A New Apparatus for Control of Temperature and Humidity in Leaf and Small Plant Chambers

A new electronic system for effecting feedback control of temperaturefrom 3 to 50 °C and humidity from approximately 15 to 99per cent is described. This system uses thermistor sensors tocontrol, through electronic control units, the temperature ofwater baths which determined the temperature and humidity ofthe airstream passing through leaf or small plant chambers.The control has been found to be precise, and the temperatureand humidity can each be reset from a separate single controlknob.     

A Cell-Permeable Fluorescent Polymeric Thermometer for Intracellular Temperature Mapping in Mammalian Cell Lines

Changes in intracellular temperatures reflect the activity of the cell. Thus, the tool to measure intracellular temperatures could provide valuable information about cellular status. We previously reported a method to analyze the intracellular temperature distribution using a fluorescent polymeric thermometer (FPT) in combination with fluorescence lifetime imaging microscopy (FLIM). Intracellular delivery of the FPT used in the previous study required microinjection. We now report a novel FPT that is cell permeable and highly photostable, and we describe the application of this FPT to the imaging of intracellular temperature distributions in various types of mammalian cell lines. This cell-permeable FPT displayed a temperature resolution of 0.05°C to 0.54°C within the range from 28°C to 38°C in HeLa cell extracts. Using our optimized protocol, this cell-permeable FPT spontaneously diffused into HeLa cells within 10 min of incubation and exhibited minimal toxicity over several hours of observation. FLIM analysis confirmed a temperature difference between the nucleus and the cytoplasm and heat production near the mitochondria, which were also detected previously using the microinjected FPT. We also showed that this cell-permeable FPT protocol can be applied to other mammalian cell lines, COS7 and NIH/3T3 cells. Thus, this cell-permeable FPT represents a promising tool to study cellular states and functions with respect to temperature.     

A Mobile Apparatus for Measuring Leaf Photosynthesis in the Field

An apparatus is described that has been designed to measurethe gas exchange of individual cereal leaves in the field, usinginfrared gas analysis. Measurements on leaves of winter wheatshowed that there was true saturation of net photosynthesisat high irradiances but as irradiance decreased there was aproportional decrease in net photosynthesis. Measurements madeon wheat plants grown in a controlled environment, for comparison,showed the photosynthesis–light response curve to be betterfitted by a rectangular hyperbola. This dissimilar responseof photosynthesis to irradiancc for plants grown in controlledenvironments indicates the possible problems of extrapolationfrom the laboratory to the field.     

An Automatic Device for Measuring Leaf Area

The design and construction of a digital device, which measuresthe area of leaves by scanning with a rapidly moving spot oflight, is described.     

A Comparison between Conductive and Infrared Devices for Measuring Mean Skin Temperature at Rest,during Exercise in the Heat,and Recovery

PurposeSkin temperature assessment has historically been undertaken with conductive devices affixed to the skin. With the development of technology, infrared devices are increasingly utilised in the measurement of skin temperature. Therefore, our purpose was to evaluate the agreement between four skin temperature devices at rest, during exercise in the heat, and recovery.MethodsMean skin temperature (T-sk) was assessed in thirty healthy males during 30 min rest (24.0 ± 1.2°C, 56 ± 8%), 30 min cycle in the heat (38.0 ± 0.5°C, 41 ± 2%), and 45 min recovery (24.0 ± 1.3°C, 56 ± 9%). T-sk was assessed at four sites using two conductive devices (thermistors, iButtons) and two infrared devices (infrared thermometer, infrared camera).ResultsBland–Altman plots demonstrated mean bias ± limits of agreement between the thermistors and iButtons as follows (rest, exercise, recovery): -0.01 ± 0.04, 0.26 ± 0.85, -0.37 ± 0.98°C; thermistors and infrared thermometer: 0.34 ± 0.44, -0.44 ± 1.23, -1.04 ± 1.75°C; thermistors and infrared camera (rest, recovery): 0.83 ± 0.77, 1.88 ± 1.87°C. Pairwise comparisons of T-sk found significant differences (p < 0.05) between thermistors and both infrared devices during resting conditions, and significant differences between the thermistors and all other devices tested during exercise in the heat and recovery.ConclusionsThese results indicate poor agreement between conductive and infrared devices at rest, during exercise in the heat, and subsequent recovery. Infrared devices may not be suitable for monitoring T-sk in the presence of, or following, metabolic and environmental induced heat stress.     

