Identification of protein related to dietary vitamin B3 deficiency in Mediterranean fruit fly larvae.

Mediterranean fruit fly (medfly), Ceratitis capitata, is among the most destructive agricultural pest. The sterile insect technique (SIT) can effectively control medfly populations. To rear healthy medflies for the purpose of SIT, it is essential to supplement B vitamins in the diet. However, the function of the dietary B vitamins in C. capitata larvae is not known. With the microscopic analysis, several organs in the head were examined and the spiracle formation and sensory organs were normally formed between the niacin-supplied and niacin-absent groups. However, formation of the ocular depression was differently developed between the two groups, although the hypostomal sclerite was formed properly. These results signify that niacin deficiency maybe interrupt development of medfly larvae ocular depression. Proteomic analyses using LC MS/MS detected a total of 1845 proteins in two flies. A total of 607 of the 1845 proteins were overexpressed and one third (598 proteins) were downexpressed in the niacin-deficient larvae, while about one third were similarly expressed. Overexpressed proteins in the niacin-deficient larvae included ryanodine receptor 44 F, intergrin-PS, spalt-major homeotic protein, and chiffon protein. One of important overexpressed proteins was optomotor-blind protein in relation to wing development in the niacin-deficient medfly larvae.

Construction of Highly Efficient Carbazol-9-yl-Substituted Benzimidazole Bipolar Hosts for Blue Phosphorescent Light-Emitting Diodes: Isomer and Device Performance Relationships.

Four isomers of carbazol-9-yl-substituted 1,2-diphenylbenzimidazole at 7, 6, 5, and 4 positions, named as 1-CbzBiz, 2-CbzBiz, 3-CbzBiz, and 4-CbzBiz, respectively, have been synthesized. Instead of having an identical molecular weight, the CbzBiz's have their glass transition temperatures ( Tg) spanning a large range from 53 to 90 °C. Their Tg and melting point ( Tm) basically obey the Boyer-Kauzmann rule ( Tg = g· Tm with g ≈ 0.7) on the absolute temperature scale (in kelvins). However, while 1-CbzBiz and 4-CbzBiz demonstrate a small g value of 0.66, which is significantly smaller than other common organic glass molecules, 3-CbzBiz shows an unexpectedly high g value of 0.73, implying higher intermolecular interactions in the glass. These CbzBiz's are suitable hosts for bis[2-(4,6-difluorophenyl)pyridinato- C2, N](picolinato)iridium(III) (FIrpic) in phosphorescence organic light-emitting diodes (PhOLEDs). The optimized PhOLED of indium tin oxide/4,4'-cyclohexylidenebis[ N, N-bis(4-methylphenyl)benzenamine]/4Cbz/4-CbzBiz-FIrpic(15%)/diphenylbis(4-(pyridin-3-yl)phenyl)silane/LiF-Al shows a maximum brightness of 18 760 cd/m2, a current efficiency of 64.1 cd/A, and the external quantum efficiency of 30.9%.

Comparative Proteomic Profiling between Each of Two Consecutive Developmental Stages of the Solanum Fruit Fly, Bactrocera latifrons (Hendel).

The Solanum fruit fly, Bactrocera latifrons (Hendel), has a complex life cycle including multiple stages (egg, larva, pupa, and adult). Understanding the details of "what", "when", "where", "why", and "how" many hundred thousand proteins operate in this insect, interact, and express between each two consecutive developmental stages at molecular level not only can expand our knowledge, but also lead to the development of novel fruit fly control techniques. We tried to find what, when, and where in this study. Why and how will be presented in upcoming papers. We conducted a proteome profiling using 2-D gel electrophoresis and mass spectrometry. Samples of 3-day-old eggs, 1- and 10-day-old larvae, 1- and 10-day-old pupae, 1- and 9-day-old females and males of B. latifrons were used. A custom peptide database, derived from the de novo B. latifrons whole genome assembly was used for peptide identification. Differentially expressed proteins (DEPs) with significant fold expression and protein functions between two consecutive developmental stages were identified, annotated, described, and listed in gel images and/or charts. With this foundational information, we are not only providing valuable information, but also any impacts due to the biotic or abiotic environmental factors can be identified and manipulated, and lead to further research on gene editing and biomarker discovery.

