Design, synthesis and biological evaluation of novel modified dual-target shikonin derivatives for colorectal cancer treatment.

Warburg effect provides energy and material essential for tumor proliferation, the reverse of Warburg effect provides insights into the development of a novel anti-cancer strategy. Pyruvate kinase 2 (PKM2) and pyruvate dehydrogenase kinase 1 (PDK1) are two key enzymes in tumor glucose metabolism pathway that not only contribute to the Warburg effect through accelerating aerobic glycolysis, but also serve as druggable target for colorectal cancer (CRC). Considering that targeting PKM2 or PDK1 alone does not seem to be sufficient to remodel abnormal glucose metabolism and achieve significant antitumor activity, a series of novel benzenesulfonyl shikonin derivatives were designed to regulate PKM2 and PDK1 simultaneously. By means of molecular docking and antiproliferative screen, we found that compound Z10 could act as the combination of PKM2 activator and PDK1 inhibitor, thereby significantly inhibited glycolysis that reshaping tumor metabolism. Moreover, Z10 could inhibit proliferation, migration and induce apoptosis in CRC cell HCT-8. Finally, the in vivo anti-tumor activity of Z10 was evaluated in a colorectal cancer cell xenograft model in nude mice and the results demonstrated that Z10 induced tumor cell apoptosis and inhibited tumor cell proliferation with lower toxicity than shikonin. Our findings indicated that it is feasible to alter tumor energy metabolism through multi-target synergies, and the dual-target benzenesulfonyl shikonin derivative Z10 could be a potential anti-CRC agent.

Somatic Mutation Analysis in Salix suchowensis Reveals Early-Segregated Cell Lineages.

Long-lived plants face the challenge of ever-increasing mutational burden across their long lifespan. Early sequestration of meristematic stem cells is supposed to efficiently slow down this process, but direct measurement of somatic mutations that accompanies segregated cell lineages in plants is still rare. Here, we tracked somatic mutations in 33 leaves and 22 adventitious roots from 22 stem-cuttings across eight major branches of a shrub willow (Salix suchowensis). We found that most mutations propagated separately in leaves and roots, providing clear evidence for early segregation of underlying cell lineages. By combining lineage tracking with allele frequency analysis, our results revealed a set of mutations shared by distinct branches, but were exclusively present in leaves and not in roots. These mutations were likely propagated by rapidly dividing somatic cell lineages which survive several iterations of branching, distinct from the slowly dividing axillary stem cell lineages. Leaf is thus contributed by both slowly and rapidly dividing cell lineages, leading to varied fixation chances of propagated mutations. By contrast, each root likely arises from a single founder cell within the adventitious stem cell lineages. Our findings give straightforward evidence that early segregation of meristems slows down mutation accumulation in axillary meristems, implying a plant "germline" paralog to the germline of animals through convergent evolution.

Plant volatiles mediate evolutionary interactions between plants and tephritid flies and are evolutionarily more labile than non-volatile defenses.

Studies show that plant defenses influence the host-use of herbivores and tend to be evolutionarily more labile than herbivore traits (e.g. feeding preferences). However, all previous studies have focused exclusively on non-volatile plant defenses thereby overlooking the roles of plant volatiles. We hypothesized that volatiles are equally important determinants of herbivore host-use and are evolutionarily more labile than herbivore traits. To test these hypotheses, the following experiments were conducted. We identified the volatiles and non-volatiles of 17 Asteraceae species and measured their relative contents. We also used a highly resolved bipartite trophic network of the 17 host species and 20 herbivorous (pre-dispersal seed predator) tephritid fly species to determine the evolutionary interactions between plants and herbivores. The chemical data showed that interspecific similarity in volatiles-but not non-volatiles and phylogenetic distance-significantly accounted for the herbivore community across the plant species; this implies that plant volatiles-but not non-volatile compounds and species identity-dictate plant-tephritid fly interactions. Moreover, we observed phylogenetic signal for non-volatiles but not for volatiles; therefore closely related herbivores do not necessarily use closely related host species with similar non-volatiles, but do tend to attack plants producing similar volatiles. Thus, plant volatiles are evolutionarily more labile than non-volatiles and herbivore traits associate with host use. These results show that the interactions between plants and herbivores are evolutionary asymmetric, shed light on the role of plant volatiles in plant-herbivore interactions, and highlight the need to include data for both volatiles and non-volatiles when investigating plant-animal interactions.

