Rice physical defenses and their role against insect herbivores.

MAIN CONCLUSION: Understanding surface defenses, a relatively unexplored area in rice can provide valuable insight into constitutive and induced defenses against herbivores. Plants have evolved a multi-layered defense system against the wide range of pests that constantly attack them. Physical defenses comprised of trichomes, wax, silica, callose, and lignin, and are considered as the first line of defense against herbivory that can directly affect herbivores by restricting or deterring them. Most studies on physical defenses against insect herbivores have been focused on dicots compared to monocots, although monocots include one of the most important crops, rice, which half of the global population is dependent on as their staple food. In rice, Silica is an important element stimulating plant growth, although Silica has also been found to impart resistance against herbivores. However, other physical defenses in rice including wax, trichomes, callose, and lignin are less explored. A detailed exploration of the morphological structures and functional consequences of physical defense structures in rice can assist in incorporating these resistance traits in plant breeding and genetic improvement programs, and thereby potentially reduce the use of chemicals in the field. This mini review addresses these points with a closer look at current literature and prospects on rice physical defenses.

Local adaptation to continuous mowing makes the noxious weed Solanum elaeagnifolium a superweed candidate by improving fitness and defense traits.

The role of disturbance in accelerating weed growth is well understood. While most studies have focused on soil mediated disturbance, mowing can also impact weed traits. Using silverleaf nightshade (Solanum elaeagnifolium), a noxious and invasive weed, through a series of field, laboratory, and greenhouse experiments, we asked whether continuous mowing influences growth and plant defense traits, expressed via different avenues, and whether they cascade into offspring. We found that mowed plants produced significantly less number of fruits, and less number of total seeds per plant, but had higher seed mass, and germinated more and faster. When three herbivores were allowed to feed, tobacco hornworm (Manduca sexta) caterpillars, gained more mass on seedlings from unmowed plants, while cow pea aphid (Aphis craccivora), a generalist, established better on mowed seedlings; however, leaf trichome density was higher on unmowed seedlings, suggesting possible negative cross talk in defense traits. Texas potato beetle (Leptinotarsa texana), a co-evolved specialist on S. elaeagnifolium, did not show any differential feeding effects. We also found that specific root length, an indicator of nutrient acquisition, was significantly higher in first generation seedlings from mowed plants. Taken together, we show that mowing is a selective pressure that enhances some fitness and defense traits and can contribute to producing superweeds.

Inbreeding in Solanum carolinense alters floral attractants and rewards and adversely affects pollinator visitation.

PREMISE: 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.

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.

Host plant defense produces species-specific alterations to flight muscle protein structure and flight-related fitness traits of two armyworms.

Insects manifest phenotypic plasticity in their development and behavior in response to plant defenses, via molecular mechanisms that produce tissue-specific changes. Phenotypic changes might vary between species that differ in their preferred hosts and these effects could extend beyond larval stages. To test this, we manipulated the diet of southern armyworm (SAW; Spodoptera eridania) and fall armyworm (FAW; Spodoptera frugiperda) using a tomato mutant for jasmonic acid plant defense pathway (def1), and wild-type plants, and then quantified gene expression of Troponin t (Tnt) and flight muscle metabolism of the adult insects. Differences in Tnt spliceform ratios in insect flight muscles correlate with changes to flight muscle metabolism and flight muscle output. We found that SAW adults reared on induced def1 plants had a higher relative abundance (RA) of the A isoform of Troponin t (Tnt A) in their flight muscles; in contrast, FAW adults reared on induced def1 plants had a lower RA of Tnt A in their flight muscles compared with adults reared on def1 and controls. Although mass-adjusted flight metabolic rate showed no independent host plant effects in either species, higher flight metabolic rates in SAW correlated with increased RA of Tnt A Flight muscle metabolism also showed an interaction of host plants with Tnt A in both species, suggesting that host plants might be influencing flight muscle metabolic output by altering Tnt This study illustrates how insects respond to variation in host plant chemical defense by phenotypic modifications to their flight muscle proteins, with possible implications for dispersal.

Exposure to polyphenol-rich purple corn pericarp extract restricts fall armyworm (Spodoptera frugiperda) growth.

