9 research outputs found
Molecular and Historical Aspects of Corn Belt Dent Diversity
Tens-of-thousands of open-pollinated cultivars of corn (Zea mays L.) are being maintained in germplasm banks. Knowledge of the amount and distribution of genetic variation within and among accessions can aid end users in choosing among them. We estimated molecular genetic variation and looked for influences of pedigree, adaptation, and migration in the genetic makeup of conserved Corn-Belt Dent-related germplasm. Plants sampled from 57 accessions representing Corn-Belt Dents, Northern Flints, Southern Dents, plus 12 public inbreds, were genotyped at 20 simple sequence repeat (SSR) loci. For 47 of the accessions, between 5 and 23 plants per accession were genotyped (mean = 9.3). Mean number of alleles per locus was 6.5 overall, 3.17 within accessions, and 3.20 within pooled inbreds. Mean gene diversity was 0.53 within accessions and 0.61 within pooled inbreds. Open-pollinated accessions showed a tendency toward inbreeding (FIS = 0.09), and 85% of genetic variation was shared among them. A Fitch-Margoliash tree strongly supported the distinctiveness of flint from dent germplasm but did not otherwise reveal evidence of genetic structure. Mantel tests revealed significant correlations between genetic distance and geographical (r = 0.54, P= 0.04) or maturity zone (r = 0.33, P = 0.03) distance only if flint germplasm was included in the analyses. A significant correlation (r = 0.76, P \u3c 0.01) was found between days to pollen shed and maturity zone of accession origin. Pedigree, rather than migration or selection, has most influenced the genetic structure of the extant representatives of the open-pollinated cultivars at these SSR loci
A rapid live-cell ELISA for characterizing antibodies against cell surface antigens of \u3ci\u3eChlamydomonas reinhardtii\u3c/i\u3e and its use in isolating algae from natural environments with related cell wall components
Background: Cell walls are essential for most bacteria, archaea, fungi, algae and land plants to provide shape, structural integrity and protection from numerous biotic and abiotic environmental factors. In the case of eukaryotic algae, relatively little is known of the composition, structure or mechanisms of assembly of cell walls in individual species or between species and how these differences enable algae to inhabit a great diversity of environments. In this paper we describe the use of camelid antibody fragments (VHHs) and a streamlined ELISA assay as powerful new tools for obtaining mono-specific reagents for detecting individual algal cell wall components and for isolating algae that share a particular cell surface component.
Results: To develop new microalgal bioprospecting tools to aid in the search of environmental samples for algae that share similar cell wall and cell surface components, we have produced single-chain camelid antibodies raised against cell surface components of the single-cell alga, Chlamydomonas reinhardtii. We have cloned the variable-region domains (VHHs) from the camelid heavy-chain-only antibodies and overproduced tagged versions of these monoclonal-like antibodies in E. coli. Using these VHHs, we have developed an accurate, facile, low cost ELISA that uses live cells as a source of antigens in their native conformation and that requires less than 90 minutes to perform. This ELISA technique was demonstrated to be as accurate as standard ELISAs that employ proteins from cell lysates and that generally require \u3e24 hours to complete. Among the cloned VHHs, VHH B11, exhibited the highest affinity (EC50 \u3c 1 nM) for the C. reinhardtii cell surface. The live-cell ELISA procedure was employed to detect algae sharing cell surface components with C. reinhardtii in water samples from natural environments. In addition, mCherry-tagged VHH B11 was used along with fluorescence activated cell sorting (FACS) to select individual axenic isolates of presumed wild relatives of C. reinhardtii and other Chlorphyceae from the same environmental samples.
Conclusions: Camelid antibody VHH domains provide a highly specific tool for detection of individual cell wall components of algae and for allowing the selection of algae that share a particular cell surface molecule from diverse ecosystems
Limitations and future perspectives for satellite-based soil carbon monitoring
Soil organic carbon (SOC) plays a crucial role in terrestrial C storage and ecosystem services. Agricultural management practices have the potential to increase C inputs and reduce its losses. However, uniform standard protocols for measuring, monitoring, and assessing changes using remote sensing is lacking for SOC in the scientific literature. In this discussion paper, we present techniques for collecting and analyzing ground samples and employing remote sensing to quantify SOC, along with its limitations and future perspectives. Our analysis identified a number of key limitations to advancing the science for remotely sensed terrestrial C in croplands including i) lack of consensus in sampling depth and density, ii) the absence of a standard (or universally accepted) laboratory procedure and statistical methodology, and iii) lack of details on imagery pre-processing or information on the spectral properties of the targeted soils. Establishing standard protocols for ground-truth data collection and remote sensing approaches, as well as a knowledge of the impacts of diverse soil types, land uses, and landscapes on C assessment, are all required to enhance the accuracy and reliability of future SOC assessments