Role of castor plant phenolics on performance of its two herbivores and their impact on egg parasitoid behaviour

Plants are reported to reconfigure their metabolism and generate secondary metabolites as a defence response towards herbivore attack. In this study we aimed to evaluate the anti-nutritive and defensive properties of phenolic compounds that were enhanced in Castor, Ricinus communis L. plant due to herbivore damage. Through HPLC studies it was observed that damage caused by two pests, Achaea janata L. and Spodoptera litura F. on the castor plant resulted in quantitative enhancement of syringic, coumaric, cinnamic and vanillic acids. These phenolic compounds showed considerable impact on insect’s feeding and their toxic nature towards the herbivores was determined by estimating three detoxification mid-gut enzymes, glutathione-s-transferase, carboxyl esterase and β-glucosidase. In the ovipositional and orientation bioassays with the egg parasitoid Trichogramma chilonis Ishii, syringic and coumaric acids had shown good kairomonal activity, thus defining a new role of these chemicals in indirect plant defence. Thus, this study demonstrates a quantitative association between plant phenolic accumulation and the elevated reproductive performance of the herbivore natural enemies, by illustrating the role of plant phenolics in both the direct and indirect induced plant defences.     

PHYSIOLOGICAL EFFECTS OF RESVERATROL AND COUMARIC ACID ON TWO MAJOR GROUNDNUT PESTS AND THEIR EGG PARASITOID BEHAVIOR

Groundnut, Arachis hypogea L., is one of the plant species that synthesizes phenolic compounds, resveratrol and coumaric acid. They are induced as a defense strategy in plant in response to feeding lepidopterans. The present study investigated the role of resveratrol and coumaric acid in producing antiherbivore effects as a direct defense against two major groundnut pests, Spodoptera litura F. and Amsacta albistriga W., and in indirect defense by attracting the egg parasitoid Trichogramma chilonis Ishii under laboratory conditions. The phenolic compounds deterred the feeding of both pests and caused reduction in the larval weights in a dose‐dependent manner in leaf disk bioassays. Antioxidant mechanisms of larvae fed with these phenols were measured by estimating the activities of superoxide dismutase (SOD), ascorbate peroxidase (APOX), and catalase (CAT). Enzyme activities increased significantly in experimental larvae, more so in resveratrol‐treated than in coumaric acid treated larvae. Feeding larvae with resveratrol and coumaric acid resulted in enhanced activities of detoxifying enzymes, carboxyl esterase (EST), and glutathione‐S‐transferase (GST) in the midgut tissues of both species, indicating the toxic nature of these compounds. Trichogramma chilonis was more attracted toward coumaric acid treatments in Y‐olfactometer tests than to resveratrol. This study contributes to the understanding of the roles of resveratrol and coumaric acid in direct as well as indirect defense, we infer a possible utilization of these compounds in alternate measures of groundnut pest control in future.     

Uptake of phenolic compounds from plant foods in human intestinal Caco-2 cells

In continuation of our studies on the bioaccessibility of phenolic compounds from food grains as influenced by domestic processing, we examined the uptake of phenolics from native/sprouted finger millet (Eleucine coracana) and green gram (Vigna radiata) and native/heat-processed onion (Allium cepa) in human Caco-2 cells. Absorption of pure phenolic compounds, as well as the uptake of phenolic compounds from finger millet, green gram, and onion, was investigated in Caco-2 monolayer model. Transport of individual phenolic compounds from apical compartment to the basolateral compartment across Caco-2 monolayer was also investigated. Sprouting enhanced the uptake of syringic acid from both these grains. Open-pan boiling reduced the uptake of quercetin from the onion. Among pure phenolic compounds, syringic acid was maximally absorbed, while the flavonoid isovitexin was least absorbed. Apparent permeability coefficient P_(app) of phenolic compounds from their standard solutions was 2.02 × 10^?6 cm/s to 8.94 × 10^?6 cm/s. Sprouting of grains enhanced the uptake of syringic acid by the Caco-2 cells. Open-pan boiling drastically reduced the uptake of quercetin from the onion. The permeability of phenolic acids across Caco-2 monolayer was higher than those of flavonoids.     

