Sensitivity of the mitotic cells of barley (Hordeum vulgare L.) to insecticides on various stages of cell cycle

In this study, the genotoxic effects of insecticides in different phases of cell cycle were investigated in the mitotic cells of barley (Hordeum vulgare L.). The seeds of H. vulgare L. Var. Karan 4 were treated with different concentrations (0.05%, 0.1% and 0.5%) of insecticides alphamethrin (AM) and monocrotophos (MP) for 6 h after synchronization of cells in G₁, S and G₂ phase of cell cycle with the help of various presoaking durations (7 h, 17 h and 27 h.). Positive and negative control was run parallel in the form of ethyl methane sulphonate (EMS) and distilled water, respectively. The data indicate that higher dose of alphamethrin and monocrotophos produce toxicity, chromosomal aberrations and mitotic aberrations in Hordeum vulgare L. The present study indicates the genotoxic potential of the insecticides AM and MP and it also showed that S-phase of cell cycle was more sensitive compared to the G₁ and G₂ phases.     

Azadirachtin-A and tetrahydroazadirachtin-A concentrates: preparation, LC-MS characterization and insect antifeedant/IGR activity against Helicoverpa armigera (Hübner)

A 60% azadirachtin-A concentrate has been obtained through repeated precipitation with hexane from a methanolic solution of a 20% concentrate. Azadirachtin-A (90%) has been obtained by medium-pressure liquid chromatography of the 60% concentrate with an RP-18 column and a methanol + water (1 + 1 by volume) solvent system. Catalytic hydrogenation of the 60 and 90% azadirachtin concentrates yielded the corresponding tetrahydroazadirachtin concentrates. Dihydroazadirachtin and tetrahydroazadirachtin formed during the first 5 h of hydrogenation were identified by LC-ESI-MS on the basis of their unique mass fragmentation pattern. The efficacy of tetrahydroazadirachtin concentrates in inhibiting the feeding and growth of Helicoverpa armigera (Hübner) larvae has been compared with that of azadirachtin concentrates. They were in general more active and deterred larvae from feeding at all concentrations. Tetrahydroazadirachtin-A (90%) and azadirachtin-A (90%) with respective IC(50) values of 280 and 390 mg L(-1) were effective as insect growth regulators, while tetrahydroazadirachtin-A (90%) displayed higher antifeedant activity (AI(50) 14 mg L(-1)) against the test insect.     

Haematological and biochemical responses of freshwater teleost fish Cyprinus carpio (Actinopterygii: Cypriniformes) during acute and chronic sublethal exposure to lindane

The impact of acute and sublethal toxicity of a synthetic organochlorine pesticide lindane on some haematological and biochemical parameters of a freshwater fish Cyprinus carpio was estimated. The median lethal concentration of lindane for 24 h was 0.38 ppm. During acute treatment (24 h), hemoglobin (Hb), hematocrit (Hct) and erythrocyte (RBC) values were decreased, whereas leucocyte (WBC) count increased in the pesticide treated fish. The hematological indices like mean cellular volume (MCV), mean cellular hemoglobin (MCH) and mean cellular hemoglobin concentration (MCHC) were decreased when compared to control group. Biochemical profiles like plasma glucose and protein levels were increased in lindane exposed fish showing a percent increase of 50.36 and 3.98 at the end of 24 h treatment. However, glycogen content in liver and muscle were decreased showing a percent decrease of 21.70 and 1.74. In sublethal treatment (1/10th of LC50 24 h value, 0.038 ppm), RBC count was decreased whereas WBC count increased in the pesticide treated fish throughout the study period (25 days). Hb and Hct values were decreased up to 10th day and after that recovered showing a significant increase in the rest of the study period. Similarly, a biphasic response was observed in the value of MCV, MCH and MCHC. Plasma glucose level was significantly increased while plasma protein level decreased throughout the study period. Further, glycogen level in muscle and liver were showed a mixed trend. In the present study, the pesticide lindane caused alterations on haematological and biochemical parameters of C. carpio and these alterations can be used as non specific biomarkers in pesticide contaminated aquatic ecosystem.     

Ritacestus gen. n. (Cestoda: Proteocephalidea) and redescription of R. ritaii comb. n., a parasite of Rita rita (Siluriformes) in India

A new genus, Ritacestus, is proposed to accommodate Ritacestus ritaii (Verma, 1926) comb. n. (syn. Proteocephalus ritaii), a parasite of the catfish Rita rita (Hamilton) in India. The new genus, which is placed in the Gangesiinae, is characterized by (i) a small, subspherical scolex formed by four large lobes separated from one another by longitudinal grooves, with a large, widely oval to pyriform rostellum-like apical organ, larger than suckers and possessing an apical hemispherical depression; (ii) paramuscular and cortical position of some vitelline follicles (most follicles are situated medullary); (iii) ventral and dorsal bands of vitelline follicles usually uninterrupted ventral to terminal genitalia and reaching to the posterior margin of proglottides; (iv) the vagina always anterior to the cirrus-sac; (v) a large size of the body (length up to 51 cm); and (vi) development of the uterus of type 2. In its morphology, especially shape of the scolex and apical organ, and paramuscular and cortical position of some vitelline follicles, Ritacestus resembles Postgangesia Akhmerov, 1969, but differs in the presence of a genital atrium (both genital pores of Postgangesia are separate), the anterior position of the vagina (almost always posterior in the latter genus), position of vitelline follicles in cross sections (dorsal and ventral bands in Ritacestus versus only a lateral band in the latter genus), and dorsal excretory canals indistinguishable in mature and gravid proglottides of R. ritaii (well developed in Postgangesia spp.). The type and only species of the genus, R. ritaii, is redescribed on the basis of new material from the type host from the Ganges River basin in India and its neotype is designated.     

