Rapid backward movement of anaphase chromosome whose kinetochore fibers were cut by ultraviolet microbeam irradiation

Summary— kinetochore spindle fibers in meiosis I and II grasshopper spermatocytes were cut with a heterochromatic ultraviolet (UV) microbeam converging on the specimen to form a slit-shaped microspot 1.5 × 8 μm or 3 × 8 μm. A total exposure of 3 × 10^?8 joules per μm² was administered within 0.8–2.4 s, which was sufficient for severing. The cells were observed with a high extinction polarizing microscope or phase contrast optics and a record made by time-lapse video microscopy, continuously before, during and after the irradiation. When kinetochore fibers were irradiated i anaphase with UV, an area of reduced birefringence (ARB) was produced at the exposed site. The newly created + ends of the microtubules rapidly disassembled poleward, at a constant speed of 17 μm/min. The — ends at the edge of ARB also depolymerized at a slower rate. When a kinetochore fiber was cut with UV in early anaphase at which time its associated chromosome had not disjoined from the partner chromosome, the chromosome of the irradiated kinetochore fiber moved rapidly back to its partner. The speed during this movement was faster than the normal poleward chromosome movement in anaphase by an order to magnitude or more. When a kinetochore and its associated kinetochore fiber were included in the irradiation are, the effects were more pronounced than the effects of irradiation on a kinetochore fiber alone; the direction of the line connecting the irradiated half-bivalent with the partner half-bivalent deviated so much from the longitudinal axis of the original spindle with time that the division assumed a tripolar figure.     

Inhibition of ATP-regulated K+-channels by a photoactivatable ATP-analogue in mouse pancreatic β-cells

The effects of a photoactivatable (DMNPE-caged) ATP-analogue on ATP-regulated K⁺-channels (K_ATP-channel) in mouse pancreatic β-cells were investigated using the inside-out patch configuration of the patch-clamp technique. The caged precursor caused a concentration-dependent reduction of channel activity with a K_i of 17 μM; similar to the 11 μM obtained for standard Mg-ATP. The small difference in the blocking capacity between the precursor and ATP is probably the reason why no change in channel activity was observed upon photolysis of the caged molecule and liberation of ATP. It is suggested that the part of the ATP molecule involved in the blocking reaction of the K_ATP-channel is not sufficiently protected in DMNPE-caged ATP making this compound unsuitable for studying the rapid kinetics of ATP-induced K_ATP-channel inhibition.     

CO2 uptake by the cultivated hemiepiphytic cactus, Hylocereus undatus

The climate of the native tropical forest habitats of Hylocereus undatus, a hemiepiphytic cactus cultivated in 20 countries for its fruit, can help explain the response of its net CO2 uptake to environmental factors. Under wet conditions, about 85% of the total daily net CO₂ uptake occurs at night via Crassulacean acid metabolism, leading to a high water‐use efficiency. Total daily net CO₂ uptake is reduced 57% by only 10 days of drought, possibly involving stomatal closure induced by abscisic acid produced in the roots, which typically occupy a small substrate volume. Total daily net CO₂ uptake for H. undatus is maximal at day/night air temperatures of 30/20°C, optimal temperatures that are higher than those for desert cacti but representative of ambient temperatures in the tropics; its total daily net CO₂ uptake becomes zero at day/night air temperatures of 42/32°C. Stem damage occurs at 45°C for H. undatus, whose photosynthetic cells show little acclimation to high temperatures compared with other cacti and are also sensitive to low temperatures, ‐1.5°C killing half of these cells. Consistent with its shaded habitat, total daily net CO2 uptake is appreciable at a total daily PPF of only 2 mol m2 day' and is maximal at 20 mol m^?2 day^?1, above which photoinhibition reduces net CO₂ uptake. Net CO₂ uptake ability, which is highly correlated with stem nitrogen and chlorophyll contents, changes only gradually (halftimes of 2–3 months) as the concentration of applied N is changed. Doubling the atmospheric CO₂ concentration raises the total daily net CO₂ uptake of H. undatus by 34% under optimal conditions and by even larger percentages under adverse environmental conditions.     