Effect of host feeding on life history traits of <Emphasis Type="Italic">Tamarixia radiata</Emphasis>, parasitoid of the Asian citrus psyllid, <Emphasis Type="Italic">Diaphorina citri</Emphasis>

The parasitoid Tamarixia radiata (Hymenoptera: Eulophidae) is being used for the biological control of the Asian citrus psyllid (ACP), Diaphorina citri (Hemiptera: Liviidae). The parasitoid is strongly synovigenic, as it is born with very few mature eggs. Synovigenic insects need to feed on host haemolymph to mature additional eggs, and are able to resorb mature eggs to allocate resources toward maintenance. We investigated the effect of host feeding on parasitism, longevity and egg load dynamics, and estimated egg maturation and resorption rates. Although host feeding does not increase survival or longevity, it results in increased parasitization rates when parasitoids are seven days old, and that a single host meal leads to an average gain of three eggs. We discuss the importance of these data to predict the foraging and parasitization behavior of T. radiata in the field, and to potentially improve current efforts to control ACP.     

<Emphasis Type="Italic">Synechocystis</Emphasis> PCC 6803 cells heterologously expressing bacterial tyrosine ammonia lyase can use exogenous tyrosine for <Emphasis Type="Italic">p</Emphasis>-coumaric acid production

Phototrophic cyanobacteria may be considered as an alternative host for producing numerous bioactive compounds. We demonstrate that the Synechocystis PCC 6803 expressing tyrosine ammonia-lyase from Rhodobacter sphaeroides under Ptrc1O promoter produce p-coumaric acid at a rate three times higher than that under Ptrc1Ocore promoter, accounting for 18.4 ± 1.5 μg of p-coumaric acid per 10⁸ cells (0.36 mg L^?1). Additionally, our study is the first report to show the biotransformation of tyrosine to p-coumaric acid reaching a maximum 2.4-fold increase when 0.5 mM tyrosine was supplemented to the growth medium. Liquid chromatography-mass spectrometry analysis revealed the occurrence of diverse patterns of metabolites under different concentrations of supplemented tyrosine, suggesting that it is used in additional metabolic pathways.     

Phenolic compounds profiling in shake flask and bioreactor system cell cultures of <Emphasis Type="Italic">Scrophularia striata</Emphasis> Boiss

Variation in levels of phenolic acids and flavonoids in Scrophularia striata Boiss. cells cultured in both shake flask and bioreactor in vitro systems, was studied at different growth phases. Four phenolic acids (cinnamic, salicylic, coumaric, and caffeic acid), one stilbenoid (resveratrol), and seven flavonoids (diosmin, rutin, kaempferol, catechin, myricetin, quercetin, and luteolin) were analyzed by high-performance liquid chromatography with photodiode array detection. Production of phenolics in the bioreactor was higher than in shake flasks. Catechin was the most abundant flavonoid in both culture systems, while quercetin, which was detected only in the bioreactor, was the lowest amount represented (32.82 μg g^?1 DW). Resveratrol accumulation in bioreactor cultures was 59.84-fold higher than that in shake flasks. Moreover, hierarchical clustering analysis based on Pearson’s correlation coefficient confirmed a positive correlation between the growth phase and some metabolites. The flavonoid accumulation increased with the cells’ physiological age in the bioreactor. Principal component analysis showed that the time course of induction of phenolic acids, flavonoids, and a stilbenoid (resveratrol) was significantly correlated. These findings highlight the capacity of S. striata for large-scale production of desired phenolics using a bioreactor system.     

Bioactive sesquiterpene,plasticizer, and phenols from the fungal endophytes of <Emphasis Type="Italic">Polygonum chinense</Emphasis> L.

