The holobiont concept before Margulis.

In recent years, Lynn Margulis has been credited in various articles as the person who introduced the concept of holobiont into biology in the early 1990s. Today, the origin of evolutionary studies on holobionts is closely linked to her name. However, Margulis was not the first person to use this concept in its current context. That honor goes to the German theoretical biologist Adolf Meyer-Abich, who introduced the holobiont concept nearly 50 years before her (in 1943). Although nearly completely forgotten today, in the 1940-60s he developed a comprehensive theory of evolutionary change through "holobiosis." It had a surprisingly modern outlook, as it not only addressed tenets of today's evolutionary developmental biology (evo-devo), like the origin of form and production of variation, but also anticipated key elements of Margulis' later endosymbiotic theory. As the holobiont concept has become an important guiding concept for organizing research, labeling conferences, and publishing articles on host-microbiota collectives and hologenomes, the field should become aware of the independent origin of this concept in the context of holistic biology of the 1940s.

An optimized protocol for the preparation of oxygen-evolving thylakoid membranes from Cyclotella meneghiniana provides a tool for the investigation of diatom plastidic electron transport.

BACKGROUND: The preparation of functional thylakoid membranes from diatoms with a silica cell wall is still a largely unsolved challenge. Therefore, an optimized protocol for the isolation of oxygen evolving thylakoid membranes of the centric diatom Cyclotella meneghiniana has been developed. The buffer used for the disruption of the cells was supplemented with polyethylene glycol based on its stabilizing effect on plastidic membranes. Disruption of the silica cell walls was performed in a French Pressure cell and subsequent linear sorbitol density gradient centrifugation was used to isolate the thylakoid membrane fraction. RESULTS: Spectroscopic characterization of the thylakoids by absorption and 77 K fluorescence spectroscopy showed that the photosynthetic pigment protein complexes in the isolated thylakoid membranes were intact. This was supported by oxygen evolution measurements which demonstrated high electron transport rates in the presence of the artificial electron acceptor DCQB. High photosynthetic activity of photosystem II was corroborated by the results of fast fluorescence induction measurements. In addition to PSII and linear electron transport, indications for a chlororespiratory electron transport were observed in the isolated thylakoid membranes. Photosynthetic electron transport also resulted in the establishment of a proton gradient as evidenced by the quenching of 9-amino-acridine fluorescence. Because of their ability to build-up a light-driven proton gradient, de-epoxidation of diadinoxanthin to diatoxanthin and diatoxanthin-dependent non-photochemical quenching of chlorophyll fluorescence could be observed for the first time in isolated thylakoid membranes of diatoms. However, the ∆pH, diadinoxanthin de-epoxidation and diatoxanthin-dependent NPQ were weak compared to intact diatom cells or isolated thylakoids of higher plants. CONCLUSIONS: The present protocol resulted in thylakoids with a high electron transport capacity. These thylakoids can thus be used for experiments addressing various aspects of the photosynthetic electron transport by, e.g., employing artificial electron donors and acceptors which do not penetrate the diatom cell wall. In addition, the present isolation protocol yields diatom thylakoids with the potential for xanthophyll cycle and non-photochemical quenching measurements. However, the preparation has to be further refined before these important topics can be addressed systematically.

Earth's earliest non-marine eukaryotes.

