Fruiting at High Temperature and Its Genetic Control in the Basidiomycete Flammulina velutipes

Of 10 geographic strains of Flammulina velutipes, 4 were found capable of fruiting at 22°C (Fr_H) rather than at the typical 15°C (Fr_L). Crosses made between Fr_H and Fr_L monokaryons were never observed to fruit at 22°C. However, some hybrids did fruit at the intermediate temperature of 18°C when grown on appropriate substrates, indicating incomplete dominance of the low-temperature requirement. Analysis of progeny of five Fr_H × Fr_L crosses indicated that a minimum of two genes appears to control the requirement for fruiting at ≤15°C. The genes are not closely linked to either incompatibility locus.     

Molecular and genetic analysis of factors controlling host range in Agrobacterium tumefaciens

Summary We have investigated the factors which contribute to the host specificity of a tumor inducing plasmid of Agrobacterium, pTiAg162, which confers a narrow host range. Determinants both within the T-DNA and virulence regions contribute to host specificity. Within the T-DNA a defective cytokinin biosynthetic gene limits host range. Nucleotide sequence analysis revealed a large deletion in the 5 coding region of this gene when compared with the homologous gene from the wide host range tumor inducing plasmid, pTiA6. Introduction of the wide host range cytokinin biosynthesis gene into the T-DNA of the limited host range strain expanded the host range and suppressed the rooty morphology of tumors incited by the limited host range strain. Two genes from the virulence region of the wide host range plasmid, designated virA and virC, must also be introduced into the limited host range strain in order to restore a wide host range phenotype. The wide host range strain is avirulent on some cultivars of Vitis plants on which the limited host range strain induces tumors. This avirulence is apparently due to a hypersensitive response in which infected plant cells are killed at the site of inoculation. Mutations within the virC locus of the wide host range plasmid prevented the hypersensitive response and allowed the formation of tumors by the wide host range strain.     

Initial stages in the course of adventitious bud formation on embryos of Picea abies

Adventitious buds on embryos of Picea abies (L.) Karst. developed after a pulse treatment with 250 μ M benzyladenine (BA) of pH 5.5 for 2 h. Light and temperature regimes were not critical during the initial stages. Adventitious buds developed faster after a pulse treatment and the variation among different experiments was lower compared to when the embryos were cultured on media supplemented with BA. Various stages of the differentiation of adventitious buds were identified: stage 1 - appearance of meristematic centres (approximately the first two weeks); stage 2 - development of adventitious bud primordia (approximately the third week); stage 3 - adventitious bud development (from approximately the 4th to the 8th week). This system may be used for further studies on bud differentiation.     

Initiation and development of wound tissue and roots on hypocotyl cuttings of Pinus sylvestris in vitro

The rooting of hypocotyl cuttings from 20-day-old seedlings of Pinus sylvestris L. cultured in vitro is discussed. About 40% of the cuttings cultured on medium lacking activated charcoal produced roots during the first two months. When activated charcoal was added to the medium, either root formation (75% formed roots) or wound tissue growth (95% formed large wound tissues) was stimulated in different experiments. These large wound tissues did not develop any roots. The anatomical changes in the basal part of the cuttings were similar during the first two weeks in all the cuttings studied. A vascular cylinder composed of short tracheids with many pores developed. Thereafter the differentiation process became varied. The amount of wound tissue produced and the time for rooting differed among the cuttings. Tracheid nests which were in contact with the vascular system in the hypocotyl via short tracheids were observed after three weeks. Subsequently, roots developed from the tracheid nests. The longer root formation was delayed, the larger the wound tissue became.
Short tracheids were found close to the wound tissue surface. Their ability to adsorb nutrients and water is discussed.     

Limited-host-range plasmid of Agrobacterium tumefaciens: molecular and genetic analyses of transferred DNA.