A Microflow Respirometer for Measuring the Oxygen Consumption of Small Aquatic Organisms

A microrespiration device is decribed which uses a Clark electrode to measure the oxygen consumption or production of small and microscopic aquatic organisms in an open flow system. The construction and working principles of the device, which can measure oxygen consumptions as low as 0.5 nl · h⁻¹, are described. The design of the apparatus permits parallel measurements under identical conditions with a single electrode. The device can be matched to various sizes of animal and oxygen consumption rates by means of specimen chambers of different volumes (6 μl, 35 μl, 140 μl) and a variable water flow rate. The microflow respiration device has been used successfully to measure the respiration of zooplankton and meiobenthos organisms as well as protozoans and has also been used successfully on board a research vessel.     

An Apparatus for Measuring Volumes of Small Objects

An apparatus for measuring volumes of small objects such as tissue blocks is described. The apparatus measures volumes by fluid displacement and consists of a micropipette adapted to fit the mouth of an Erleiuneyer flask, a Luer adaptor fused to the side of the flask, and a glass syringe. When assembled with fluid enclosed, the fluid rises to a low level in the micropipette. Withdrawal of fluid into the syringe lowers the fluid level below the mouth of the flask. The micropipette is raised, the object to be measured is placed in the flask, and the micropipette is joined to the flask again. Fluid returned to the flask from the syringe rises to a higher level in the micropipette. The difference between the two fluid levels equals the volume of the object measured.

This apparatus gives reproducible measurements and can be calibrated for absolute volume determination. It is inexpensive to construct and easy to use.     

A Cellophane-Strip Technique for Culturing Tissue in Multipurpose Culture Chambers

A new technique for the cultivation of living tissues in the multipurpose culture chamber is described. This procedure employs strips of cellophane as the agent for anchoring tissue explants to the coverslip walls of the chamber and disposes of the time-honored plasma-clot technique. The primary advance embodied in this procedure lies in the fact that cells emigrating from so-cultured explants manifest themselves in a highly differentiated manner comparable to the cells of origin, whereas the outgrowth from the same types of tissue in plasma clots results in a more undifferentiated type of growth. Comparisons of outgrowths from embryonic thyroid, bone, and muscle (chicken) are photographically documented, and attention is called to certain cytochemical methods which further corroborate the differentiated quality obtained with the cellophane-strip technique.     

A New Method for Measuring the Permeability of Plant Cell Membranes Using Epidermis-free Leaf Discs

A method has been developed for measuring the uptake and effluxof radiochemicals by higher plant cells, using epidermis-freeleaf discs. The advantages and disadvantages of this methodare discussed and some physical characteristics of the tissuedescribed. The relative permeability (urea=1) of tobacco leafcells to nonelectrolytes was thiourea 2.3, ethylene glycol 39,-amino isobutyric acid 1.9. N-methylpyridinium chloride accumulatedin the tissue according to the Nernst Distribution, by a processwhich was temperature dependent.     

叶面积指数田间测量中有限长度平均法的改进

叶面积指数(LAI)的田间测量是生态和农业等领域的常规工作之一, 测量方法分为直接测量和间接测量, 间接测量中有一类方法基于数字相机照片提取冠层孔隙率, 再用有限长度平均法同时估算LAI和聚集指数。然而, 有限长度平均法自提出以来缺少进一步的发展, 在有限长度的样线/样方上应用比尔定律的方式具有理论缺陷, 可能造成无效值或高估LAI。从模拟的训练数据中提取经验公式以取代比尔定律进行样线/样方的LAI估算, 提高了有限长度平均法的精度和鲁棒性。进一步分析在一定精度需求下对样线/样方大小和数量的要求, 对于非均匀样地, 提出样线长度为8倍等效叶片边长、样方边长为3倍等效叶片边长的推荐设置。在基于数字相机照片提取非均匀样地LAI的应用中, 使用样方采样比样线采样更为适宜。     

