Computer-Assisted Image Analysis of Plant Growth, Thigmomorphogenesis, and Gravitropism

A nonintrusive auxonometric system, based on the DARWIN image processor (Telewski et al. 1983 Plant Physiol 72: 177-181), is described and demonstrated in the analysis of gravitropism and thigmomorphogenesis in corn seedlings (Zea mays). Using this system, growth and bending of regularly shaped plants or organs can be quickly and accurately measured without, in any way, interfering with the plant. Furthermore, the growth and bending curves are automatically plotted. Thigmomorphogenesis in the aerial part of corn seedlings involves growth promotion at a low force load and growth retardation at higher force loads. The time courses of the two kinds of response are somewhat different, with retardation occurring immeditely after mechanical perturbation and growth promotion taking somewhat longer to begin. Gravitropic experiments show that when dark-grown corn seedlings are placed on their side in the light, the resulting curvature is due to two consecutive morphological mechanisms. In the first instance, lasting for about 15 minutes, the elongation of the bottom edge of the plant accelerates, while the elongation of the top edge remains constant. After that, for the next 1.75 hours, the elongation of the top edge decelerates and stops while that of the bottom edge remains constant at the increased rate for most of the period. The measurements taken from both experiments at relatively high resolution (0.08-0.1 millimeter) show that the growth curves are not smooth but show many small irregularities which may or may not involve micronutations.     

Monoclonal antibodies to epitopes on different regions of the 200 00 dalton neurofilament protein : Probes for the geometry of the filament

Neurofilaments in mammalian nervous tissues have three subunit proteins. These subunit proteins have apparent molecular masses of 200 (NF200), 150 (NF150) and 68 (NF68) kD. Biochemical assembly studies have indicated that the NF68 protein forms the core of the filament and that the other two proteins are associated proteins. Electron microscopy immunolocalization studies have been performed previously on isolated filaments and on filaments from neurons in culture, and have confirmed the localization of NF68 as a core filament protein and NF200 as a peripheral protein. We have raised two monoclonal antibodies to the NF200 components. Using immunogold labelled protein A, we have been able to localize these antibodies to tissue sections of adult cerebellum at the EM level. With this method, we have found that one of the monoclonal antibodies (NF2) shows a linear arrangement of gold particles directly on the filament, whereas the second monoclonal antibody (NF111) reacts with the filaments to give a periodic arrangement of gold particles. By immunoblotting against chymotryptic fragments of the NF200 protein, we have found that the mAB-NF111 reacts solely with a 160 kD piece, whereas the other monoclonal antibody reacts with both the 160 kD piece and the 40 kD piece. The latter piece was shown to be associated to the filament by binding studies with iodinated NF68. Thus the EM localization studies and the biochemical studies indicate that the two monoclonal antibodies react with different parts of the NF200 molecule, one binding to a part of the molecule which is located closer to the filament, and one to a more peripheral part of the molecule.     

Novel extracellular enzymes (ligninases) of Phanerochaete chrysosporium

Abstract 3 New spectrophotometric enzyme assays were developed for the study of microbial lignin-degrading enzymes. The conversion of 2-methoxy-3-phenylbenzoic acid to 2-hydroxy-3-phenylbenzoic acid led to the discovery of an extracellular, aromatic methyl ether demethylase produced by the white-rot fungus Phanerochaete chrysosporium . The conversion of methyl 2-hydroxy-3-phenylbenzoate to 2-hydroxy-3-phenylbenzoic acid allowed the identification of an extracellular, aromatic methyl ester esterase produced by this fungus. The Phanerochaete sp. also excreted an enzyme complex that oxidized 4-(4-hydroxy-3-methoxyphenyl)-3-buten-2-one, probably to aliphatic products. All 3 novel enzyme activities were produced together with, and probably comprise a part of, the Phanerochaete ligninolytic enzyme complex. Unlike previously known ligninases, these enzymes did not oxidize 3,4-dimethoxybenzyl alcohol. All 3 were H₂O₂-dependent and were activated by Mn²⁺ ions.     

The effects of crude oil on the growth of Spartinaalterniflora Loisel. and Spartina cynosuroides (L.) Roth

Greenhouse studies were conducted to examine the effects of crude oil on the growth of Spartinaalterniflora Loisel. and S. cynosuroides (L.) Roth from North Carolina. The way in which crude oil came into contact with the plant tissue and/or substratum was an important factor in determining the responses of both species to oil pollution. Plants recovered from a single application of oil to aerial tissue with relatively little impact on productivity. The presence of an oil layer on the surface of an overlying layer of water had little impact on existing aerial portions of S. alterniflora plants; however, regrowth following harvest was completely inhibited. Incorporation of oil into the substratum significantly reduced aerial productivity and regrowth of S. alternflora and S. cynosuroides. Observations suggest that decreased productivity and regrowth may have been caused by decreased root and rhizome growth. Regrowth potential of S. alterniflora grown in oiled substratum was greater in fine-textured marsh substratum than in sand substratum.     