A tumor-inducing (Ti) plasmid from a strain of Agrobacterium tumefaciens that induces tumors on only a limited range of plants was characterized and compared with the Ti plasmids from strains that induce tumors on a wide range of plants. Whereas all wide-host-range Ti plasmids characterized to date contain closely linked oncogenic loci within a single transferred DNA (T-DNA) region, homology to these loci is divided into two widely separated T-DNA regions on the limited-host-range plasmid. These two plasmid regions, TA-DNA and TB-DNA, are separated by approximately 25 kilobases of DNA which is not maintained in the tumor. The TA-DNA region resembles a deleted form of the wide-host-range TL-DNA and contains a region homologous to the cytokinin biosynthetic gene. However, a region homologous to the two auxin biosynthetic loci of the wide-host-range plasmid mapped within the TB-DNA region. These latter genes play an important role in tumor formation because mutations in these loci result in a loss of virulence on Nicotiana plants. Furthermore, the TB-DNA region alone conferred tumorigenicity onto strains with an intact set of vir genes. Our results suggest that factors within both the T-DNA and the vir regions contribute to the expression of host range in Agrobacterium species. There was a tremendous variation among plants in susceptibility to tumor formation by various A. tumefaciens strains. This variation occurred not only among different plant species, but also among different varieties of plants within the same genus.     

Transient enhancement of serum somatomedin levels prior to weaning of growth-impaired vasopressin-deficient Brattleboro rats

Somatomedin serum levels of congenitally vasopressin-deficient Brattleboro rats were determined postnatally between day 1 and 55, and compared with heterozygous control values. Assays were performed with a radioimmunoassay of insulin-like growth factor 1 (IGF-1). A transient enhancement of immunoreactive IGF-1 levels between day 8 and 21 of age and a reduction in adulthood was found. This observation shows that the early growth impairments of the Brattleboro mutant are not due to a deficiency of IGF-1.     

Antibodies to spiroperidol and their anti-idiotypes as probes for studying dopamine receptors

Spiroperidol was covalently conjugated to bovine serum albumin (BSA). Conjugated spiroperidol was almost as efficient as free spiroperidol in its binding capacity to dopamine receptor. Antibodies to spiroperidol were produced in rabbits following repeated immunizations with the conjugate of spiroperidol and BSA. The obtained antibodies have an apparent K_D of 0.02 nM for [³H]-spiroperidol. These antibodies bind also to other butyrophenones with IC₅₀ values three to four orders of magnitude higher than the IC₅₀ obtained with unlabeled spiroperidol. Antibodies were purified from anti-spiroperidol sera by affinity chromatography. Anti-idiotypic antibodies were raised in rabbits by immunization with the purified anti-spiroperidol antibodies. Some rabbits produced anti-idiotypic antibodies which bind to rat and calf striatum.     

Monoclonal antibodies modify acetylcholine-induced ionic channel properties in cultured chick myoballs

Summary Monoclonal antibodies directed against the cholinergic binding site of the acetylcholine receptor were found to alter the ion channel properties in cultured chick myoballs. Time and dose dependent reduction in acetylcholine sensitivity was observed. Noise analysis experiments indicated a decrease in the mean single channel conductance and an increase in the mean single channel open time.     

The effect of osmotic `shock'' on the swelling pattern and respiratory control of rat-liver mitochondria

1. Rat-liver mitochondria suspended in 0.25m-sucrose were exposed for a few seconds to strongly hypo-osmotic conditions, and then the osmolarity of the medium was raised again to 0.25 with the aid of tris chloride (osmotic ;shock'). 2. Mitochondria after hypo-osmotic pretreatment lost their capacity for slow energy-dependent swelling in iso-osmotic tris buffer and showed no respiratory control. 3. Swelling could be induced in the ;shocked' mitochondria by ATP but not by addition of respiratory substrates. 4. It was shown that cytochrome c is lost from ;shocked' mitochondria when they come into contact with the tris buffer present in the assay medium, and that the changes observed in the pattern of swelling, as well as in respiratory control, are directly connected with this loss of cytochrome c. 5. The results of the investigation are discussed with regard to the role of cytochrome c in swelling and respiratory control.