Effects of biotic and abiotic factors on the temporal dynamic of bat-fruit interactions

Mutualistic interactions between animals and plants vary over time and space based on the abundance of fruits or animals and seasonality. Little is known about this temporal dynamic and the influence of biotic and abiotic factors on the structure of interaction networks. We evaluated changes in the structure of network interactions between bats and fruits in relation to variations in rainfall. Our results suggest that fruit abundance is the main variable responsible for temporal changes in network attributes, such as network size, connectance, and number of interactions. In the same way, temperature positively affected the abundance of fruits and bats. An increase in temperature and alterations in rainfall patterns, due to human induced climate change, can cause changes in phenological patterns and fruit production, with negative consequences to biodiversity maintenance, ecological interactions, and ecosystem functioning.     

Evaluation of mycotoxins,mycobiota, and toxigenic fungi in selected medicinal plants of Khyber Pakhtunkhwa,Pakistan

Medicinal plants are used worldwide to treat a variety of ailments. Due to the provenance of medicinal plants, they are subjected to contamination by moulds, which may be responsible for spoilage and production of mycotoxins. The investigation was designed to throw light on mycological and mycotoxicological status of some medicinal plants from Pakistan and the result showed 30 % and 26.7 % samples were contaminated with aflatoxins and ochratoxin A, respectively. Mould contamination was present in 90 % samples, of which 70 % exceeded the permissible limits. Opium poppy, licorice root, and Indian rennet were most contaminated samples. The predominant moulds found were Aspergillus flavus, Aspergillus niger, Aspergillus parasiticus, and Penicillium spp. and 31 % of the 47 isolates tested were found to be toxigenic. The findings indicate that the contamination in the medicinal plants may contribute to adverse human health problems. This information would prove helpful for regulatory agencies to establish limits for these contaminants in medicinal plants and will explore ways for export of herbal products to countries where more stringent permissible limits of mycotoxins exist. The study is first of its kind in the country reporting natural occurrence of mycotoxins in medicinal plants in Pakistan.     

Gossypol toxicity and detoxification in Helicoverpa armigera and Heliothis virescens

Gossypol is a polyphenolic secondary metabolite produced by cotton plants, which is toxic to many organisms. Gossypol's aldehyde groups are especially reactive, forming Schiff bases with amino acids of proteins and cross-linking them, inhibiting enzyme activities and contributing to toxicity. Very little is known about gossypol's mode of action and its detoxification in cotton-feeding insects that can tolerate certain concentrations of this compound. Here, we tested the toxicity of gossypol and a gossypol derivative lacking free aldehyde groups (SB-gossypol) toward Helicoverpa armigera and Heliothis virescens, two important pests on cotton plants. Larval feeding studies with these two species on artificial diet supplemented with gossypol or SB-gossypol revealed no detectable toxicity of gossypol, when the aldehyde groups were absent. A cytochrome P450 enzyme, CYP6AE14, is upregulated in H. armigera feeding on gossypol, and has been claimed to directly detoxify gossypol. However, using in vitro assays with heterologously expressed CYP6AE14, no metabolites of gossypol were detected, and further studies suggest that gossypol is not a direct substrate of CYP6AE14. Furthermore, larvae feeding on many other plant toxins also upregulate CYP6AE14. Our data demonstrate that the aldehyde groups are critical for the toxicity of gossypol when ingested by H. armigera and H. virescens larvae, and suggest that CYP6AE14 is not directly involved in gossypol metabolism, but may play a role in the general stress response of H. armigera larvae toward plant toxins.     

Biochemical and Physiological Responses of <i>Arabidopsis thaliana</i> Leaves to Moderate Mechanical Stimulation