Rare plant species are at a disadvantage when both herbivory and pollination interactions are considered in an alpine meadow.

Rare plant species often suffer less damage than common species because of positive density-dependent herbivory, and it has been suggested that this 'rare species advantage' fosters plant species coexistence. However, it is unknown whether rare species have an advantage when pollination interactions are also considered. We hypothesized that a 'positive density-dependent pollination success' across plant species would result in common plants experiencing higher seed set rates compared to rare species, and that positive density-dependent effects would negate or even override the positive density-dependent damage due to herbivory resulting in higher seed loss rates in common plant species. We tested this hypothesis by concurrently examining a plant-predispersal seed predator system and a plant-pollinator system for 24 Asteraceae species growing in an alpine meadow community (Sichuan Province, China). Having previously reported a positive density-dependent effect on seed loss rates due to seed predators, we here focus on the density-dependent effects on pollination success by investigating pollinator species richness, visitation frequencies and seed set rates for each plant species. We also estimated the seed output rate of each plant species as the product of seed set rate and the rate of surviving seeds (i.e. 1 - the seed loss rate). Consistent with our hypothesis, a positive density-dependent effect was observed for pollinator species richness, visitation frequencies and seed set rates across plant species. Moreover, the positive effect overrode the negative density-dependent effect of herbivores on seed production, such that common species tended to have a higher seed output rate than rare species (i.e. we observed a 'rare species disadvantage'). These results indicate that the low seed output rate of rare species might result from a pollination limitation, and that both mutualistic and antagonistic interactions should be examined simultaneously to fully understand plant species coexistence in local communities.

Linkage between species traits and plant phenology in an alpine meadow.

Plant phenology differs largely among coexisting species within communities that share similar habitat conditions. However, the factors explaining such phenological diversity of plants have not been fully investigated. We hypothesize that species traits, including leaf mass per area (LMA), seed mass, stem tissue mass density (STD), maximum plant height (Hmax), and relative growth rate in height (RGRH), explain variation in plant phenology, and tested this hypothesis in an alpine meadow. Results showed that both LMA and STD were positively correlated with the onset (i.e., beginning) and offset (i.e., ending) times of the four life history events including two reproductive events (flowering and fruiting) and two vegetative events (leafing and senescing). In contrast, RGRH was negatively correlated with the four life phenological events. Moreover, Hmax was positively correlated with reproductive events but not with vegetative events. However, none of the eight phenological events was associated with seed size. In addition, the combination of LMA and STD accounted for 50% of the variation in plant phenologies. Phylogenetic generalized least squares analysis showed plant phylogeny weakened the relationships between species traits vs. phenologies. Phylogeny significantly regulated the variation in the ending but not the beginning of phenologies. Our results indicate that species traits are robust indicators for plant phenologies and can be used to explain the diversity of plant phenologies among co-occurring herbaceous species in grasslands. The findings highlight the important role of the combination of and trade-offs between functional traits in determing plant phenology diversity in the alpine meadow.

Domestic honeybees affect the performance of pre-dispersal seed predators in an alpine meadow.

Flowering plants interact simultaneously with mutualistic pollinators and antagonistic herbivores such that plant-mediated interactions between pollinators and herbivores must exist. Although the effects of herbivores on pollinator behavior have been investigated extensively, the effect of pollinators on herbivore performance has seldom been explored. We hypothesized that insect pollinators could improve the survival and growth of pre-dispersal seed predators by increasing seed production. We tested this hypothesis along three transects radiating from well-established apiaries in an alpine meadow by supplementing pollination in sites close to and distant from apiaries and subsequently examining seed production of the dominant nectariferous plant species Saussurea nigrescens (Asteraceae) and the performance of three dominant pre-dispersal seed predators (tephritid fly species). Pollen supplementation (1) significantly increased seed set and mass of developed seed per capitulum (i.e., flowerhead) in the distant but not the close sites, (2) did not change the survival and growth rates of the smaller-bodied species (Tephritis femoralis and Campiglossa nigricauda) at either site, but (3) improved the performance of the larger-bodied seed predator (Terellia megalopyge) at distant sites but not close sites. In addition, the larger-bodied tephritid fly showed higher infestation rates and relative abundance in the close sites than in the distant sites, whereas the smaller-bodied species had lower relative abundances in the close sites and similar infestation rates in both site types. These observations demonstrate contrasting effects of plant mutualists on the performance of antagonists with potential consequences for population sizes of insect herbivores.

