YEAR IN REVIEW 1996: LIFE-SCIENCES

BOTANY The publication in June of Botany for the Next Millennium by the Botanical Society of America (BSA) was a landmark in the history of botany. The report, the result of a collaboration of the BSA and nine other scientific societies that represented diverse interests ranging from mycology and lichenology to taxonomy and chemical ecology, established a framework for identifying research and educational goals, priorities, and opportunities in the botanical sciences on the eve of the 21st century. One set of goals addressed what the report called vigor of the profession and suggested actions that would help maintain botany's record of achievement and discovery. A second set addressed continuity of the profession and included actions designed to attract future botanists and support their training. A third set spoke to integration into the community in order to broaden the relevance of botanical knowledge beyond academia. The report was needed because, although plants and plant products were ubiquitous in daily life, botany was underemphasized in biology curricula at all levels of education. The brief (54-page), easily readable publication was expected to spur improvements in botanical education and help focus research efforts in the coming years. Within the space of eight months, the U.S. journal Science produced two special issues focusing on subjects important to botany. One reviewed recent discoveries in plant biotechnology, and the other dealt with aspects of developmental biology. By 1995 plant biotechnology had become an emerging and rapidly expanding field in which the techniques of the molecular biologist were being applied to practical problems in plant science. Reports in one of the special issues included descriptions of plants that had been genetically engineered to synthesize specific substances that stimulate the animal immune system, with the eventual goal of making plant-produced vaccines against human diseases. There was even speculation that vaccines could be built into plants that then would be eaten as part of a normal diet. Other reports described ways in which biotechnology was helping to transform the concept of plant-based raw materials. For instance, researchers engineered cotton plants to produce natural fibres incorporating some of the desired characteristics of synthetic polyesters. Other potential products from engineered plants included naturally produced biodegradable plastics and industrial lubricants. Among the more promising studies were those aimed at a better understanding of plant disease-resistance mechanisms and the pertinent genes so that plants might be better equipped to protect themselves against the wide array of disease organisms that attack them. At the very least, such studies could lead to novel strategies for plant-disease control and less reliance on traditional chemical pesticides. No matter how complex the form of a mature organism, all sexually reproducing organisms, after fertilization, begin as single cells called zygotes, which are nearly spherical in shape. In higher plants, once fertilization has been achieved, the zygote begins to differentiate into an embryo. Polarity, the development of recognizably different ends, begins with the first cell division. Although the precise genetic and molecular mechanisms that result in embryo formation and polarity were still poorly understood, major insights were gained from recent studies of plant embryo mutants and others that used molecular approaches. These were summarized in the Science special issue on development by Robert Goldberg and collaborators of the University of California, Los Angeles. Taken together, the research suggested that the structure of a developing plant embryo is modular, with several regions forming independently. Parasitism, an association between two different organisms in which one benefits at the expense of the other, is widespread in nature. Mutualism, an association in which both organisms benefit, is also widespread although less commonly noticed. Mutualism is a characteristic of a large group of plantlike organisms called lichens, which develop as an association between an alga and a fungus. The alga photosynthesizes and produces food for the fungus, which in turn absorbs water and perhaps supplies other benefits to the algal partner. It had been accepted by many that lichen associations probably started as parasitic associations, which in the course of evolution transformed into mutualistic ones. A collaborative study among scientists of the Smithsonian Institution, Washington, D.C., Graz (Austria) University, and the University of Stockholm challenged that idea. By comparing DNA sequences taken from different fungus species, including lichen-derived fungi, and using the relationships to construct a fungal family tree, they showed that the alga-fungus association originated independently at least five times and involved very different groups of fungi whose lifestyles range from benign to parasitic. On the basis of such results, the authors found no support for the frequently expressed notion that mutualism must begin with