Efficiency of using electric toothbrush as an alternative to a tuning fork for artificial buzz pollination is independent of instrument buzzing frequency.

BACKGROUND: Breeding programs and research activities where artificial buzz-pollinations are required to have primarily relied upon using tuning forks, and bumble bees. However, these methods can be expensive, unreliable, and inefficient. To find an alternative, we tested the efficiency of pollen collection using electric toothbrushes and compared it with tuning forks at three vibration frequencies-low, medium, and high and two extraction times at 3 s and 16 s- from two buzz-pollinated species (Solanum lycopersicum and Solanum elaeagnifolium). RESULTS: Our results show that species, and extraction time significantly influenced pollen extraction, while there were no significant differences for the different vibration frequencies and more importantly, the use of a toothbrush over tuning fork. More pollen was extracted from S. elaeagnifolium when compared to S. lycopersicum, and at longer buzzing time regardless of the instrument used. CONCLUSIONS: Our results suggest that electric toothbrushes can be a viable and inexpensive alternative to tuning forks, and regardless of the instrument used and buzzing frequency, length of buzzing time is also critical in pollen extraction.

Tobacco Hornworm (Manduca sexta) Oral Secretion Elicits Reactive Oxygen Species in Isolated Tomato Protoplasts.

Plants are under constant attack by a suite of insect herbivores. Over millions of years of coexistence, plants have evolved the ability to sense insect feeding via herbivore-associated elicitors in oral secretions, which can mobilize defense responses. However, herbivore-associated elicitors and the intrinsic downstream modulator of such interactions remain less understood. In this study, we show that tobacco hornworm caterpillar (Manduca sexta) oral secretion (OS) induces reactive oxygen species (ROS) in tomato (Solanum lycopersicum) protoplasts. By using a dye-based ROS imaging approach, our study shows that application of plant-fed (PF) M. sexta OS generates significantly higher ROS while artificial diet-fed (DF) caterpillar OS failed to induce ROS in isolated tomato protoplasts. Elevation in ROS generation was saturated after ~140 s of PF OS application. ROS production was also suppressed in the presence of an antioxidant NAC (N-acetyl-L-cysteine). Interestingly, PF OS-induced ROS increase was abolished in the presence of a Ca2+ chelator, BAPTA-AM (1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid). These results indicate a potential signaling cascade involving herbivore-associated elicitors, Ca2+, and ROS in plants during insect feeding. In summary, our results demonstrate that plants incorporate a variety of independent signals connected with their herbivores to regulate and mount their defense responses.

Bees and thrips carry virus-positive pollen in peach orchards in South Carolina, United States.

Prunus necrotic ringspot virus (PNRSV) and prune dwarf virus (PDV) are pollen-borne viruses of important stone fruit crops, including peaches, which can cause substantial yield loss. Although both horizontal and vertical (i.e., seed) transmission of both viruses occurs through pollen, the role of flower-visiting insects in their transmission is not well understood. Bees and thrips reportedly spread PNRSV and PDV in orchards and greenhouse studies; however, the field spread of PNRSV and PDV in peach orchards in the southeastern United States is not explored. We hypothesized that bees and thrips may facilitate virus spread by carrying virus-positive pollen. Our 2-yr survey results show that 75% of captured bees are carrying virus-positive pollen and moving across the orchard while a subsample of thrips were also found virus positive. Based on morphology, Bombus, Apis, Andrena, Eucera, and Habropoda are the predominant bee genera that were captured in peach orchards. Understanding the role of bees and thrips in the spread of PNRSV and PDV will enhance our understanding of pollen-borne virus ecology.

Examining the Role of Buzzing Time and Acoustics on Pollen Extraction of Solanum elaeagnifolium.

Buzz pollination is a specialized pollination syndrome that requires vibrational energy to extract concealed pollen grains from poricidal anthers. Although a large body of work has examined the ecology of buzz pollination, whether acoustic properties of buzz pollinators affect pollen extraction is less understood, especially in weeds and invasive species. We examined the pollination biology of Silverleaf nightshade (Solanum elaeagnifolium), a worldwide invasive weed, in its native range in the Lower Rio Grande Valley (LRGV) in south Texas. Over two years, we documented the floral visitors on S. elaeagnifolium, their acoustic parameters (buzzing amplitude, frequency, and duration of buzzing) and estimated the effects of the latter two factors on pollen extraction. We found five major bee genera: Exomalopsis, Halictus, Megachile, Bombus, and Xylocopa, as the most common floral visitors on S. elaeagnifolium in the LRGV. Bee genera varied in their duration of total buzzing time, duration of each visit, and mass. While we did not find any significant differences in buzzing frequency among different genera, an artificial pollen collection experiment using an electric toothbrush showed that the amount of pollen extracted is significantly affected by the duration of buzzing. We conclude that regardless of buzzing frequency, buzzing duration is the most critical factor in pollen removal in this species.

Cascading effects of polyphenol-rich purple corn pericarp extract on pupal, adult, and offspring of tobacco hornworm (Manduca sexta L.).

A major bottleneck in the commercialization of plant-based pest management compounds is that the extraction methods are complex, time-consuming, and even highly expensive. Using a recently developed inexpensive extraction and quantification methodology to isolate polyphenols (including anthocyanins and condensed tannins) from purple corn pericarp, we examined their effects on Manduca sexta, a common insect herbivore. Following up on our previous work which demonstrated the negative impacts of polyphenol-rich extract on larval stages, we further examined whether there are any cascading effects on subsequent life stages (pupal and adult) including any possible transgenerational effects. Our results show that polyphenol-rich purple corn extract-fed caterpillars had significantly lower pupal mass and survival. Moreover, adult moths also had lower mass when eclosed from caterpillars reared on the extract diet. To test whether there were any transgenerational effects, we allowed male and female adults fed on purple corn extract diet and control diet to mate and lay eggs in a full factorial experiment. We found that purple corn extract-fed adult pair laid a lower number of eggs compared to other treatments. In addition, we also found that second instar M. sexta caterpillars hatched from eggs laid by any mating combination with at least one parent reared on purple corn extract gained significantly lower mass compared to caterpillars with both parents reared on the control diet. Taken together, our results show that there are cascading negative effects for feeding purple corn pericarp extract on pupal, adult, and second generation of M. sexta, reaffirming its potential application as a cost-effective and environmentally friendly pest deterrent.

Polyphenol-Rich Purple Corn Pericarp Extract Adversely Impacts Herbivore Growth and Development.

Plant secondary metabolites such as terpenes, phenolics, glycosides, and alkaloids play various functional roles including pigmentation, foliar and floral volatile synthesis, hormonal regulation, and direct and indirect defenses. Among these, phenolic compounds are commonly found in plants, but vary in the distribution of their specific compounds among plant families. Polyphenols, including anthocyanins and tannins, are widely distributed and have been well documented for their roles- primarily in plant pigmentation and also in plant defenses. However, commercialization of such compounds for use in insect pest management is severely hampered by expensive, inefficient, and time-consuming extraction protocols. Using a recently developed inexpensive and easy extraction method using the byproducts of pigmented (purple) corn processing, we examined whether the crude pericarp extract rich in polyphenols can affect the growth and development of tobacco hornworm (Manduca sexta L.) caterpillars. Our findings show that purple corn pericarp extract negatively affected M. sexta egg hatching and larval mass gain and prolonged developmental time compared to regular yellow corn extract or an artificial control diet. We also found that these effects were more severe during the early stages of caterpillar development. These results conclusively demonstrate that purple corn pericarp, an inexpensive by-product of the corn milling industry, is a valuable product with excellent potential as an insect antifeedant.