Plant-mediated resource partitioning by coexisting parasitoids.

Although it has been frequently suggested that resource partitioning of species coexisting at the same trophic level can be mediated by interactions with species at non-adjacent trophic levels, empirical evidence supporting this claim is scarce. Here we demonstrate that plants may mediate resource partitioning for two parasitoids that share the same herbivorous host. The tephritid fly Tephritis femoralis is the primary pre-dispersal seed predator of two Asteraceae species, Saussurea nigrescens and Anaphalis flavescens, both of which dominate the plant community in the alpine meadows of the Tibetan Plateau. Field surveys and molecular barcoding analyses showed that the identity of the fly's main predator depended on the plant in which the fly developed. Tephritid flies that developed in S. nigrescens were preyed upon mainly by the parasitoid wasp Pteromalus albipennis, while the parasitoid Mesopolobus sp. was the main predator of flies that developed in A. flavescens. Microcosm experiments revealed that P. albipennis could not exploit the host flies within the capitula of A. flavescens due to food limitation (capitula are too small), while Mesopolobus sp. could not exploit the host flies within the capitula of S. nigrescens due to its inability to reach the host with its ovipositor (capitula are too large). Such bottom-up control of plant species traits may facilitate the coexistence of parasitoid wasps sharing a common host in this system. We suggest that interactions between non-adjacent trophic levels may potentially promote species coexistence and diversity in biological communities.

Chlorophyll fluorescence tracks seasonal variations of photosynthesis from leaf to canopy in a temperate forest.

Accurate estimation of terrestrial gross primary productivity (GPP) remains a challenge despite its importance in the global carbon cycle. Chlorophyll fluorescence (ChlF) has been recently adopted to understand photosynthesis and its response to the environment, particularly with remote sensing data. However, it remains unclear how ChlF and photosynthesis are linked at different spatial scales across the growing season. We examined seasonal relationships between ChlF and photosynthesis at the leaf, canopy, and ecosystem scales and explored how leaf-level ChlF was linked with canopy-scale solar-induced chlorophyll fluorescence (SIF) in a temperate deciduous forest at Harvard Forest, Massachusetts, USA. Our results show that ChlF captured the seasonal variations of photosynthesis with significant linear relationships between ChlF and photosynthesis across the growing season over different spatial scales (R2  = 0.73, 0.77, and 0.86 at leaf, canopy, and satellite scales, respectively; P < 0.0001). We developed a model to estimate GPP from the tower-based measurement of SIF and leaf-level ChlF parameters. The estimation of GPP from this model agreed well with flux tower observations of GPP (R2  = 0.68; P < 0.0001), demonstrating the potential of SIF for modeling GPP. At the leaf scale, we found that leaf Fq '/Fm ', the fraction of absorbed photons that are used for photochemistry for a light-adapted measurement from a pulse amplitude modulation fluorometer, was the best leaf fluorescence parameter to correlate with canopy SIF yield (SIF/APAR, R2  = 0.79; P < 0.0001). We also found that canopy SIF and SIF-derived GPP (GPPSIF ) were strongly correlated to leaf-level biochemistry and canopy structure, including chlorophyll content (R2  = 0.65 for canopy GPPSIF and chlorophyll content; P < 0.0001), leaf area index (LAI) (R2  = 0.35 for canopy GPPSIF and LAI; P < 0.0001), and normalized difference vegetation index (NDVI) (R2  = 0.36 for canopy GPPSIF and NDVI; P < 0.0001). Our results suggest that ChlF can be a powerful tool to track photosynthetic rates at leaf, canopy, and ecosystem scales.

