Salicylic Acid Induces Changes in Mango Fruit that Affect Oviposition Behavior and Development of the Oriental Fruit Fly,Bactrocera dorsalis

The Oriental fruit fly, Bactrocera dorsalis (Hendel) is an important quarantine pest around the globe. Although measures for its control are implemented worldwide through IPM and male annihilation, there is little effect on their population. Hence, there is a need for new strategies to control this minacious pest. A strategy that has received negligible attention is the induction of ‘natural plant defenses’ by phytohormones. In this study, we investigated the effect of salicylic acid (SA) treatment of mango fruit (cv. Totapuri) on oviposition and larval development of B. dorsalis. In oviposition choice assays, gravid females laid significantly less eggs in SA treated compared to untreated fruit. Headspace volatiles collected from SA treated fruit were less attractive to gravid females compared to volatiles from untreated fruit. GC-MS analysis of the headspace volatiles from SA treated and untreated fruit showed noticeable changes in their chemical compositions. Cis-ocimene and 3-carene (attractants to B. dorsalis) were reduced in the headspace volatiles of treated fruit. Further, reduced pupae formation and adult emergence was observed in treated fruit compared to control. Increased phenol and flavonoid content was recorded in treated fruit. We also observed differential expression of anti-oxidative enzymes namely catalase (CAT), polyphenoloxidase (PPO) and peroxidase (POD). In summary, the results indicate that SA treatment reduced oviposition, larval development and adult emergence of B. dorsalis and suggest a role of SA in enhancing mango tolerance to B. dorsalis.     

Anaesthetic Tricaine Acts Preferentially on Neural Voltage-Gated Sodium Channels and Fails to Block Directly Evoked Muscle Contraction

Movements in animals arise through concerted action of neurons and skeletal muscle. General anaesthetics prevent movement and cause loss of consciousness by blocking neural function. Anaesthetics of the amino amide-class are thought to act by blockade of voltage-gated sodium channels. In fish, the commonly used anaesthetic tricaine methanesulphonate, also known as 3-aminobenzoic acid ethyl ester, metacaine or MS-222, causes loss of consciousness. However, its role in blocking action potentials in distinct excitable cells is unclear, raising the possibility that tricaine could act as a neuromuscular blocking agent directly causing paralysis. Here we use evoked electrical stimulation to show that tricaine efficiently blocks neural action potentials, but does not prevent directly evoked muscle contraction. Nifedipine-sensitive L-type Ca_v channels affecting movement are also primarily neural, suggesting that muscle Na_v channels are relatively insensitive to tricaine. These findings show that tricaine used at standard concentrations in zebrafish larvae does not paralyse muscle, thereby diminishing concern that a direct action on muscle could mask a lack of general anaesthesia.     

Cloning and characterization of differentially expressed genes of internal breakdown in mango fruit (Mangifera indica)

Internal breakdown in mango fruits has become a major concern in recent years. This disorder renders the fruits unfit for human consumption. The overall loss due to this disorder is about 35-55%. Environmental and physiological factors like high temperature, humidity, respiration and low transpiration rates have been attributed to cause spongy tissue due to reduced loss of heat from fruits. Biochemical studies have shown that there is a reduction in pH, total soluble solids, ascorbic acid, total sugars and carotenoids, low reducing and non-reducing sugar contents, lower amylase and invertase activities and high acid and starch content in spongy tissue affected pulp. There are no reports on molecular studies to determine changes in gene expression in these tissues. The present study was conducted using PCR based subtractive hybridization and RNA gel blot analysis of a few selected genes. The latter showed a higher expression of catalase, ubiquitin, alcohol dehydrogenase, coproporphyrinogen oxidase and keratin associated protein. A lower expression of ribosomal gene, fructose bisphosphate aldolase and cysthathionine gamma synthase was also noticed in spongy tissue. Biochemical studies indicated a lower amylase activity and a lower content of the total and reducing sugars in spongy tissue as compared to healthy tissue. Analyses of results indicate that oxidative stress may be one of the causes for formation of spongy tissue, which affects the expression of many genes. The role of these genes in spongy tissue formation is discussed.     

Toxicological aspects of pendimethalin induced activities of certain oxidising and hydrolytic enzymes in the germinating seedlings of maize (Zea mays L.)

