutional hypothesis which has been proposed that a homologous recombination event involving gene F of quite divergent ancestral lineages should be included in the history of the microvirid family.     

Involvement of the skeletal Renin-Angiotensin system in age-related osteoporosis of ageing mice

The local tissue-specific renin-angiotensin system (RAS) was identified. The aim of this study was to investigate the role of local bone RAS in the osteoporosis of aging mice. Twelve-month-old and two-month-old male mice were respectively assigned to the ageing and young groups. The tibias and femurs were collected for an analysis of histomorphology, bone mass, and gene and protein expression. H&E staining and micro-CT measurement showed a loss of the trabecular bone network and decrease of bone mineral density in the proximal tibial metaphysis of the aged mice. The PCR results indicated the significant up-regulation of renin and angiotensinogen (AGT) mRNA expression in both the tibia and femur of the ageing mice. Western blotting data showed that the tibial angiotensin II protein expression was significantly increased in the ageing group. The enhancement of renin and AGT expression in the bone tissue resulted in the increased production of angiotensin II which plays an important role in the pathology of age-related osteoporosis.     

Validating thermal inactivation of Salmonella spp. in fresh and aged chicken litter

Our results revealed that a 7-log reduction of Salmonella can be achieved by exposing fresh chicken litter for 80.5 to 100.8, 78.4 to 93.1, and 44.1 to 63 min at 70, 75, and 80°C, respectively, depending on initial moisture contents. However, the aged chicken litter requires more heat treatment.     

Mitochondrial transfer from aged adipose‐derived stem cells does not improve the quality of aged oocytes in C57BL/6 mice

Female fertility declines dramatically over the age of 35 due to age‐related decreases in oocyte quality and quantity. Although mitochondrial transfer promises to be a technology that can improve the quality of such age‐impaired oocytes, the ideal mitochondrial donor remains elusive. In the present study, we aimed to identify whether aged adipose‐derived stem cells constitute an excellent mitochondrial donor that would improve the quality of aged mouse oocytes. We showed that aging significantly impaired the mitochondrial function in mouse oocytes, but did not significantly affect the mitochondrial function of adipose‐derived stem cells. However, the mitochondrial transfer from aged adipose‐derived stem cells did not mitigate the poor fertilization and embryonic development rates of aged oocytes.