Inheritance of resistance to Bacillus thuringiensis Cry1Ac toxin in Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) from India

BACKGROUND: The cotton bollworm, Helicoverpa armigera (Hübner), is one of the most serious insect pests of cotton. It has developed resistance to almost all groups of chemical insecticides because of their intensive use. The failure of insecticides to control H. armigera has been a strong incentive for the adoption of transgenic cotton (Bt cotton). However, the value of Bt could be diminished by widespread resistance development to Bt toxins in insect populations. Therefore, understanding the genetic basis of resistance is essential for developing and implementing strategies to delay and monitor resistance. RESULTS: A resistant strain designated as BM‐R was obtained from the cross of adults from Bathinda () and Muktsar (), Punjab, India, which showed the highest survival (60.68%) and LC₅₀ value (1.396 µg mL^?1 diet). Similarly, a laboratory‐maintained strain from Hoshiarpur, Punjab, showed maximum susceptibility to Cry1Ac toxin with the lowest LC₅₀ value (0.087 µg mL^?1), and was designated as HP‐S. The genetic purity of both strains was confirmed by RAPD profile analysis at each generation, and genetic similarity reached more than 90% after the third generation. Continuous maintenance of the resistant BM‐R strain on Cry1Ac resulted in an increase in LC₅₀ from 0.531 µg mL^?1 in F₀ to 4.28 µg mL^?1 in F₁₄ and 7.493 µg mL^?1 in F₁₉, while the LC₅₀ values for HP‐S larvae on diet without Cry1Ac increased to 0.106 and 0.104 µg mL^?1, which lay within the fiducial limits of the baseline LC₅₀ value. The mode of inheritance of resistance was elucidated through bioassay response of resistant, susceptible heterozygotes and backcross progeny to Cry1Ac incorporated in semi‐synthetic diet. CONCLUSION: Based on dominance, degree of dominance and backcross values, resistance was inferred to be polygenic, autosomal and inherited as a recessive trait. Copyright © 2011 Society of Chemical Industry     

H2O2 degradation is suppressed in kumquat leaves infected with Xanthomonas axonopodis pv. citri

We found in a previous study that after leaves of kumquat [Fortunella margarita (Lour.) Swingle] cv ‘Nagami’ were inoculated with Xanthomonas axonopodis pv. citri (Xac), total superoxide dismutase activity (SOD) increased to promote higher H₂O₂ concentrations that coincided with a 4-fold decline in Xac populations ( Kumar et al., 2011a). The objective of the current study was to determine how activities and isoforms of important enzymes that catabolize H₂O₂, specifically catalase (CAT), ascorbate peroxidase (APOD), and the Class III peroxidases (POD) that are located in the apoplast, change in infected kumquat leaves to affect concentration and compartmentalization of H₂O₂. DAB (3,3-diaminobenzidine) staining of the Xac-infected leaves confirmed higher overall concentration of H₂O₂ as in our earlier study. One day after inoculation (dai), APOD activity declined below the controls and declines steadily up to 10 dai when the experiment was terminated. CAT activity was similar to the controls until 4 dai then declined rapidly to about 60% the activity of the controls by 6 dai, after which it remained fairly constant until 10 dai. There were 4 CAT isoforms in control leaves and 5 isoforms in infected leaves. The CAT-1 isoform band was much smaller in infected plants than the control at all sampling times. The CAT-3 isoform band disappeared at 10 dai. The CAT-5 isoform band, which was not observed in control leaves, appeared only at 4 dai in infected leaves. POD activity of infected leaves increased above the controls starting 1 dai and reached a maximum of about 3-fold higher than the controls 8 dai after which it declined. Two POD isoforms were detected in control and infected plants. This study demonstrated that the higher accumulation of H₂O₂ in kumquat leaves infected with Xac was promoted during pathogenesis first by the suppression of APOD activity and later by suppression of CAT activity. We propose that the higher SOD and lower APOD and CAT activities in the symplast contributed H₂O₂ substrate for the higher POD activity in the apoplast, which is known to be involved in plant defense against pathogens.     

Genetic diversity and population structure analysis of isolates of the rice false smut pathogen Ustilaginoidea virens in India

Genetic diversity assessment and population structure analysis are essential for characterization of pathogens and their isolates. Markers are essential tools for exploring genetic variation among the isolates. False smut of rice caused by Ustilaginoidea virens, formerly Villosiclava virens, is a major emerging disease of rice in India. A high level of variability is observed at the field level, but no information is available from India on genetic diversity and population structure. This is the first report of genetic diversity and population structure of U. virens from India that included 63 isolates distributed across the vast geographical area of eastern and north-eastern India (18.9 to 26.7°N and 82.6 to 94.2°E). Seventeen RAPDs and 14 SSRs were identified as polymorphic and a total of 140 alleles were detected across the populations. The average number of alleles per locus for each primer was 4.5. All the isolates were grouped into two major clusters, with partial geographical segregation that was supported by principal coordinate analysis. Mantel test suggested genetic distance within the isolates increased with increasing geographical distance. Analysis of molecular variation showed more genetic variation within populations and less among populations. This outcome will help in understanding genetic diversity of U. virens from eastern and north-eastern India and in planning effective management strategies.