The existence of a terrestrial Precambrian (more than 542 Myr ago) biota has been largely inferred from indirect chemical and geological evidence associated with palaeosols, the weathering of clay minerals and microbially induced sedimentary structures in siliciclastic sediments. Direct evidence of fossils within rocks of non-marine origin in the Precambrian is exceedingly rare. The most widely cited example comprises a single report of morphologically simple mineralized tubes and spheres interpreted as cyanobacteria, obtained from 1,200-Myr-old palaeokarst in Arizona. Organic-walled microfossils were first described from the non-marine Torridonian (1.2-1.0 Gyr ago) sequence of northwest Scotland in 1907. Subsequent studies found few distinctive taxa-a century later, the Torridonian microflora is still being characterized as primarily nondescript "leiospheres". We have comprehensively sampled grey shales and phosphatic nodules throughout the Torridonian sequence. Here we report the recovery of large populations of diverse organic-walled microfossils extracted by acid maceration, complemented by studies using thin sections of phosphatic nodules that yield exceptionally detailed three-dimensional preservation. These assemblages contain multicellular structures, complex-walled cysts, asymmetric organic structures, and dorsiventral, compressed organic thalli, some approaching one millimetre in diameter. They offer direct evidence of eukaryotes living in freshwater aquatic and subaerially exposed habitats during the Proterozoic era. The apparent dominance of eukaryotes in non-marine settings by 1 Gyr ago indicates that eukaryotic evolution on land may have commenced far earlier than previously thought.

An early Ediacaran assemblage of macroscopic and morphologically differentiated eukaryotes.

The deep-water Avalon biota (about 579 to 565 million years old) is often regarded as the earliest-known fossil assemblage with macroscopic and morphologically complex life forms. It has been proposed that the rise of the Avalon biota was triggered by the oxygenation of mid-Ediacaran deep oceans. Here we report a diverse assemblage of morphologically differentiated benthic macrofossils that were preserved largely in situ as carbonaceous compressions in black shales of the Ediacaran Lantian Formation (southern Anhui Province, South China). The Lantian biota, probably older than and taxonomically distinct from the Avalon biota, suggests that morphological diversification of macroscopic eukaryotes may have occurred in the early Ediacaran Period, perhaps shortly after the Marinoan glaciation, and that the redox history of Ediacaran oceans was more complex than previously thought.

Using FTIR spectroscopy for rapid determination of lipid accumulation in response to nitrogen limitation in freshwater microalgae.

In this study Fourier transform infrared micro-spectroscopy (FTIR) was used to determine lipid and carbohydrate content over time in the freshwater microalgae Chlamydomonas reinhardtii and Scenedesmus subspicatus grown in batch culture in limiting concentrations of nitrogen (N). Both algae exhibited restricted cell division and increased cell size following N-limitation. FTIR spectra of cells in N-limited media showed increasing lipid:amide I and carbohydrate:amide I ratios over time. The use of lipid- and starch-staining dyes confirmed that the observed ratio changes were due to increased lipid and carbohydrate synthesis. These results demonstrate rapid metabolic responses of C. reinhardtii and S. subspicatus to changing nutrient availability, and indicate the efficiency of FTIR as a reliable method for high-throughput determination of lipid induction.

Effect of DL-malic acid supplementation on feed intake, methane emission, and rumen fermentation in beef cattle.

The objective of this study was to determine the effect of dietary concentration of dl-malic acid (MA) on DMI, CH(4) emission, and rumen fermentation in beef cattle. Two Latin square experiments were conducted. In Exp. 1, six beef heifers (19 +/- 1 mo old) were assigned in a duplicated Latin square to 1 of 3 dietary concentrations of MA on a DMI basis (0%, MA-0; 3.75%, MA-3.75; or 7.5%, MA-7.5) over 3 periods. In Exp. 2, four rumen-fistulated steers (48 +/- 1 mo old) were assigned to 1 of 4 dietary concentrations of MA (0%, MA-0; 2.5%, MA-2.5; 5.0%, MA-5.0; or 7.5%, MA-7.5) on a DMI basis, over 4 periods. Both experimental diets consisted of grass silage and pelleted concentrate (containing MA). Silage was fed ad libitum once daily (a.m.), whereas concentrate was fed twice daily (a.m. and p.m.) with the aim of achieving a total DMI of 40:60 silage:concentrate. In both Exp. 1 and 2, experimental periods consisted of 28 d, incorporating a 13-d acclimatization, a 5-d measurement period, and a 10-d washout period. In Exp. 1, enteric CH(4), feed apparent digestibility, and feed intake were measured over the 5-d measurement period. In Exp. 2, rumen fluid was collected on d 16 to 18, immediately before (a.m.) feeding and 2, 4, 6, and 8 h thereafter. Rumen pH was determined and samples were taken for protozoa count, VFA, and ammonia analysis. Enteric CH(4) emissions were estimated by using the sulfur hexafluoride tracer technique and feed apparent digestibility was estimated by using chromic oxide as an external marker for fecal output. In Exp. 1, increasing dietary MA led to a linear decrease in total DMI (P < 0.001) and total daily CH(4) emissions (P < 0.001). Compared with the control diet, the greatest concentration of MA decreased total daily CH(4) emissions by 16%, which corresponded to a 9% reduction per unit of DMI. Similarly, in Exp. 2, inclusion of MA reduced DMI in a linear (P = 0.002) and quadratic (P < 0.001) fashion. Increasing dietary MA led to a linear decrease in molar proportion of acetic (P = 0.004) and butyric acids (P < 0.001) and an increase in propionic acid (P < 0.001). Ruminal pH tended to increase (P = 0.10) with increasing dietary MA. Dietary inclusion of MA led to a linear (P = 0.01) decrease in protozoa numbers. Increasing supplementation with MA decreased CH(4) emissions, but DMI was also decreased, which could have potentially negative effects on animal performance.