Mechanical stimulation of plants can be caused by various abiotic and biotic environmental factors. Apart from the negative consequences, it can also cause positive changes, such as acclimatization of plants to stress conditions. Therefore, it is necessary to study the physiological and biochemical mechanisms underlying the response of plants to mechanical stimulation. Our aim was to evaluate the response of model plant Arabidopsis thaliana to a moderate force of 5 N (newton) for 20 s, which could be compared with the pressure caused by animal movement and weather conditions such as heavy rain. Mechanically stimulated leaves were sampled 1 h after exposure and after a recovery period of 20 h. To study a possible systemic response, unstimulated leaves of treated plants were collected 20 h after exposure alongside the stimulated leaves from the same plants. The effect of stimulation was assessed by measuring oxidative stress parameters, antioxidant enzymes activity, total phenolics, and photosynthetic performance. Stimulated leaves showed increased lipid peroxidation 1 h after treatment and increased superoxide dismutase activity and phenolic oxidation rate after a 20-h recovery period. Considering photosynthetic performance after the 20-h recovery period, the effective quantum yield of the photosystem II was lower in the stimulated leaves, whereas photochemical quenching was lower in the unstimulated leaves of the treated plants. Nonphotochemical quenching was lower in the stimulated leaves 1 h after treatment. Our study suggested that plants sensed moderate force, but it did not induce pronounced change in metabolism or photosynthetic performance. Principal component analysis distinguished three groups–leaves of untreated plants, leaves analysed 1 h after stimulation, while stimulated and unstimulated leaves of treated plants analysed 20 h after treatment formed together the third group. Observed grouping of stimulated and unstimulated leaves of treated plants could indicate signal transduction from the stimulated to distant leaves, that is, a systemic response to a local application of mechanical stimuli.     

Cuticular wax variation in the tomato (Solanum lycopersicum L.), related wild species and their interspecific hybrids

The tomato (Solanum lycopersicum L.) is one of the world's most important vegetable crop species. Among the many tomato accessions available, only a few are tolerant to abiotic stresses, which are responsible for the majority of the crop losses worldwide. Wild tomato species are then secondary gene pool in the breeding of more resistant tomato cultivars. In the current study, the composition of leaf cuticular waxes from fourteen tomato accessions, including S. lycopersicum, Solanum pennellii, Solanum pimpinellifolium, and their interspecific hybrids was studied in order to select the most adequate chemotaxonomic markers. Total cuticular wax load of S. pennellii plants was much higher than in the other plant species. Hydrocarbons were usually the most abundant wax components, followed by minor quantities of triterpenes and other compounds. Interspecific hybrids showed intermediate wax characteristics. The amount and composition of surface waxes were not correlated with the abiotic stress tolerance in S. lycopersicum. The composition of the hydrocarbon fraction was the least variable both within a single accession and between all the plants studied. Based on the results, cuticular hydrocarbons are proposed as potential chemotaxonomic markers in the classification of tomato and related species.     

The biotrophy-associated secreted protein 4 (BAS4) participates in the transition of Magnaporthe oryzae from the biotrophic to the necrotrophic phase

The physiological and metabolic processes of host plants are manipulated and remodeled by phytopathogenic fungi during infection, revealed obvious signs of biotrophy of the hemibiotrophic pathogen. As we known that effector proteins play key roles in interaction of hemibiotrophic fungi and their host plants. BAS4 (biotrophy-associated secreted protein 4) is an EIHM (extrainvasive hyphal membrane) matrix protein that was highly expressed in infectious hyphae. In order to study whether BAS4 is involved in the transition of rice blast fungus from biotrophic to necrotrophic phase, The susceptible rice cultivar Lijiangxintuanheigu (LTH) that were pre-treated with prokaryotic expression product of BAS4 and then followed with inoculation of the blast strain, more serious blast disease symptom, more biomass such as sporulation and fungal relative growth, and lower expression level of pathogenicity-related genes appeared in lesion of the rice leaves than those of the PBS-pretreated-leaves followed with inoculation of the same blast strain, which demonstrating that BAS4 invitro changed rice defense system to facilitate infection of rice blast strain. And the susceptible rice cultivar (LTH) were inoculated withBAS4-overexpressed blast strain, we also found more serious blast disease symptom and more biomass also appeared in lesion of leaves inoculated with BAS4-overexpressed strain than those of leaves inoculated with the wild-type strain, and expression level of pathogenicity-related genes appeared lower in biotrophic phase and higher in necrotrophic phase of infection, indicating BAS4 maybe in vivo regulate defense system of rice to facilitate transition of biotrophic to necrotrophic phase. Our data demonstrates that BAS4 in vitro and in vivo participates in transition from the biotrophic to the necrotrophic phase of Magnaporthe oryzae.     