parasitism. (PHILIP D. REID) This updates the articles biosphere; conservation; plant. MARINE BIOLOGY Studies reported from the Finnish research vessel Aranda in 1995 demonstrated a reversal of a recent trend detected in the Gotland Basin of the Baltic Sea toward reduced oxygen and increased hydrogen sulfide concentrations in the water of the basin. In 1993 a major inflow of North Sea water had occurred, and it was thought that the event created favourable preconditions for even small subsequent inflows to increase oxygen concentration drastically in the basin. Another report focusing on northern Europe presented the results of a comprehensive analysis of the effects of offshore gas and oil exploration and production on bottom-living animals of the Norwegian continental shelf. The study showed that oil-based drilling fluid resulted in severe depletion of key species, some of which serve as food for bottom-living fish. Whereas replacement organisms were abundant, they were too small and too deeply burrowing to serve as substitute food for fish. In work in the U.S., researchers reported that the oyster population in the Maryland portion of Chesapeake Bay had fallen 50-fold since the early 1900s. The decline was blamed on habitat destruction and overfishing, not on worsening water quality and disease, as had been previously thought. Phytoplankton is the plantlike part of the community of the generally minute, drifting organisms called plankton that live at or near the water's surface. Japanese workers showed for the first time that phytoplankton growth can be inhibited by cell-to-cell contact with another organism. The phytoplanktonic flagellate Gyrodinium instriatum was observed to be killed by contact with a species of Heterocapsa, one of the dinoflagellates responsible for the toxic blooms called red tides that occasionally discolour the ocean. A recently described dinoflagellate, Pfiesteria piscicida, was implicated as a causative agent of major fish kills in the southeastern U.S. Pfiesteria responded to as-yet-unidentified substances secreted from fish schools by producing toxins that in laboratory assays proved lethal to 19 species of native and exotic finfish and shellfish. U.S. scientists reported a severe decline (80% since 1951) in the biomass of zooplankton, the animal-like component of plankton, in the ocean off southern California. The decline was correlated with a rise in ocean surface temperatures of more than 1.5 C (2.7 F) in some areas. The warming was thought to have caused zooplankton reduction, and a consequent decline in abundance of fish and some birds in the region, by slowing cold-water upwellings that replenish the surface waters with nitrates and other nutrients. In a U.K. study miniature acoustic transponders wrapped in bait were released on the seafloor from an autonomous vehicle at depths of 1,500-4,000 m (4,900-13,100 ft) in the Porcupine Sea Bight of the North Atlantic. Photographs of deep-sea fish taking the baits and sonar records of signals from the swallowed transponders showed that the fish moved out of range of the sonar in three to nine hours, which indicated that they had no home range or territorial behaviour. New threats to the survival of the so-called living fossil known as the coelacanth (Latimeria chalumnae) were reported. In its protected habitats around the island of Grande Comore in the Comoros, the rare lobe-finned fish was being illegally taken by local fishermen who were unable to move their motorized canoes out from shore beyond the designated coelacanth-protection zone. Maltese studies revealed the first occurrence in the Mediterranean Sea of "imposex," a phenomenon observed in marine snails whereby females became masculinized and unable to reproduce. This threat to species survival was caused by tributyl tin, until recently a commonly used antifoulant compound in marine paints. Such findings continued to spur searches for less toxic antifoulants, including compounds made naturally by marine organisms. The presence of such a natural antifoulant was suggested in studies in the U.K. of the egg cases of a shark species, the dogfish Scyliorhinus canalicula, which were found to survive cleanly in the sea for as long as 300 days before hatching. Similar properties of resistance to marine fouling were also reported for ascidians, marine animals commonly called sea squirts. In Australia the large herbivorous marine mammals known as dugongs (Dugong dugon) were seen to practice "cultivation grazing." They fed intensively in large herds in a manner that favoured rapidly growing, nutritious algae such as Halophila ovalis. As a result, those algal species thrived at the expense of slower growing, normally dominant species such as Zostera capricorni, which the dugongs favoured less. A female green turtle (Chelonia mydas) was fitted with a radio transponder and tracked by satellite in the South China Sea from its nesting beach to its normal foraging grounds more than 600 km (370 mi) away. The final 475-km (295-mi) leg of the journey brought the turtle directly to its goal with pinpoint accuracy, the animal maintaining constant speed and direction by day and night. The