Screening of crude plant extracts as feeding deterrents of the wireworm, Melanotus communis

Extracts from 78 plant species (24 families) were screened for antifeedant activity against the corn wireworm, Melanotus communis Gyll. (Coleoptera: Elateridae). Five extracts from four families significantly reduced wireworm feeding damage in a series of choice feeding tests. Two extracts, Asclepias tuberosa and Hedera helix, exhibited exceptional levels of feeding deterrency. An x-ray technique indicated that wireworms burrowed indiscriminately between soil containing either of these extracts and surrounding untreated soil, but were found more frequently in the untreated areas.

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Résumé L'aptitude à empêcher la prise de nourriture de Melanotus communis Gyll. a été recherchée dans les extraits de 78 plantes appartenant à 24 familles. Cinq extraits de 4 familles ont réduit significativement les dégâts alimentaires lorsqu'il y avait choix alimentaire. Deux extraits — Asclepias tuberosa et Hedera helix — présentaient des niveaux exception-nellement élevés de dissuasion de l'alimentation. Une technique aux rayons X a montré que les taupins fouillaient indistinctement le sol contenant de ces extraits ou le sol avoisinant non traité, mais on les recontrait plus fréquemment dans le sol non traité.

     

Cross habituation to feeding deterrents and acceptance of a marginal host plant byPieris rapae larvae

Sensitivity of the cabbage butterfly,Pieris rapae L. to feeding deterrents was compared for larvae reared on different food sources under laboratory conditions. Since cabbage-reared larvae normally reject nasturtium,Tropaeolum majus L., the effects of previous exposure to allelochemicals on larval acceptance or rejection of this plant were also examined. When compared with cabbage-reared larvae, nasturtium-reared larvae were less sensitive to feeding deterrents including cymarin, erysimoside and 2-O-β-d-glucosyl cucurbitacin E. Nasturtium-reared larvae were insensitive to chlorogenic acid, which was deterrent to cabbage-reared larvae. Feeding by larvae reared on a wheat germ diet was not deterred by these compounds. The results indicate that dietary experience can extensively affect larval sensitivity to feeding deterrents and that cross habituation of larvae to deterrents occurs in response to certain chemical constituents of nasturtium and wheat germ diet. Digitoxin, however, proved to be an exception. Larvae reared on either nasturtium or wheat germ diet were as sensitive to digitoxin as those reared on cabbage. Previous results have shown that rejectionof nasturtium by cabbage-reared larvae is due to the presence of strong feeding deterrents in this plant. However, more than 50% of 2nd instar larvae reared from neonate on cabbage leaves treated with strophanthidin, cymarin, erysimoside, digitoxigenin and digitoxin accepted nasturtium as a food source. 2-O-β-d-glucosyl cucurbitacin E, 2-O-β-d-glucosyl cucurbitacin I and rutin were also active in causing larvae to feed on nasturtium. Thus dietary exposure to unrelated plant chemicals can profoundly affect insect acceptance of a plant that contains feeding deterrents.     

Spontaneous plant regeneration in transformed roots and calli from Tylophora indica: changes in morphological phenotype and tylophorine accumulation associated with transformation by Agrobacterium rhizogenes

We examined the effects of genetic transformation by Agrobacterium rhizogenes on the production of tylophorine, a phenanthroindolizidine alkaloid, in the Indian medicinal plant, Tylophora indica. Transformed roots induced by the bacterium grew in axenic culture and produced shoots or embryogenic calli in the absence of hormone treatments. However, hormonal treatment was required to regenerate shoots in root explants of wild type control plants. Transformed plants showed morphological features typically seen in transgenic plants produced by A. rhizogenes, which include, short internodes, small and wrinkled leaves, more branches and numerous plagiotropic roots. Plants regenerated from transformed roots showed increased biomass accumulation (350–510% in the roots and 200–320% in the whole plants) and augmented tylophorine content (20–60%) in the shoots, resulting in a 160–280% increase in tylophorine production in different clones grown in vitro.     