Potential detoxification of gossypol by UDP-glycosyltransferases in the two Heliothine moth species Helicoverpa armigera and Heliothis virescens

The cotton bollworm Helicoverpa armigera and the tobacco budworm Heliothis virescens are closely related generalist insect herbivores and serious pest species on a number of economically important crop plants including cotton. Even though cotton is well defended by its major defensive compound gossypol, a toxic sesquiterpene dimer, larvae of both species are capable of developing on cotton plants. In spite of severe damage larvae cause on cotton plants, little is known about gossypol detoxification mechanisms in cotton-feeding insects. Here, we detected three monoglycosylated and up to five diglycosylated gossypol isomers in the feces of H. armigera and H. virescens larvae fed on gossypol-supplemented diet. Candidate UDP-glycosyltransferase (UGT) genes of H. armigera were selected by microarray studies and in silico analyses and were functionally expressed in insect cells. In enzymatic assays, we show that UGT41B3 and UGT40D1 are capable of glycosylating gossypol mainly to the diglycosylated gossypol isomer 5 that is characteristic for H. armigera and is absent in H. virescens feces. In conclusion, our results demonstrate that gossypol is partially metabolized by UGTs via glycosylation, which might be a crucial step in gossypol detoxification in generalist herbivores utilizing cotton as host plant.     

Determining tipping points in aquatic ecosystems: The case of biodiversity and chlorophyll α relations in fish pond systems

The management of biodiversity in aquatic ecosystems requires knowing the state of water quality linked to regime shifts in various taxonomic groups. We examine this question by studying the fish ponds in the Dombes region, France. These waterbodies are characterized by a high diversity of species. High levels of nutrients due to certain fish farming practices may cause significant eutrophication leading to loss in biodiversity and a shift from high coverage of aquatic vegetation to phytoplankton dominance may also be observed. The aim of this study is to assess tipping points, thresholds for effect, along a gradient of chlorophyll α in different taxonomic groups: aquatic vascular plants, phytoplankton, dragonflies and aquatic macro-invertebrates. Tipping points are analyzed with three different statistical methods: a method which evaluates tipping points with a difference in the mean (TMEAN), a second method which evaluates tipping point by comparing the mean and linear regressions before and after the tipping point (FSTAT) and third a method which evaluates linear regressions with a pivotal tipping point (SEGMENTED). We also compare tipping points for the different taxonomic groups using five different diversity indices: Observed richness, Jackknife first order, Fisher's alpha, Simpson index and Evenness.Our results show that there is an important variation in tipping points following the three statistical methods, but the SEGMENTED is the best method for evaluating tipping points. We observe a high difference of tipping point values for the different taxonomic groups depending on the diversity indices used. Jackknife first order has a better performance to evaluate a eutrophic change according to the diversity than the other indices.In all taxonomic groups, aquatic vascular plants are the most impacted by the chlorophyll α and almost all their tipping points are observed around 60 μg/L chlorophyll α concentrations. No significant relationship is found between chlorophyll α and phytoplankton diversity, while the two other groups, dragonflies and macro-invertebrates, are both impacted by the chlorophyll α but their relevant tipping points are situated in higher values than aquatic vascular plants.     

Root distribution in an Amazonian seasonal forest as derived from δ13C profiles

Carbon isotope ratios of the main stem in trees, saplings, and seedlings were correlated with their main stem diameter in an Amazonian seasonal forest. This correlation became the basis of using carbon isotope ratios of roots from various levels of the soil profile in order to determine root distribution from emergent, canopy and subcanopy trees, saplings and herbaceous understorey plants. It was observed that the distribution of roots in the soil profile is horizontally and vertically heterogeneous. Pockets of roots from saplings or herbaceous understorey plants were found as deep as 4 m and pockets of roots from emergent trees were found as shallow as 1 m depth.     

