Relationship of vector insert size to homologous integration during transformation of Neurospora crassa with the cloned am (GDH) gene

Summary We used lambda and plasmid vectors containing the am ⁺ gene in an insert of from 2.7 to 9.1 kb, to transform am point mutant and deletion strains. A total of 199 transformants were examined with the potential to yield am ^– transformants by homologous recombination. When we used vectors that had 9.1 kb of homology with the chromosomal DNA, 30% of the transformants obtained were the result of homologous recombination regardless of whether the vector was a lambda molecule, a circular plasmid, or a plasmid that had been linearized prior to transformation. When vectors with up to 5.1 kb of homology were used, very few transformants (1 of 89 tested) resulted from homologous recombination. Of a sample of 29 ectopic integration events obtained by transformation with the 9.1 kb fragment cloned in a vector, 18 included a major part (usually almost all) of both arms of lambda with the entire Neurospora 9.1 kb insert between them. Four included only long arm sequence together with an adjacent segment of the insert containing the am gene. The remaining seven were the result of multiple integrations. There was no evidence of circularization of the vector prior to integration. All transformants that had multiple copies of the am gene appeared to be subject to the RIP process, which causes multiple mutations in duplicated sequences during the sexual cycle.     

Apparent phytotoxicity of mononuclear hydroxy-aluminum to four dicotyledonous species

It has long been assumed that Al³⁺ is an important rhizotoxic ion in acid soils around the world, but the toxicity of Al³⁺ relative to mononuclear hydroxy-Al [AlOH²⁺ and Al(OH)⁺₂] has been examined in detail only for an Al-sensitive wheat variety ( Triticum aestivum L. cv. Tyler). That plant appears to be sensitive to Al³⁺ but not to AlOH²⁺ and Al(OH)⁺₂. New experiments, and reanalyses of previously published experiments, provide evidence that dicotyledonous species may be sensitive to mononuclear hydroxy-Al and that Al³⁺ may be nontoxic, or less toxic, to those plants. Despite these consistently measured differences between wheat and the dicotyledons, the determination of relative toxicities (Al³⁺ vs mononuclear hydroxy-Al) may be an intractable problem. Because of hydrolysis equilibria, (AlOH²⁺) and (Al(OH)⁺₂) are equivalent to (Al³⁺)k₁(H⁺)⁻¹ and (l³⁺)k₂(H⁺)⁻², respectively, in which k₁ and k₂ are the first and second hydrolysis constants (braces denote activities). Thus, any expression of root elongation as a function of mononuclear hydroxy-Al can be alternatively expressed as a function of (Al³⁺) and (H⁺). Toxicity attributed to mononuclear hydroxy-Al may actually be Al³⁺ toxicity that increases as pH rises (i.e. Al³⁺ toxicity ameliorated by H⁺).     

Phytophagous insects enhance nitrogen flux in a desert creosotebush community

Summary We tested the hypothesis that herbivorous insects on desert shrubs contribute to short-term nitrogen cycling, and increase rates of nitrogen flux from nutrient rich plants. Creosotebush (Larrea tridentata) shrubs were treated with different combinations of fertilizer and water augmentations, resulting in different levels of foliage production and foliar nitrogen contents. Foliage arthropod populations, and nitrogen in canopy dry throughfall, wet throughfall and stemflow were measured to assess nitrogen flux rates relative to arthropod abundances on manipulated and unmanipulated shrubs over a one-month period during peak productivity. Numbers and biomass of foliage arthropods were significantly higher on fertilized shrubs. Sap-sucking phytophagous insects accounted for the greatest numbers of foliage arthropods, but leaf-chewing phytophagous insects represented the greatest biomass of foliage arthropods. Measured amounts of bulk frass (from leaf-chewing insects) were not significantly different among the various treatments. Amounts of nitrogen from dry and wet throughfall and stemflow were significantly greater under fertilized shrubs due to fine frass input from sap-sucking insects. Increased numbers and biomass of phytophagous insects on fertilized shrubs increased canopy to soil nitrogen flux due to increased levels of herbivory and excrement. Nitrogen excreted by foliage arthropods accounted for about 20% of the total one month canopy to soil nitrogen flux, while leaf litter accounted for about 80%.     

