Paclobutrazol- and Hardening-Induced Thermotolerance of Wheat: Are Heat Shock Proteins Involved?

The present report describes the effects of paclobutrazol andheat hardening treatments on the protein synthesis patternsin imbibing and germinating wheat seedlings (Triticum aestivumL. cv Frederick) during heat stress. A heat hardening treatmentgiven during the imbibition period induced the transient expressionof 118, 90, 70 and 18 kDa heat shock proteins (HSPs). However,the hardening and paclobutrazol treatments did not enhance thethermotolerance of imbibed seeds or etiolated seedlings. Bycontrast, the hardening and paclobutrazol treatments enhancedthe thermotolerance of light-grown seedlings. While, both hardenedand unhardened control seedlings synthesized several HSPs duringa high temperature stress period, these proteins were not synthesizedby the paclobutrazol-treated, light-grown seedlings. Thus, HSPsynthesis during heat shock may have been a manifestation ofstress perception by the seedlings and may not have mediatedthe thermotolerance induced by the triazole treatments. Sincedifferential thermotolerance was only apparent in light-grownseedlings, it is suggested that chloroplasts may be requiredfor the expression of paclobutrazol- and hardening-induced thermoprotection.Additional evidence indicating that chloroplasts are an importantsite of injury during high temperature stress was obtained fromchlorophyll fluorescence measurements. (Received July 11, 1994; Accepted October 26, 1994)     

Interspecific competition between larvae is not an important strncturing force in mixed communities of Rana and Bufo on an English sand-dune system

We investigated the roles of competition and predation in the structuring of mixed anuran communities ( Rana temporaria and Bufo bufo ) in an English sand-dune system. Rana temporaria breeds earlier in spring than B. bufo. but the latter species was numerically superior in all ponds at the study site. We measured survival and growth rates of larvae in three duplicate sets of natural ponds. In one set we removed B. bufo spawn, leaving only R. temporaria^: in a second sel we removed R. temporaria spawn, leaving only B. bufo: and in a third set we left boih together. The third set also contained substantial numbers of predatory fish ( Perea fluviatilis and Gasterosteus aeuleatus ). Rana temporaria survival was lower than that of B, bufo in all ponds, on average by sixfold. Rana temporaria fared better on its own than in the mixed-species ponds, but lower survival in the latter ponds was probably due to fish predation rather than to Inierspecific competition. By eonlrast, B. bufo survival was sevenfold higher in mixed-species compared with single-species ponds, probably because they are distasteful to fish and thus benefited indirectly from fish-predation of invertebrates, Protothieca rieharsi. a mediator of interference competition between anuran larvae under laboratory conditions, was present in some of the dune ponds but was not correlated with larval growth inhibition. Taken together the evidence implicated predation of larvae and intraspecific competition, rather than interspecific competition, as the dominant structuring forces in this community.     

Stage-specific nitrogen metabolism in developing carrot somatic embryos

The physiology of individual somatic embryo developmental stages otDaucus carota L. was examined by in vivo nuclear magnetic resonance (NMR) spectroscopy, amino acid analysis and ¹⁴C-labeling. ¹⁵N NMR spectroscopy was used to examine the uptake and incorporation of ¹⁵N isotopically labeled inorganic nitrogen sources. NMR spectra of proembryogenic masses (PEMs) contained resonances for histidine, amino sugars, glutamine, arginine, urea, alanine. α-amino nitrogen, serine, aliphatic amines and several unknowns. Similar resonances were found in various embryo developmental stages. However, resonances for arginine and aliphatic amines peaked during globular and torpedo stages and substantially decreased in germinating stage embryos. The dominant resonances observed in non-embryogenic cells and germinating embryos were glutamine and α-amino nitrogen. Amino acid analysis of the various embryo stages showed that glutamate, glutamine and arginine were the major contributors to the soluble amino acid profiles. During development, glutamate and glutamine continued to increase in concentration whereas arginine and its related metabolites (i.e. ornithine and y-aminobutyric acid [GABA]) were biphasic; increasing in globular and torpedo stage embryos and decreasing in germinating embryos. Carbon-14 labeling indicated that labeled glutamine pools in non-embryogenic and germinating embryos were greatest compared to other embryo stages, whereas labeled GABA pools were greatest in globular and torpedo stage embryos. Taken together, these data indicate that the physiology of each embryo developmental stage is distinct. They also suggest that during somatic embryo development, a switch takes place in metabolism whereby the glutamine synthetase/glutamate synthase (GS/GOGAT) pathway is predominant in non-embryogenic cells and germinating stage embryos. Furthermore, during early to mid-embryo development (PEMs, globular and torpedo stage embryos), metabolism utilizing the omithine cycle is enhanced and predominant.     

Disruption of keratin filaments in embryonic epithelial cell types.

The murine keratins Endo B and Endo A, which are homologs of the human keratins K18 and K8, constitute the intermediate filaments (IFs) that are found in all simple epithelia of the adult and in the first epithelial derivatives of the early embryo. The cellular role of simple epithelial keratins in development and differentiation was investigated by inducing filament collapse in HR9 endoderm and F9 embryonal carcinoma cells in which mutant Endo B protein was constitutively expressed. By immunolocalization techniques a perturbation of the keratin network was revealed as well as concomitant disruption of vimentin IFs and displacement of surface desmosomal proteins, demonstrating an intimate structural association of Endo B/A filaments with these cellular components. In aggregates of differentiating F9 cells displaying altered Endo A/B IFs, the formation of a compact, polarized visceral endoderm layer was significantly compromised. These results indicate that an intact keratin network influences the three-dimensional formation of cell-cell or cell-substratum contacts in embryonic visceral endoderm.     