Influence of incubation temperature on productivity and quality of Spodoptera litura nucleopolyhedrovirus

Mass production of baculoviruses by in vivo methods is influenced by several factors like larval age at virus treatment, virus concentration and the incubation temperature. The larval age at virus treatment and virus concentration should be synchronized to result in insect death at a fully grown larval stage to maximize the productivity. Since temperature influences both the growth of the larvae and replication of the virus, we evaluated the influence of incubation temperature on mass production of Spodoptera litura nucleopolyhedrovirus (SpltNPV) at four different temperature regimes viz., 25, 30, and 35 °C and room temperature by diet-surface contamination method. Incubation of early fifth instar larvae dosed with 3932.4 polyhedral inclusion bodies (PIB)/mm² at 25 °C enhanced the virus productivity to 6.623 × 10¹¹ PIB yield/100 inoculated larvae, while it was only 1.779 × 10¹¹ at 35 °C. The disease progression in the virus treated larvae was slow with median lethal time (LT₅₀) of 208 h at 25 °C as compared to 136 h at 35 °C. In spite of the slow death which means lower production cycles/year, the productivity/year was higher at 25 °C than at other temperatures. The SpltNPV produced at 25 °C was also found to be of superior quality in terms of low bacterial contaminants than at 35 °C. Neonate larval bioassay conducted with viruses produced at different temperature treatments revealed that they were similar in their activity and virulence. Hence our results indicate that maintenance of the SpltNPV production facility at 25 °C would enhance both the virus productivity and quality.     

Corylomyces: a new genus of Sordariales from plant debris in France

The new genus Corylomyces, isolated from the surface of a hazelnut (Corylus avellana) in the French Pyrenees, is described, illustrated and compared with morphologically similar taxa. It is characterised by tomentose, ostiolate ascomata possessing long necks composed of erect to sinuose hairs, and one- or two-celled, opaque, lunate to reniform ascospores. Analyses of the SSU and LSU fragments rDNA gene sequences support its placement in the Lasiosphaeriaceae (Sordariales).     

Synthesis and evaluation of bioactive naphthoquinones from the Brazilian medicinal plant, Tabebuia avellanedae

A series of naphthoquinones based on the naphtho[2,3-b]furan-4,9-dione skeleton such as (−)-5-hydroxy-2-(1′-hydoxyethyl)naphtho[2,3-b]furan-4,9-dione (1) and its positional isomer, (−)-8-hydroxy-2-(1′-hydoxyethyl)naphtho[2,3-b]furan-4,9-dione (2), which are secondary metabolites found in the inner bark of Tabebuia avellanedae, were stereoselectively synthesized and their biological activities were evaluated in conjunction with those of their corresponding enantiomers. Compound 1 exhibited potent antiproliferative effect against several human tumor cell lines, but its effect against some human normal cell lines was much lower than that of mitomycin. On the other hand, its enantiomer (R)-1 was less active toward the above tumor cell lines than 1. The antiproliferative effect of 2 against all tumor cell lines was significantly reduced. These results indicated the presence of the phenolic hydroxy group at C-5 is of great important for increasing antiproliferative effect. In addition, 1 also showed higher cancer chemopreventive activity than 2, while there were no significant differences between 1 and 2 in antimicrobial activity. Both compounds displayed modest antifungal and antibacterial activity (Gram-positive bacteria), whereas they were inactive against Gram-negative bacteria.     

skippy, a retrotransposon from the fungal plant pathogen Fusarium oxysporum

A retrotransposon from the fungal plant pathogen Fusarium oxysporum f. sp. lycopersici has been isolated and characterized. The element, designated skippy (skp) is 7846 by in length, flanked by identical long terminal repeats (LTR) of 429 by showing structural features characteristic of retroviral and retrotransposon LTRs. Target-site duplications of 5 bp were found. Two long overlapping open reading frames (ORF) were identified. The first ORF, 2562 by in length, shows homology to retroviral gag genes. The second ORF, 3888 bp in length, has homology to the protease, reverse transciptase. RNase H and integrase domains of retroelement pol genes in that order. Sequence comparisons and the order of the predicted proteins from skippy indicate that the element is closely related to the gypsy family of LTR-retrotransposons. The element is present in similar copy numbers in the two races investigated, although RFLP analysis showed differences in banding patterns. The number of LTR sequences present in the genome is higher than the number of copies of complete elements, indicating excision by homologous recombination between LTR sequences.     

Antibacterial efficacy of Withania somnifera (ashwagandha) an indigenous medicinal plant against experimental murine salmonellosis

In the present study, we evaluated the antibacterial activity of ashwagandha [Withania somnifera L. Dunal (Solanaceae; root and leaves)], an Indian traditional medicinal plant against pathogenic bacteria. Both aqueous as well as alcoholic extracts of the plant (root as well as leaves) were found to possess strong antibacterial activity against a range of bacteria, as revealed by in vitro Agar Well Diffusion Method. The methanolic extract was further subfractionated using various solvents and the butanolic sub-fraction was found to possess maximum inhibitory activity against a spectrum of bacteria including Salmonella typhimurium. Moreover, in contrast to the synthetic antibiotic (viz. chloramphenicol), these extracts did not induce lysis on incubation with human erythrocytes, advocating their safety to the living cells. Finally, the antibacterial efficacy of the extracts isolated from plant (both root and leaves) was determined against experimental salmonellosis in Balb/C mice. Oral administration of the aqueous extracts successfully obliterated salmonella infection in Balb/C mice as revealed by increased survival rate as well as less bacterial load in various vital organs of the treated animals.     

