Effect of Nitrate Concentration on the Root: Shoot Ratio in Dactylis glomerata L. and on the Kinetics of Growth in the Vegetative Phase

The vegetative growth of Dactylis glomerata L. in sand was studiedunder controlled light, temperature, and nutritional conditions.Plants were daily supplied with three nutrient solutions ofdifferent nitrate concentrations (10^–2, 10^–3 and2 x 10^–4 mol I^–1). For each concentration, growthobeyed an exponential law between the fourth and seventh weeksafter sowing. The time constant of the exponential was the samefor the shoot as for the root, and showed no significant variationwith nitrate concentration. The kinetic results and the strong dependence of the root: shootratio on nitrate concentration are discussed on the basis ofThornley's model. Hypothesizing that the molecular mechanismsof nitrate absorption are independent of the nitrate concentrationof the nutrient solution, we derived a relationship betweenthe root: shoot ratio and nitrate concentration. This relationshipwas found to be compatible with the experimental results. Dactylis glomerata L., vegetative phase, kinetics of growth, root: shoot equilibrium, nitrate absorption     

Biflavonoids as novel antituberculosis agents

A series of naturally occurring and synthetic biflavonoids was evaluated for inhibitory activity against Mycobacterium tuberculosis H37Rv (Mtb). Compounds 6, 24, and 25 demonstrated 96, 95, and 87% inhibition, respectively, at a screening concentration of 12.5 microg/mL. The type of linkage and the presence of methoxy- and nitro-substituents in biflavonoids may contribute to the observed inhibitory activity. The results of this study represent the discovery of biflavonoids as a potential new class of antituberculosis agent.     

Genetic transformation of green bean callus via Agrobacterium mediated DNA transfer

Summary Kanamycin resistant callus was produced from leaf disc or hypocotyl expiants of green bean (Phaseolus vulgaris L.) when cultured on a defined medium containing 50 mg/l kanamycin after 4 days of co-cultivation with Agrobacterium tumefaciens strain EHA101 containing the binary vector pKYLX71GUS. The presence of neomycin phosphotransferase II (NPT-II) in crude cellular extracts from the kanamycin resistant callus was confirmed by ELISA. The expression of the ß-glucuronidase (GUS) reporter gene was confirmed by histochemical and fluorimetric analyses. Southern blot border analysis confirmed the integration of the foreign DNA. In addition to the evidence obtained from Southern analysis, the absence of Agrobacterium in the transformed callus cultures was confirmed by microscopic observation and through test cultures. Using the above protocol, bean callus cultures were also transformed with a bean chalcone synthase promoter-GUS fusion. These cultures, when treated with the elicitor glutathione, showed higher levels of GUS expression than the unelicited callus clumps.     

Field tests with insecticides and insect growth regulators to control insect pests of cowpea and its effects on certain beneficial insects

A small‐plot tests were conducted on cowpea, Vigna ungiculata (L.) Walp, to determine the effectiveness of 14 selected insecticides representing four insecticide classes (organophosphates, carbamates, pyrethroids, and insect growth regulators, IGRs), and four insecticide/IGR mixtures on cowpea insect pests and its effects on certain beneficial insects. By day 3 after treatement, the insecticides phenthoate, isoxathion, cyanophos, carbaryl, and cypermethrin used at sub‐recommended rates reduced the leafhopper, Empoasca spp., populations by >83% than those in the control. On that day, all insecticide treatments significantly reduced the cowpea aphid, Aphis craccivora, numbers below that of the control. The prothiofos, isoxathion, pirimicarb, and fenpropathrin treatments provided continuing control to both leafhoppers and aphids through day 21 after spraying. It seems that most insecticide treatments were not effective in whitefly, Bemisia tabaci, control. The best control of the whitefly immatures was obtained by day 3 after spraying in plots received thiodicarb (76%) and fenpropathrin (60%). All selected insecticides and rates used had very low residual effect against B. tabaci immatures. By day 3 posttreatment, the carbaryl (2.02 kg/ha) caused completely protection for cowpea pods frMw blue pea butterfly, Lampides boeticus, larvae. The IGRs, flufenoxuron and Dowco‐439, applied alone and those applied in mixtures with insecticides, carbaryl/teflubenzuron, chlorpyrifos/XRD‐473, and methomyl/ flufenoxuron, exhibited satisfactory control (>81%) to butterfly larvae on day 7 posttreatment. All treatments did not exhibit a considerable residual activity against L. boeticus larvae through 2–3 weeks followed spray. Most insecticides applied at the higher rates used significantly reduced the numbers of limabean pod borer, Etiella zinckenella, larvae found in cowpea pods collected on day 7 after application. The IGRs, flufenoxuron and Dowco‐439, applied alone, at 0.119 kg/ha, or in mixtures, methomyl/flufenoxuron (0.167 kg/ha) and chlorpyrifos/Dowco‐439 (0.911 kg/ha) caused >73% control of E. zinckenella larvae through day 21 posttreatment. The best control of cowpea weevil, Callosobru‐chus spp., adults (>83%), on day 3 after spraying, was obtained in treatments of cyanophos (1.19 kg/ha), prothiofos (1.43 kg/ha), flufenoxuron (0.119 kg/ha), and chlorpyrifos/Dowco‐439 (0.911 kg/ha). The IGR flufenoxuron applied alone or in mixture, methomyl/flufenoxuron (0.164 kg/ ha) exhibited satisfactory residual activity against Callosobruchus adults through day 21 posttreatment. Percentage seed damage by the larvae of cowpea weevils was significantly lower in most treatments than that of control. Populations of insect predators found in all treated plots were extremely reduced than those found in untreated plots. By day 21 after spraying, the IGRs flufenoxuron, XRD‐473, and Dowco‐439, applied at the low rate of 0.071 kg/ha, seemed to be less effect against insect predators than other insecticides used.     