Studies on the biosynthesis and metabolism of the phytoalexin lubimin and related compounds in Datura stramonium L.

Arachidonic acid, cellulase, CuSO₄, a sonicate of Phytophthora infestans mycelium and a spore suspension of Penicillium chrysogenum all elicited the formation of the sesquiterpenoid phytoalexins lubimin, 3-hydroxylubimin and rishitin in fruit cavities of Datura stramonium. 3-Hydroxylubimin was the predominant phytoalexin formed after treatment of the fruits with arachidonic acid, cellulase and the P. infestans preparation. Copper sulphate was a potent elicitor of lubimin but not 3-hydroxylubimin. The fungus P. chrysogenum metabolized lubimin and 3-hydroxylubimin to 15-dihydrolubimin and 3-hydroxy-15-dihydrolubimin respectively, both in fruit cavities inoculated with spores of this fungus and in pure culture. The 15-dihydrolubimin formed in the fruits by the fungus was further metabolized (by the fruits) to both isolubimin and 3-hydroxy-15-dihydrolubimin. The precursor-product relationships between all of the subject compounds was investigated by feeding experiments with ³H-labelled compounds. 2-Dehydro-[15-³H₁]lubimin was rapidly and efficiently incorporated into lubimin and may be the direct precursor of lubimin in planta. 3-Hydroxy[2-³H₁]lubimin was incorporated into the nor-eudesmane rishitin but 10-epi-3-hydroxy[2-³H₁]lubimin was not. An updated scheme for the biosynthesis and metabolism of lubimin and related compounds in infected tissues of solanaceous plants is presented.We thank Mr Vic Swetez for the provision of plant material, Mrs Margaret Huffee for technical assistance, Dr David Ewing for help with obtaining NMR spectra, and the Agricultural and Food Research Council for financial support.     

Social relationships between adult male and female rhesus macaques: II. non-sexual affiliative behaviour

The social relationships of adult male rhesus macaques (Macaca mulatta) in Group I of the Cayo Santiago colony were studied over a period of 14 months. Relationships were found in which adult males and sexually mature females were persistently close to one another in nonsexual contexts, both within and across seasons. Males of long tenure and high dominance rank tended to have more female partners, and more persistent relationships, than more recent immigrants of lower rank. Correlations with length of tenure were stronger than those with dominance rank. Closely-related females tended to have persistent relationships with the same male. In most cases the female was primarily responsible for maintaining proximity in non-sexual contexts. Dyads which were persistently within 5 m of each other in the birth season were more likely to form a consortship in the subsequent mating season than those which had a brief relationship. A similar tendency was apparent in the 1 m data but it was not statistically significant. There was no association between persistent proximity during the birth season and the occurrence of long, or multiple consortships, nor with the maintenance of proximity or direction of grooming between consort partners. The pattern of consortships was not closely related to the formation of persistent relationships in the subsequent birth season. Females occasionally received protection from their male partners and, in some cases, spent more time in the feeding corral with them than did other females. Affiliative relationships can be very enduring and may have long-term benefits that were not apparent during the study period.     

E infα supk transgene in B10 mice suppresses the development of myasthenia gravis

Mice bearing the H-2 ^bhaplotype are susceptible to the development of experimental autoimmune myasthenia gravis (EAMG), induced by acetylcholine receptor (AChR) autoimmunity. One of the genes influencing EAMG susceptibility has been mapped to the A ^blocus of the major histocompatibility complex, and the A chain has been implicated in the pathogenesis. Mice of the H-2 ^bhaplotype, including C57BL/10 (B10), have a genomic deletion of the E gene and therefore fail to express the E molecule on their cell surface. To test the hypothesis that failure to express the cell surface E molecule in B10 mice contributes to EAMG pathogenesis, E _inf ^supk transgenic B10 mice expressing the T molecule were examined. Expression of the E molecule in E _inf ^supk transgenic B10 mice partially prevented the development of EAMG.