Food plant specialization and feeding efficiency in the tent caterpillars Malacosoma disstria and M. americanum

Larvae of Malacosoma americanum (F.) an oligophagous species that feeds primarily on Prunus and other rosaceous trees, were compared to larvae of the more highly polyphagous congener M. disstria Hb., with respect to the efficiency of utilization of the foliage of a common host plant, Prunus serotina Ehrh. We obtained values similar to those reported for other Lepidoptera for the commonly used measures of the fraction of ingested food that was assimilated, and for the growth in dry weight per unit of food ingested or assimilated. Moreover, the two species did not differ in any measure of efficiency. Our results are compatible with the conclusion that specialized phytophagous insects do not use their host plants with greater physiological efficiency than do generalized insects.

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Zusammenfassung Raupen von Malacosoma americanum, einer oligophagen Art, die sich vor allem auf Prunus und andern baumartigen Rosaceen. entwickelt, wurden mit Raupen der polyphageren Verwandten M. disstria in Hinblick auf die Verwertung der Blätter ihres gemeinsames Wirtes Prunus serotina verglichen. Wirerhielten ähnliche Werte wie sie früher für andere Lepidopteren publiziert worden waren und zwar in Bezug auf die üblichen Messwerte, Anteil verwertete Nahrung, Wachstum in Trockengewicht pro Einheit gefressene oder verwertete Nahrung. Zudem unterschieden sich die beiden Arten in keiner Masszahl für Effizienz. Unsere Resultate sind im Einklang mit der Folgerung, das spezialisierte phytophage Insekten ihre Wirtspflanzen nicht besser ausnützen als as polyphage Arten tun.

     

Genetic transformation of Tylophora indica with Agrobacterium rhizogenes A4: growth and tylophorine productivity in different transformed root clones

We have developed an efficient transformation system for Tylophora indica, an important medicinal plant in India, using Agrobacterium rhizogenes strains LBA9402 and A4 to infect excised leaf and stem explants and intact shoots at different sites. The induction of callus and transformed roots was dependent on the bacterial strain, explant type and inoculation site used. Transformed roots were induced only in explants infected with A. rhizogenes strain A4, while an optimal transformation frequency of up to 60% was obtained with intact shoots inoculated at the nodes. The presence of the left-hand transferred DNA (T_L-DNA) in the genome of T. indica roots induced by A. rhizogenes was confirmed by PCR amplification of the rooting locus genes of A. rhizogenes. Root growth and the production of tylophorine, the major alkaloid of the plant, varied substantially among the nine root clones studied. Both parameters increased over time in liquid cultures, with maximum biomass and tylophorine accumulation occurring within 4–6 weeks of growth in fresh medium. Interestingly, in liquid culture, the culture medium also accumulated tylophorine up to concentrations of 9.78±0.21 mg l^–1.     

Antioxidant efficacy of iridoid and phenylethanoid glycosides from the medicinal plant Teucrium chamaedris in cell-free systems

Twelve glycosides, seven iridoids and five phenylethanoids, have been isolated from leaf and root methanolic extracts of Wall Germander (Teucrium chamaedrys), a Mediterranean species historically used as a medicinal plant. Among them, three iridoid and one phenylethanoid glycosides have been isolated and characterized for the first time. All of the structures have been elucidated on the basis of their spectral data, especially 1D and 2D NMR experiments.The antioxidative properties of pure metabolites, as well as of crude organic extracts of the plant, have been analyzed on the basis of their DPPH radical scavenging capability. The antioxidant capacity in cell-free systems of the isolated metabolites was carried out by measuring their capabilities to inhibit the synthesis of thiobarbituric acid reactive species in assay media using as oxidable substrates a vegetable fat and the pentose sugar 2-deoxyribose and to prevent oxidative damage of the hydrosoluble bovine serum albumin (BSA) protein. Phenylethanoid glycosides resulted efficacious DPPH radical, while iridoid glycosides prevent massively the 2-deoxyribose and BSA oxidations in assay media.     

