Sucrose-phosphate synthase activity and yield analysis of tomato plants transformed with maize sucrose-phosphate synthase

Sucrose synthesis is a major element of the interactions between photosynthesis and plant growth and development. Tomato (Lycopersicon esculentum Mill. cv. UC82B) plants transformed with maize sucrose-phosphate synthase (SPS; EC 2.3.1.14) expressed from either a ribulose-1,5-bisphosphate carboxylase-oxygenase (Rubisco) small subunit promoter (SSU) or the cauliflower mosaic virus 35S promoter (35S) were used to study effects of increased sucrose synthesis rates on plant growth. The plants were grown in growth chambers, field plots, and open-top chambers. The 35S plants had a 2 to 3-fold increase in young-leaf SPS activity, a 10 to 20-fold increase in young-root SPS activity and no increase in young-fruit SPS activity. The leaf SPS activity in one of the 35S lines fell to control levels by two months of age. The SSU plants had a 4 to 5-fold increase in leaf SPS activity and no significant increase in root or young-fruit SPS activity. One 35S line, which maintained high leaf SPS activity throughout development, yielded 70–80% more than controls at both normal and elevated CO₂ in open-top chambers in the field and 20–30% more than controls in two additional field trials. The other 35S line and the two SSU lines either yielded less or did not differ from controls under several growth conditions. Since only one of four transformed lines showed an increase in yield, we can not yet conclude that increased leaf SPS activity leads to increased yield. However, increased leaf SPS activity appears to result in increased fruit sugar content since all three lines with increased leaf SPS usually also had increased fruit sugars. Received: 18 November 1996 / Accepted: 22 January 1997     

Vessel contents during transpiration - embolisms and refilling

A test was made of the previous unexpected observation that embolized vessels were refilled during active transpiration. The contents of individual vessels in petioles of sunflower plants were examined, after snap-freezing at 2-h intervals during a day's transpiration, in the cryo-scanning electron microscope, and assessed for the presence of liquid or gas (embolism) contents. Concurrent measurements were made of irradiance, leaf temperature, transpiration rate, and leaf water potential (by pressure chamber). Up to 40% of the vessels were already embolized by 0900 (transpiration rate ~5 _g_cm-2_s-1, water potential about -300 J/kg), and the proportion declined to a minimum (as low as 4%) at 1500. This was the time of highest transpiration rate (~25 _g_cm-2_s-1) and most negative water potential (-600 to -700 J/kg). Images of vessels with mixed gas and liquid contents showed water being extruded through pits in the walls of the vessels to refill them. The data indicate that: (1) the water columns are weak and break under quite small tensions; (2) embolisms are repaired by refilling the vessels with water on a short time scale (minutes) throughout the day; (3) the vigor of this refilling process is adjusted by the plant on a longer time scale (hours) to the intensity of the water stress; (4) the pressure chamber balance pressure (P) does not measure tension in the vessels; (5) P is also not a measure of water stress (as measured by vessel embolization); and (6) P is a measure of the plant's response to water stress, i.e., a measure of the vigor of the refilling process. The test confirms the previous observations and negates all the assumptions and evidences of the Cohesion Theory. The data are fully consistent with the Compensating Pressure Theory, which predicted the relations demonstrated in this experiment. Using the assumptions of that theory it is easy to outline a simple mechanism by which the refilling of vessels might be achieved by reverse osmosis, and the adjustment in (3) might be achieved by osmoregulation in the starch sheath.     

