Inbreeding depression is well documented in flowering plants and adversely affects a wide range of fitness-related traits. Recent work has begun to explore the effects of inbreeding on ecological interactions among plants and other organisms, including insect herbivores and pathogens. However, the effects of inbreeding on floral traits, floral scents, and pollinator visitation are less well studied. METHODS: Using inbred and outbred maternal families of horsenettle (Solanum carolinense, Solanaceae), we examined the effects of inbreeding on traits associated with pollinator attraction and floral rewards. Specifically, we measured corolla size, counted pollen grains per flower, and analyzed floral volatile emissions via gas chromatography and mass spectrometry. We also examined pollinator visitation to experimental arrays of flowering inbred and outbred plants under field conditions. RESULTS: Compared to those of outbred plants, flowers of inbred plants exhibited reduced corolla size and pollen production, as well as significantly reduced emission of the two most abundant volatile compounds in the floral blend. Furthermore, bumblebees-the main pollinators of horsenettle-discriminated against inbred flowers in the field: bees were more likely to make initial visits to flowers on outbred plants, visited outbred flowers more often overall, and spent more time on outbred flowers. CONCLUSIONS: These results show that inbreeding can (1) alter floral traits that are known to mediate pollinator attraction; (2) reduce the production of floral rewards (pollen is the sole reward in horsenettle); and (3) adversely affect pollinator visitation under field conditions.

Premise Plant maternal effects on offspring phenotypes are well documented. However, little is known about how herbivory on maternal plants affects offspring fitness. Furthermore, while inbreeding is known to reduce plant reproductive output, previous studies have not explored whether and how such effects may extend across generations. Here, we addressed the transgenerational consequences of herbivory and maternal plant inbreeding on the reproduction of Solanum carolinense offspring. Methods Manduca sexta caterpillars were used to inflict weekly damage on inbred and outbred S. carolinense maternal plants. Cross‐pollinations were performed by hand to produce seed from herbivore‐damaged outbred plants, herbivore‐damaged inbred plants, undamaged outbred plants, and undamaged inbred plants. The resulting seeds were grown in the greenhouse to assess emergence rate and flower production in the absence of herbivores. We also grew offspring in the field to examine reproductive output under natural conditions. Results We found transgenerational effects of herbivory and maternal plant inbreeding on seedling emergence and reproductive output. Offspring of herbivore‐damaged plants had greater emergence, flowered earlier, and produced more flowers and seeds than offspring of undamaged plants. Offspring of outbred maternal plants also had greater seedling emergence and reproductive output than offspring of inbred maternal plants, even though all offspring were outbred. Moreover, the effects of maternal plant inbreeding were more severe when plant offspring were grown in field conditions. Conclusions This study demonstrates that both herbivory and inbreeding have fitness consequences that extend across generations even in outbred progeny.

Background Transfer RNA (tRNA) gene predictions are complicated by challenges such as structural variation, limited sequence conservation and the presence of highly reiterated short interspersed sequences (SINEs) that originally derived from tRNA genes or tRNA-like transcription units. Annotation of “tRNA genes” in sequenced genomes generally have not been accompanied by experimental verification of the expression status of predicted sequences. Results To address this for mouse tRNA genes, we have employed two programs, tRNAScan-SE and ARAGORN, to predict the tRNA genes in the nuclear genome, resulting in diverse but overlapping predicted gene sets. From these, we removed known SINE repeats and sorted the genes into predicted families and single-copy genes. In particular, four families of intron-containing tRNA genes were predicted for the first time in mouse, with introns in positions and structures similar to the well characterized intron-containing tRNA genes in yeast. We verified the expression of the predicted tRNA genes by microarray analysis. We then confirmed the expression of appropriately sized RNA for the four intron-containing tRNA gene families, as well as the other 30 tRNA gene families creating an index of expression-verified mouse tRNAs. Conclusions These confirmed tRNA genes represent all anticodons and all known mammalian tRNA structural groups, as well as a variety of predicted “rogue” tRNA genes within families with altered anticodon identities.

Resource sharing within clonal plant networks can occur via the translocation of water, nutrients, and photoassimilates through rhizomes and stolons. Similar mechanisms may mediate the sharing of information (e.g., about herbivory or other environmental stressors) among ramets via molecular or biochemical signals. The storage of such information in belowground structures could facilitate the transmission of appropriate phenotypic responses across growing seasons in perennial species. However, few previous studies have explored the potential transfer of ecologically relevant information within such networks. This study addresses the effects of foliar herbivory and belowground overwintering on the growth and flowering, physical defenses, and herbivore resistance in the clonally spreading species Solanum carolinense L. (Solanaceae). We used rhizomes from inbred and outbred plants that were repeatedly exposed to feeding damage by Manduca sexta L. (Sphingidae) caterpillars and rhizomes from undamaged control plants. These rhizomes were either planted immediately or exposed to overwintering conditions and allowed to produce new ramets (rhizomatous offshoots). We then assessed offshoot emergence, flowering, physical defense traits, and herbivore performance. Relative to controls, offshoots of herbivore-damaged plants exhibited greater spine and trichome densities, and reduced performance of M. sexta larvae. However, they also emerged and flowered significantly later, and produced fewer flowers than offshoots of undamaged plants. Inbreeding also negatively affected offshoot emergence, flowering, trichome production, and herbivore resistance. These effects of parental herbivory were more pronounced in outbred offshoots, indicating that inbreeding may compromise the trans-seasonal induction of plant defenses. Finally, exposure to overwintering conditions increased trichome production and reduced caterpillar performance on offshoots. Together, these results show that induced defenses can be transmitted through rhizomes and affect offshoot growth, flowering, defensive traits, and herbivore resistance. They also document fitness-related costs associated with defense induction in offshoots and suggest that the transfer of defenses across seasons can be compromised by inbreeding.

Virus infection can increase drought tolerance of infected plants compared to non-infected plants; however, the mechanisms mediating virus-induced drought tolerance remain unclear. In this study, we demonstrate turnip mosaic virus (TuMV) infection increases Arabidopsis thaliana survival under drought compared to uninfected plants. To determine if specific TuMV proteins mediate drought tolerance, we cloned the coding sequence for each of the major viral proteins and generated transgenic A. thaliana that constitutively express each protein. Three TuMV proteins, 6K1, 6K2, and NIa-Pro, enhanced drought tolerance of A. thaliana when expressed constitutively in plants compared to controls. While in control plants drought induced transcripts related to abscisic acid (ABA) biosynthesis and ABA levels, there were no changes in ABA or related transcripts in plants expressing 6K2 under drought conditions compared to well-watered. Expression of 6K2 also conveyed drought tolerance in another host plant, Nicotiana benthamiana, when expressed using a virus over expression construct. In contrast to ABA, 6K2 expression enhanced salicylic acid (SA) accumulation in both Arabidopsis and N. benthamiana. These results suggest 6K2-induced drought tolerance is mediated through increased SA levels, and SA-dependent induction of plant secondary metabolites, osmolytes, and antioxidants that convey drought tolerance.