Efficient screening and breeding of Bacillus thuringiensis subsp. kurstaki for high toxicity against Spodoptera exigua and Heliothis armigera

Spodoptera exigua is one of the most renowned agricultural pest insects and relatively insensitive to Bacillus thuringiensis subsp. kurstaki strains which are widely used commercial products to control lepidopterans such as Heliothis armigera. In the current study, we have developed a new and efficient approach to screen and breed a B. thuringiensis subsp. kurstaki strain exhibiting high toxicity against S. exigua while retaining its high toxicity against H. armigera. UV and diethyl sulfate methods were used for mutagenesis, followed by an agar plug plate diffusion assay for preliminary screening of Zwittermicin A over-producing mutants, from which we obtained a mutant strain, designated here as B. thuringiensis subsp. kurstaki D1-23, with high toxicity against S. exigua. The toxicity of D1-23 against S. exigua and H. armigera was improved by 115.4 and 25.9%, respectively, compared to its parental commercial strain BMB005.     

Effect of Bacillus thuringiensis naturally colonising Brassica campestris var. chinensis leaves on neonate larvae of Pieris brassicae

The feeding of neonate larvae of Pieris brassicae (Order Lepidoptera) on leaves of brassica plants that had been colonised by Bacillus thuringiensis resulted in the death of 35% of the population within 72 h. The bacteria multiplied in the cadavers, resulting in an increase of about 50-fold compared to the living insects. Surviving insects showed no ill effects during the time of the study. There was negligible multiplication of B. thuringiensis in the frass.     

Nomenclatural changes resulting from the transfer of tropical African Sarcostemma to Cynanchum ( Apocynaceae : Asclepiadoideae )

Summary  Four species of tropical African Sarcostemma are transferred to Cynanchum together with two subspecies of S. viminale. In addition, Sarcostemma mulanjense is reduced to subspecific rank under C. viminale.     

Baculovirus-mediated Expression of a Gene for Trehalase of the Mealworm Beetle, Tenebrio molitor, in Insect Cells, SF-9, and Larvae of the Cabbage Armyworm, Mamestra brassicae

It is of interest to understand what kinds of physiological and biochemical changes occur in insects if the homeostasis of trehalose in the hemolymph is disrupted by the infection with a recombinant baculovirus containing a secretory-trehalase gene. For this purpose, two recombinant non-occluded Autographa california multicapsid nucleopolyhedroviruses (AcMNPVs), vTREVL and vERTVL, containing a trehalase cDNA of the mealworm beetle, Tenebrio molitor, were constructed. The trehalase cDNA was inserted in the sense orientation downstream of the polyhedrin promoter for vTREVL, and in the anitsense orientation for vERTVL. The active trehelase of T. molitor was found outside of cells when SF-9 cells or larvae of the cabbage armyworm, Mamestra brassicae, were infected with vTREVL. In the hemolymph of vTREVL-infected larvae, expression of the active trehelase was followed by disappearance of trehalose and appearance of glucose. However, the mortality time of virus-infected 5th instar larvae increased in the following order: AcMNPV C6 (wild-type virus) ≤ vERTVL < vTREVL. The symptoms (the browning and liquefying of the host body) of NPV infection were moderated considerably in vTREVL-infected larvae.     

Insect antifeedant activity of clerodane diterpenes and related model compounds

A comprehensive compilation of all test results on the insect antifeedant activity of clerodane diterpenes and related model compounds is reported. To increase the compatibility of data from different sources, some of the results reported in the literature have been converted into a standardized form. The compounds were sorted into groups according to the different types of sidechain attached to C-9. Despite the wealth of information, collected in 15 tables, it remains difficult to assign importance to separate structural elements in relation to the observed antifeedant activity. A detailed analysis of the structure-activity relationships could not be presented, but some interesting trends can be distinguished based on the structures of the strongest antifeedants. The compilation covers the literature up to December 2001.     

Genes from Bacillus thuringiensis entomocidus 60.5 coding for insect-specific crystal proteins

Summary Five crystal protein genes have been isolated from DNA of Bacillus thuringiensis entomocidus 60.5, an isolate selected for its high toxicity against Spodoptera littoralis and Spodoptera exigua. Two of these genes belong to a family of well-described crystal protein genes. The toxic properties of the corresponding proteins are similar to those of isolate kurstaki HD1. The other three genes belong to gene families not described before. One of these genes codes for a protein product exhibiting a high degree of specificity towards Spodoptera species, explaining the high toxicity of isolate entomocidus 60.5 against these species. This gene product is much less toxic against larvae of Heliothis virescens and Pieris brassicae. Its coding sequence is separated from a supposed fourth crystal protein gene by a stretch of DNA of 3 kb. The crystal protein encoded by the fifth gene is mainly active against P. brassicae. Homology between the crystal protein genes is limited to the central region of the coding sequences, including the proteolytic cleavage site, except for the first two genes between which homology is extensive.