There is a constant need for novel antibiotic and antioxidant sources due to the ever-increasing resilience of pathogens and the occurrence of chronic diseases. The natural sources of these agents have advantages due to lower production cost, structural variation, and uses of active compounds for pharmaceutical uses. The microbes living in planta termed “endophytes” are alternative sources of host bioactive compounds. In this study, ten endophytic fungi were isolated from Polygonum chinense L. and identified by sequencing of the internal transcribed spacer regions. The fungal strains were fermented and the ethyl acetate extracts were evaluated for antimicrobial and antioxidant capacities. Almost 80% of the endophytes showed antibacterial potency against one or more pathogenic bacteria. Among all strains, Penicillium canescens showed broad-spectrum antimicrobial activity against gram-positive and gram-negative pathogens as well as significant antioxidative and DNA protective capacities. The strain Fusarium chlamydosporum displayed significant anti-radical (126.8?±?6.7 μg/ml) and ferric reducing (84.7?±?2.1 mg AA/g dry extract) capacities. The bio-autography, chromatography, and mass spectroscopy analyses of P. canescens extract revealed the presence of sesquiterpene (germacrene), plasticizer (phthalic acid ester) along with phenolic acids, flavonoid (quercetin), and short chain hydrocarbons. The secondary metabolites of F. chlamydosporum were identified with phenolic acids as bioactive compounds by chromatography and mass spectroscopy. This study indicates P. chinense endophytes as potential sources of antimicrobial and antioxidant compounds for novel drug discovery.     

Effect of <Emphasis Type="Italic">Fopius arisanus</Emphasis> oviposition experience on parasitization of <Emphasis Type="Italic">Bactrocera dorsalis</Emphasis>

Fopius arisanus (Sonan) (Hymenoptera: Braconidae) is an important fruit fly parasitoid, successfully introduced in programs of classical biological control around the world. One aspect of its reproductive biology that has received increasing attention is the role of learning on parasitization by individual females. In this study, we examined the effect of previous oviposition opportunities on the reproductive success and progeny sex ratio of F. arisanus on eggs of Bactrocera dorsalis Hendel (Diptera: Tephritidae). Our results indicate that the proportion of eggs parasitized and parasitoid yield both increase with increasing experience, as acquired by individual females via previous oviposition opportunities. These effects are shown to be unrelated to parasitoid age out to three weeks, which we found not to have an effect on parasitization, yield, or sex ratio. We discuss the implications of our results on mass rearing and also in terms of the efficiency of this biological control agent in the field.     

<Emphasis Type="Italic">Fusarium</Emphasis> Infection Causes Phenolic Accumulations and Hormonal Disorders in <Emphasis Type="Italic">Orobanche</Emphasis> spp.

The physiological effects of Fusarium oxysporum on in-root parasitic weed, Orobanche spp. (broomrape) with references to change in plant hormones and secondary plant constituents were investigated. The levels of IAA, GA, ABA and JA in the experimental group were significantly lower than those in the control group, while the level of SA was higher in the experimental group. In secondary metabolic studies, the quantities of various phenols were measured in the two groups and catechin, syringic acid and p-coumaric acid amounts were significantly higher in the experimental group than in the control group, unlike gallic acid which have a lower amount. Consequently, in the light of all data, it was concluded that Fusarium oxysporum (1) causes heavy hormonal disorder, (2) triggered only SA-mediated defense and (3) induced intensively accumulation of phenolic substances in orobanche. Fusarium oxysporum causes lethal physiological damage on Orobanche spp.     