Bioactive compounds such as polyphenols in plants have been well studied for their potential insecticidal activities. These are considered as safe alternatives against chemical pesticides because of their lower persistence in environment, lower toxicity to humans and other organisms. However, they are present constitutively in lower amounts in plants and have to undergo complicated extraction methods - hampering their commercial exploitation in pest management. Using an inexpensive extraction method developed to recover polyphenol-rich liquid extract from purple corn pericarp, we recently documented that this extract has anti-feeding effects that cascade from larval to adult stages in a model herbivore tobacco hornworm (Manduca sexta). However, M. sexta does not feed on corn or any other major crops other than the species in the nightshade family (Solanaceae). In this study, we explored the same idea but using a generalist and common herbivore on corn, the fall armyworm (Spodoptera frugiperda). We found that purple corn pericarp extract inhibited the larval growth and development as well as negatively affected the pupal stages of S. frugiperda. However, unlike on M. sexta, time to complete larval life cycle was unaffected. Our findings confirm the toxicity of this extract on a generalist, economically important herbivore, but also suggest potential species-specific effects that should be explored further.

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.

Sorghum 3-Deoxyanthocyanidin Flavonoids Confer Resistance against Corn Leaf Aphid.

In this study we examined the role of sorghum flavonoids in providing resistance against corn leaf aphid (CLA) Rhopalosiphum maidis. In sorghum, accumulation of these flavonoids is regulated by a MYB transcription factor, yellow seed1 (y1). Functional y1 alleles accumulate 3-deoxyflavonoids (3-DFs) and 3-deoxyanthocyanidins (3-DAs) whereas null y1 alleles fail to accumulate these compounds. We found that significantly higher numbers of alate CLA adults colonized null y1 plants as compared to functional y1 plants. Controlled cage experiments and pairwise choice assays demonstrated that apterous aphids preferred to feed and reproduce on null y1 plants. These near-isogenic sorghum lines do not differ in their epicuticular wax content and were also devoid of any leaf trichomes. Significantly higher mortality of CLA was observed on artificial aphid diet supplemented with flavonoids obtained from functional y1 plants as compared to null y1 plants or the relevant controls. Our results demonstrate that the proximate mechanism underlying the deleterious effects on aphids is y1-regulated flavonoids which are important defense compounds against CLA.

Leaf trichomes affect caterpillar feeding in an instar-specific manner.

Leaf trichomes play well-established roles in defense against insect herbivores, both as a physical barrier that impedes herbivore movement and by mediating chemical defenses. However, little work has examined how different trichome types influence herbivory by herbivores at different stages of development. We examined whether caterpillar instar and trichome type (glandular or non-glandular) affected the ability of the specialist herbivore caterpillar Manduca sexta to initiate feeding on 11 Solanaceous species exhibiting variation in the density and type of leaf trichomes. Our results suggest that non-glandular trichomes are far more effective than glandular trichomes in deterring the initiation of feeding by first- and second-instar caterpillars. Meanwhile, neither glandular nor non-glandular trichomes significantly affected the ability of third-instar caterpillars to commence feeding. These findings suggest that while non-glandular trichomes deter feeding initiation by early instar caterpillars, the contribution of both trichomes on later instars may depend on effects after feeding initiation.

Field data on plant growth and insect damage on the noxious weed Solanum eleaegnifolium in an unexplored native range.

In this data article, we provide a novel data set on plant growth, insect damage levels, and herbivore community of the noxious and invasive weed Solanum eleaegnifolium (Solanaceae). The data is collected from disturbed and un-disturbed urban populations of the species from one of its unexplored native range in Southern United States (South Texas). The data include plant height measurements, insect damage levels, GPS coordinates of the populations, and their disturbance status. Additional data includes the number of chewing herbivore (specialist herbivore Texas potato beetle (Leptinotarsa texana; Chrysomelidae), their eggs, and any lepidopteran caterpillars found on the plants.

Plant spines deter herbivory by restricting caterpillar movement.