Changes in Leaf Morphology and Composition with Future Increases in CO<Subscript>2</Subscript> and Temperature Revisited: Wheat in Field Chambers

Whether leaf morphology is altered by future increases in atmospheric CO₂ and temperature has been reexamined over 3 years in wheat grown in field chambers at two levels of nitrogen supply. Flag leaf fresh and dry mass, area, volume, and ratios of these parameters, as well as the contents of water, chlorophyll, nonstructural carbohydrates, and nitrogen compounds have been determined at anthesis and 14 days later. High CO₂ decreased rather than increased, as reported in the literature, leaf mass per area and leaf density, and increased water content per area and per volume and water percentage. Warmer temperatures also decreased leaf mass per area, but did not affect density or water per area or per volume, whereas they increased water percentage. Nitrogen supply did not change CO₂ and temperature effects on leaf morphology. Nonstructural carbohydrates increased and nitrogen compounds decreased in elevated CO₂, and the sum of these compounds decreased with warmer temperatures. These changes in composition did not account for modifications of leaf morphology. We conclude that increases in atmospheric CO₂ and temperature after leaf initiation can decrease leaf mass per area, and elevated CO₂ can also decrease leaf density, due to decreases in leaf structural compounds. The functional significance of these changes is probably a decrease in photosynthetic capacity per unit leaf area.     

Bean Leaf Expansion in Relation to Temperature

When dwarf Phaseolus vulgaris plants were grown in a controlledenvironment at 20, 25, 30, and 35° C, expansion of the primaryleaves occurred in two phases with an intermediate lag. Varyingrates and duration of expansion were involved, leading to greatestfinal areas at the two intermediate temperatures. Dry weightsof the leaves and leaf areas were similary influenced by temperature,except that the initial rates of increase continued for a longerperiod for weights than for areas. The rates of cell divisionand final numbers of cells were similar from 25 to 35° C,but both were decreased at 20° C. Final cell sizes were,on the other hand, decreased only at the highest temperature.The time trends of cell expansion varied greatly with temperature. Leaf expansion is discussed as a possible consequence of substratesupply, which may be determined by temperature in a number ofways. Cell division and cell expansion are not considered tobe joint direct determinants of leaf expansion. Temperatureinfluences division, with two consequences; the rate interactswith substrate supply to determine size of cells, and finalcell number affects potential leaf area. Cell size is regardedas being secondary to numbers of cells and total material available,although some factors can vary cell size independently of substrate,e.g. water status. An important control of leaf growth, until the attainment ofabout half the final area, may be exercised by way of the leaf.Subsequently, intra-plant competition is likely to dominate.     

Diurnal Changes in Citrus Leaf Thickness, Leaf Water Potential and Leaf to Air Temperature Difference

Diurnal changes in leaf water potential and leaf thickness ofwell-watered citrus trees were found to be highly correlated.Midday decreases in leaf thickness of about 30–35 µm reflected midday decreases in leaf water potential of about1.1–1.3 MPa from predawn values. Leaf water potentialwas also correlated with changes in leaf-to-air temperaturedifference and ambient vapour pressure deficit. Leaf thicknessas well as leaf to air temperature difference could possiblybe used to monitor leaf water status continuously as an indicatorof citrus tree water stress.     

A Mechanistic Niche Model for Measuring Species' Distributional Responses to Seasonal Temperature Gradients

Niche theory is central to understanding how species respond geographically to climate change. It defines a species'' realized niche in a biological community, its fundamental niche as determined by physiology, and its potential niche—the fundamental niche in a given environment or geographic space. However, most predictions of the effects of climate change on species'' distributions are limited to correlative models of the realized niche, which assume that species are in distributional equilibrium with respect to the variables or gradients included in the model. Here, I present a mechanistic niche model that measures species'' responses to major seasonal temperature gradients that interact with the physiology of the organism. I then use lethal physiological temperatures to parameterize the model for bird species in North and South America and show that most focal bird species are not in direct physiological equilibrium with the gradients. Results also show that most focal bird species possess broad thermal tolerances encompassing novel climates that could become available with climate change. I conclude with discussion of how mechanistic niche models may be used to (i) gain insights into the processes that cause species to respond to climate change and (ii) build more accurate correlative distribution models in birds and other species.     