Improvement of bioactive compound accumulation in adventitious root cultures of an endangered plant species, <Emphasis Type="Italic">Oplopanax elatus</Emphasis>

Efficiently culturing adventitious roots (ARs) has become an alternative route for the protection and utilization of endangered plant resources. In the present study, to improve accumulation of bioactive compounds (polysaccharides, phenolics, and flavonoids) in AR cultures of endangered plant species—Oplopanax elatus—effects of methyl jasmonate (MeJA) and salicylic acid (SA) were investigated. The optimal concentration of MeJA was 200 μM and SA was 100 μM for enhancement of polysaccharide, phenolic, and flavonoid contents. In addition, MeJA (200 μM) was more suitable than SA (100 μM) for polysaccharide and flavonoid production, but both elicitors were equally favorable for phenolic production. During AR bioreactor culture, MeJA was as an elicitor to study the effect of its addition time and contact time. Contents of polysaccharides, phenolics, and flavonoids increased when MeJA was added to culture medium after 40 days of culture, but the increased degree was lower and the AR biomass significantly inhibited. However, when MeJA was added to culture medium after 30 days of culture, polysaccharide, phenolic, and flavonoid contents dramatically increased without AR biomass decrease; the maximum productivity of three bioactive compounds was found on day 8 after the MeJA treatment. Therefore, a novel elicitation method during bioreactor culture of O. elatus ARs was established in the present study, the method could be applied to commercial production of O. elatus products in the future.     

Thermal tolerance of potential <Emphasis Type="Italic">Trichogramma</Emphasis> strains for mass-production and paddy field release in the Greater Mekong Subregion

In order to identify suitable Trichogramma strains for mass production and successful control of key lepidopteran pests in paddy fields in the Greater Mekong Subregion, the effects of high temperatures on wasp life history traits were compared among three strains of Trichogramma chilonis Ishii and two strains of Trichogramma ostriniae Pang et Chen, collected from paddy fields in the region. At a rearing temperature of 25 °C, life history traits differed significantly among the five strains tested and the three T. chilonis strains and a T. ostriniae strain were of high performance. When female wasps were exposed to higher temperatures commonly encountered in the region, negative effects were observed on key life history parameters of adult females and their offspring at 34 °C, which became even more serious at 37 °C. In particular, the two T. ostriniae strains were not able to successfully develop to adulthood at all at 37 °C while for the T. chilonis strains adult emergence was significantly reduced. In addition, the emerged offspring females lived only around one day and no parasitism was observed. When the three T. chilonis strains were exposed to 37 °C for 4 to 12 h at prepupal and pupal stages, reflecting heat shocks that the released Trichogramma may experience in the field, adult emergence was significantly reduced after an exposure time of 12 h. In summary, the three T. chilonis strains show a relatively high potential for incorporation in a biological control program in the target region. Our results also highlight that tests at both rearing and field temperatures are necessary in selection of potential Trichogramma strains for an inundative release program where there is significant difference between rearing and field temperatures.     

In search of secondary plants to enhance the efficiency of cabbage seed weevil management

The infestation rate and parasitoid communities of Ceutorhynchus obstrictus (Marsham) (Coleoptera: Curculionidae) were assessed on seven spring sown brassicaceous plant species to find potential secondary plants that might help increase the parasitism rates of this serious oilseed pest. Over the three-year study, the average infestation rate of pods by C. obstrictus remained below 10 % for each plant species. Despite the low pest abundance, C. obstrictus was parasitized by hymenopteran parasitoids on all plant species, except on Eruca vesicaria subsp. sativa ((Mill.) Thell.). Parasitism rates were remarkably high: between 33.7 and 70.8 % on average and peaked at 94.7 % on Raphanus sativus (L.) var. oleiformis (Pers.). Not only was the parasitism rate high on R. sativus, but it also had a different parasitoid species composition consisting mainly of egg parasitoids (Mymaridae), while on the other plant species larval parasitoids (Pteromalidae) dominated. These findings are important for planning new sustainable pest management approaches.     