Laboratory evaluation on a potential birth control diet for fruit fly sterile insect technique (SIT).

Sterile insect technique (SIT) is one of the most effective fruit fly control technologies. Irradiation has been used to sterilize male fruit flies before release to the field to compete with the wild males for females. Imagine an environmental and cost effective method using a rearing diet that can make insects sterile indefinitely, by feeding for 7days before release. This could replace costly irradiation process. A potential birth control diet was evaluated on fertility, mating, survival, and protein analysis for fruit fly species in Hawaii. Insects were continuously fed an agar diet with lufenuron (LFN) for 7d after emergence and then switched to a control diet to simulate the actual field condition. The influence on egg hatch was dose dependent. With dose of 2-4mg/g in the diet, egg hatch from LFN-fed was almost 100% suppressed for 24 experimental days if adults of Ceratitia capitate (Widemann), Bactrocera dorsalis (Hendel), and B. latifrons (Hendel) continued to feed on LFN diet. B. cucurbitae (Coquillett) was not affected by LFN. However, egg hatch from LFN fed B. latifrons and B. dorsalis were suppressed for at least 2weeks after switching to the control diet at 7d. Egg hatch did not recover >4% up to 24d. Proteome analysis revealed that ABD-4 protein was under expressed by 70-83% on LFN fed females and males of B. latifrons and B. dorsalis while Pbprp2 protein was significantly over expressed by 6-12 fold on LFN fed males only. These two proteins were not expressed in C. capitata and B. cucurbitae. Therefore, this report focused more on B. latifrons and B. dorsalis. This finding suggested a great potential for one alternative to sterilize fruit flies for SIT without irradiation.

LARVAL X-RAY IRRADIATION INFLUENCES PROTEIN EXPRESSION IN PUPAE OF THE ORIENTAL FRUIT FLY, BACTROCERA DORSALIS.

The sterile insect technique (SIT) was developed to eradicate the new world screwworm from the southern United States and Mexico, and became a component of many area-wide integrated pest management programs, particularly useful in managing tephritid fruit flies. SIT is based on the idea of rearing and sterilizing male pests, originally by ionizing radiation, and then releasing into field, where they compete for and mate with wild females. Mating with sterile males leads to reduced fecundity to lower pest populations. There are concerns with the use and distribution of radioisotopes for SIT programs, which have led to developing X-ray irradiation protocols to sterilize insects. We considered the possibility that X-ray irradiation exerts sublethal impacts aside form sterilizing insects. Such effects may not be directly observable, which led us to the hypothesis that X-ray irradiation in one life stage creates alterations in biological fitness and protein expression in the subsequent stage. We tested our hypothesis by irradiating larvae of Bactrocera dorsalis. There are two major points. One, exposing larvae to X-ray treatments led to reduced adult emergence, fecundity, fertility, and flight capacity from the corresponding pupae and emerged adults. Two, the X-ray treatments led to substantial expression changes in 27 pupal proteins. We assorted the 67 spots representing these proteins into three groups, metabolism, development, and structure. Our interpretation is these X-ray induced changes in biological performance and protein expression indicate their adult counterparts may be disabled in their abilities to successfully compete for and mate wild females in native habitats.

Pyrrolo-[3,2-b]pyrroles for Photochromic Analysis of Halocarbons.

Dramatic photochromic-change of 2,5-bis(triphenylamine)-substituted N,N'-diphenylpyrrolo-[3,2-b]pyrrole (1) with halocarbons provides an effective route for halocarbon analysis with the naked eye. The visual detection range can reach as low as 10(-4) ∼ 10(-5) M (1-10 ppm) in CH3CN. This method can also be applied for detection of CHCl3 in water. Fabrication of a disposable paper test cartridge along with using a camera flash as the light source allows on-site halocarbon detection in seconds. Quantitative analysis for CHCl3 and CH2Cl2 have also been demonstrated.

Pupal X-ray irradiation influences protein expression in adults of the oriental fruit fly, Bactrocera dorsalis.