The spines of flowering plants are thought to function primarily in defence against mammalian herbivores; however, we previously reported that feeding by Manduca sexta caterpillars on the leaves of horsenettle plants (Solanum carolinense) induces increased development of internode spines on new growth. To determine whether and how spines impact caterpillar feeding, we conducted assays with three Solanaceous plant species that vary in spine numbers (S. carolinense, S. atropurpureum and S. aethiopicum) and also manipulated spine numbers within each species. We found that M. sexta caterpillars located experimentally isolated target leaves much more quickly on plants with experimentally removed spines compared with plants with intact spines. Moreover, it took caterpillars longer to defoliate species with relatively high spine numbers (S. carolinense and particularly Satropurpureum) compared with S. aethiopicum, which has fewer spines. These findings suggest that spines may play a significant role in defence against insect herbivores by restricting herbivore movement and increasing the time taken to access feeding sites, with possible consequences including longer developmental periods and increased vulnerability or apparency to predators.

Non-glandular trichomes of Solanum carolinense deter feeding by Manduca sexta caterpillars and cause damage to the gut peritrophic matrix.

Plant trichomes constitute a first line of defence against insect herbivores. The pre- and post-ingestive defensive functions of glandular trichomes are well documented and include direct toxicity, adhesion, antinutrition and defence gene induction. By contrast, the defensive functions of non-glandular trichomes are less well characterized, although these structures are thought to serve as physical barriers that impede herbivore feeding and movement. We experimentally varied the density of stellate non-glandular trichomes in several ways to explore their pre- and post-ingestive effects on herbivores. Larvae of Manduca sexta (Sphingidae) initiated feeding faster and gained more weight on Solanum carolinense (Solanaceae) leaves having lower trichome densities (or experimentally removed trichomes) than on leaves having higher trichome densities. Adding trichomes to artificial diet also deterred feeding and adversely affected caterpillar growth relative to controls. Scanning electron and light microscopy revealed that the ingestion of stellate trichomes by M. sexta caterpillars caused extensive damage to the peritrophic membrane, a gut lining that is essential to digestion and pathogen isolation. These findings suggest that, in addition to acting as a physical barrier to deter feeding, trichomes can inhibit caterpillar growth and development via post-ingestive effects.

Inbreeding compromises host plant defense gene expression and improves herbivore survival.

Inbreeding commonly occurs in flowering plants and often results in a decline in the plant's defense response. Insects prefer to feed and oviposit on inbred plants more than outbred plants--suggesting that selecting inbred host plants offers them fitness benefits. Until recently, no studies have examined the effects of host plant inbreeding on insect fitness traits such as growth and dispersal ability. In a recent article, we documented that tobacco hornworm (Manduca sexta L.) larvae that fed on inbred horsenettle (Solanum carolinense L.) plants exhibited accelerated larval growth and increased adult flight capacity compared to larvae that fed on outbred plants. Here we report that M. sexta mortality decreased by 38.2% when larvae were reared on inbred horsenettle plants compared to larvae reared on outbreds. Additionally, inbred plants showed a notable reduction in the average relative expression levels of lipoxygenease-D (LoxD) and 12-oxophytodienoate reductase-3 (OPR3), two genes in the jasmonic acid signaling pathway that are upregulated in response to herbivore damage. Our study presents evidence that furthers our understanding of the biochemical mechanism responsible for differences in insect performance on inbred vs. outbred host plants.

Plant inbreeding and prior herbivory influence the attraction of caterpillars (Manduca sexta) to odors of the host plant Solanum carolinense (Solanaceae).

PREMISE OF THIS STUDY: The mediation of plant-insect interactions by plant odors has been studied extensively, but most previous work has focused on documenting the role of constitutive and herbivore- or pathogen-induced plant volatiles as foraging cues for insect herbivores and their natural enemies. Relatively little work has explored genotypic variation in plant-odor profiles within species, and few studies have addressed the perception and use of olfactory cues by lepidopteran larvae or other herbivores during feeding. METHODS: We examined the effects of plant breeding (inbred vs. outbred individuals) and plant exposure to prior herbivory on the preferences of caterpillars (Manduca sexta) for odors of Solanum carolinense in leaf-disc and whole-plant choice assays. KEY RESULTS: Second- and third-instar larvae of M. sexta clearly and consistently preferred undamaged over herbivore-damaged plants of both breeding types and also consistently preferred inbred over outbred plants that had the same damage status. Similar preferences were observed even when plants were covered with bridal-veil cloth to mask visual cues, demonstrating that olfactory cues influence larval preferences. CONCLUSIONS: The observed preferences are consistent with our previous findings regarding the constitutive and induced volatile profiles of inbred and outbred horsenettle plants and their effects on plant-herbivore interactions. They furthermore correspond to differences in host-plant quality predicted by previous work and, thus, suggest that naive larvae of M. sexta can accurately assess aspects of host-plant quality via olfactory cues perceived at a distance.

