Coilin mediates m6A RNA methylation through phosphorylation of METTL3.

MicroRNAs (miRNAs) are a class of noncoding RNAs that regulate gene expression. An important step in miRNA biogenesis occurs when primary miRNAs are bound and cleaved by the microprocessor to generate precursor miRNAs. Regulation at this step is essential and one such regulator includes m6A RNA methylation, an RNA modification found on primary miRNAs that is installed by METTL3 and bound by hnRNPA2B1. Our lab has recently discovered that the Cajal body marker protein coilin also participates in miRNA biogenesis and hypothesized that coilin may be influencing miRNA biogenesis through m6A RNA methylation. Here we report that coilin suppression reduces m6A on primary Let7a and miR-21. We also found that coilin suppression reduced the protein expression of hnRNPA2B1 and METTL3. We observed an interaction between coilin and ectopically expressed METTL3 and found that coilin suppression reduced the nucleoplasmic portion of METTL3 and blunted ectopic METTL3 phosphorylation. Finally, coilin suppression disrupted the greater METTL3 complex with cofactors METTL14 and WTAP. Collectively, our work has uncovered a role for coilin in mediating m6A RNA methylation and provides an avenue by which coilin participates in miRNA biogenesis.

Effect of Corolla Slitting and Nectar Robbery by the Eastern Carpenter Bee (Hymenoptera: Apidae) on Fruit Quality of Vaccinium corymbosum L. (Ericales: Ericaceae).

Eastern carpenter bees, Xylocopa virginica (L.) (Hymenoptera: Apidae), are among the most abundant native bee visitors to highbush blueberry, Vaccinium corymbosum L., flowers in the northeastern United States, and they sometimes display corolla-slitting behavior to rob nectar. We studied foraging behavior of X. virginica on 14 blueberry cultivars in an experimental planting in Rhode Island, and assessed factors related to slitting frequency, and the effects of slitting on fruit set and blueberry quality. Among 14 cultivars in bloom, an average of 35% (range 16-67%) of flowers were slit in 2017, and 39% (range 20-62%) in 2018. Factors that affected the proportion of corollas slit included cultivar, anther length, flower volume, and number of days in bloom at or above 15°C. Corolla slitting did not affect fruit set. Average weight and percent soluble solids of fruit resulting from slit and non-slit corollas did not differ significantly in two early- ('Bluehaven', 'Earliblue'), two mid- ('Collins', 'Bluecrop'), and two late-season ('Herbert', 'Lateblue') ripening cultivars in 2017. In 2018, average fruit weight and percent soluble solids resulting from slit and non-slit flowers did not differ significantly in most cultivars, but slit corollas resulted in berries with greater mass in two cultivars, 'Bluehaven' and 'Collins'. 'Collins' fruit from non-slit corollas had a significantly higher percentage of soluble solids at maturity than fruit from slit corollas in 2018. Corolla slitting and nectar robbery by X. virginica did not have a significant negative effect on fruit quality under the described growing conditions and pollinator community.

Eastern Carpenter Bee (Hymenoptera: Apidae): Nest Structure, Nest Cell Provisions, and Trap Nest Acceptance in Rhode Island.

Analysis of pollen provisions in Xylocopa virginica (L.) nests in southern Rhode Island showed that this species produced pollen loaves from 21 different genera of plants in 2016, 19 in 2017, and 39 in 2018. Antirrhinium majus L. (garden snapdragon) pollen was the most common type collected in all three years (21.4%). Overall, wind-pollinated tree pollen comprised 22.1% of all pollen loaves. Blueberry pollen was a minor component of pollen loaves (0.1%), despite abundant blueberry plants nearby. Mean values of X. virginica nest measurements (tunnel length 15.4 ± 1.2 cm, width 15.0 ± 0.5 mm, and cell length 17.7 ± 0.3 mm) were similar to those reported in previous studies. Only 2 of the 216 trap nests deployed in 2017 were occupied by 11 X. virginica bees (9 females and 2 males). However, 17 nests contained 230 Osmia taurus Smith, 6 nests contained 73 O. cornifrons (Radoszkowski), and 1 nest contained 8 O. lignaria Say. Thirty-four nests (15.7%) were occupied by 151 grass-carrying wasps, Isodontia sp. and 6 vespid wasps occupied three nests (1.4%) in 2017. In 2018, 4 of 96 trap nests were occupied by carpenter bees. Understanding the nesting and foraging habits of X. virginica will help us to manage natural populations for pollination services.

Analysis of vertebrate vision in a 384-well imaging system.

Visual impairment affects 253 million people worldwide and new approaches for prevention and treatment are urgently needed. While small molecules with potential beneficial effects can be examined in various model systems, the in vivo evaluation of visual function remains a challenge. The current study introduces a novel imaging system for measuring visually-guided behaviors in larval zebrafish. The imaging system is the first to image four 96-well plates with a single camera for automated measurements of activity in a 384-well format. In addition, it is the first system to project moving visual stimuli and analyze the optomotor response in the wells of a 96-well plate. We found that activity is affected by tricaine, diazepam and flumazenil. Surprisingly, diazepam treatments induce a loss of visual responses, at concentrations that do not affect activity or induce hyperactivity. Overall, our studies show that the developed imaging system is suitable for automated measurements of vertebrate vision in a high-throughput format.