Biotic regionalization and climatic controls in western North America

New methods of weather analysis accompanied by microhabitat ‘bioassays’ have been applied in several case studies to demonstrate effects of atmospheric processes on patterns of community composition and structure and potential species evolution. Average spatial and seasonal airmass dynamics which determine regional and elevational patterns of relative microhabitat favorability, were found to vary between a recent global warming trend (ca 1900 to 1940) and the subsequent global cooling trend (ca 1940 to 1970). These apparently systematic spatial and temporal shifts in weather were related to plant establishment patterns and community composition and structure. The proposed causal mechanisms function, in part, through regional shifts in microhabitat size. These effects are similar to larger scale, longer term shifts deduced from the late Quaternary fossil record. By modifying the spatial approach, month-to-month and year-to-year variability of weather has been examined for the last 130 years at individual points in southwestern North America. Three climatic regimes (the end of the Little Ice Age, the recent warming trend and the recent cooling trend) exhibited distinct year-to-year patterns of weather that can be related to the establishment of different kinds of plants (e.g., C₄ grasses versus C₃ shrubs). Oscillations between different temporal climatic regimes appear to promote the episodic establishment of different life forms, but not necessarily their local extinction. The two methods of weather analysis have been combined in a regional assessment of climatic controls of different biomes in space and time with a primary focus on the Chihuahuan desert. Natural ecotones between the Chihuahuan desert and neighboring biomes are clearly related to large scale airmass dynamics associated with seasonal oscillations in jetstream position. The weather patterns controlling ecotonal positions result from seasonal topographic influences on the general circulation of the atmosphere. The apparent stability of these patterns allows causal hypotheses of biogeographic dynamics and the evolution of physiological traits and life history characteristics.     

High-cell-density fermentation studies of a recombinantE. coli that expresses atrial natriuretic factor

Summary Studies are presented on the fermentation of recombinantEscherichia coli that express rat atrial natriuretic factor (ANF) as a fusion protein. Our objective was to achieve high cell density while maintaining ANF expression at the same level as observed in shake flasks. Improved fermentation conditions included: maintaining glucose concentrations at 1 g/l, using an enriched medium, adding concentrates of medium throughout the fermentation, and blending oxygen for adequate aeration. Cell densities of 12 g/l (dry weight) were achieved, which represented a 10-fold increase over non-improved conditions, while maintaining ANF levels at 7 mg/g of dry cell mass. When galactose was used as an initial carbon source or as a feed supplement, there was a 2-3-fold increase in the expression of ANF from these high-cell-density fermentations. The recombinant ANF was biologically active.     

Microbial-invertebrate interactions and potential for biotechnology

Conclusion As the interactions between marine invertebrates and their bacterial commensals and symbionts are better understood, the application of biotechnology will enhance both environmental and economic benefit. In the immediate future, marine bacteria, either selected or genetically engineered, will play a significant role in enhancing the development of selected invertebrates in aquaculture and in the field. Luck may also favor discovery of mechanisms to suppress the development of biofouling species, perhaps by making it possible to coat submerged surfaces with bacterial films designed to repell larvae and/or interfere with larval morphogenesis. In any case, the future is appealing.     

Temporal relationship between the deposition and microbial degradation of lignocellulosic detritus in a Georgia salt marsh and the Okefenokee Swamp

Temperature dependence and seasonal variations in rates of microbial degradation of the lignin and polysaccharide components of specifically radiolabeled lignocelluloses were determined in sediment and water samples from a Georgia salt marsh and the nearby Okefenokee Swamp. Although temperature regimes in the two ecosystems were similar, rates of mineralization ofSpartina alterniflora lignocellulose in salt marsh sediments increased eightfold between winter and summer, whereas rates of mineralization of lignocellulose from an analogous freshwater macrophyte,Carex walteriana, in Okefenokee sediments increased only twofold between winter and summer. Temperature was the major factor influencing seasonal variations in rates of lignocellulose degradation in both environments. At any given temperature, no substantial differences in lignocellulolytic potential were observed with sediment samples collected at each season. In both ecosystems, the bulk of the lignocellulosic detritus was not degraded at the time of its peak deposition during the fall and winter. Instead, the periods of maximal decomposition occurred during the following spring and summer. These results suggest that periods of maximal nutrient regeneration from the mineralization of lignocellulosic detritus coincide with periods of highest primary production, and that, depending on hydrologic conditions, significant horizontal transport of essentially intact lignocellulosic material is possible due to the lag period between deposition and microbial degradation.