The oriental fruit fly, Bactrocera dorsalis, is a pest of fruit in the Asia-Pacific region and also, due to quarantine restrictions, a threat to California fruit production. Area-wide suppression of B. dorsalis integrated several approaches including the sterile insect technique (SIT). SIT involves exposing juveniles to gamma radiation and releasing sterile males in substantial numbers, where they successfully compete for wild females. The resulting infertile eggs lead to reduction of the pest populations. Although these protocols are well documented, arising issues about the international transport and distribution of radioactive products is creating difficulties in use of radioactive sources for sterilizing radiation. This led to a shift toward use of X-ray irradiation, which also sterilizes male and female insects. However, use of X-ray technologies is in its infancy and there is virtually no information on the effects of irradiation, other than sterilization, at the physiological and molecular levels of fruit fly biology. We posed the hypothesis that sterilizing male oriental fruit flies via radiation treatment also influences protein expression in the flies. We found that exposing pupae to X-ray irradiation impacted expression of 26 proteins in adult females and 31 proteins in adult males. Seven proteins (glyceraldehyde-3-phosphate dehydrogenase, fructose-bisphosphate aldolase, larval cuticle protein 2, sarcoplasmic calcium-binding protein alpha-B and A chains, general odorant-binding protein 99b, polyubiquitin, and protein disulfide-isomerase) were impacted in both sexes. Some of the proteins act in central energy-generating and in pheromone-signal processing pathways; we infer that males sterilized by X-ray irradiation may be enfeebled in their ability to compete with wild males for females in nature.

A qPCR-based method for detecting parasitism of Fopius arisanus (Sonan) in oriental fruit flies, Bactrocera dorsalis (Hendel).

BACKGROUND: Parasitism detection and species identification are necessary in fruit fly biological control. Currently, release of mass-reared Fopius arisanus is practiced worldwide, as it is effective in controlling Bactrocera dorsalis and Ceratitis capitata. To detect and assess parasitism in parasitoid mass-rearing colonies and parasitism levels in field populations across all life stages of hosts, the development of a rapid, specific and sensitive method is important. RESULTS: A species-specific probe was designed for F. arisanus, as well as a universal tephritid probe. Utilizing rapid DNA extraction techniques coupled with quantitative-PCR, a simple and fast assay has been developed to detect parasitism of F. arisanus that is sensitive enough to detect the parasitoid across all developmental stages, including a single egg per host egg or 0.25 ng of parasitoid DNA in 40 ng of host DNA. The qPCR methods also detect a higher parasitism rate when compared with rearing-based methods where parasitism rate is based on wasp emergence and where unemerged wasps are not included. CONCLUSION: This method is a rapid, sensitive and specific technique to determine the parasitism rate of F. arisanus across all life stages of B. dorsalis, which will be useful to predict parasitoid output from mass rearing and evaluate the outcome of pest suppression after mass release in the field.

Tunable electrofluorochromic device from electrochemically controlled complementary fluorescent conjugated polymer films.

The fluorescent behavior of the electrofluorochromic devices (Type I) of greenish-yellow emitting P1 and blue emitting P2 can be reversibly switched between the nonfluorescent (oxidized) state and the fluorescent (neutral) state with a superb on/off ratio of 23.8 and 21.9, respectively. Moreover, a tunable electrofluorochromic device (Type II) based on two P1 and P2 polymeric layers that are coated individually on two independent ITO electrodes shows switchable blue-white-(greenish-yellow) multifluorescence states.

Dietary lufenuron reduces egg hatch and influences protein expression in the fruit fly Bactrocera latifrons (Hendel).

Lufenuron (LFN), a chitin synthase inhibitor, impacts the fertility of Ceratitis capitata, Bactrocera dorsalis, B. cucurbitae, and B. latifrons. We posed the hypothesis that LFN curtails egg hatch in the solanaceous fruit fly, B. latifrons. In this study, newly emerged virgin adults were sexed and fed for 12 days with varying concentrations of LFN-laced agar diets until sexual maturation. Eggs were collected from 12-d-old adults and the egg hatch was assessed. Egg hatch decreased in adults reared on LFN-treated diets. LFN-treated media did not influence fertility after one gender was reared on experimental and the other on control media before mating. Exposure to LFN-treated medium after mating led to reduced egg hatch. We infer that LFN is not a permanent sterilant, and reduced egg hatch depends on continuous exposure to dietary LFN after mating. Proteomic analysis identified two differentially expressed proteins, a pheromone binding protein and a chitin binding protein, between adults maintained on LFN-treated and control diets. Expression of two genes encoding chitin synthase 2, and chitin binding protein, was altered in adults exposed to dietary LFN. LFN treatments also led to increased expression of two odorant binding proteins one in females and one in males. We surmise these data support our hypothesis and provide insight into LFN actions.