best explanation for such precise orientation seemed to be a geomagnetic compass similar to that previously reported in birds, honeybees, and other animals. (ERNEST NAYLOR) This updates the articles crustacean; fish; mollusk. MOLECULAR BIOLOGY Mitochondria, Aging, and Disease. Aptly called the powerhouses of the cell, mitochondria are the organelles responsible for most of the cell's respiration and energy production. They possess their own DNA that is distinct from that which makes up the chromosomes of the cell nucleus. In 1995 more evidence of the importance of this "other genome" came to light as researchers continued studying mitochondrial genetics and its role in aging and disease. About the size and shape of bacilli, mitochondria are bound by two membranes, as are some bacteria. Indeed, it is likely that mitochondria arose from an ancient symbiosis between a bacterium and a primitive amoeba-like cell. As befits an association that has lasted a billion years, there have been accommodations. For example, the bacterial symbiont lost its cell wall, which was superfluous in the protected environment provided by the host cell. Also, its inner membrane became corrugated, increasing greatly in surface area to accommodate the extra molecular machinery required for meeting the energy needs of the host cell. The host cell, for its part, took over most of the biochemical chores for the symbiont's replication and maintenance. The host benefited from the ability of the symbiont to trap the abundant energy released during aerobic respiration--the oxygen-dependent breakdown of foodstuff molecules--while the symbiont benefited from the stable environment and nutrients supplied by its host. The association has proved highly successful, as evidenced by its numerous and diverse progeny, which constitute all macroscopic life on Earth. There are hundreds of mitochondria in the average cell. Perhaps as a relic of its symbiotic origin, each mitochondrion retains a bit of its own DNA, which codes for 13 different proteins and 24 different RNA molecules that assist in protein synthesis. It retains the ability to replicate its DNA and make its proteins, which are essential components of its energy-producing and energy-trapping functions. Both egg and sperm cells contain mitochondria. During fertilization in humans and in nearly all other animal species, however, the mitochondria of the sperm are not incorporated into the fertilized egg. Consequently, mitochondrial genes are transmitted to offspring only by the mother. DNA, the repository of the genetic information of the cell, is not a perfectly stable storage medium, and changes can creep in for a variety of reasons. Cells go to great lengths to minimize such changes and to repair those that do occur. Yet some changes persist and, if they are transmitted to progeny, are the cause of mutations, which often are deleterious. Mitochondrial DNA is at greater risk of mutation than is nuclear DNA. The reasons remained to be fully understood, but it was clear that damage accumulates in mitochondrial DNA 10-20 times faster. Such damage, as investigators were learning, is involved in senescence--i.e., the biological changes related to aging--and disease. A decrease in the usable energy available to cells and tissues as they age would necessarily undermine their function, and a decline in mitochondrial integrity would certainly curtail that energy supply. When researchers looked for evidence to support the idea that mitochondrial integrity declines with age, they found it. Mitochondria isolated from aged animals were seen to be enlarged, full of cavities (vacuoles), and lacking in the degree of inner-membrane corrugation seen in the mitochondria of young animals. Senescent mitochondria also were fragile and less likely to survive the isolation procedure itself, which means that the most severely affected mitochondria were likely underrepresented in the observations. In spite of the difficulty in isolating senescent mitochondria, scientists detected a number of age-related losses of function, including less-efficient coupling of respiration to the production of useful energy and a decline in the activities of enzymes crucial to respiration. Furthermore, they found that mitochondrial DNA from aged animals contain a variety of genetic damage, which can reduce or destroy mitochondrial function. How is it that the cell can tolerate such damage at all? The answer lies in the large numbers of mitochondria in each cell and of DNA molecules in each mitochondrion. Damage thus has a graded effect, with a little cumulative damage causing little loss of function for the cell and more damage causing more loss. Different tissues are dependent to different degrees on the metabolic energy production by mitochondria and will reflect to different degrees the cumulative damage to their mitochondria. Whereas damage to mitochondrial DNA in somatic (nonreproductive) cells may be a problem for the individual, it will not be passed on to offspring. On the other hand, damage that occurs in egg cells may or may not be transmitted to progeny. Why is this the case? The fertilized egg contains