Approaches to breeding for salinity tolerance - a case study on Porteresia coarctata

Cereals are the world's major source of food for human nutrition. Among these, rice (Oryza sativa) is the most prominent and represents the staple diet for more than two-fifths (2.4 billion) of the world's population, making it the most important food crop of the developing world (Anon., 2000a). Rice production in vast stretches of coastal areas is hampered due to high soil salinity. This is because rice is a glycophyte and it does not grow well under saline conditions. In order to increase rice production in these areas there is a need to develop rice varieties suited to saline environments. Research has shown that Porteresia coarctata, a highly salt tolerant wild relative of rice growing in estuarine soils, is an important material for transferring salt tolerant characteristics to rice. It is quite possible that Porteresia may be used as a parent for evolving better and truly salt resistant varieties. The inadequate results and the difficulties associated with conventional breeding techniques necessitate the use of the tools of crop biotechnology in unravelling some of the characteristics of Porteresia that have been highlighted in this report. In view of the limited resources available for increasing salinity tolerance to the breeders to wild rice germplasm, Porteresia is undoubtedly one of the key source species for elevating salinity tolerance in cultivated rice.     

Effect of salinisation of soil on growth and macro- and micro-nutrient accumulation in seedlings of Acacia catechu (Mimosaceae)

The effects of salinisation of soil on Acacia catechu (Mimosaceae) were studied by means of emergence and growth of seedlings and pattern of mineral accumulation. A mixture of chlorides and sulphates of Na, K, Ca and Mg was added to the soil and salinity was maintained at 4.1, 6.3, 8.2,10.1 and 12.2 dSm⁻¹. A negative relationship between proportion of seed germination and salt concentration was obtained. Seedlings did not emerge when soil salinity exceeded 10.1 dSm⁻¹. Results suggested that this tree species is salt tolerant at the seed germination stage. Seedlings survived and grew up to soil salinity of 10.1 dSm⁻¹, which suggests that this species is salt tolerant at the seedling stage too. Elongation of stem and root was retarded by increasing salt stress. Among the tissues, young roots and stem were most tolerant to salt stress and were followed by old roots and leaves, successively. Leaf tissue exhibited maximum reduction in dry mass production in response to increasing salt stress. However, production of young roots and death of old roots were found to be continuous and plants apparently use this process as an avoidance mechanism to remove excess ions and delay onset of ion accumulation in this tissue. This phenomenon, designated “fine root turnover”, is of importance to the mechanisms of salt tolerance. Plants accumulated Na in roots and were able to regulate transfer of Na ions to leaves. Stem tissues were a barrier for translocation of Na from root to leaf. Moreover, K was affected in response to salinity; it rapidly decreased in root tissues with increased salinisation. Nitrogen content decreased in all tissues (leaf, stem and root) in response to low water treatment and salinisation of soil. Phosphorus content significantly decreased, while Ca increased in leaves as soil salinity increased. Changes in tissue and whole plant accumulation patterns of the other elements tested, as well as possible mechanisms for avoidance of Na toxicity in this tree species during salinisation, are discussed.     

Physiological and Molecular Basis of Susceptibility and Tolerance of Rice Plants to Complete Submergence

Rice plants are much damaged by several days of total submergence.The effect can be a serious problem for rice farmers in therainfed lowlands of Asia, and runs contrary to a widespreadbelief amongst plant biologists that rice is highly tolerantof submergence. This article assesses the characteristics ofthe underwater environment that may damage rice plants, examinesvarious physiological mechanisms of injury, and reviews recentprogress achieved using linkage mapping to locate quantitativetraits loci (QTL) for tolerance inherited from a submergence-tolerantcultivar FR13A. Progress towards identifying the gene(s) involvedthrough physical mapping of a dominant tolerance locus on chromosome9 is also summarized. Available physiological evidence pointsaway from responses to oxygen shortage as being inextricablyinvolved in submergence injury. An imbalance between productionand consumption of assimilates is seen as being especially harmful,and is exacerbated by strongly accelerated leaf extension andleaf senescence that are ethylene-mediated and largely absentfrom FR13A and related cultivars. DNA markers for a major QTLfor tolerance are shown to be potentially useful in breedingprogrammes designed to improve submergence tolerance.     

Antioxidant and Pigment Composition during Autumnal Leaf Senescence in Woody Deciduous Species Differing in their Ecological Traits