Diet-induced over-expression of flightless-I protein and its relation to flightlessness in Mediterranean fruit fly, Ceratitis capitata.

The Mediterranean fruit fly (medfly), Ceratitis capitata is among the most economically important pests worldwide. Understanding nutritional requirement helps rearing healthy medfly for biocontrol of its population in fields. Flight ability is a high priority criterion. Two groups of medfly larvae were reared with two identical component diets except one with fatty acids (diet A) and another without it (diet B). Adults from larvae reared on diet B demonstrated 20±8% of normal flight ability, whereas those from larvae reared on diet A displayed full flight ability of 97±1%. Proteomes were profiled to compare two groups of medfly pupae using shotgun proteomics to study dietary effects on flight ability. When proteins detected in pupae A were compared with those in pupae B, 233 and 239 proteins were, respectively, under- and over-expressed in pupae B, while 167 proteins were overlapped in both pupae A and B. Differential protein profiles indicate that nutritional deficiency induced over-expression of flightless-I protein (fli-I) in medfly. All proteins were subjected to Ingenuity Pathway Analysis (IPA) to create 13 biological networks and 17 pathways of interacting protein clusters in human ortholog. Fli-I, leucine-rich repeat (LRR)-containing G protein-coupled receptor 2, LRR protein soc-2 and protein wings apart-like were over-expressed in pupae B. Inositol-1,4,5-trisphosphate receptor, protocadherin-like wing polarity protein stan and several Wnt pathway proteins were under-expressed in pupae B. These results suggest down-regulation of the Wnt/wingless signaling pathway, which consequently may result in flightlessness in pupae B. The fli-I gene is known to be located within the Smith-Magenis syndrome (SMS) region on chromosome 17, and thus, we speculate that nutritional deficiency might induce over-expression of fli-I (or fli-I gene) and be associated with human SMS. However, more evidence would be needed to confirm our speculation.

Resveratrol modifies tephritid fruit fly response to radiation but not nutritional stress.

PURPOSE: Resveratrol (3,5,4'-trihydroxystilbene) is a polyphenol compound found in many plants and fruits that has antioxidant and radioprotective properties. Two model invertebrates, Bactrocera dorsalis (oriental fruit fly) and B. cucurbitae (melon fly) (Diptera: Tephritidae), were studied to determine if the addition of resveratrol to an artificial diet could modify their response to radiation and nutritional stress. MATERIALS AND METHODS: Resveratrol at concentrations of 0, 50, 100, or 200 µM of was incorporated into a liquid larval fruit fly diet. Third instars were treated with: (i) A radiation dose of 30 Gy (radiation stress), (ii) a wheat germ oil-deficient diet (nutritional stress), or (iii) left untreated as a control. RESULTS: The addition of resveratrol to the diet partially mitigated the adverse effects of radiation on several life history parameters. In B. cucurbitae, a significantly higher 49-53% of adults could fly when 50-200 µM resveratrol was added to the diet compared with 32% in irradiated flies reared without resveratrol. B. cucurbitae egg hatch in irradiated insects improved significantly from 46 to 66% with the addition of 50 µM resveratrol. In irradiated B. dorsalis, adult emergence was significantly improved from 12 to 29% with the addition of 100 µM resveratrol. Resveratrol did not mediate any of the negative effects of a wheat germ oil-deficient diet in either species. CONCLUSION: Resveratrol has potential as a means to partially mitigate the adverse effects of radiation treatment under the conditions tested. This study is the first to show that resveratrol can have radioprotective effects in invertebrates.

Dietary wheat germ oil influences gene expression in larvae and eggs of the oriental fruit fly.