Artificial asymmetric warming reduces nectar yield in a Tibetan alpine species of Asteraceae.

BACKGROUND AND AIMS: Asymmetric warming is one of the distinguishing features of global climate change, in which winter and night-time temperatures are predicted to increase more than summer and diurnal temperatures. Winter warming weakens vernalization and hence decreases the potential to flower for some perennial herbs, and night warming can reduce carbohydrate concentrations in storage organs. This study therefore hypothesized that asymmetric warming should act to reduce flower number and nectar production per flower in a perennial herb, Saussurea nigrescens, a key nectar plant for pollinators in Tibetan alpine meadows. METHODS: A long-term (6 years) warming experiment was conducted using open-top chambers placed in a natural meadow and manipulated to achieve asymmetric increases in temperature, as follows: a mean annual increase of 0·7 and 2·7 °C during the growing and non-growing seasons, respectively, combined with an increase of 1·6 and 2·8 °C in the daytime and night-time, respectively, from June to August. Measurements were taken of nectar volume and concentration (sucrose content), and also of leaf non-structural carbohydrate content and plant morphology. KEY RESULTS: Six years of experimental warming resulted in reductions in nectar volume per floret (64·7 % of control), floret number per capitulum (8·7 %) and capitulum number per plant (32·5 %), whereas nectar concentration remained unchanged. Depletion of leaf non-structural carbohydrates was significantly higher in the warmed than in the ambient condition. Overall plant density was also reduced by warming, which, when combined with reductions in flower development and nectar volumes, led to a reduction of ∼90 % in nectar production per unit area. CONCLUSIONS: The negative effect of asymmetric warming on nectar yields in S. nigrescens may be explained by a concomitant depletion of leaf non-structural carbohydrates. The results thus highlight a novel aspect of how climate change might affect plant-pollinator interactions and plant reproduction via induction of allocation shifts for plants growing in communities subject to asymmetric warming.

Parasitoid wasps indirectly suppress seed production by stimulating consumption rates of their seed-feeding hosts.

In parasitoid-herbivore-plant food chains, parasitoids may be simultaneously linked with both herbivore hosts and plants, as occurs when herbivores attacked by parasitoids continue to consume plants although they are destined to die. This peculiar property may cause parasitoids to confer a differential trophic cascading effect on plants than that known for typical predators. We hypothesized that larval koinobiont parasitoids would confer an immediate negative effect on plant seed production by stimulating consumption of their seed-predator hosts. We tested this hypothesis in an alpine parasitic food chain of plant seeds, pre-dispersal seed predators (tephritid fly larvae) and koinobiont parasitoids using field observations, a field experiment and a microcosm study. We first compared observed seed production in (i) non-infected capitula, (ii) capitula infected only by seed predators (tephritid flies) and (iii) capitula infected by both seed predators and their parasitoids in five Asteraceae species. Consistent with our hypothesis, seed loss in the capitula with both seed predators and parasitoids was significantly greater than in the capitula infested only by seed predators. This effect was replicated in a controlled field experiment focusing on the most common parasitoid-seed predator-plant interaction chain in our system, in which confounding factors (e.g. density and phenology) were excluded. Here, we show that parasitoids indirectly decreased plant seed production by changing the behaviour of seed predators. In a microcosm study, we show that larval parasitoids significantly extended the growth period and increased the terminal size of their host tephritid maggots. Thus, parasitoids suppressed plant seed production by stimulating the growth and consumption of the fly maggots. In contrast to the typical predator-induced trophic cascade, we highlight the significance of parasitoids indirectly decreasing plant fitness by stimulating consumption by seed predators. Future studies on trophic interactions should consider the net effect of both increased consumption by seed predators and their death after development of parasitoids.

The sign of cascading predator effects varies with prey traits in a detrital system.