Estimation of Cochlodinium polykrikoides growth potential using a dialysis membrane.

We developed a test to measure the growth potential of C. polykrikoides using a dialysis membrane and artificial seawater. Nitrite nitrogen and inorganic phosphorus in the medium were almost completely removed when the medium was dialyzed against artificial seawater for five or more 6-hour cycles using a dialysis membrane (Spectrum's Spectra/Por 7 Membrane) with a molecular-weight cut-off of 50,000, regardless of the presence of C. polykrikoides. The phytoplankton grew well even after dialysis. To estimate the growth potential of C. polykrikoides, a minimum initial concentration of > 100 cells/ml is required. Methods using short-term starvation culturing of C. polykrikoides to measure growth potential were determined to be ineffective; instead, controlled tests using artificial seawater are recommended. The dialysis membrane used in this study can also be employed to measure the algal growth potential of other phytoplankton species.

The impact of cleaner processing on nutrient availability in the bleached kraft industry.

The pulp and paper industry has invested heavily over recent years in cleaner processing, to reduce losses and minimise its impact on the environment. Over the past fifteen years, a New Zealand integrated bleached kraft mill has undergone a comprehensive programme of upgrades to increase production, reduce water consumption and streamline its biological treatment process. Whilst the overall discharge of contaminants from the site decreased, the treatment system performance did not show a concurrent improvement as may have been expected. Reduced BOD removal, low dissolved oxygen levels, and poor solids settlability were symptomatic of phosphorus limitation in the aerated lagoon treatment system. The wastewater entering the system was found to be phosphorus limited at a BOD:P ratio of 100:0.2. Mono-ammonium-phosphate was supplemented, at approximately 30 kg P/d, to raise the phosphorus levels to a BOD:P ratio of 100:0.3. Treatment efficiencies improved very quickly after phosphorus dosage, with a 50% reduction in BOD and TSS discharge, a significant increase in dissolved oxygen levels, and improved BOD removal (85% to 93%). This case study demonstrates that whilst more closed operation can result in reduced discharge of organic loads, there may be negative impacts on the availability of nutrients for balanced biological growth.

Differential responses of lichen symbionts to enhanced nitrogen and phosphorus availability: an experiment with Cladina stellaris.

