Flavone-associated resistance of two Lemna species to duckweed weevil attack.

Lemna perpusilla and Lemna minor are free-floating plants that often live in the same habitat. However, little is known about how they differ in response to herbivore attacks. In this study, we examined the species-specific resistance of two Lemna species to the duckweed weevil, Tanysphyrus lemnae. The female adults of T. lemnae preferred to lay eggs on L. perpusilla over L. minor. In addition, the larvae of T. lemnae performed better when fed on L. perpusilla than on L. minor. To understand the physiological basis of species-specific resistance in the two Lemna species, we measured the amounts of jasmonic acid (JA), phytosterols, and flavonoids. Attacks by duckweed weevils increased the levels of JA in the two Lemna species, but these levels did not differ significantly between the two species. Interestingly, the levels of flavones (isoorientin, vitexin, and isovitexin) in L. minor species were higher than those in L. perpusilla. The in vitro bioassay showed that three flavones significantly decreased the survival rate of duckweed weevil larvae. Although L. perpusilla was less resistant to duckweed weevil attack compared to L. minor, L. perpusilla grew faster than L. minor regardless of the duckweed weevil attack. These results suggest that these two Lemna species have different defense strategies against the duckweed weevil.

Jasmonate regulates plant resistance to Pectobacterium brasiliense by inducing indole glucosinolate biosynthesis.

Pectobacterium brasiliense (P. brasiliense) is a necrotrophic bacterium that causes the soft rot disease in Brassica rapa. However, the mechanisms underlying plant immune responses against necrotrophic bacterial pathogens with a broad host range are still not well understood. Using a flg22-triggered seedling growth inhibition (SGI) assay with 455 Brassica rapa inbred lines, we selected six B. rapa flagellin-insensitive lines (Brfin2-7) and three B. rapa flagellin-sensitive lines (Brfs1-3). Brfin lines showed compromised flg22-induced immune responses (oxidative burst, mitogen-activated protein kinase (MAPK) activation, and seedling growth inhibition) compared to the control line R-o-18; nevertheless, they were resistant to P. brasiliense. To explain this, we analyzed the phytohormone content and found that most Brfin lines had higher P. brasiliense-induced jasmonic acid (JA) than Brfs lines. Moreover, MeJA pretreatment enhanced the resistance of B. rapa to P. brasiliense. To explain the correlation between the resistance of Brfin lines to P. brasiliense and activated JA signaling, we analyzed pathogen-induced glucosinolate (GS) content in B. rapa. Notably, in Brfin7, the neoglucobrassicin (NGBS) content among indole glucosinolates (IGS) was significantly higher than that in Brfs2 following P. brasiliense inoculation, and genes involved in IGSs biosynthesis were also highly expressed. Furthermore, almost all Brfin lines with high JA levels and resistance to P. brasiliense had higher P. brasiliense-induced NGBS levels than Brfs lines. Thus, our results show that activated JA-mediated signaling attenuates flg22-triggered immunity but enhances resistance to P. brasiliense by inducing indole glucosinolate biosynthesis in Brassica rapa. This study provides novel insights into the role of JA-mediated defense against necrotrophic bacterial pathogens within a broad host range.

MSD2-mediated ROS metabolism fine-tunes the timing of floral organ abscission in Arabidopsis.