about 200,000 molecules of mitochondrial DNA. In the early stages of development, however, cells divide without replicating their mitochondrial DNA; consequently, the number of copies per cell falls dramatically. Each cell destined to give rise to different tissues in the developing embryo thus receives a relatively small number of molecules of mitochondrial DNA. If that DNA is seriously defective, the result is death; if it is moderately defective, the result is transmission of a genetic disease. The first mitochondrial disease was described in the 1960s by investigators who were attempting to understand the symptoms of an extremely emaciated, weak, feverish patient who consumed enormous amounts of food and liquids and sweated profusely. Her basal metabolic rate was nearly double the normal value. After eliminating hyperthyroidism as the possible cause, the investigators realized that her symptoms might be explained by "uncoupled" mitochondria--that is, mitochondria in which respiration was liberating energy from foodstuffs but not trapping it in metabolically useful form. Indeed, when mitochondria from muscle tissue of the patient were isolated, they were found to be uncoupled. This pioneering discovery opened the door to the field of mitochondrial diseases. Other investigators followed the lead, linking mitochondrial defects with maladies such as Kearns-Sayre syndrome, chronic external opthalmoplegia, and myoclonic epilepsy and ragged red-fibre disease. In 1995 medical science knew of about 120 mitochondrial diseases. As expected, they are maternally inherited, and they tend to affect specific tissues because of the different dependence of various tissues on mitochondrial energy production. Many make their appearance only later in life because the defects that accumulate with age add to those that are inherited and must reach a critical threshold level before symptoms appear. Knowledge of the cause of a problem often leads to a solution. Consequently, researchers were optimistic that their growing appreciation of the complexities of mitochondrial genetics would eventually produce practical benefits. (IRWIN FRIDOVICH) Telomeres: New Beginnings from Old Ends. A major distinction between prokaryotes, or bacteria, and eukaryotes, or so-called higher organisms made of nucleated cells, is the manner in which they arrange the DNA of their genetic endowment, or genome. Bacteria generally maintain their genomes as circular molecules, whereas animals and plants maintain their nuclear genomes as collections of linear molecules, called chromosomes. Although a linear architecture has its benefits, it also presents problems--perhaps most notably, what to do about the ends. The trouble with having ends is at least twofold. First, free ends on DNA molecules are notoriously unstable; they degrade chemically and undergo recombination much more often than their non-end, protected counterparts. Second, the DNA polymerase enzyme that is responsible for replicating the nuclear genome during cell proliferation has difficulty copying the very ends of DNA molecules, so without special precaution the end molecular sequences tend to be lost in the copies. To circumvent the problems, eukaryotic cells cap their chromosomes at both ends with specialized structures called telomeres. New evidence gathered in a number of laboratories suggests that telomeres and the enzyme or enzymes that create and maintain them play key roles in cellular aging and the immortalization of cells so often associated with cancer. The first telomere was isolated in the 1970s from the single-celled ciliated protozoan Tetrahymena thermophila. It was found to consist of 50-70 tandem copies of the short DNA base sequence TTGGGG (T is the base thymine; G is guanine). Both the structure and sequence were considered peculiar at the time, but subsequent work with many different organisms served to validate and extend the observations. For example, all mammals examined as of 1995, including humans, carry the repeated sequence TTAGGG (A is adenine) in their telomeres. Indeed, not only do telomeric sequences from different organisms look alike, but they also may function alike. This was first demonstrated when linear pieces of DNA carrying ciliate-derived telomeres were put into cells of yeast, an extremely distant relative of ciliates. The ends of the DNA remained stable; in other words, the ciliate telomeres worked in yeast. With time, however, the ciliate-derived telomeric sequences eroded and were replaced by the corresponding yeast sequences. The gradual erosion of the ciliate-derived telomeric sequences suggested that telomeres do not escape the fate of unprotected DNA ends during cell replication; they simply buffer the loss. Indeed, telomeric sequences made of multiple repeats are, in retrospect, very logical; such sequences can be at least partially sacrificed without losing genetic information. That the eroded ciliate sequences were replaced by yeast counterparts indicated that the repeats are not only expendable but also renewable by means of a cellular activity that is independent of the sequence of existing repeats. That activity was found to