Pollen-mediated gene flow in managed fields of lowbush blueberry

Pollen-mediated gene flow was analyzed in two managed fields of lowbush blueberry (Vaccinium angustifolium Ait.) in Maine. Seedlings derived from open-pollinated crosses of two mother plants, one from each of the two fields, were genotyped using simple sequence repeat markers. The mother plants, the four to five nearest neighbour plants, and a group of 22–23 plants further away from the site of fruit collection were genotyped as well. The paternity of 70 seedlings produced by the mother plants, 35 from each field, was analyzed using FAMOZ to determine if each seedling was most likely a result of a self-cross, a cross with one of the nearest neighbours, or a cross from outside the nearest neighbourhood. Approximately 91%–97% of the seedlings appeared to result from crosses with plants outside the nearest neighbourhood, whereas 0% appeared to have resulted from self-crosses. This suggests that the primary pollinators of lowbush blueberry, native bees and honey bees, routinely collect and move pollen from distances greater than the adjacent neighbours of the plants receiving the pollen.The accepted manuscript in pdf format is listed with the files at the bottom of this page. The presentation of the authors' names and (or) special characters in the title of the manuscript may differ slightly between what is listed on this page and what is listed in the pdf file of the accepted manuscript; that in the pdf file of the accepted manuscript is what was submitted by the author

Design and sampling plan optimization for RT-qPCR experiments in plants : a case study in blueberry

The qPCR assay has become a routine technology in plant biotechnology and agricultural research. It is unlikely to be technically improved, but there are still challenges which center around minimizing the variability in results and transparency when reporting technical data in support of the conclusions of a study. There are a number of aspects of the pre- and post-assay workflow that contribute to variability of results. Here, through the study of the introduction of error in qPCR measurements at different stages of the workflow, we describe the most important causes of technical variability in a case study using blueberry. In this study, we found that the stage for which increasing the number of replicates would be the most beneficial depends on the tissue used. For example, we would recommend the use of more RT replicates when working with leaf tissue, while the use of more sampling (RNA extraction) replicates would be recommended when working with stems or fruits to obtain the most optimal results. The use of more qPCR replicates provides the least benefit as it is the most reproducible step. By knowing the distribution of error over an entire experiment and the costs at each step, we have developed a script to identify the optimal sampling plan within the limits of a given budget. These findings should help plant scientists improve the design of qPCR experiments and refine their laboratory practices in order to conduct qPCR assays in a more reliable-manner to produce more consistent and reproducible data. [This Document is Protected by copyright and was first published by Frontiers. All rights reserved. it is reproduced with permission.