Abstract: Photoprotection mechanisms have been studied during autumnal senescence in sun and shade leaves of woody plants with different ecological characteristics and senescence patterns. Three of them belonging to the same family, Betulaceae: the shade‐intolerant and early successional species (Betula alba L.), the shade‐tolerant and late successional species (Corylus avellana L.), and an N‐fixing tree with low N resorption efficiency (Alnus glutinosa L.). The other two species: a shade‐intolerant (Populus tremula L.) and a shade‐tolerant (Cornus sanguinea L.), were chosen because of their ability to accumulate anthocyanins during autumnal leaf senescence. The study of plants with different ecological strategies allowed us to establish general trends in photoprotection mechanisms during autumnal senescence, when nutrient remobilisation occurs, but also during whole leaf ontogeny. We have not found a clear relationship between shade tolerance and the level of photoprotection; the main difference between both groups of species being the presence of α‐carotene in shade leaves of shade‐tolerant species. Preceding autumn, nitrogen resorption started in mid‐summer and occurred in parallel with a slight and continuous ascorbate, chlorophyll and carotenoid degradation. However, the ascorbate pool remained highly reduced and lipid oxidation did not increase at this time. Contrasting with ascorbate, α‐tocopherol accumulated progressively in all species. Only during the last stages of senescence was chlorophyll preferentially degraded with respect to carotenoids, leading to the yellowing of leaves, except in A. glutinosa in which a large retention of chlorophyll and N took place. Senescing leaves were characterised, except in C. sanguinea, by a relative increase in the proportion of de‐epoxidised xanthophylls: zeaxanthin, antheraxanthin and lutein. The light‐induced accumulation of anthocyanins in C. sanguinea could play an additional protective role, compensating for the low retention of de‐epoxidised xanthophylls. These different strategies among deciduous species are consistent with a role for photoprotective compounds in enhancing nitrogen remobilization and storage for the next growing season.     

Adaptation to salinity inHordeum jubatum L. populations studied using reciprocal transplants

Tillers and seedlings ofHordeum jubatum L. from three sites with contrasting salinity regimes in central Saskatchewan, Canada were reciprocally transplanted in order to examine the tolerance of populations of this species to salinity and related habitat factors. Survival, growth and fecundity of the three populations were controlled more by transplant site characteristics than by genetic differences, i.e. differences among populations at a site tended to be smaller than differences among sites. Survival, growth and reproduction of all three populations were best at the non-saline site. The population originating at the non-saline site showed the poorest growth in the two saline habitats, but still had substantial salt tolerance. Fecundity was greatest when the populations were grow at their site of origin.     

Early needle senescence and thinning of the crown structure of Picea abies as induced by chronic SO2 pollution. II. Field data basis, model results and tolerance limits

Field data on the sulphur and cation budget of growing Norway spruce canopies (Picea abies [L.] Karst.) are summarized. They are used to test a spruce decline model capable of quantifying effects of chronic SO₂ pollution on spruce forests. At ambient SO₂ concentrations, acute SO₂ damage is rare, but exposure to polluted air produces reversible thinning of the canopy structure with a half-time of a few years. Canopy thinning in the spruce decline model is highest (i) at elevated SO₂ pollution, (ii) in the mountains, (iii) at unfertilized sites with poor K⁺, Mg²⁺ or Zn²⁺ supply, (iv) at low spruce litter decomposition rates, and (v) acidic, shallow soils at high annual precipitation rates in the field and vice versa. Model application using field data from Würzburg (moderate SO₂ pollution, alkaline soils, no spruce decline) and from the Erzgebirge (extreme SO₂ pollution, acidic soils in the mountains, massive spruce decline) predicts canopy thinning by 2–11% in Würzburg and by 45–70% in the Erzgebirge. The model also predicts different SO₂-tolerance limits for Norway spruce depending on the site elevation and on the nutritional status of the needles. If needle loss of more than 25% (damage class 2) is taken to indicate ‘real damage’ exceeding natural variances, then for optimum soil conditions SO₂ tolerance limits range from (27.3 ± 7.4) μg m^?3 to (62.6 ± 16.5) μg m^?3. For shallow and acidic soils, SO₂ tolerance limits range from (22.0 ± 5.5) μg m^?3 to (37.4 ± 7.5) μ m^?3. These tolerance limits, which are calculated on an ecophysiological data basis for Norway spruce are close to epidemiological SO₂-toIerance limits as recommended by the IUFRO, UN-ECE and WHO. The observed statistical regression slope of the plot (damaged spruce trees vs. SO₂-pollution) in west Germany is confirmed by modelling (6% error). Model application to other forest trees allows deduction of the observed sequence of SO₂-sensitivity: Abies > Picea > Pinus > Fagus > Quercus. Thus, acute phytotoxicity of SO₂ seems not to be involved in ‘forest decline’. Chronic SO₂-pollution induces massive canopy thinning of Abies alba and Picea abies only at unfavourable sites, where natural stress factors and secondary effects of SO₂pollution act together to produce tree decline.     

Improved ethanol tolerance and production in strains of Clostridium thermocellum

Two Clostridium thermocellum strains were improved for ethanol tolerance, to 5% (v/v), by gradual adaptation and mutation. The best mutant gave an ethanol yield of 0.37 g/g substrate, with a growth yield 1.5 times more than its parent. Accumulation of acids and reducing sugars by the mutant strain with 5% (v/v) ethanol was lower than that of the parent strain with 1.5% (v/v) ethanol.