be carried out by a most unusual enzyme, named telomerase, that consists of both RNA and protein. Subsequent experiments involving the ciliate Tetrahymena revealed that if telomerase is inactivated (by a mutation, for example), the telomeres in the mutant cells grow shorter and shorter; moreover, the single-celled organisms, which normally do not have a finite life span, eventually die. In other words, in the absence of functional telomerase, the length of a cell's telomeres appears to have an inverse relationship to its age. The mortality of human beings and most other living creatures is a characteristic not only of the body as a whole but also of most of the body's cells, even cultured cells. Although aging is clearly a complex process that likely reflects the interactions of many genes and environmental factors, a variety of recent observations imply a role for telomeres and telomerase in the replicative senescence of cells. For example, it was found that proliferating cultured human fibroblasts, which normally die after a finite number of divisions, lack telomerase activity; as they age, their telomeres become gradually, and ultimately profoundly, shorter. Similar observations were made for somatic cells functioning normally in the body; in other words, chromosomes from the cells of younger people tend to have longer telomeres, while those from older people tend to have shorter ones. This is consistent with the general observation that cells taken from younger donors tend to live longer in culture than do cells from older donors. Indeed, in a study of cultured fibroblasts from 31 donors ranging from newly born to 93 years in age, investigators saw a striking correlation between initial telomere length in the donated cells and ultimate proliferative capacity. In addition, they found that telomeres from fibroblasts donated by Hutchinson-Gilford progeria patients, who experience abnormally rapid aging, were unusually short. In contrast to the findings in somatic cells, studies of normal human and nonhuman cells that are naturally "immortal," namely, germ, or reproductive cells, revealed that the telomeres of those cells appear to be stable with time. For example, telomeres from normal human sperm do not shorten with age, which suggests that some mechanism such as telomerase activity maintains telomere length. Consistent with this idea, telomerase activity was directly observed in frogs' eggs. All this information taken together suggests that telomerase activity is generally absent from normal mortal cells, which thus experience replicative telomere shortening, but is found in normally immortal cells, such as protozoans or germ cells. If this is true, what then of normally mortal cells that become immortalized, such as cancer cells? The potential role of telomeres and telomerase in cancer has been revealed in at least two ways. First, experiments were conducted in which normal cultured cells, which lacked telomerase activity, were exposed to oncogenic (cancer-inducing) DNA sequences derived from tumour viruses. Whereas most of the cells died after a number of replications, some continued to divide without limit, presumably owing to transformation by the viral DNA. Under examination the telomeres of the newly immortalized populations were found to be stable with time, and telomerase activity was observed. The results suggested that the switching on of telomerase activity, presumably by activation of one or more otherwise silent genes, might be part of the process whereby normal cells are transformed into cancer cells. A second line of information came from studies of cells taken directly from tumours removed from patients. Again, in sets of matched cells derived either from tumours or from the corresponding normal tissues, telomerase activity was found in tumour cells but not in normal cells. These studies offered considerable food for thought about how telomere shortening interacts with other factors involved in the aging process, for both replicating and nonreplicating cells. On a much more practical note, they pointed to telomerase as a possible new target for anticancer drugs. Indeed, during the year researchers worked to develop specific and effective inhibitors against the enzyme, following the logic that if telomerase can be inactivated in tumour cells, the cells may become mortal again and eventually die. Because most normal cells already lack telomerase activity, a truly specific inhibitor should cause them little if any harm. (JUDITH FRIDOVICH-KEIL) See also Mathematical and Physical Sciences: Chemistry. This updates the articles cancer; cell; disease; heredity; biological development; reproduction. PALEONTOLOGY Paleontological discoveries during the year shed light on the phylogeny of living and extinct organisms. Neil H. Shubin of the University of Pennsylvania and Farish A. Jenkins, Jr., of Harvard University reported the discovery of the fossilized remains of a jumping frog from the Early Jurassic Kayenta formation of Arizona. The species, which the investigators named Prosalirus bitis, lived about 190 million years ago. Although the fossil retained some of the primitive characters