Maize (Zea mays L.) variety NAC-6002 grains were exposed to a range of field relevant concentrations (0.5, 1.0, 2.5, 5.0 and 10.0 ppm) of formulated grade of pendimethalin. The seedlings were maintained in Hogaland and Arnon's nutrient media for up to 15 days to study the toxicity of pendimethalin on germination and activities of certain oxidizing and hydrolytic enzymes using embryonic axis and endosperms of 4, 6, 8, 10 and 15-day-old germinating maize seedlings. The percent germination, length of radicle and plumule decreased significantly with increasing pendimethalin concentrations. The α-amylase activity in the treated seedlings decreased with increasing concentration of pendimethalin in both endosperm and embryonic axis of maize seedling, when compared with control in all the days of observations. The catalase, peroxidase and polyphenoloxidase activity was high in both cotyledons and embryonic axis of treated sets at all time intervals of seedling during early growth over the period of 15 day when compared to control. However, catalase and peroxidase activity was decreased and polyphenoloxidase activity was higher in treated seedlings at higher pendimethalin concentrations. The present study therefore indicates that herbicidical stress brings constraints in the physiological events of seed germination and enzyme activity.     

Prokaryotic expression of the thyrotropin receptor and identification of an immunogenic region of the protein using synthetic peptides

Graves' disease is characterized by hypersecretion of thyroid hormones due to binding of autoantibodies to the thyrotropin receptor (TSHR). In order to study immunological aspects of the TSHR we expressed the extracellular domain of the rat TSHR (ETSHR) as a fusion protein with beta-galactosidase in a prokaryotic system. The identity of this ETSHR-fusion protein was confirmed by Western blot, using antibodies to synthetic peptides derived from TSHR. Patients' sera reacted to a significantly greater extent with the affinity purified ETSHR relative to control sera. Similarly, sera from patients with Graves' disease displayed significant reactivity with only one of five peptides, RH2 (residues 352-366), when compared with normal sera. These data, together with the predicted hydrophilicity of the peptide RH2, suggest that amino acids 352-366 which lie within one of the unique regions of the extracellular domain of the TSHR may be important for antibody binding.     

Effects of photo-oxidative loss of FeNa2EDTA and of higher iron supply on chlorophyll content, growth and propagation rate in triploid watermelon cultures

Summary Triploid watermelon (Citrullus lanatus [Thunb.] Matsum. & Nakai) Arka Manik cultures exhibited a general pale color and yellowing of lower leaves by 3–4 wk following subculturing to fresh medium. Protecting the medium from light by covering the surface with sterile aluminum foil significantly improved leaf color and chlorophyll content by 50–60%. Raising the concentration of FeNa₂EDTA (Fe-EDTA) in the light-exposed medium from the normal level (100 μM) to 150 or 200 μM significantly increased the leaf chlorophyll two- to threefold, suggesting that the light-induced effect was mainly attributable to photo-oxidation of Fe-EDTA. Fe-EDTA supplementation also resulted in better expansion and lobe formation of upper leaves, a reduction in internode length, an increase in the number of nodes, and a significant improvement in propagation rate. There was also a reduction in axillary sprout growth at elevated Fe-EDTA levels, indicating a more prominent apical dominance effect and a reduction in the number and vigor of tendrils. Increasing the Fe-EDTA level in the medium to 150 μM is suggested for routine micropropagation of triploid watermelon, in order to compensate for the photo-oxidative loss of Fe-EDTA.     

Neuroendocrine-induced synthesis of bone marrow-derived cytokines with inflammatory immunomodulating properties

Although cytokines and other soluble regulators of immunity are known to be involved in hematopoiesis, little is known about the signals that induce the synthesis of those mediators locally. Based on recent studies linking the neuroendocrine hormone thyrotropin [thyroid-stimulating hormone (TSH)] to immune cell function in other tissues, we investigated the capacity of TSH to activate cytokine responses from bone marrow cells. These studies reveal that stimulation of the TSH receptor on bone marrow cells-using highly purified or recombinant TSH or by direct stimulation with anti-TSH receptor antibodies-rapidly induces the synthesis of cytokines from bone marrow cells that are classically used in the regulation of inflammatory responses. Of 13 cytokines screened for activity by ELISA or by RNase protection assays for gene expression, IL-6, IFN-beta, TNFalpha, TNFbeta, TGFbeta2, and lymphotoxin-beta responses were reproducibly induced by TSH within 2-3 h of stimulation. Intracellularly, TSH stimulation of bone marrow cells caused rapid increases in cAMP levels and induced the phosphorylation of the Jak2 protein kinase, thereby defining a novel G-protein-coupled receptor/cytokine synthesis pathway. These findings demonstrate that TSH can serve as a primary inductive signal of cytokine production by bone marrow cells.