The timely removal of end-of-purpose flowering organs is as essential for reproduction and plant survival as timely flowering. Despite much progress in understanding the molecular mechanisms of floral organ abscission, little is known about how various environmental factors are integrated into developmental programmes that determine the timing of abscission. Here, we investigated whether reactive oxygen species (ROS), mediators of various stress-related signalling pathways, are involved in determining the timing of abscission and, if so, how they are integrated with the developmental pathway in Arabidopsis thaliana. MSD2, encoding a secretory manganese superoxide dismutase, was preferentially expressed in the abscission zone of flowers, and floral organ abscission was accelerated by the accumulation of ROS in msd2 mutants. The expression of the genes encoding the receptor-like kinase HAESA (HAE) and its cognate peptide ligand INFLORESCENCE DEFICIENT IN ABSCISSION (IDA), the key signalling components of abscission, was accelerated in msd2 mutants, suggesting that MSD2 acts upstream of IDA-HAE. Further transcriptome and pharmacological analyses revealed that abscisic acid and nitric oxide facilitate abscission by regulating the expression of IDA and HAE during MSD2-mediated signalling. These results suggest that MSD2-dependent ROS metabolism is an important regulatory point integrating environmental stimuli into the developmental programme leading to abscission.

Ontogeny-dependent effects of elevated CO2 and watering frequency on interaction between Aristolochia contorta and its herbivores.

Effects of environmental change on plants can differ due to sequential changes in their life-history strategies (i.e., ontogenetic variations). The fitness of herbivorous insects by physiological changes of the host plant could be affected depending on their diet breadth. However, little is known regarding the combinational effects of plant ontogeny and climate change on plant-herbivore interactions. This study examined the plant ontogeny-dependent effects of climate change on the interaction between a host plant (Aristolochia contorta), its specialist herbivore (Sericinus montela), and a generalist herbivore (Spodoptera exigua). Plants were grown under a factorial design of two distinct CO2 concentrations (ambient, 400 ppm; elevated, 560 ppm) and two watering frequencies (control, once a week; increased, twice a week). Plant ontogeny ameliorated the effects of climate change by altering its defensive traits, where nutrient-related factors were cumulatively affected by climate change. Herbivore performance was assessed at three different plant ontogenetic stages (1st-year juvenile, 1st-year senescence, and 2nd-year juvenile). Elevated CO2 levels reduced the growth and survival of the specialist herbivore, whereas increased watering frequency partially alleviated this reduced performance. Generalist herbivore performance slightly increased under elevated CO2 levels with progressing ontogenetic stages. The effects of climate change, both elevated CO2 and increased watering frequency were weaker in 2nd-year juveniles than in 1st-year juveniles. Elevated CO2 levels detrimentally affected the nutritional quality of A. contorta leaves. The effects of climate change on both specialist and generalist herbivore performance differed as plant ontogenetic stage proceeded. Increased growth rates and survival of the generalist herbivore at the latter ontogenetic stage might negatively affect the population dynamics of a specialist herbivore. This study suggests that biases are possible when the plant-herbivore interaction under a changing environment is predicted from a singular plant ontogenetic stage.

Effect of Soybean Volatiles on the Behavior of the Bean Bug, Riptortus pedestris.

The bean bug, Riptortus pedestris, is a polyphagous insect that feeds primarily on leguminous plants, especially soybean (Glycine max). Although the bean bug is an economically important pest of soybean, little is known about how the insect locates soybean fields. In this study, we examined the electroantennogram responses of R. pedestris to soybean volatiles and examined the behavioral responses of the adult bean bugs. R. pedestris adults were attracted more to their host-plant soybean, even when physical contact was absent, than to air or a non-host plant. Accordingly, we hypothesized that R. pedestris can recognize soybean through a plant's volatile organic compounds (VOCs). Five VOCs were identified from intact soybean plants at the vegetative stage: (Z)-3-hexen-1-ol, (Z)-3-hexenyl acetate, 4-ethylbenzaldehyde, α-farnesene, and methyl salicylate. Response spectra of the antennae to these volatiles clearly showed that both male and female R. pedestris can detect soybean volatiles. The adult bean bugs did not show behavioral orientation to any individual compounds but showed significant orientation to a particular blend of synthetic soybean volatiles when tested under laboratory conditions. In the field, this soybean volatile blend did not significantly attract the bean bugs, but it did interact synergistically with the aggregation pheromone to attract the bean bugs. These results highlight the role of host plant volatiles in the sensory ecology of R. pedestris and help explain colonization pattern of the bean bugs in soybean fields.