of earlier amphibians, certain features of the pelvic girdle, legs, and feet were clearly indicative of saltatory, or leaping, locomotion. The investigators concluded that the species unquestionably belonged within the Anura, the frog and toad order, and was the order's earliest known member. The question of whether dinosaurs were endothermic or ectothermic (warm-blooded or cold-blooded) has long been an issue of intense debate. The physiology of endothermic animals enables them to maintain a nearly constant body temperature in the face of varying environmental conditions and, as a result, to maintain high levels of activity. Among ectothermic animals, on the other hand, body temperature and, consequently, level of activity vary significantly under changing conditions. Although dinosaurs originally had been thought to be ectothermic, like living reptiles, some paleontologists had argued, largely on the basis of skeletal characteristics, that they were endothermic, like birds and mammals. John Ruben of Oregon State University and his former student Willem Hillenius took a stride toward possibly resolving the issue. They proposed that the key lay in the respiratory turbinates, which are found in the nasal passages of living mammals and birds but are absent in living reptiles. These bony plates, which in life are covered with a thin membrane, provide a large surface of exposed tissue. The tissue and the blood that it contains are cooled by inhaled air and then are reheated as warm air is exhaled, thus providing an important mechanism for maintaining body temperature. Because the turbinates are delicate, they are seldom preserved in fossils. According to Ruben and Hillenius, however, the turbinates are attached to a distinct ridge of bone in the nasal passage, which the investigators identified in fossil mammals dating back to the origin of the class in the Jurassic Period some 160 million years ago. On the other hand, they found no such evidence in the dinosaur fossils that they examined. Opponents argued that dinosaurs still may have had turbinates or may have evolved other mechanisms of thermoregulation. Whatever the outcome of the debate over dinosaurs, the findings of Ruben and Hillenius were an important contribution to scientific knowledge of the physiology of fossil vertebrates. Paleontologists have long been puzzled by the so-called Cambrian explosion some 540 million years ago, when most of the great phyla that were to dominate the subsequent history of life appeared, in geologic terms, quite suddenly. Whereas many Precambrian fossils were known, none of them contributed much to an understanding of the remarkable increase in animal diversity at the start of the Cambrian Period. During the year a report drawn from knowledge of developmental biology offered an explanation for the lack of any evidence that would presage the dramatic events that were to occur at the beginning of the Cambrian. E. Davidson and R.A. Cameron of the California Institute of Technology and K. Peterson of the University of California, Los Angeles, argued that before such wide diversification could occur, organisms had to overcome a barrier that limited their size and complexity by restricting to about 10 the number of times a fertilized egg could divide. According to the authors, the solution lay in the appearance of cells, of a kind found in the embryos of some organisms living today, that are not destined to develop into a particular kind of tissue. It was those cells, the authors contended, that may have provided an opportunity for the evolution of much larger and more complex organisms. Davidson and his colleagues concluded that this crucial development would have taken place in soft-bodied, embryolike animals, which almost never would have left any trace, hundreds of millions of years before the Cambrian explosion. Clusters of fossil dinosaur eggs presumed to be the remains of nests have been known for decades. A discovery in south central Mongolia, however--an example of the recent continuing Chinese-U.S. cooperation--appeared to demonstrate that dinosaurs not only laid eggs in clutches but tended to them as well. In a press conference at the American Museum of Natural History, New York City, it was announced that the skeletal remains of an oviraptor, a toothless predaceous dinosaur about the size of an ostrich, were found positioned with at least 15 eggs in such a way as to suggest that the animal had been sitting on the eggs when it was killed in some catastrophic event, perhaps a sandstorm, in the late Cretaceous Period about 80 million years ago. The eggs, which measured about 18 cm (7 in) long, were found arranged in a circular pattern under the skeleton. Mark Norell of George Washington University, Washington, D.C., who found the specimen, pointed out that, judging from the skeleton, oviraptors were more closely related to birds than to other meat-eating dinosaurs--which perhaps accounted for the birdlike brooding behaviour. (DAVID B. KITTS) See also Earth and Space Sciences: Geology and Geochemistry; The Environment; Libraries and Museums. This updates the articles evolution, theory of; geochronology.