Theory and experiments show that the nature of 'green' trophic cascades, between predators, herbivores and plants, varies with several key species traits: predator hunting mode and predator and prey habitat domains. Meanwhile, 'brown' cascades between predators, environment-modifying detritivores and plants have been largely overlooked and the roles of species traits, particularly prey traits, in determining the nature of these cascades remains unclear. We hypothesize that, in predator-detritivore-plant interaction chains, the burrowing ability of plant-facilitating detritivores determines their response to predators and thus the sign of indirect effect transmitted. In the dung-decomposer food web of an alpine meadow, we predicted that in the presence of above-ground predacious beetles: (i) non-burrowing detritivores will suffer mortality due to predation and transmit negative indirect effects to plants, whereas (ii) burrowing detritivores will escape predation by retreating deeper into the soil, transmitting positive indirect effects to plants. In support of predictions, experiments showed that a single species of predacious beetle (i) reduced the density of the non-burrowing species and indirectly reduced dung loss rate, soil nutrient concentrations and plant biomass, but (ii) drove the burrowing species deeper, indirectly improved soil conditions and increased plant biomass. These results show that the burrowing ability of a detritivore can determine whether it transmits a negative indirect effect mediated by a reduction in its density, or a positive indirect effect mediated by its behavioural response to predation risk. We call for further tests of our detritivore-trait hypothesis in different regions and ecosystems to further develop a general trait-based framework for trophic cascades in detrital food webs. We further advance the general hypothesis that the locomotion traits of prey species (e.g. burrowing/flying ability) may help explain their behavioural response to predation risk and the nature of indirect effect they transmit from predators to plants.

Cascading effects of predator-detritivore interactions depend on environmental context in a Tibetan alpine meadow.

Studies of grazing food webs show that species traits can interact with environmental factors to determine the strength of trophic cascades, but analogous context dependencies in detrital food webs remain poorly understood. In predator-detritivore-plant interaction chains, predators are expected to indirectly suppress plant biomass by reducing the density of plant-facilitating detritivores. However, this outcome can be reversed where above-ground predators drive burrowing detritivores to lower soil levels, strengthening their plant-facilitating effects. Here, we show that these trait-mediated indirect interactions further depend on environmental context in a Tibetan alpine meadow. In our study system, undulating topography generates higher (dry soil) patches interspersed with lower (wet soil) patches. Because the ability of detritivores to form deep burrows is likely to be limited by oxygen availability in low patches (wet soil), we hypothesized that (i) burrowing detritivores would undergo a vertical habitat shift, allowing them to more effectively avoid predation, in high - but not low - patches, and (ii) this shift would transmit positive effects of predators to plants in high patches by improving conditions in the lower soil layer. We tested these hypotheses using complementary field and glasshouse experiments examining whether the cascading effects of above-ground predatory beetles (presence/absence) on the density and behaviour of tunnel-forming detritivorous beetles, soil properties, and plant growth varied with patch type (low/high). Results revealed that predatory beetles did not reduce the density of detritivores in either patch type but had context-dependent trait-mediated effects, increasing the tunnelling depth of detritivores, improving soil conditions and ultimately increasing plant biomass in the high but not low patches. This study adds to an emerging predictive framework linking predators to plants in detritus food webs, demonstrating that these indirect interactions depend not just on the relative habitat domains of predators and prey, but also on environmental conditions that can predictably constrain the behavioural response of detritivores to predation risk.

Trade-offs between light interception and leaf water shedding: a comparison of shade- and sun-adapted species in a subtropical rainforest.

Species in high-rainfall regions have two major alternative approaches to quickly drain off water, i.e., increasing leaf inclination angles relative to the horizontal plane, or developing long leaf drip tips. We hypothesized that shade-adapted species will have more pronounced leaf drip tips but not greater inclination angles (which can reduce the ability to intercept light) compared to sun-adapted species and that length of leaf drip tips will be negatively correlated with photosynthetic capacity [characterized by light-saturated net photosynthetic rates (Amax), associated light compensation points (LCP), and light saturation points (LSP)]. We tested this hypothesis by measuring morphological and physiological traits that are associated with light-interception and water shedding for seven shade-adapted shrub species, ten sun-adapted understory shrub species, and 15 sun-adapted tree species in a subtropical Chinese rainforest, where mean annual precipitation is around 1,600 mm. Shade-adapted understory species had lower LMA, Amax, LSP, and LCP compared to understory or canopy sun-adapted species; their leaf and twig inclination angles were significantly smaller and leaf drip tips were significantly longer than those in sun-adapted species. This suggests that shade-adapted understory species tend to develop pronounced leaf drip tips but not large leaf inclination angles to shed water. The length of leaf drip tips was negatively correlated with leaf inclination angles and photosynthetic capacity. These relationships were consistent between ordinary regression and phylogenetic generalized least squares analyses. Our study illustrates the trade-offs between light interception and leaf water shedding and indicates that length of leaf drip tips can be used as an indicator of adaptation to shady conditions and overall photosynthetic performance of shrub species in subtropical rainforests.