Digestive proteolysis in the Colorado potato beetle,Leptinotarsa decemlineata: Activity-based profiling and imaging of a multipeptidase network

The Colorado potato beetle (CPB), Leptinotarsa decemlineata, is a major pest of potato plants, and its digestive system is a promising target for development of pest control strategies. This work focuses on functional proteomic analysis of the digestive proteolytic enzymes expressed in the CPB gut. We identified a set of peptidases using imaging with specific activity-based probes and activity profiling with selective substrates and inhibitors. The secreted luminal peptidases were classified as: (i) endopeptidases of cathepsin D, cathepsin L, and trypsin types and (ii) exopeptidases with aminopeptidase (cathepsin H), carboxypeptidase (serine carboxypeptidase, prolyl carboxypeptidase), and carboxydipeptidase (cathepsin B) activities. The proteolytic arsenal also includes non-luminal peptidases with prolyl oligopeptidase and metalloaminopeptidase activities. Our results indicate that the CPB gut employs a multienzyme network of peptidases with complementary specificities to efficiently degrade ingested proteins. This proteolytic system functions in both CPB larvae and adults and is controlled mainly by cysteine and aspartic peptidases and supported by serine and metallopeptidases. The component enzymes identified here are potential targets for inhibitors with tailored specificities that could be engineered into potato plants to confer resistance to CPB.     

Know your ABCs: Characterization and gene expression dynamics of ABC transporters in the polyphagous herbivore Helicoverpa armigera

Polyphagous insect herbivores are adapted to many different secondary metabolites of their host plants. However, little is known about the role of ATP-binding cassette (ABC) transporters, a multigene family involved in detoxification processes. To study the larval response of the generalist Helicoverpa armigera (Lepidoptera) and the putative role of ABC transporters, we performed developmental assays on artificial diet supplemented with secondary metabolites from host plants (atropine-scopolamine, nicotine and tomatine) and non-host plants (taxol) in combination with a replicated RNAseq experiment. A maximum likelihood phylogeny identified the subfamily affiliations of the ABC transporter sequences. Larval performance was equal on the atropine-scopolamine diet and the tomatine diet. For the latter we could identify a treatment-specific upregulation of five ABC transporters in the gut. No significant developmental difference was detected between larvae fed on nicotine or taxol. This was also mirrored in the upregulation of five ABC transporters when fed on either of the two diets. The highest number of differentially expressed genes was recorded in the gut samples in response to feeding on secondary metabolites. Our results are consistent with the expectation of a general detoxification response in a polyphagous herbivore. This is the first study to characterize the multigene family of ABC transporters and identify gene expression changes across different developmental stages and tissues, as well as the impact of secondary metabolites in the agricultural pest H. armigera.     

Evidence for growth ofSporothrix schenckii on dead but not on living sphagnum moss

When clinical isolates ofSporothrix schenckii were inoculated onto the apices of living or dead sphagnum moss plants maintained under growth chamber conditions, populations of the fungus, assessed by standard dilution plate methods, increased swiftly up to about 70-fold on moist, dead plants but did not increase on the live moss. Light and scanning electron microscopy revealed fungal growth and sporulation on and within dead plants, but no evidence of either on live plants. These data provide indirect support for the contention thatS. schenckii does not grow on living sphagnum in bogs, but rather that sporotrichosis epidemics associated with sphagnum moss are likely to result from contamination of the dead plants at some point(s) in the chain of events during or after harvest. One practical implication of our results is that precautions should be taken to insure that sphagnum moss is stored dry and that it is not wetted any sooner than necessary before use. We also report here improvement of the Mycose isolation medium by an increase in cycloheximide from 400 to 800 mg/l, chloramphenicol from 50 to 250 mg/l, and the addition of rifampicin at 20 mg/l.     

Isolation of NBS-LRR RGAs from invasive Wedelia trilobata and the calculation of evolutionary rates to understand bioinvasion from a molecular evolution perspective

Supports for the molecular evolution of host–pathogen interactions on enemy release hypothesis are rare. According to the theory of plant immunity and the coevolution of hosts and pathogens, we hypothesized that the evolutionary rate (d_N/d_S) of resistance genes (R-genes) in invasive plants would be greater than in non-invasive plants, assuming that based on the enemy release hypothesis, the former would suffer less selection stress from co-evolutionary specialist pathogens. To test our hypothesis, we isolated and analyzed the conserved nucleotide-binding sites (NBS) of resistance gene analogues (RGAs) of an invasive weed, Wedelia trilobata (WTRGA). We then used the information in GenBank to compare the d_N/d_S of the NBS R-gene/RGAs in invasive and homologous non-invasive plants. Three W. trilobata NBS RGA sequences were obtained, belonging to the Toll/Interleukin-1 receptor (TIR) (WTRGA1 and WTRGA2) and non-TIR subclasses (WTRGA3). Compared with the homologous non-invasive plants, the invasive plants showed a significantly greater d_N/d_S for TIR NBS R-gene/RGAs (p < 0.0001), supporting our hypothesis. Future research should include an examination of R-genes/RGAs from more invasive plants on a population level to understand diversity and R-gene functions in invasive plant species, as well as to explore how disease resistance allows plants to adapt to changing pathogen stresses.     