Changes in animal nutrition, particularly essential dietary components, alter global gene expression patterns. Our goal is to identify molecular markers that serve as early indicators of the quality of insect culture media. Markers of deficient culture media will increase the efficiency of developing optimal systems for mass rearing beneficial insects and some pest species because decisions on culture media quality can be made without waiting through one or several life cycles. The objective of our current study is to discover molecular markers of essential dietary lipid deficiency in the oriental fruit fly, Bactrocera dorsalis. We reared groups of fruit flies separately on media either devoid of or supplemented with wheat germ oil (WGO) and analyzed gene expression in third instar larvae and F(1) eggs using 2D electrophoresis. Gel densitometry revealed significant changes in expression levels of genes encoding eight proteins in larvae and 22 proteins in eggs. We identified these proteins by using mass spectrometry (MALDI TOF/TOF) and bioinformatic analyses of the protein sequences. Among these, we identified one gene encoding the receptor of activated C Kinase 1 (RACK1) that increased in expression by 6.8-fold in eggs from adults that were reared as larvae on media supplemented with WGO. RACK1 is an essential component of at least three intracellular signal transduction pathways, making it a good molecular marker candidate of lipid deficiency in fruit flies and possibly many other insect species.

Wheat germ oil in larval diet influences gene expression in adult oriental fruit fly.

Culture medium supplemented with wheat germ oil (WGO) causes physiological reactions, such as increased fecundity and mobility, in some insects. Although the impact of WGO on insect physiology is important, the mechanisms of these actions are poorly understood. In this paper, we test the hypothesis that the addition of WGO to medium developed for larval oriental fruit flies modulates gene expression in the corresponding adults. We separately reared larvae of Bactrocera dorsalis on diets lacking or supplemented with WGO, and analyzed for expressed proteins in the resulting adult males and females by 2D-electrophoresis. Analysis of the gels revealed significant changes in expression levels of >70 proteins, 64 of which were identified by mass spectrometric analysis on MALDI-TOF/TOF. Apparent changes in expression levels for 6 of these proteins were confirmed by quantitative real-time PCR, showing that the changes in mRNA expression were reflected in changes in protein expression. These findings support the hypothesis that one mechanism of WGO actions in insect nutrition is the modulation of gene expression.

Insecticidal activity of basil oil, trans-anethole, estragole, and linalool to adult fruit flies of Ceratitis capitata, Bactrocera dorsalis, and Bactrocera cucurbitae.

Basil oil and its three major active constituents (trans-anethole, estragole, and linalool) obtained from basil (Oscimum basilicum L.) were tested on three tephritid fruit fly species [Ceratitis capitata (Wiedemann), Bactrocera dorsalis (Hendel), and Bactrocera cucurbitae (Coquillett)] for insecticidal activity. All test chemicals acted fast and showed a steep dose-response relationship. The lethal times for 90% mortality/knockdown (LT90) of the three fly species to 10% of the test chemicals were between 8 and 38 min. The toxic action of basil oil in C. capitata occurred significantly faster than in B. cucurbitae but slightly faster than in B. dorsalis. Estragole acted faster in B. dorsalis than in C. capitata and B. cucurbitae. Linalool action was faster in B. dorsalis and C. capitata than in B. cucurbitae. trans-Anethole action was similar to all three species. Methyl eugenol acted faster in C. capitata and B. cucurbitae than in B. dorsalis. When linalool was mixed with cuelure (attractant to B. cucurbitae male), its potency to the three fly species decreased as the concentration of cuelure increased. This was due to linalool hydrolysis catalyzed by acetic acid from cuelure degradation, which was confirmed by chemical analysis. When methyl eugenol (B. dorsalis male attractant) was mixed with basil oil, trans-anethole, estragole, or linalool, it did not affect the toxicity of basil oil and linalool to B. dorsalis, but it did significantly decrease the toxicity of trans-anethole and estragole. Structural similarity between methyl eugenol and trans-anethole and estragole suggests that methyl eugenol might act at a site similar to that of trans-anethole and estragole and serve as an antagonist if an action site exists. Methyl eugenol also may play a physiological role on the toxicity reduction.

Wheat germ oil and its effects on a liquid larval rearing diet for oriental fruit flies (Diptera: Tephritidae).