Effects of Agitating Culture Condition on the Growth,Metabolic and Carotenoid Profiles of <Emphasis Type="Italic">Lemna paucicostata</Emphasis>

The effects of agitating culture conditions on the growth, metabolic and carotenoid profiles of Lemna paucicostata were investigated in this study using gas chromatography–mass spectrometry and liquid chromatography–mass spectrometry. Lemna paucicostata cultivated under static and agitating conditions showed no significant difference in total plant growth. Various alcohols, amino acids, fatty acids, organic acids, sugars, and phenolic compounds were identified in the sample. The higher relative yields of ferulic acid and GABA were obtained under the agitating condition on day 28, whereas that of coumaric acid was higher under the static condition on day 42. Significantly higher relative yields of antheraxanthin and lutein were achieved under agitation condition than static condition on day 42. In addition, fucoxanthinol was identified for the first time in L. paucicostata culture. The results of this study suggest that static and agitating culture conditions, as well as optimal harvest timing, should be selected according to the target products including ferulic acid, GABA, coumaric acid, antheraxanthin and lutein in L. paucicostata culture.     

Development of <Emphasis Type="Italic">Cleruchoides noackae</Emphasis>, an egg-parasitoid of <Emphasis Type="Italic">Thaumastocoris peregrinus</Emphasis>, in eggs laid on different substrates,with different ages and post-cold storage

Cleruchoides noackae Lin and Huber (Hymenoptera: Mymaridae) is an egg parasitoid of Thaumastocoris peregrinus Carpintero and Dellapé (Hemiptera: Thaumastocoridae). The parasitism and development of C. noackae was studied in T. peregrinus eggs of different ages, laid on eucalyptus leaves or paper towel and stored at 5 °C. The emergence, sex ratio and development of C. noackae and hatched nymphs of T. peregrinus were evaluated. This parasitoid had an emergence rate higher than 60% from zero to one, one to two, and two to three-day old eggs and lower than 10% for those 3–4 and 4–5 days old. The female proportion was 78% and the egg-adult period for C. noackae was 19.5 days. The use of T. peregrinus eggs up to three days old, laid on paper towel and stored at 5 °C for 14 days did not affect the biological parameters of C. nockae and should be used for mass rearing of this parasitoid.     

Attraction of <Emphasis Type="Italic">Telenomus podisi</Emphasis> to volatiles induced by <Emphasis Type="Italic">Euschistus heros</Emphasis> in three different plant species

Specialized natural enemies that forage for polyphagous hosts need to locate hosts on different plants. Telenomus podisi (Hymenoptera: Platygastridae) is a stink bug egg parasitoid with a preference for Euschistus heros (Hemiptera, Pentatomidae), a polyphagous species. The aim of this study was to evaluate the induction of defences in three E. heros host plants: maize (Zea mays), sunflower (Helianthus annuus) and pigeon pea (Cajanus cajan). We hypothesized that E. heros damage to these three plants enhances the attraction of the parasitoid T. podisi as has been observed in other systems. Using Y-tube olfactometer bioassays, we tested parasitoid responses to combinations of the following odour sources: clean air, undamaged plants and plants damaged by stink bug feeding. Volatiles were collected by means of dynamic headspace collection and analysed by gas chromatography coupled to mass spectrometry. T. podisi did not distinguish odours from undamaged plants against air for any of the three plant species. For maize, the parasitoid preferred the odour from herbivore-damaged plants over both clean air and undamaged plants. For sunflower, the parasitoid only preferred the odour of herbivore-damaged plants over the odour of undamaged plants. For pigeon pea, no preferences were observed. Quantitative differences in the volatile profile of damaged and undamaged plants were observed in each plant species. We conclude that sunflower and maize plants, when damaged by E. heros, release volatiles that attract the parasitoid T. podisi; the parasitoid appears to use a different blend composition to distinguish herbivore-damaged plants of each species.     