Chloroplast tRNAAsp: nucleotide sequence and variation of in vivo levels during plastid maturation

Two chloroplast tRNA^Asp species from barley were purified by chromatography on benzoylated DEAE-cellulose and sequenced. They differ in the modification at position 34, where queuosine (Q) is present in one of the species. The same chromatographic procedure yielded only one tRNA^Glu species, corroborating the assumption that the same tRNA^Glu species participates in both protein and chlorophyll biosynthesis. The level of tRNA^Glu remains unchanged after light treatment of etiolated seedlings, whereas the amount of tRNA^Asp decreases to about 50% relative to the level of dark-grown plants.     

Effects of Different Spectra from LED on the Growth,Development and Reproduction of <i>Arabidopsis thaliana</i>

Light is the major source of energy for plants and as such has a profound effect on plant growth and development. Red and blue lights have been considered to best drive photosynthetic metabolism and are beneficial for plant growth and development, and green light was seen as a signal to slow down or stop. In this study, Arabidopsis thaliana (Arabidopsis) was used to investigate the effects of red, blue and green lights on the growth and development of plants from seed germination to seeding. Results demonstrated that red light showed a promotion effect but blue light a prohibition one in most stages except for the flowering time in which the effect of each light was just reversed. When mixed with red or blue light, green light generally at least partially cancelled out the effects caused by each of them. Results also showed that the same number of photons the plant received could cause different effects and choosing the right combination of different color of lights is essential in both promoting the growth and development of plants and reducing the energy consumption of lighting in plant factory.     

Essential oil yield,composition, bioactivity and leaf morphology of Juniperus oxycedrus L. from Bulgaria and Serbia

Juniperus oxycedrus L. (Cupressaceae Bartlett) is widely distributed in countries with a Mediterranean climate. All plant parts contain highly aromatic essential oil (EO) and recently there have been efforts to introduce it as a cultivated crop. The species is known for its large morphological and chemical variation and its debatable taxonomic status. This study aimed to (1) compare content, composition, and antimicrobial activity of J. oxycedrus EO samples from plants growing in Bulgaria and Serbia, and (2) quantify morphological variations of leaves. Тhe EO content (yield) in dried juniper leaves varied from 0.06% (Кopaonik, Serbia) to 0.24% (Markovo, Bulgaria). We identified 51 EO constituents, belonging to monoterpenes, sesquiterpenes, and diterpenes. The class monoterpenes (monoterpene hydrocarbons and oxygenated monoterpenes) were the predominant compounds, representing 38.6–65.4% of the total EO, consisting primarily of α-pinene, limonene, sabinene, β-pinene, and β-myrcene. In addition, α-pinene was the major oil constituent in plants from all locations. Sesquiterpenes (sesquiterpene hydrocarbons and oxygenated sesquiterpenes) were the second largest class of constituents, which represented 19.3% tо 33.6% of the total EO. γ-Elemene was found only in the EO of J. oxycedrus from Bulgaria, while a high concentration of α-curcumene was found only in samples from Serbia (7.5–7.8%). Significant differences in antimicrobial activity of the EO were found in bacterial strains Bacillus cereus and Streptococcus pneumoniae. There was no significant difference among the mean leaf width of the six combinations location x sex, and the overall leaf mean width was 1.24 mm. However, there was a significant difference between the mean leaf lengths. In this study, none of the studied populations had a higher concentration of limonene than of α-pinene, indicating that the flora of the two countries include J. oxycedrus and not the previously reported J. deltoides. The results revealed significant variation in EO profile that may contribute to the development of new cultivars of J. oxycedrus.