Pith-specific lignification in Nicotiana attenuata as a defense against a stem-boring herbivore.

Plants have developed tissue-specific defense strategies in response to various herbivores with different feeding habits. Although defense responses to leaf-chewing insects have been well studied, little is known about stem-specific responses, particularly in the pith, to stem-boring herbivores. To understand the stem-specific defense, we first conducted a comparative transcriptomic analysis of the wild tobacco Nicotiana attenuata before and after attack by the leaf-chewing herbivore Manduca sexta and the stem borer Trichobaris mucorea. When the stem-boring herbivore attacked, lignin-associated genes were upregulated specifically in the inner parenchymal cells of the stem, the pith; lignin also accumulated highly in the attacked pith. Silencing the lignin biosynthetic gene cinnamyl alcohol dehydrogenase enhanced the performance of the stem-boring herbivore but had no effect on the growth of the leaf-chewing herbivore. Two-dimensional nuclear magnetic resonance results revealed that lignified pith contains feruloyltyramine as an unusual lignin component in the cell wall, as a response against stem-boring herbivore attack. Pith-specific lignification induced by the stem-boring herbivore was modulated by both jasmonate and ethylene signaling. These results suggest that lignin provides a stem-specific inducible barrier, protecting plants against stem-boring insects.

Submergence deactivates wound-induced plant defence against herbivores.

Flooding is a common and critical disaster in agriculture, because it causes defects in plant growth and even crop loss. An increase in herbivore populations is often observed after floods, which leads to additional damage to the plants. Although molecular mechanisms underlying the plant responses to flooding have been identified, how plant defence systems are affected by flooding remains poorly understood. Herein, we show that submergence deactivates wound-induced defence against herbivore attack in Arabidopsis thaliana. Submergence rapidly suppressed the wound-induced expression of jasmonic acid (JA) biosynthesis genes, resulting in reduced JA accumulation. While plants exposed to hypoxia in argon gas exhibited similar reduced wound responses, the inhibitory effects were initiated after short-term submergence without signs for lack of oxygen. Instead, expression of ethylene-responsive genes was increased after short-term submergence. Blocking ethylene signalling by ein2-1 mutation partially restored suppressed expression of several wound-responsive genes by submergence. In addition, submergence rapidly removed active markers of histone modifications at a gene locus involved in JA biosynthesis. Our findings suggest that submergence inactivates defence systems of plants, which would explain the proliferation of herbivores after flooding.

Strigolactone signaling regulates specialized metabolism in tobacco stems and interactions with stem-feeding herbivores.