Insect root feeders incur negative density-dependent damage across plant species in an alpine meadow.

Although herbivores are well known to incur positive density-dependent damage and mortality, thereby likely shaping plant community assembly, the response of belowground root feeders to changes in plant density has seldom been addressed. Locally rare plant species (with lower plant biomass per area) are often smaller with shallower roots than common species (with higher plant biomass per area) in competition-intensive grasslands. Likewise, root feeders are often distributed in the upper soil layers. We hypothesized, therefore, that root feeders would incur negative density (biomass)-dependent damage across plant species. To test this hypothesis, we investigated the diversity and abundance of plant and root feeder species in an alpine meadow and determined the diet of the root feeders using metabarcoding. Across all species, root feeder load decreased with increasing aboveground plant biomass, root biomass, and total plant biomass per area, indicating a negative density dependence of damage across plant species. Aboveground plant biomass per area increased with increasing individual plant biomass and root depth per area across species, suggesting that rare plant species were smaller in size and had shallower root systems compared to common plant species. Both root biomass per area and root feeder biomass per area decreased with soil depth, but the root feeder biomass decreased disproportionately faster compared to root biomass with increasing root depth. Root feeder load decreased with increasing root depth but was not correlated with the feeding preference of root feeder species. Moreover, the prediction derived from a random process incorporating vertical distributions of root biomass and root feeder biomass significantly accounted for interspecific variation in root feeder load. In conclusion, the data indicate that root feeders incur negative density-dependent damage across plant species. On this basis, we suggest that manipulative experiments should be conducted to determine the effect of the negative density-dependent damage on plant community structure and that different types of plant-animal interactions should be concurrently examined to fully understand the effect of plant density on overall herbivore damage across plant species.

Harnessing the power of microbes: Enhancing soybean growth in an acidic soil through AMF inoculation rather than P-fertilization.

The low phosphorus (P) availability of acidic soils severely limits leguminous plant growth and productivity. Improving the soil P nutritional status can be achieved by increasing the P-content through P-fertilization or stimulating the mineralization of organic P via arbuscular mycorrhizal fungi (AMF) application; however, their corresponding impacts on plant and soil microbiome still remain to be explored. Here, we examined the effects of AMF-inoculation and P-fertilization on the growth of soybean with different P-efficiencies, as well as the composition of rhizo-microbiome in an acidic soil. The growth of recipient soybean NY-1001, which has a lower P-efficiency, was not significantly enhanced by AMF-inoculation or P-fertilization. However, the plant biomass of higher P-efficiency transgenic soybean PT6 was significantly increased by 46.74%-65.22% through AMF-inoculation. Although there was no discernible difference in plant biomass between PT6 and NY-1001 in the absence of AMF-inoculation and P-fertilization, PT6 had approximately 1.9-2.5 times the plant biomass of NY-1001 after AMF-inoculation. Therefore, the growth advantage of higher P-efficiency soybean was achieved through the assistance of AMF rather than P-fertilization in available P-deficient acidic soil. Most nitrogen (N)-fixing bacteria and some functional genes related to N-fixation were abundant in endospheric layer, as were the P-solubilizing Pseudomonas plecoglossicida, and annotated P-metabolism genes. These N-fixing and P-solubilizing bacteria were positive correlated with each other. Lastly, the two most abundant phytopathogenic fungi species accumulated in endospheric layer, they exhibited positive correlations with N-fixing bacteria, but displayed negative interactions with the majority of the other dominant non-pathogenic genera with potential antagonistic activity.