Rapid,robotic, small‐scale protein production for NMR screening and structure determination

Three‐dimensional protein structure determination is a costly process due in part to the low success rate within groups of potential targets. Conventional validation methods eliminate the vast majority of proteins from further consideration through a time‐consuming succession of screens for expression, solubility, purification, and folding. False negatives at each stage incur unwarranted reductions in the overall success rate. We developed a semi‐automated protocol for isotopically‐labeled protein production using the Maxwell‐16, a commercially available bench top robot, that allows for single‐step target screening by 2D NMR. In the span of a week, one person can express, purify, and screen 48 different ¹⁵N‐labeled proteins, accelerating the validation process by more than 10‐fold. The yield from a single channel of the Maxwell‐16 is sufficient for acquisition of a high‐quality 2D ¹H‐¹⁵N‐HSQC spectrum using a 3‐mm sample cell and 5‐mm cryogenic NMR probe. Maxwell‐16 screening of a control group of proteins reproduced previous validation results from conventional small‐scale expression screening and large‐scale production approaches currently employed by our structural genomics pipeline. Analysis of 18 new protein constructs identified two potential structure targets that included the second PDZ domain of human Par‐3. To further demonstrate the broad utility of this production strategy, we solved the PDZ2 NMR structure using [U‐¹⁵N,¹³C] protein prepared using the Maxwell‐16. This novel semi‐automated protein production protocol reduces the time and cost associated with NMR structure determination by eliminating unnecessary screening and scale‐up steps.     

Annual growth of Cassiope tetragona as a proxy for Arctic climate: developing correlative and experimental transfer functions to reconstruct past summer temperature on a millennial time scale

Annual growth of the polar evergreen shrub Cassiope tetragona on Svalbard was evaluated as a proxy for Arctic summer temperatures. Transfer functions were derived from temperature‐growth correlations of shoots and from a temperature‐growth response, obtained from experimental warming using open top chambers (OTC) in high Arctic tundra vegetation at Isdammen approximately 1.5 km southeast of Longyearbyen, Svalbard (78°N, 15 E) and in Longyeardalen, 3 km west of Isdammen from 2004 to 2006. Air temperatures, monitored throughout the summer months, were 1.3 °C higher inside the OTCs than in the control plots. Annual stem growth was measured by tagging stems and leaves, and in the lab with shoots harvested from OTCs and control plots. Annual growth parameters assessed were leaf production, sum of length and weight of individual leaves, and stem length increment derived from leaf scar distances and the distances between wintermarksepta in the stem. Wintermarksepta are formed at the end of the summer growth period when the pith is narrowing and consist of dense and dark tissue ( Fig. 1b ). The variation of annual growth in a 34‐year site chronology (based on Cassiope shoots from the surroundings of the OTCs and control plots) correlated strongly with the mean summer temperature on Svalbard. The number of leaf pairs, leaf length and stem length also increased in the OTC warmed plots in the second and third year of warming. Transfer functions were derived from the temperature‐annual growth correlations from a single shoot from Longyeardalen, from the cross‐dated Isdammen site chronology and from the growth response to experimental warming. Based on leaf scar distances and distances between wintermarksepta of well‐preserved subfossil shoots in arctic tundra soil, annual stem length increase was assessed for the layers of a soil core collected at the Isdammen site. Based on the derived transfer functions summer temperature of the period relating to the 15 cm deep tundra soil core layer, radiocarbon dated at 4230±40 bp , may have been 3.0 °C lower than the present‐day 6.2 °C value. These results indicate that the transfer functions can be used to reconstruct past temperatures, beyond the time range of instrumental temperature and ice core records of Svalbard.

Figure 1 Open in figure viewer PowerPoint (a) Morphology of shoot of Cassiope tetragona collected May 26, 2007, Longyeardalen. Removal of the front row of leaves shows four leaf pairs of the 2006 summer, the leaf primordium for the 2007 growing season and the stem length increase summer 2006. (b) Wintermarksepta, darker colored than the pith tissue, indicating the winterperiod in a longitudinal section of an air‐dried shoot collected August 26, 2006, Longyeardalen.     

New Pedal Remains of Megaladapis and Their Functional Significance. Am J Phys Anthropol 100:115–139

Erratum: Wunderlich RE, Simons EL, Jungers WL. 1996. New Pedal Remains of Megaladapis and Their Functional Significance. Am J Phys Anthropol 100:115–139.