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Accumulation of Alkaloids in Different Tall Fescue KY31 Clones Harboring the Common Toxic IEpichloë coenophiala/I Endophyte under Field Conditions
Tall fescue (Lolium arundinaceum) is a highly adaptable forage, pasture and turf grass that is grown on over 14 M ha in the eastern half of the United States and in other temperate regions of the world. A significant factor in adaptability, productivity and stand persistence is in part due to the presence of an intercellular, seed-transmissible, endophytic fungus, Epichloë coenophiala. Epichloë endophytes have been shown to produce a number of alkaloid compounds only in planta, some that are beneficial in repelling insects, while others are toxic to animals. The goal of this work was to monitor the level of the ergot and loline (classified as pyrrolizidine) alkaloid accumulation in individual plants to determine the plant genotype contribution to alkaloid concentrations. The experimental design consisted of sixteen tall fescue KY31 clones in a space-planted, replicated trial over three years. Our results demonstrated that while changes in the alkaloid concentrations for each plant/endophyte genotype were observed over the three years, the overall alkaloid levels remained relatively constant when compared to other plant/endophyte genotypes combinations in the field. Additionally, overall levels of the ergot and loline alkaloid accumulation did not vary in the same way over the three years. Since the E. coenophiala endophyte genotype was the same across all clones, our results indicate that it is the plant genotype that is responsible for determining alkaloid levels in each plant, and suggest that the signal(s) from the plant to the endophyte may not be the same for ergot and loline alkaloid production.
Journal Article
The ABCs of gene cloning
Clear and concise, this easy-to-use book offers an introductory course on the language of gene cloning, covering microbial, plant, and mammalian systems. It presents the nuts and bolts of gene cloning in a well-organized and accessible manner. Part I of this book outlines the essentials of biology and genetics relevant to the concept of gene cloning. Part II describes common techniques and approaches of gene cloning, ranging from the basic mechanics of DNA manipulation, vector systems, process transformation, to gene analysis. Part III & IV present application technologies of major impact in agriculture, biomedicine, and related areas. The ABCs of Gene Cloning, Third Edition contains updates including a tutorial chapter on gene-vector construction, methodologies on exome sequencing in finding disease genes, revised topics on gene therapy and whole genome sequencing, new developments for gene targeting and genome editing, as well as the current state of next generation sequencing. With more than 140 illustrations, this new edition provides an invaluable text for students and anyone who have interest in gaining proficiency in reading and speaking the language of gene cloning.
Book
Correction: A MultiSite Gateway Toolkit for Rapid Cloning of Vertebrate Expression Constructs with Diverse Research Applications
[This corrects the article DOI: 10.1371/journal.pone.0159277.].
Journal Article
De-extinction : the science of bringing lost species back to life
In the twenty-first century, because of climate change and other human activities, many animal species have become extinct, and many others are at risk of extinction. Once they are gone, we cannot bring them back or can we?
Book
Geno‐ and phenotypic correlates of virologic response to the attachment inhibitor BMS‐626529 in an 8‐day monotherapy study of its prodrug BMS‐663068
Background
Administered as monotherapy for 8 days, BMS‐663068, the prodrug of the attachment inhibitor BMS‐626529, demonstrated significant reductions in plasma HIV‐1 RNA. Although baseline IC50>100 nM to BMS 626529 correlated with a poor virologic response, BMS‐663068 did not appear to select for BMS‐626529 resistance on population sequencing or phenotyping.
Methods
Genotypic population analyses of baseline samples from non‐responders identified amino acid changes that could potentially encode for reduced susceptibility to BMS‐626529. Reverse genetics of functional envelope clones confirmed changes responsible for this in phenotypic assays. Additional genotypic, phenotypic and reverse genetic assays were performed on samples from responders to probe the context dependence of the identified substitutions.
Results
The gp120 M426L substitution was the major change associated with reduced virologic response (present in 5 of 6 non‐responders) and high BMS‐626529 IC50 (present in virus from 6 of 7 subjects with IC50>100 nM). The remaining non‐responder virus sample contained a S375M substitution that encoded reduced susceptibility. However, the M426L substitution was also identified in two responders, one with reduced susceptibility (IC50 6300 nM) and another with low IC50 (38 nM). A series of functional clones from 4 samples (including 2 responders with resistance mutations on population genotyping) were analyzed for susceptibility to BMS‐626529. Variability of susceptibility of clones (37‐246 fold) was higher than variability observed with other entry inhibitors (enfuvirtide, 6‐9 fold; maraviroc, 3‐9 fold). In the responder subject with M246L, all functional clones contained M426L and susceptibility varied by 246‐fold, suggesting that susceptibility is highly context dependent. One of the responders contained viruses of either tropism. Clones of R5‐ or X4‐tropic viruses from this individual exhibited the same variable range of susceptibility to BMS‐626529.
Conclusions
gp120 substitutions M426L and S375M were found to be strongly, albeit not exclusively, associated with low susceptibility to BMS‐626529 and a lack of virologic response to its prodrug, BMS‐663068. Functional clones derived from single individuals exhibited 2‐3 log10 variability in susceptibility to the agent, regardless of tropism, suggesting that susceptibility can be highly context‐dependent.
Journal Article
Scientists are cloning ferrets to try to save the species
Noreen and her sister Antonia are two endangered black footed ferrets that were cloned from cells frozen nearly four decades ago.
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