Predatory beetles facilitate plant growth by driving earthworms to lower soil layers.

Theory suggests that predators of soil-improving, plant-facilitating detritivores (e.g. earthworms) should suppress plant growth via a negative tri-trophic cascade, but the empirical evidence is still largely lacking. We tested this prediction in an alpine meadow on the Tibetan Plateau by manipulating predatory beetles (presence/absence) and quantifying (i) direct effects on the density and behaviour of earthworms; and (ii) indirect effects on soil properties and above-ground plant biomass. In the absence of predators, earthworms improved soil properties, but did not significantly affect plant biomass. Surprisingly, the presence of predators strengthened the positive effect of earthworms on soil properties leading to the emergence of a positive indirect effect of predators on plant biomass. We attribute this counterintuitive result to: (i) the inability of predators to suppress overall earthworm density; and (ii) the predator-induced earthworm habitat shift from the upper to lower soil layer that enhanced their soil-modifying, plant-facilitating, effects. Our results reveal that plant-level consequences of predators as transmitted through detritivores can hinge on behaviour-mediated indirect interactions that have the potential to overturn predictions based solely on trophic interactions. This work calls for a closer examination of the effects of predators in detritus food webs and the development of spatially explicit theory capable of predicting the occurrence and consequences of predator-induced detritivore behavioural shifts.

Moderate Grazing Promotes Arthropod Species Diversity in an Alpine Meadow.

Livestock grazing is an important tool used in grassland land management practices. Studies have substantially addressed the effect of grazing on plant species diversity, revealing that moderate grazing increases plant species diversity. However, few studies have dealt with the relationship between grazing and arthropod species diversity, which remains unclear. Here, we hypothesize that moderate grazing promotes arthropod species diversity because arthropods are directly or indirectly dependent on plant diversity. In this study, we conducted a two-year plant and arthropod survey from 2020 to 2021 at four levels of grazing intensity, i.e., nongrazing (as a control), light grazing, moderate grazing, and heavy grazing, of the long-term grazing experiment starting in 2016. The data show that plant species diversity peaked in the moderate grazing treatment, and herbivore species diversity was positively correlated with plant species diversity (and hence peaked in the moderate grazing treatment). Moderate grazing promoted parasitoid species diversity, which was positively correlated with herbivore species diversity. However, predator species diversity did not significantly differ among the four treatments. In addition, saprophage species diversity decreased, whereas coprophages increased with increasing grazing levels, such that species richness (but not species diversity of detritivores statistically) was highest in the moderate grazing treatment. Consequently, the species diversity of arthropods as a whole peaked at the moderate grazing level, a phenomenology that is consistent with the intermediate disturbance hypothesis. Considering that moderate grazing has been found to increase plant species diversity, facilitate soil carbon accumulation, and prevent soil erosion, we suggest that moderate grazing would maximize multi-functional ecosystem services.

Mycorrhizae Enhance Soybean Plant Growth and Aluminum Stress Tolerance by Shaping the Microbiome Assembly in an Acidic Soil.

