Prochymosin expression in Bacillus subtilis

Prochymosin (PC) sequence was cloned in Bacillus subtilis using two kinds of plasmid constructions. In plasmid pSM316 the cDNA was inserted to obtain the intracellular expression of the enzyme. The enzyme turned out to be expressed in an insoluble form which could be converted to native enzyme under proper denaturing and refolding conditions. The levels of intracellular expression of PC were further enhanced by modifying the 5' region of the gene in a way that a two-cistron expression system was created. For the PC secretion, the cDNA was fused to the subtilisin leader sequence and expressed under the control of the B. subtilis neutral protease promoter. A properly folded PC was secreted by the cells, although to low levels.     

Interacting effects of increased tannin levels on leaf-mining insects

Tannin-rich extracts were painted on leaves of Q. emoryi (Fagaceae) that contained first instar larvae of a leaf miner, Cameraria sp. (Lepidoptera: Gracillariidae, Agrifoliella group), to test the role of tannins in increasing parasitism, while controlling for changes in protein content and physical damage. Miners from leaves that were treated with tannins had higher rates of parasitism by hymenopterans, but decreased mortality from other causes. The negative and positive effects of tannins balanced since overall larval survivorship was equivalent for leaf miners on tannin-enriched and control leaves. The tannin-rich extract also affected larval development because pupal weight was decreased for leaf miners in treated leaves. Tannins can have both adverse and beneficial effects on these phytophagous insects.

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Résumé Des extraits riches en tanins ont été étalés sur des feuilles de Quercus emoryi contenant des chenilles de ler stade de Cameraria sp., pour contrôler l'influence du tanin dans l'accroissement du parasitisme sans modifier la teneur en protéines ni provoquer de dommages physiques. Les mineuses de feuilles traitées avaient des taux plus élevés de parasitisme par les hyménoptères, mais une diminution des autres causes de mortalité. Les effets négatifs et positifs des tanins se sont équilibrés, puisque la survie des mineuses était la même pour les feuilles traitées que pour les témoins. Les extraits riches en tanin ont agi sur le développement larvaire puisque le poids des chrysalides des mineuses de feuilles traitées était plus faible. Les tanins peuvent ainsi avoir des effets nuisibles et bénéfiques sur ces insectes phytophages.

     

横断山齿突蟾属一新种(两栖纲:锄足蟾科)

1982年在横断山区考察期间,发现四川木里截族自治县的齿突蟾Scutiger标本在第二性征、趾蹼等特征与已知的各种齿突蟾均不相同,描述为新种木里齿突蟾Scutiger muliensis。     

Identification of Tn4430, a transposon of Bacillus thuringiensis functional in Escherichia coli

Summary The mobile genetic element Tn4430, originating from the gram-positive bacterium, Bacillus thuringiensis, and previously described as the Th-sequence, is the first transposon isolated from the genus Bacillus. In the present work a gene (APH-III) conferring resistance to kanamycin was inserted into this 4.2 kb transposon. Transposition experiments showed that Tn4430APH-III could transpose in the gram-negative host Escherichia coli when its insertion functions were supplied by an intact copy of Tn4430. By transposing Tn4430APH-III directly onto pBR322, it was possible to determine the nucleotide sequence of the terminal inverted repeats of Tn4430 and of the target DNA site. Identical 38 bp in inverted orientation are situated at each end of the transposon and there is a direct duplication of 5 bp at the insertion site. Thus, it is clear that Tn4430 is closely related to the transposons belonging to the Tn3 family (class II elements).     

The spiders of a citrus grove in israel and their role as biocontrol agents ofCeroplastes floridensis [Homoptera: Coccidae]

A 12 month survey was carried out of the spider population in a commercial citrus grove at Kibbutz Afeq in Northern Israel. The spiders collected from grapefruit foliage and ground cover were identified; young stages were reared to maturity and then identified.Chiracanthium mildei L. Koch represented 52% of all spiders captured during the year andTheridion sp. accounted for 34%. In the undercover,Gnaphosidae spiders represented 43% of the total spiders captured in pitfall traps andLycosidae 35%. A field experiment was carried out to evaluate the effectivness of spiders in biological control of the scaleCeroplastes floridensis Mask. in the citrus ecosystem. Wherever spiders were undisturbed on tree branches, populations ofC. floridensis were not able to develop to a level sufficient to cause economic damage. The increase in the number of scales was minimal: from 47 initially to 56. There was no damage to leaves nor was honeydew or sooty mold observed. During the same time period, on the 3 branches from which spiders had been eliminated, the number of scales increased seven-fold from 44 to 309. Leaves were heavily infested with sooty mold fungi that developed on the honeydew produced by the scales. Contribution from the Agricultural Research Organization, Neve Ya'ar, Regional Experiment Station. P.O. Haifa 31999, Israel. No. 1314-E 1984, series.     