BACKGROUND AND AIMS: Lichens can be both nitrogen- (N) and phosphorous- (P) limited and thus may be susceptible to nutrient enrichment. Nutrient enrichment with N and P may have differing impacts on the lichen structure because of different physiological responses of fungal and algal partners to these nutrients. The hypothesis was tested that the differential responses of lichen symbionts to enhanced availability of N and P is reflected in the lichen thallus structure and the wall-to-wall interface between the algal and fungal cells. METHODS: Lichen cushions of Cladonia stellaris were treated with one P and two N concentrations alone and in combination that yielded total depositions of approx. 300 (moderate) and 1000 (high) mg N m(-2) and 100 (high) mg P m(-2) over an experiment lasting 14 weeks. The effects of N and P inputs on the relative volumes of fungal and algal cell in the medullary tissue and on the thallus structure were studied using light microscopy. The interface between algal and fungal cell walls was examined using transmission electron microscopy. KEY RESULTS: The influence of excess P on the lichen thallus structure was stronger than that of additional N. Addition of P reduced the N : P ratio in podetia, the proportion of the medullary layer volume occupied by the algal cells, the thallus volume occupied by the internal lumen, and the algal cell-wall area covered by fungal hyphae. CONCLUSIONS: Ecologically realistic changes in the availability of key macronutrients can alter the growth of symbionts. Reduction in the proportion of photobiont cells indicates that the application of P either stimulates fungal hyphal growth in the medullary tissue or impairs the cell division of the algal cells. The results suggest that both the N and P availability and thallus N : P ratio affect the growth rates of lichen symbionts.

[Distribution of microcystin-LR and anatoxin-A and their influence factors in Dianshan Lake during summer and autumn].

Six different sampling locations in Dianshan Lake were selected to study the distributions of algaes and algae toxins and their influencing factors in summer and autumn. Total nitrogen (TN), total phosphorous (TP), Fe, chlorophyla(Chla) and COD in water samples were determined. At the same time, water temperature (TEM), pH value, illumination, turbidity degree (TUR) of water body were recorded. In addition, the total number and attribute of algae cell and the content of microcystin-LR(MC-LR) and anatoxin-A(ANTX-A) were checked. The results showed that the pollution of both toxins was most serious in July. The MC-LR were mostly distributed in Wangxiang, but the ANTX-A were mostly distributed in Jishuigang. Generalized estimating equations (GEE) regression analysis showed that total number of algae cell was significantly correlated with TN, TP and Chla (P < 0.05). The distribution of MC-LR was significantly affected by total number of algae cells and TEM (P < 0.01), but TUR mainly affected the distributions of ANTX-A(P < 0.01). There was weak correlation between both toxins (P < 0.05). In short, there was a serious pollution of MC-LR, but ANTX-A pollution was light. The factors influencing the distributions of both algae toxins were not completely consistent.

Biotechnological potential of immobilized algae for wastewater N, P and metal removal: a review.

This presentation comprises a review on the use of immobilized algae for wastewater nitrogen, phosphorus and metal removal purposes. Details of the use of immobilized algae, the techniques of immobilization and the effects of immobilization on cell function are included. Particularly relevant in their use for heavy metal removal from wastewaters; upon enriching the biomass in metal, can be recoverd, thereby providing economic advantages. The use of immobilized microalgae in these processes is very adequate and offers significant advantages in bioreactors. The future of this area of algal cell biotechnology is considered.

[Evaluation of cell nitrogen and phosphorus quotas in accumulative cultures of chlorococcous algae]. / Otsenka kletochnykh kvot azota i fosfora v nakopitel'nykh kul'turakh khlorokokkovykh vodoroslei.

A conceptual model of cell quota was proposed and the parameters were determined that characterize the quota dynamics during the development of algae. Based on the experimental and empirical-statistical procedures of analysis, the basal parameters of cell quotas of nitrogen and phosphorus were obtained for four species of chlorococcous algae. The data were also obtained on a variability of nitrogen and phosphorus quotas during the development of algae, as well as coefficients of excessive accumulation. The phosphorus quota changed within the limits of three orders of magnitude and the nitrogen quota, within the limits of one order. The highest coefficients of excessive accumulation were 52 and 1.8 for phosphorus and nitrogen, respectively.

Sperm entry induces polarity in fucoid zygotes.