Vessel contents of leaves after excision - a test of Scholander's assumption

A test was attempted of the assumption that, when a leaf is cut, the xylem still contains water under tension beyond the first vessel cross walls. This assumption enabled Scholander to argue that the balance pressure in his pressure chamber measured the tension in water columns in the vessels before cutting. The numbers of embolized vessels were counted, after rapid freezing of petiole and midrib samples of sunflower leaves, in the cryo-scanning electron microscope. Counts were made on leaves still attached to the plant and at intervals after cutting from the plant (up to 16 min) during a short spring day's transpiration. The lengths of vessels in the leaves, measured by latex particle perfusion, showed that 8% of vessels in the mid-petioles and 0% in the midribs should be opened by cutting. The changing percentages of embolized vessels (E) with time showed that: (1) in intact plants E was close to zero until midday when it rose to ~40%, and then fell progressively to near zero by 1600; (2) in excised leaves there was no detectable change in E immediately after cutting, and, in all but two time courses, no change as large as the 8% of opened vessels within 16 min; (3) but briefly, when E was high (midday), it rose further after cutting to a plateau (_E = 30-40%) in 4 min. From this rate of emptying, the estimated maximum pressure difference between vessels and parenchyma was of the order of 0.05-0.2 MPa (0.5 to 2 bar) at this time. (4) All these changes occurred in the petioles 1 h before they were found in the midribs. The test failed because the expected large pressure difference between vessels and parenchyma was not present. Further, the embolized vessels were refilled at the time of peak transpiration, which would be impossible with any substantial tension in the vessels. Because these results contradict the whole basis of the Cohesion Theory, a second experiment was carried out to test them, and is reported in a companion paper.     

Effects of elevated atmospheric CO2 on fine root length and distribution in an oak-palmetto scrub ecosystem in central Florida

Atmospheric CO₂ concentration is rising and it has been suggested that a portion of the additional carbon is being sequestered in terrestrial vegetation and much of that in below-ground structures. The objective of the present study was to quantify the effects of elevated atmospheric CO₂ on fine root length and distribution with depth with minirhizotrons in an open-top chamber experiment in an oak-palmetto scrub ecosystem at Kennedy Space Centre, Florida, USA. Observations were made five times over a period of one and a half years in three ambient chambers (350 p.p.m. CO₂), three CO₂ enriched chambers (700 p.p.m. CO₂), and three unchambered plots. Greater root length densities were produced in the elevated CO₂ chambers (14.2 mm cm^?2) compared to the ambient chambers (8.7 mm cm^?2). More roots may presumably lead to more efficient acquisition of resources. Fine root abundance varied significantly with soil depth, and there appeared to be enhanced proliferation of fine roots near the surface (0–12 cm) and at greater depth (49–61 cm) in the elevated CO₂ chambers. The vertical root distribution pattern may be a response to availability of nutrients and water. More studies are needed to determine if increased root length under CO₂ enriched conditions actually results in greater sequestering of carbon below ground.     

Soil CO2 efflux of two silver birch clones exposed to elevated CO2 and O3 levels during three growing seasons

In the present open‐top chamber experiment, two silver birch clones (Betula pendula Roth, clone 4 and clone 80) were exposed to elevated levels of carbon dioxide (CO₂) and ozone (O₃), singly and in combination, and soil CO₂ efflux was measured 14 times during three consecutive growing seasons (1999–2001). In the beginning of the experiment, all experimental trees were 7 years old and during the experiment the trees were growing in sandy field soil and fertilized regularly. In general, elevated O₃ caused soil CO₂ efflux stimulation during most measurement days and this stimulation enhanced towards the end of the experiment. The overall soil respiration response to CO₂ was dependent on the genotype, as the soil CO₂ efflux below clone 80 trees was enhanced and below clone 4 trees was decreased under elevated CO₂ treatments. Like the O₃ impact, this clonal difference in soil respiration response to CO₂ increased as the experiment progressed. Although the O₃ impact did not differ significantly between clones, a significant time × clone × CO₂× O₃ interaction revealed that the O₃‐induced stimulation of soil respiration was counteracted by elevated CO₂ in clone 4 on most measurement days, whereas in clone 80, the effect of elevated CO₂ and O₃ in combination was almost constantly additive during the 3‐year experiment. Altogether, the root or above‐ground biomass results were only partly parallel with the observed soil CO₂ efflux responses. In conclusion, our data show that O₃ impacts may appear first in the below‐ground processes and that relatively long‐term O₃ exposure had a cumulative effect on soil CO₂ efflux. Although the soil respiration response to elevated CO₂ depended on the tree genotype as a result of which the O₃ stress response might vary considerably within a single tree species under elevated CO₂, the present experiment nonetheless indicates that O₃ stress is a significant factor affecting the carbon cycling in northern forest ecosystems.     