Isolation of spore-forming bacilli from mosquitoes in natural breeding habitats in Iraq

New isolates of spore-forming bacilli from larvae and pupae of 3 species of mosquitoes are recorded in central Iraq.Bacillus sphaericus Meyer and Neide was isolated fromCulex pipiens (L.) larva.Bacillus carotarum Koch andBacillus cereus Frankland & Frankland were isolated fromTheobaldia longiareolata (Macquart) pupae.Bacillus laterosporus Laubach andBacillus thuringiensis (H 18) were isolated fromAedes caspius (Pallas) larvae. In addition, unidentifiedBacillus spp. were isolated fromCx. pipiens, T. longiareolata andAe. caspius larvae. Examination of soil samples collected from mosquito natural breeding habitats revealed isolates ofB. cereus, Bacillus thuringiensis H4a 4b; H 12 and H 16 and an unidentifiedBacillus sp.

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Résumé Des souches bactériennes sporogènes sont isolées de moustiques qui se trouvent dans la région centrale de l'Irak. Les résultats obtenus sont les suivants:Bacillus sphaericus Meyer & Neide [Culex pipiens (L.), larve],Bacillus carotarum Koch etBacillus cereus Frankland & Frankland [Theobaldia longiareolata (Macquart), nymphe],Bacillus laterosporus Laubach etBacillus thuringiensis (H 18) [Aedes caspius (Pallas), larvel]. L'examen des larves deCulex pipiens. T. longiareolata etAe. caspius, ainsi que l'analyse des échantillons du sol prélevés dans la région montrent la présence deBacillus cereus, Bacillus thuringiensis H4a 4b; H12 plus H16 et d'autresBacillus non identifiés.

     

Comparative study of methods for the quantification of fungal spores at the leaf surfaces ofPopulus xeuramericana

Summary The removal of fungal spores, urediniospores ofMelampsora medusae and conidia ofPestalozzia sp., from the leaf surfaces ofPopulus xeuramericana (Dode) Guinier cv. I-488 was assessed using three cultural techniques conventionally employed in phylloplane studies. The method of removal and the original density of spore deposition, but not the interaction of these factors, were significant determinants of variability in spore removal. Irrespective of the original density of deposition, the leaf print method was the most, and the leaf washing technique the least, efficient means of spore removal from the leaf surface. Factors which could contribute to this difference in efficiency are discussed.     

Purification and characterization of thermostable leucine dehydrogenase from Bacillus stearothermophilus

Leucine dehydrogenase (l-leucine: NAD⁺ oxidoreductase, deaminating, EC 1.4.1.9) has been purified to homogeneity from a moderate thermophilic bacterium, Bacillus stearothermophilus. Am improved method of preparative slab gel electrophoresis was used effectively to purify it. The enzyme has a molecular mass of about 300,000 and consists of six subunits with identical molecular mass (Mr, 49,000). The enzyme does not lose its activity by heat treatment at 70° C for 20 min, and incubation in the pH range of 5.5–10.0 at 55° C for 5 min. It is stable in 10 mM phosphate buffer (pH 7.2) containing 0.01% 2-mercaptoethanol at over 1 month, and is resistant to detergent and ethanol treatment. The enzyme catalyzes the oxidative deamination of branched-chain l-amino acids and the reductive amination of their keto analogs in the presence of NAD⁺ and NADH, respectively, as the coenzymes. The pH optima are 11 for the deamination of l-leucine, and 9.7 and 8.8 for the amination of -ketoisocaproate and -ketoisovalerate, respectively. The Michaelis constants were determined: 4.4 mM for l-leucine, 3.3 mM for l-valine, 1.4 mM for l-isoleucine and 0.49 mM for NAD⁺ in the oxidative deamination. The B. stearothermophilus enzyme shows similar catalytic properties, but higher activities than that from Bacillus sphaericus.Dedicated to Prof. Dr. G. Drews on the occasion of his 60th birthday