Plants are attacked by herbivores, which often specialize on different tissues, and in response, have evolved sophisticated resistance strategies that involve different types of chemical defenses frequently targeted to different tissues. Most known phytohormones have been implicated in regulating these defenses, with jasmonates (JAs) playing a pivotal role in complex regulatory networks of signaling interactions, often generically referred to as "cross talk." The newly identified class of phytohormones, strigolactones (SLs), known to regulate the shoot architecture, remain unstudied with regard to plant-herbivore interactions. We explored the role of SL signaling in resistance to a specialist weevil (Trichobaris mucorea) herbivore of the native tobacco, Nicotiana attenuata, that attacks the root-shoot junction (RSJ), the part of the plant most strongly influenced by alterations in SL signaling (increased branching). As SL signaling shares molecular components, such as the core F-box protein MORE AXILLARY GROWTH 2 (MAX2), with another new class of phytohormones, the karrikins (KARs), which promote seed germination and seedling growth, we generated transformed lines, individually silenced in the expression of NaMAX2, DWARF 14 (NaD14: the receptor for SL) and CAROTENOID CLEAVAGE DIOXYGENASE 7 (NaCCD7: a key enzyme in SL biosynthesis), and KARRIKIN INSENSITIVE 2 (NaKAI2: the KAR receptor). The mature stems of all transgenic lines impaired in the SL, but not the KAR signaling pathway, overaccumulated anthocyanins, as did the stems of plants attacked by the larvae of weevil, which burrow into the RSJs to feed on the pith of N. attenuata stems. T. mucorea larvae grew larger in the plants silenced in the SL pathway, but again, not in the KAI2-silenced plants. These phenotypes were associated with elevated JA and auxin (indole-3-acetic acid [IAA]) levels and significant changes in the accumulation of defensive compounds, including phenolamides and nicotine. The overaccumulation of phenolamides and anthocyanins in the SL pathway-silenced plants likely resulted from antagonism between the SL and JA pathway in N. attenuata. We show that the repressors of SL signaling, suppressor of max2-like (NaSMXL6/7), and JA signaling, jasmonate zim-domain (NaJAZs), physically interact, promoting NaJAZb degradation and releasing JASMONATE INSENSITIVE 1 (JIN1/MYC2) (NaMYC2), a critical transcription factor promoting JA responses. However, the increased performance of T. mucorea larvae resulted from lower pith nicotine levels, which were inhibited by increased IAA levels in SL pathway-silenced plants. This inference was confirmed by decapitation and auxin transport inhibitor treatments that decreased pith IAA and increased nicotine levels. In summary, SL signaling tunes specific sectors of specialized metabolism in stems, such as phenylpropanoid and nicotine biosynthesis, by tailoring the cross talk among phytohormones, including JA and IAA, to mediate herbivore resistance of stems. The metabolic consequences of the interplay of SL, JA, and IAA signaling revealed here could provide a mechanism for the commonly observed pattern of herbivore tolerance/resistance trade-offs.

Shoot phytochrome B modulates reactive oxygen species homeostasis in roots via abscisic acid signaling in Arabidopsis.

Underground roots normally reside in darkness. However, they are often exposed to ambient light that penetrates through cracks in the soil layers which can occur due to wind, heavy rain or temperature extremes. In response to light exposure, roots produce reactive oxygen species (ROS) which promote root growth. It is known that ROS-induced growth promotion facilitates rapid escape of the roots from non-natural light. Meanwhile, long-term exposure of the roots to light elicits a ROS burst, which causes oxidative damage to cellular components, necessitating that cellular levels of ROS should be tightly regulated in the roots. Here we demonstrate that the red/far-red light photoreceptor phytochrome B (phyB) stimulates the biosynthesis of abscisic acid (ABA) in the shoots, and notably the shoot-derived ABA signals induce a peroxidase-mediated ROS detoxification reaction in the roots. Accordingly, while ROS accumulate in the roots of the phyb mutant that exhibits reduced primary root growth in the light, such an accumulation of ROS did not occur in the dark-grown phyb roots that exhibited normal growth. These observations indicate that mobile shoot-to-root ABA signaling links shoot phyB-mediated light perception with root ROS homeostasis to help roots adapt to unfavorable light exposure. We propose that ABA-mediated shoot-to-root phyB signaling contributes to the synchronization of shoot and root growth for optimal propagation and performance in plants.

What happens in the pith stays in the pith: tissue-localized defense responses facilitate chemical niche differentiation between two spatially separated herbivores.