The use of the Cole/Hurlbert C8 association coefficient in inverse ecological classification

Abstract. Inverse classification is routinely used in vegetation surveys to produce groups of sociologically similar species. However, no classification methods have been proposed specifically for this purpose, nor has any evaluation been made of the suitability of existing methodsforthe purpose. Anewvariant of Cluster Analysis is introduced, i. e. using the Cole/Hurlbert association measure C₈ as coefficient, and named for convenience Cole Cluster Analysis. This method, and four standard ones, were used on saltmarsh vegetational data from New Zealand. Ecophysiological data were obtained from salt-tolerance experiments. These data, and distributional information, were used as independent criteria against which to compare the inverse vegetation classifications. Information Analysis did not prove suitable for inverse analysis in this test. Nor did Cluster Analysis with the Simple Matching Coefficient or with Jaccard's coefficient. Indicator Species Analysis was more suitable, but the new Cole Cluster Analysis seemed the most effective on these data.     

Pyrimidine deoxyribonucleotide metabolism during maturation and germination of white spruce (Picea glauca) somatic embryos: metabolic fate of C-labelled cytidine, deoxycytidine and thymidine

Pyrimidine deoxyribonucleotide metabolism was investigated during maturation and germination of white spruce somatic embryos by following the metabolic fate of [2‐¹⁴C]cytidine, [2‐¹⁴C]deoxycytidine and [2‐¹⁴C]thymidine. The de-novo pathway of deoxyribonucleotides was estimated indirectly, by the ability of the tissue to incorporate cytidine into DNA after conversion to dCTP. The salvage pathway was estimated by the utilization of labelled cytidine, deoxycytidine and thymidine for synthesis of deoxyribonucleotides and nucleic acids. Utilization of cytidine for DNA synthesis, via the de novo pathway, was always lower than that observed for RNA throughout the course of the experiment. Incorporation of cytidine into RNA was found to occur either directly, after conversion to CTP, mediated by the enzymes cytidine kinase, nucleoside monophosphate kinase and nucleoside diphosphate kinase, or indirectly, after conversion to UTP via uridine and UMP. Active incorporation of uridine into RNA of white spruce-cultured cells was demonstrated previously. Salvage of deoxycytidine and thymidine was operative in maturing and germinating white spruce somatic embryos, as label from both compounds was recovered in nucleotides and DNA. However, the utilization of these precursors by the cells was different. Salvage of deoxycytidine was always higher than that observed for thymidine, which was extensively catabolized to CO₂ at all stages of embryo development.     

Evaluating Restoration Success on Lyell Island, British Columbia Using Oblique Videogrammetry

In degraded ecosystems where the impact on wildlife and the destruction of natural systems is high, restoration becomes a critical component of recovery. Monitoring restoration activities plays a key role in determining end points for restoration and assessing effectiveness. Appropriate monitoring of major systems, particularly in assessing vegetation reestablishment and slope stabilization, requires a long‐term commitment to annual assessment of change and improvement over time. However, intrinsic factors built into government or public management systems, such as budgeting and staffing limitations, limit the ability for long‐term monitoring of critical restoration projects. In the research reported in this article, we devised and assessed a new remote method for assessing restoration success and tested it on restoration and monitoring requirements in Lyell Island, British Columbia. We developed a system (the oblique data fusion system [ODFS]) to extract spatial information from oblique aerial video imagery. The ODFS enables low‐cost change detection and database updates at a range of operational scales. System tests show an absolute spatial accuracy on the order of ±2.1 m. The evaluation, based on digitized historical data, ground surveys, and the ODFS‐derived data, indicates that the landslide rate (new area/year) tapered off following treatment; after 5 years it had been reduced by a factor of 3 relative to the background rate. The recovery is deemed sufficient to initiate secondary restoration tasks. The evaluation demonstrates the accuracy and utility of the ODFS for long‐term monitoring of landscape restoration efforts, particularly in remote areas. In conclusion, this new innovative method shows considerable promise for park managers.     

An Origin of Transfer (oriT) on the Conjugative Element pRS01 from Lactococcus lactis subsp. lactis ML3

Previous analysis of the Tra1 region of the conjugative element pRS01 from Lactococcus lactis subsp. lactis ML3 suggested that an origin of transfer (oriT) was present. Deletion derivatives of this cloned Tra1 region were assayed for mobilization in the presence of the wild-type pRS01 element in trans. The pRS01 oriT was localized to a 446-nucleotide segment in the intergenic region between open reading frames ltrD and ltrE. Sequence analysis of this region revealed a cluster of direct and inverted repeat structures characteristic of oriT regions associated with other conjugative systems.     

Optimization of bud induction in cotyledonary explants of Pinus canariensis

A method is described for using liquid pulsing as an alternative to the conventional induction protocol for Pinus canariensis. Using Day 0 and Day 3 explants, the best exposure time was 8 h and 4 h respectively, in a non-buffered 100 M N⁶-benzyladenine solution, followed by culture on half-strength Bornman's medium containing 3% sucrose and 0.8% Difco Bacto^R agar. With this procedure, 97% of the cotyledonary explants produced about 14 buds/explant. These results were comparable to a 14-day induction period on full-strength Bornman's medium containing 10 M N⁶-benzyladenine.