Ex vivo expansion of hematopoietic stem cells in a proliferation chamber with external stirred conditioning tank: Sequential optimization of growth factors

The insufficient number of hematopoietic stem cells (HSCs) extracted from various cell sources creates the need for the use of growth factors in the culture systems. Numerous types and concentrations of growth factors have been reported in the literature. In this study, we investigated the effect of three important hematopoietic growth factors thrombopoietin (TPO), stem cell factor (SCF), and Fms‐like tyrosine kinase‐3 ligand (Flt‐3) as well as cell‐seeding density on ex vivo expansion of human HSCs using a factorial design. Sequential optimization was then followed by methods of steepest ascent and central composite design. The optimum concentrations of growth factors were 50, 90, and 34 ng/mL for SCF, TPO, and Flt‐3, respectively, at an initial cell density of 2.5 × 10⁵ cells/mL. Effective expansion factor value (EEF) of HSCs increased considerably and revealed almost similar results when the cells were cultured in a 24‐well plate (EEF = 4.54 ± 0.43) and a proliferation chamber with an external stirred conditioning tank (PC‐ESCT; EEF = 5.1 ± 0.35) at seeding density of 2.5 × 10⁵ cells/mL after 7 days. The cells did not show considerable changes in proliferation when they were cultured in medium containing serum or in a commercial serum‐free medium at the optimum concentrations of the growth factors. The present study demonstrated the optimum condition of hematopoietic growth factors as well as the potential of PC‐ESCT for ex vivo expansion of HSCs.     

Culture of fetal mouse liver in plastic chambers: A new technique for organ culture

Summary A new technique for organ culture which uses plastic culture chambers and the advantages of the cellophane-sheet technique is described with the results of a study of cultivations of fetal mouse liver. Two chambers, each containing cells, were placed in gas permeable roller tubes and rotated at 0.1 rpm in a CO₂-air gassed incubator. The fetal mouse liver cells developed electron microscopic features similar to those of the in vivo adult liver by 9 days of cultivation. The albumin content and tyrosine aminotransferase (TAT) activity were detected in the cultivated liver. TAT activity was further induced by prednisolone. These results indicate the potential of this culture method for the study of physiological and pathological processes. This work was supported in part by a Grant-in-Aid for Cancer Research from the Ministry of Education, Science and Culture, Japan and Science Technology Agency, Japan.     

An automatic system to study the responses of respiration and photosynthesis by submerged macrophytes to environmental variables

An apparatus to measure the rates of respiration and photosynthesis of aquatic plants in water at velocities of up to 200 mm s^–1 in a closed water-flow system with partial recirculation, is described. The temperature, the light regime and the concentration of dissolved oxygen are controlled automatically. Typical results are given for Ranunculus penicillatus var. calcareus which were repeatable between the same season in different years and compared with published data.     

Tooth components of mandibular deciduous molars of Homo sapiens sapiens and Homo sapiens neanderthalensis: a radiographic study.

Tooth components of deciduous molars were measured from standardized radiographs of Homo sapiens sapiens and Homo sapiens neanderthalensis. Enamel height and width were greater in deciduous teeth of Homo sapiens sapiens than in Homo sapiens neanderthalensis and the differences were statistically significant (p less than 0.01). Dentin height showed no significant differences between the two groups, but enamel to floor of pulp chamber and pulp height and width dimensions were significantly greater in Homo sapiens neanderthalensis. Discriminant analysis carried out between groups, using deciduous tooth components, showed an accuracy of 98-100% for identification of Homo sapiens sapiens and 83-92% for identification of Homo sapiens neanderthalensis. The results obtained in this study on dental dimensions support the hypothesis of a distinct evolutionary line for Neanderthals.