Following Darwin's trail: interactions affecting the evolution of plant mating systems.

• Since the time of Charles Darwin, the variation in floral characteristics and its effects on plant mating system evolution have fascinated scientists. Recent advances in the field of genetics, molecular biology, and ecology have been very effective in addressing questions regarding mechanisms and interactions underlying the evolution of plant mating systems using various model and nonmodel species. The depth of plant mating system research reflects the complexity and diversity seen in nature, ranging from self-compatible hermaphroditic flowers to separate sexed individuals. Further, the mechanisms involved in the evolution of plant mating systems are much more diverse and differ even among closely related species. Here, as a special section, we present a suite of original papers that range from theoretical modeling to multiyear field research that address different factors affecting plant mating systems, and their effects on shaping interactions between plants, insects, and their environment.

Constitutive and herbivore-induced structural defenses are compromised by inbreeding in Solanum carolinense (Solanaceae).

PREMISE OF THE STUDY: A growing number of studies document effects of inbreeding on plant interactions with insect herbivores, including deleterious effects on direct and indirect plant defenses. However, our understanding of the specific mechanisms mediating such effects remains limited. Here we examine how inbreeding affects constitutive and induced expression of structural defenses (spines and trichomes) in common horsenettle, Solanum carolinense. • METHODS: Inbred and outbred progeny from nine maternal families of horsenettle were assigned to three treatments: control, Manduca sexta caterpillar damage, or mechanical damage. Numbers of internode spines and the density of abaxial and adaxial trichomes were assessed before and after (21 d) damage treatments. Data on internode length, flowering time, and total flower production was also collected to explore the costs of defense induction. • KEY RESULTS: Inbreeding adversely affected constitutive and induced physical/structural defenses: undamaged outbred plants produced more abaxial and adaxial leaf trichomes and internode spines than did inbred plants. Foliar damage by M. sexta larvae also induced more trichomes (on new leaves) and internode spines on outbred plants. Both inbred and outbred plants exposed to mechanical or caterpillar damage had shorter internodes than did control plants, but inbred damaged plants had longer internodes than did outbred damaged plants. Control outbred plants produced significantly more flowers than did control inbred plants or damaged plants of either breeding type. • CONCLUSIONS: Constitutive and induced structural defenses in horsenettle were negatively affected by inbreeding. Reduced flower production and internode length on damaged plants compared to controls suggests that defense induction entails significant costs.

Inbreeding in horsenettle (Solanum carolinense) alters night-time volatile emissions that guide oviposition by Manduca sexta moths.

Plant volatiles serve as key foraging and oviposition cues for insect herbivores as well as their natural enemies, but little is known about how genetic variation within plant populations influences volatile-mediated interactions among plants and insects. Here, we explore how inbred and outbred plants from three maternal families of the native weed horsenettle (Solanum carolinense) vary in the emission of volatile organic compounds during the dark phase of the photoperiod, and the effects of this variation on the oviposition preferences of Manduca sexta moths, whose larvae are specialist herbivores of Solanaceae. Compared with inbred plants, outbred plants consistently released more total volatiles at night and more individual compounds-including some previously reported to repel moths and attract predators. Female moths overwhelmingly chose to lay eggs on inbred (versus outbred) plants, and this preference persisted when olfactory cues were presented in the absence of visual and contact cues. These results are consistent with our previous findings that inbred plants recruit more herbivores and suffer greater herbivory under field conditions. Furthermore, they suggest that constitutive volatiles released during the dark portion of the photoperiod can convey accurate information about plant defence status (and/or other aspects of host plant quality) to foraging herbivores.