Toxicity and kinetics of spinosad in different developmental stages of the endoparasitoid <Emphasis Type="Italic">Hyposoter didymator</Emphasis> (Hymenoptera: Ichneumonidae) and its host <Emphasis Type="Italic">Spodoptera littoralis</Emphasis> larvae (Lepidoptera: Noctuidae)

The toxicity of spinosad was evaluated using the RaPID Assay^® Spinosad immunosorbent assay in different developmental stages of the parasitoid, Hyposoter didymator, and in its host, fourth-instar larvae of the cotton leafworm Spodoptera littoralis. Spinosad was applied directly to pupae and adults of H. didymator (ingestion or topical application) or to the immature stages of the parasitoid via the host larvae. Low amounts of spinosad were recovered from S. littoralis host larvae after topical treatment, and the compound was mainly retained in the hemolymph. Amounts of spinosad detected in third-instar larvae of H. didymator, pulled out from the hemolymph of parasitized S. littoralis larvae, were 85 pg (3.57 ng a.i./g body weight) in dead larvae, and 82 pg (3.42 ng a.i./g body weight) in alive individuals. After topical treatment of H. didymator cocoons, most of the compound was retained in the silken cocoon, preventing contamination of the pupa. Also in the parasitoid adults, relatively low amounts of spinosad were accumulated in the body overall, but half of all the insecticide recovered was found in the ovaries. The kinetic results obtained help to better understand the toxicity of spinosad in the complex S. littoralis–H. didymator, and to ascertain the compatibility between spinosad and the parasitoid for optimizing the control of lepidopteran pests.     

Acute Toxicity and Sublethal Effects of Terpenoids and Essential Oils on the Predator <Emphasis Type="Italic">Chrysoperla externa</Emphasis> (Neuroptera: Chrysopidae)

The search for new safer insecticides has increased in recent agriculture. Botanical compounds such as terpenoids and plant essential oils with insecticidal activity could represent important tools in pest management, and their risk assessment against non-target organisms is necessary since they may serve as a precursor for the synthesis of new insecticide active ingredients. For this study, the acute toxicity and sublethal effects of seven terpenoids and three essential oils with recognized insecticidal activity were evaluated on the predator Chrysoperla externa (Hagen) (Neuroptera: Chrysopidae) in laboratory bioassays. Results indicate that these compounds feature relative selectivity to the predator C. externa; however, sublethal effects on reproduction were recorded for some compounds. The phenolic monoterpenoids carvacrol and thymol were more acutely toxic than other terpenoids screened, with LD₅₀ <20,000 μg/g; however, they were less toxic than natural pyrethrins (toxicity standard) in these bioassays. Sublethal effects on fecundity and fertility were observed for R-(+)-limonene, while oregano oil only affected fecundity. The compounds evaluated here have potential to be used as insecticides and can serve as backbone for future synthesis of selective active ingredients; however, a complete risk assessment to C. externa and other non-target organisms is necessary for their incorporation in future crop protection paradigms.     

Egg Parasitoid Complex of the Corn Leafhopper, <Emphasis Type="Italic">Dalbulus maidis</Emphasis> (DeLong) (Hemiptera: Cicadellidae), in Argentina

The corn leafhopper, Dalbulus maidis (DeLong), is the most important leafhopper pest of maize, Zea mays, in the Americas. A survey of the diversity of its egg parasitoids was carried out in northwestern Argentina. During summer from 2004 to 2007, the samples were collected, using sentinel eggs of D. maidis on corn leaves, exposed in 48 cornfields. Sixteen species belonging to four families of Chalcidoidea (Hymenoptera) were identified. Among the parasitoid groups, Trichogrammatidae was the most represented family with eight species, followed by Mymaridae with six species. The mymarid Anagrus incarnatus Haliday and the trichogrammatid Pseudoligosita longifrangiata (Viggiani) were the most abundant and frequent parasitoids. The mean percentage of parasitism of D. maidis eggs was 16.4% and varied greatly among the sites, ranging from 0 to 56.7%; generally, it was higher in Yungas and lower in Monte province sites. The species richness was higher in the localities within the Yungas, with 13 parasitoid species, of which two species were dominant, comprising 83.6% of the collected individuals. Monte was the province that showed the highest diversity index (H´ = 1.62). In addition, we present information on the distribution, known host associations of each parasitoid species and an identification key to all species of egg parasitoids of D. maidis in Argentina.