Fucoid zygotes establish a rhizoid-thallus growth axis in response to environmental signals; however, these extrinsic cues are not necessary for polarization, suggesting that zygotes may have inherent polarity. The hypothesis that sperm entry provides a default pathway for polarization of zygotes cultured in the absence of environmental signals was tested, and was supported by several lines of evidence. First, an F-actin patch, a cortical marker of the rhizoid pole, formed at the sperm entry site within minutes of fertilization. Second, the sperm entry site predicted the site of polar adhesive secretion (the first morphological manifestation of the rhizoid pole) and the position of rhizoid outgrowth. Third, when fertilization was restricted to one hemisphere of the egg, rhizoid outgrowth always occurred from that hemisphere. Fourth, delivery of sperm to one location within a population of eggs resulted in polarization of both adhesive secretion and rhizoid outgrowth toward the sperm source. Finally, induction of polyspermy using low sodium seawater increased the frequency of formation of two rhizoids. Sperm entry therefore provides an immediate default axis that can later be overridden by environmental cues.

Diatom response to extremely low-frequency magnetic fields.

Reports that extremely low-frequency magnetic fields can interfere with normal biological cell function continue to stimulate experimental activity as well as investigations into the possible mechanism of the interaction. The "cyclotron resonance" model of Liboff has been tested by Smith et al. (Bioelectromagnetics 8, 215-227, 1987) using as the biological test system the diatom Amphora coffeiformis. They report enhanced motility of the diatom in response to a low-frequency electromagnetic field tuned to the cyclotron resonance condition for calcium ions. We report here an attempt to reproduce their results. Following their protocol diatoms were seeded onto agar plates containing varying amounts of calcium and exposed to colinear DC and AC magnetic fields tuned to the cyclotron resonant condition for frequencies of 16, 30, and 60 Hz. The fractional motility was compared with that of control plates seeded at the same time from the same culture. We find no evidence of a cyclotron resonance effect.

[The search for cytostatic substances in the tissues of plants of the genus Maytenus molina in in vitro culture. I. Callas culture and biological studies of its extracts]. / Poszukiwanie substancji cytostatycznych w tkankach roslin rodzaju Maytenus molina w kulturze in vitro. I. Kultura kalusa i badania biologiczne jego wyciagów.

Plant tissue cultures of Maytenus wallichiana Raju et Babu and Maytenus emarginata Ding Hou were initiated. Growth conditions of the callus and the optimum medium composition have been established. Increments of callus wet mass and dynamics of callus growth were determined. Morphological and microscopic observations were also performed. The most efficient growth of the callus, resulting in increments of its wet mass up to 6460%, was obtained on the modified Murashige and Skoog medium. Extracts of the callus were found to be inactive against microorganisms, but proved cytotoxic for lymphocytic leukaemia L 5178Y (ED50 18-48 micrograms/cm3) and the protozoon T. pyriformis (ID50 43-53 micrograms/cm3).

The utilization of inorganic and organic phosphorous compounds as nutrients by eukaryotic microalgae: a multidisciplinary perspective: part 1.

This comprehensive literature review of the phosphorus nutrition and metabolism of eukaryotic microalgae deals sequentially with (1) extracellular P-compounds available for algal utilization and growth; (2) orthophosphate uptake mechanisms, kinetics, and influence from environmental variables; (3) phosphatase-mediated utilization of organic phosphates involving multiple enzymes, induction and cellular location of repressible and irrepressible phosphatases, and their role in growth physiological processes; (4) intracellular phosphate metabolism covering diversity of phosphometabolites. ATP-linked energy regulation, polyphosphate pools and storage roles, phospholipids and phospholipases; (5) steady-state and transient-state models relating phosphate utilization to growth; (6) ecological aspects covering manifestations of phosphorus limitation, interspecific competition for phosphonutrients among microorganisms, and current views on phosphorus cycling and turnover in aquatic ecosystems. Although concentrating on the microalgae, the review often points out sounder conclusions drawn from bacteria and fungi, and includes specific macroalgae in considering certain subtopics where such algae were better investigated and provided a good basis for comparison with the microalgae.