Controlled Environment Chambers for Investigating Tree Response to Elevated CO2 and Temperature Under Boreal Conditions

A closed CO₂ and temperature-controlled, long-term chamber system has been developed and set up in a typical boreal forest of Scots pine (Pinus sylvestris L.) near the Mekrijärvi Research Station (62°47N, 30°58E, 145 m above sea level) belonging to the University of Joensuu, Finland. The main objectives of the experiment were to provide a means of assessing the medium to long-term effects of elevated atmospheric CO₂ concentration (EC) and temperature (ET) on photosynthesis, respiration, growth, and biomass at the whole-tree level and to measure instantaneous whole-system CO₂ exchange. The system consists of 16 chambers with individual facilities for controlling CO₂ concentration, temperature, and the combination of the two. The chambers can provide a wide variety of climatic conditions that are similar to natural regimes. In this experiment the target CO₂ concentration in the EC chambers was set at a fixed constant of 700 µmol mol^–1 and the target air temperature in the ET chambers to track the ambient temperature but with a specified addition. Chamber performance was assessed on the base of recordings covering three consecutive years. The CO₂ and temperature control in these closed chambers was in general accurate and reliable. CO₂ concentration in the EC chambers was within 600–725 µmol mol^–1 for 90 % of the exposure time during the "growing-season" (15 April – 15 September) and 625–725 µmol mol^–1 for 88 % of the time in the "off-season" (16 September – 14 April), while temperatures in the chambers were within ±2.0 °C of the ambient or target temperature in the "growing season" and within ±3.0 °C in the "off season". There were still some significant chamber effects. Solar radiation in the chambers was reduced by 50–60 % for 82 % of the time in the "growing season" and 55–65 % for 78 % of the time in the "off season", and the relative humidity of the air was increased by 5–10 % for 72 % of the time in the "growing season" and 2–12 % for 91 % of the time in the "off season". The crown architecture and main phenophase of the trees were not modified significantly by enclosure in the chambers, but some physiological parameters changed significantly, e.g., the radiant energy-saturated photosynthesis rate, transpiration rate, maximum photochemical efficiency of photosystem 2, and chlorophyll content.     

Leaf Cytosolic Fructose-1,6-bisphosphatase : A Potential Target Site in Low Temperature Stress

Leaf cytosolic fructose-1,6-bisphosphatase (FBPase), partially purified from both spinach (Spinacia oleracea, var Hipack) and peas (Pisum sativum, var Progress No. 9), is reversibly inactivated by exposure to low temperature. Thus, even though assays were conducted at 22°C, samples incubated at 0 to 12°C had greatly reduced activity relative to controls maintained at 22°C. Following incubation at 22°C prior to assay, the inactivated samples regained their initial activity. Chloroplast FBPase, by contrast, was unaffected by low temperature treatment. This feature as well as lack of a response of cytosolic FBPase to thioredoxins f or c_f and to chloroplast FBPase antibody indicate that the FBPase isozymes of leaves are different proteins.     

小分子RNA在植物叶发育中的调控作用

叶发育是叶原基细胞有序的分裂、生长和分化的过程,受到植物激素和多个转录因子的严格调控.近年的研究表明,在叶片发育的过程中,小分子RNA是基因调控网络的重要组分.小分子RNA通过对其中一些转录因子的抑制作用,影响其表达水平和空间分布,维持叶的正常发育.本综述介绍了小分子RNA及其靶基因调控模块在叶片发生、 叶片形状、叶子极性发育和叶子衰老等过程中的调控作用,并展望了未来研究中新方向.