Functional analysis of a complex oncogene arrangement in biotype III Agrobacterium tumefaciens strains

The ubiquitous grapevine-associated octopine/cucumopine Ti plasmids of biotype III Agrobacterium tumefaciens strains carry two T regions, TA and TB, with a complex oncogene arrangement. Within the octopine/cucumopine group, two main strain types were identified: large TA strains with a TA region resembling the TL region of the biotype I octopine strain Ach5 and small TA strains with a similar T region organization as the large TA strains but with a large internal TA deletion. Structural and functional studies of the representative large TA strain Tm4 revealed six oncogenes. Each oncogene was inserted in a disarmed vector and tested for biological activity using the corresponding oncogenes of Ach5 as standards. Five Tm4 oncogenes, TA-iaaM, T-ipt, T-6b, TB-iaaH and TB-iaaM, were shown to be active, the IS-interrupted TA-iaaH gene was inactive. To study the role of each gene in the pTiTm4 context, several single and multiple pTiTm4 mutations were constructed. It was shown that whereas TA-iaaM and TB-iaaH are essential for tumour formation on grapevine, T-ipt, T-6b and TB-iaaM are not. The avirulence of the TA-iaaM ^- mutant was shown to be due to an inhibitory effect of the T-ipt gene, since a TA-iaaM ^- /T-ipt ^- double mutant was fully virulent. We conclude that the TA-iaaM gene of large TA strains is specifically required to counteract the tumour growth inhibiting activity of the T-ipt gene. Both TA-iaaM and T-ipt are absent from the small TA strains. A model on the roles and interactions of the different oncogenes in large TA and small TA strains is presented.     

Neuromuscular development following tetrodotoxin-induced inactivity in mouse embryos

Developmental aspects of the neuromuscular system in mouse embryos chronically paralyzed in utero with tetrodotoxin (TTX) between embryonic days 14 and 18 were studied using biochemical and histological methods. The number of lumbar spinal motoneurons (MNs) was higher in inactive embryos than in controls suggesting a decreased motoneuron cell death. In association with the increase in MN number, choline acetyltransferase activity was significantly increased in both spinal cord and peripheral synaptic sites. Paralyzed muscles exhibited a decreased number of mature myofibers and the nuclei were centrally located. Creatine kinase activity was greatly decreased and total acetylcholine receptor and receptor cluster numbers per myofiber were significantly increased in paralyzed muscles. A similar pattern of changes occurs in the neuromuscular system of the mutant mouse muscular dysgenesis (mdg). However, in contrast to the mdg mutant, tetrodotoxin-treated muscles were similar to controls in their innervation pattern, in the ultrastructural aspects of the excitation–contraction coupling system (i.e., dyads and triads) and in the extent of dihydropyridine binding. Thus, neuromuscular inactivity is not sufficient to impair the pattern of muscle innervation or the appearance of either the triadic junctions or dihydropyridine receptors. These results indicate that alterations of dihydropyridine binding sites and triads in muscular dysgenesis cannot be accounted for by inactivity but rather must reflect a more primary defect involving the structural gene(s) regulating the development of one or more aspects of muscle differentiation.     

Sexual behavior of donkey jacks: influence of ejaculatory frequency and season

The courtship of 5 jacks was evaluated weekly for 12 mo. The characteristics of sexual behavior were recorded before the collection of 2 ejaculates, at 4-h intervals, into an artificial vagina in the presence of a female in either natural or induced estrus. The maximum time given to the jack to perform the ejaculatory mount was 1 h. If the jacks did not ejaculate, another attempt was made the following week. At the time of collection, the male and female remained free in a paddock (20 m2). The data presented in this study is based on results in which the first and second ejaculates occurred successively. Statistical analysis was done by a 2 x 2 factorial design in randomized blocks. The mean +/- SEM of sexual behavior characteristics for the first and second ejaculate were, respectively: time until first mount = 0.7 +/- 0.2 and 0.9 +/- 0.2 min; time until first erection = 11.9 +/- 1.1 and 11.6 +/- 1.1 min; time until ejaculation = 15.0 +/- 1.2 and 13.6 +/- 1.2 min; frequency of partial exposure of penis = 3.4 +/- 0.4 and 3.2 +/- 0.4; frequency of total exposure of penis = 1.0 + 0.2 and 1.1 +/- 0.2; frequency of flehmen responses = 6.6 +/- 0.5 and 4.6 +/-0.5; frequency of erections = 1.3 +/- 0.1 and 1.2 +/- 0.1; frequency of retreats away from the female = 1.8 +/- 0.2 and 1.0 +/- 0.2; frequency of mounts with erection but without ejaculation = 0.3 +/- 0.1 and 0.1 +/- 0.1; frequency of mounts without erection = 2.0 +/-0.1 and 1.2 +/- 0.1 ; and frequency of pelvic copulatory movements = 4.8 +/- 0.4 and 4.4 +/- 0.4. Individual differences were observed (P<0.05) for partial and total exposure, flehmen responses, mounts without erection and pelvic copulatory movements. The variables flehmen responses, retreats away from the female and mounts without erection showed significant differences (P<0.05) between ejaculates. Seasonal effects on sexual behavior characteristics were not found. However, a monthly effect was noted for flehmen responses, partial exposure of the penis, mounts without erection, retreats away from the female and pelvic copulatory movements when the 2 ejaculates were combined.     

Dynamics of colony development in the paper waspPolistes dominulus Christ (Hymenoptera,Vespidae): The influence of prey availability

This study focused on how a decrease in prey availability affected the development of aP. dominulus Christ colony. Nutritional oophagy and larval development were parameters found to be most directly affected. The more indirect effects on the growth of the nest and on offspring production were also analyzed.     

Culture analysis and external interaction models of mycelial growth

To investigate morphogenesis and in particular circularization mechanisms in young mycelia, we observe cultures of the zygomyceteMucor spinosus and develop discrete models of two-dimensional filamental branching growth. The models are based on the hypothesis that the fungus secretes a regulatory substance that diffuses into the surrounding medium and is detected by the growing hyphae. We also present a simple Markovian growth model without such a feedback, but yielding to analytical computations.     

A protein involved in co-ordinated regulation of inorganic carbon and glucose metabolism in the facultative photoautotrophic cyanobacteriumSynechocystis PCC6803

The involvement of a gene ofSynechocystis PCC6803,icfG, in the co-ordinated regulation of inorganic carbon and glucose metabolism, was established. TheicfG gene codes for a 72 kDa protein, which shows no homology with those registered in data libraries. Expression oficfG required glucose, the actual inducer probably being glucose-6-phosphate, and was independent of light and of the external inorganic carbon concentration. Mutants carrying an inactivated copy oficfG were constructed. Their growth characteristics were identical to those of the wild type under all regimes except in limiting inorganic carbon with glucose being present either before or after the transfer to the limiting conditions. These conditions completely prevented growth, both in the light and in the dark. The inhibition could be relieved by several intermediates of the tricarboxylic acid cycle. Assays of various enzymic activities related to inorganic carbon uptake and to its assimilationvia either the Calvin cycle or phosphoenolpyruvate carboxylase did not reveal the level of action of IcfG. Possible models include a blockage of the assimilation of both carbon sources in the absence of IcfG, or the inhibition of Ci incorporation route(s) essential under limiting inorganic carbon conditions, even when glucose is present, and even in the dark.