Strongly acidic soils are characterized by high aluminum (Al) toxicity and low phosphorus (P) availability, which suppress legume plant growth and nodule development. Arbuscular mycorrhizal fungi (AMF) stimulate rhizobia and enhance plant P uptake. However, it is unclear how this symbiotic soybean-AMF-rhizobial trio promotes soybean growth in acidic soils. We examined the effects of AMF and rhizobium addition on the growth of two soybean genotypes, namely, Al-tolerant and Al-sensitive soybeans as well as their associated bacterial and fungal communities in an acidic soil. With and without rhizobial addition, AMF significantly increased the fresh shoot and root biomass of Al-tolerant soybean by 47%/87% and 37%/24%, respectively. This increase in plant biomass corresponded to the enrichment of four plant growth-promoting rhizobacteria (PGPR) in the rhizospheric soil, namely, Chitinophagaceae bacterium 4GSH07, Paraburkholderia soli, Sinomonas atrocyanea, and Aquincola tertiaricarbonis. For Al-sensitive soybean, AMF addition increased the fresh shoot and root biomass by 112%/64% and 30%/217%, respectively, with/without rhizobial addition. Interestingly, this significant increase coincided with a decrease in the pathogenic fungus Nigrospora oryzae as well as an increase in S. atrocyanea, A. tertiaricarbonis, and Talaromyces verruculosus (a P-solubilizing fungus) in the rhizospheric soil. Lastly, the compartment niche along the soil-plant continuum shaped microbiome assembly, with pathogenic/saprotrophic microbes accumulating in the rhizospheric soil and PGPR related to nitrogen fixation or stress resistance (e.g., Rhizobium leguminosarum and Sphingomonas azotifigens) accumulating in the endospheric layer. IMPORTANCE Taken together, this study examined the effects of arbuscular mycorrhizal fungi (AMF) and rhizobial combinations on the growth of Al-tolerant and Al-sensitive soybeans as well as their associated microbial communities in acidic soils and concluded that AMF enhances soybean growth and Al stress tolerance by recruiting PGPR and altering the root-associated microbiome assembly in a host-dependent manner. In the future, these findings will help us better understand the impacts of AMF on rhizosphere microbiome assembly and will contribute to the development of soybean breeding techniques for the comprehensive use of PGPR in sustainable agriculture.

Parasitic flies alter the dietary preference of grasshoppers.

Parasitism is known to affect the behavior of host species to enhance parasite dispersal and transmission. However, host behavioral responses to parasitism unrelated to parasite dispersal and transmission have been much less studied. The objective of this study was to determine whether grasshopper hosts infected and uninfected with a parasitic fly (Blaesoxipha sp.) differ in terms of the nutrient content of the diet they consume. We investigated the dietary preferences of two grasshopper species (i.e. Asulconotus chinghaiensis and Chorthippus fallax) in terms of the C/N composition of plant species consumed, and determined whether this affected the egg production of unparasitized and parasitized grasshoppers by flies in a Tibetan alpine meadow. The composition of plants consumed differed significantly between the unparasitized and parasitized grasshoppers. Specifically, the abundance of N-rich legumes was lower and that of high C/N grasses was higher in the diet of the parasitized compared to the unparasitized grasshoppers. Diet N content was higher and C/N was lower in the diet of unparasitized grasshoppers, and parasitized females produced fewer eggs than their unparasitized conspecifics. Future enquiries are needed to understand the specific mechanisms underlying these dietary differences. The effects of parasites on the fitness-associated behavior of hosts should be studied more broadly to better understand parasite evolution and adaptation.

Discrepancies in rhizobacterial assembly caused by glyphosate application and herbicide-tolerant soybean Co-expressing GAT and EPSPS.

There are concerns that the innovation of genetically modified herbicide-tolerant (GMHT) plants, as well as the application of herbicide to such GMHT plants, could have an impact on ecological interactions and unintentionally harm non-targeted organisms. Consequently, we intend to use full-length 16 S rDNA amplicon sequencing to examine changes in the bacterial community in the rhizosphere of GMHT soybean (Z106) harboring 5-enolpyruvylshikimate-3-phosphate synthase and Glyphosate N-acetyltransferase genes and GMHT soybean treated with glyphosate (Z106G). Glyphosate application significantly impacted bacterial alpha diversity (species richness, and Shannon diversity). Permutational multivariate analysis of variance of beta diversity demonstrated that soil compartments and growth stages had a substantial impact on soybean rhizobacterial communities (soil compartments, growth stages, P = 0.001). Community composition revealed that Z106G soils were abundant in Taibaiella and Arthrobacter pascens at maturity, while Chryseobacterium joostei and Stenotrophomonas maltophilia predominated in Z106 soils during flowering. Nitrogen-fixing and phosphate-solubilizing microbes were found in higher proportions in the rhizosphere than in bulk soil, with Sinorhizobium being more abundant in Z106 and Bacillus and Stenotrophomonas being more prevalent in Z106G rhizosphere soils. Collectively, our findings suggest glyphosate application and glyphosate-tolerant soybean as potential regulators of soybean rhizobacterial composition.