Herbivore attack is known to elicit systemic defense responses that spread throughout the host plant and influence the performance of other herbivores. While these plant-mediated indirect competitive interactions are well described, and the co-existence of herbivores from different feeding guilds is common, the mechanisms of co-existence are poorly understood. In both field and glasshouse experiments with a native tobacco, Nicotiana attenuata, we found no evidence of negative interactions when plants were simultaneously attacked by two spatially separated herbivores: a leaf chewer Manduca sexta and a stem borer Trichobaris mucorea. T. mucorea attack elicited jasmonic acid (JA) and jasmonoyl-l-isoleucine bursts in the pith of attacked stems similar to those that occur in leaves when M. sexta attacks N. attenuata leaves. Pith chlorogenic acid (CGA) levels increased 1000-fold to levels 6-fold higher than leaf levels after T. mucorea attack; these increases in pith CGA levels, which did not occur in M. sexta-attacked leaves, required JA signaling. With plants silenced in CGA biosynthesis (irHQT plants), CGA, as well as other caffeic acid conjugates, was demonstrated in both glasshouse and field experiments to function as a direct defense protecting piths against T. mucorea attack, but not against leaf chewers or sucking insects. T. mucorea attack does not systemically activate JA signaling in leaves, while M. sexta leaf-attack transiently induces detectable but minor pith JA levels that are dwarfed by local responses. We conclude that tissue-localized defense responses allow tissue-specialized herbivores to share the same host and occupy different chemical defense niches in the same hostplant.

Trichobaris weevils distinguish amongst toxic host plants by sensing volatiles that do not affect larval performance.

Herbivorous insects use plant metabolites to inform their host plant selection for oviposition. These host-selection behaviours are often consistent with the preference-performance hypothesis; females oviposit on hosts that maximize the performance of their offspring. However, the metabolites used for these oviposition choices and those responsible for differences in offspring performance remain unknown for ecologically relevant interactions. Here, we examined the host-selection behaviours of two sympatric weevils, the Datura (Trichobaris compacta) and tobacco (T. mucorea) weevils in field and glasshouse experiments with transgenic host plants specifically altered in different components of their secondary metabolism. Adult females of both species strongly preferred to feed on D. wrightii rather than on N. attenuata leaves, but T. mucorea preferred to oviposit on N. attenuata, while T. compacta oviposited only on D. wrightii. These oviposition behaviours increased offspring performance: T. compacta larvae only survived in D. wrightii stems and T. mucorea larvae survived better in N. attenuata than in D. wrightii stems. Choice assays with nicotine-free, JA-impaired, and sesquiterpene-over-produced isogenic N. attenuata plants revealed that although half of the T. compacta larvae survived in nicotine-free N. attenuata lines, nicotine did not influence the oviposition behaviours of both the nicotine-adapted and nicotine-sensitive species. JA-induced sesquiterpene volatiles are key compounds influencing T. mucorea females' oviposition choices, but these sesquiterpenes had no effect on larval performance. We conclude that adult females are able to choose the best host plant for their offspring and use chemicals different from those that influence larval performance to inform their oviposition decisions.

Functional analysis of TiO2 nanoparticle toxicity in three plant species.

Titanium dioxide nanoparticles (nano-TiO2) are manufactured and used worldwide in large quantities. However, phytotoxicity research on nano-TiO2 has yielded confusing results, ranging from strong toxicity to positive effects. Therefore, in this research, the effects of nano-TiO2 on the germination and root elongation of seed and seedlings were studied. Additionally, the uptake and physiological responses of mature plants were investigated. Physical chemistry data were analyzed to assess the availability of nano-TiO2. Finally, a hydroponic system designed to overcome nano-TiO2 precipitation was used to reproduce the environmental conditions of actual fields. Nano-TiO2 did not have any effect on seed germination or on most of the plant species tested. Nano-TiO2 had positive effects on root elongation in some species. No physiological differences in enzyme activities or chlorophyll content were detected, even though the plants absorbed nano-TiO2. Physical chemistry data showed that nano-TiO2 agglomerated rapidly and formed particles with much bigger hydrodynamic diameters, even in distilled water and especially in a hydroponic system. Furthermore, agglomerated nano-TiO2 formed precipitates; this would be more severe in an actual field. Consequently, nano-TiO2 would not be also readily available to plants and would not cause any significant effects on plants. Our results and other reports suggest that titanium itself is not phytotoxic, even though plants absorb titanium. In conclusion, nano-TiO2 is not toxic to the three plant species, in vitro or in situ.