Wheat germ oil was added to a larval liquid diet for rearing Bactrocera dorsalis (Hendel) (Diptera: Tephritidae) to optimize fruit fly quality. Effects of various concentrations of wheat germ oil at 0.04, 0.07, 0.15, 0.30, and 0.66% and their possible mode of action were evaluated. Results suggest that addition of wheat germ oil does not affect pupal weight, larval developmental period, adult emergence, mating ability, or peak time for egg production. But there was a significant increase in pupal recovery, percentage of adult fliers, egg production, or egg hatch for larvae fed the diet with wheat germ oil compared with those reared on the liquid diet without wheat germ oil. The increase in egg hatch and fliers was dose dependent. Therefore, addition of wheat germ oil to fruit fly rearing diet is a novel way to improve fruit fly quality, especially in egg hatch, fliers, egg production, and pupal recovery.

Nicotinamide in relation to dietary nicotinic acid and nine other vitamins and larval development of Ceratitis capitata (Diptera: Tephritidae).

This study was undertaken to understand why Ceratitis capitata larvae reared on a diet fortified with nine vitamins except nicotinic acid had 100% mortality, while those reared on a 10-vitamin-free diet had 66% survival (Chang, C. L.; Li, Q. X. Ann. Entomol. Soc. Am. 2004, 97, 536-540). Our results showed that nicotinamide was detected at a level of 0.07 microg/g in second-instar larvae reared on the 10-vitamin-free diet and 0.30 microg/g in the corresponding spent diet, while it was not detected in either the larvae reared on the diet fortified with 707 microg/g of nine vitamins (nicotinic acid absent) or the corresponding spent diet. Nicotinamide was detected at concentrations of 0.13 and 0.15 microg/g in the larvae fed the diets that were fortified with 707 microg/g of nine other vitamins and 2 and 20 microg/g of nicotinic acid, respectively, but it was not found in the larvae fed the 0.2 microg/g of nicotinic acid diet. Nicotinamide was detected at concentrations of 0.44, 0.52, and 0.55 microg/g in the spent diets that were fortified with the nine vitamins (707 microg/g) and 0.2, 2, and 20 microg/g of nicotinic acid, respectively. Nicotinamide adenine dinucleotide (NAD) was in the live larvae, but not in the dead larvae. These findings indicate that dietary nicotinic acid is converted to nicotinamide, which, in turn, is used to synthesize NAD, and suggest a positive relationship between C. capitata larvae survival rates and concentrations of dietary nicotinic acid and nitcotinamide in the larvae as well as in the spent diets. The result shows that nicotinamide derived from supplemental nicotinic acid is essential for the development and survival of C. capitata larvae. Nicotinamide may be a biomarker for larval survival and development.

Lipid and protein loads in pupating larvae and emerging adults as affected by the composition of Mediterranean fruit fly (Ceratitis capitata) meridic larval diets.

The effects of sucrose and amino acid (aa) composition and concentration in meridic larval diets (e.g., partially defined at the chemical level) was examined on several parameters of Mediterranean fruit fly (Medfly) development. Lipid and protein levels of pupating larvae and emerging adults were examined. Different sucrose concentrations in the diet had small effects upon most of the development parameters. However, sucrose concentration significantly affected the ability of larvae to accumulate lipid reserves and proteins. Adults emerging from the different sucrose diets did not significantly differ in their lipid contents and protein loads. Specific deletions of aa from the diet, and general aa concentration, had a strong effect upon the parameters of development and pupating larvae lipids and proteins. Glycine-deletion was the most deleterious, followed by the deletion of all non-essential aa, and serine. High aa concentration in the diet has a detrimental effect upon development. Lipid contents in pupating larvae, and to some extent protein levels, were affected by aa manipulations in the diet. Lipid and protein loads in emerging adults were not significantly affected by aa manipulations. Based on the analysis of lipid frequency distribution it is suggested that the Medfly seems to regulate the level of lipid content in emerging adults within a certain range, regardless of the larval diet history or lipid contents. Proteins do not seem to be regulated as are lipids. These results point to an interesting and unexpected metabolic regulation of energetic resources during metamorphosis of the Medfly.