Probing biomolecular interaction forces using an anharmonic acoustic technique for selective detection of bacterial spores

Receptor-based detection of pathogens often suffers from non-specific interactions, and as most detection techniques cannot distinguish between affinities of interactions, false positive responses remain a plaguing reality. Here, we report an anharmonic acoustic based method of detection that addresses the inherent weakness of current ligand dependant assays. Spores of Bacillus subtilis (Bacillus anthracis simulant) were immobilized on a thickness-shear mode AT-cut quartz crystal functionalized with anti-spore antibody and the sensor was driven by a pure sinusoidal oscillation at increasing amplitude. Biomolecular interaction forces between the coupled spores and the accelerating surface caused a nonlinear modulation of the acoustic response of the crystal. In particular, the deviation in the third harmonic of the transduced electrical response versus oscillation amplitude of the sensor (signal) was found to be significant. Signals from the specifically-bound spores were clearly distinguishable in shape from those of the physisorbed streptavidin-coated polystyrene microbeads. The analytical model presented here enables estimation of the biomolecular interaction forces from the measured response. Thus, probing biomolecular interaction forces using the described technique can quantitatively detect pathogens and distinguish specific from non-specific interactions, with potential applicability to rapid point-of-care detection. This also serves as a potential tool for rapid force-spectroscopy, affinity-based biomolecular screening and mapping of molecular interaction networks. © 2011 Elsevier B.V.

Effects of sediment anoxia and light on turion germination and early growth of Potamogeton crispus

Potamogeton crispus is a cosmopolitan aquatic species and is widely used as a pioneer species for vegetation restoration of eutrophic lakes. However, many restoration projects applying P. crispus turions have not been successful. Earlier studies focused on effects of light and temperature on turion germination. The purpose of this study was to determine whether sediment anoxia and light interactively affected the turion germination and early growth of P. crispus. Anoxic conditions in the experiment were produced by adding sucrose to the sediment. The germination rate of the turions was 68-73% lower in the highly anoxic condition treatment than in the control. Medium light intensity (10% of natural light at the water surface) was more favorable for germination under slightly anoxic conditions than either low or high light intensity. The growth of newly-formed sprouts was also significantly inhibited by sediment anoxia. Photosynthesis and shoot biomass were reduced under sediment anoxia, whereas total chlorophyll content, root biomass, and soluble protein content were highest in the low anoxic condition treatment. Medium light improved net photosynthesis and biomass production of the sprouts. We conclude that turion germination and sprout growth can be significantly inhibited by sediment anoxia. Medium light intensity may alleviate this inhibition by anoxia, but light has little effect when sediment anoxia is severe. For the purposes of vegetation restoration, more attention should be paid to the role of sediment anoxia, and it is necessary to improve sediment and light conditions for turion germination and early growth of P. crispus in eutrophic lakes. These results will contribute to a more complete understanding of turion germination dynamics of P. crispus and will be useful for future restoration programs.

Effects of solar UV radiation on germination of conchospores and morphogenesis of sporelings in Porphyra haitanensis (Rhodophyta)

The effects of ultraviolet radiation (UVR 280-400 nm) on the germination of Porphyra haitanensis conchospores and on the growth and morphogenesis of the subsequent sporelings were investigated by culturing the released conchospores under natural sunlight from 29 September to 6 October 2005. Germination increased with time and was faster when UV-B was excluded using cut-off filters. There were significant negative effects of UV-B radiation on growth and cell division of sporelings, with decreases up to 18% for thallus length, between 6 and 18% for thallus width, up to 29% for thallus area, and between 6 and 14% for cell size as compared to PAR-controls. UV-A had a significant positive effect on morphogenesis, enhancing the formation of sporelings with cells dividing transversely; on the other hand, UV-B delayed the formation of such sporelings. We also tested the effects of solar UVR on the growth of P. haitanensis juveniles and found no significant effects. Our results indicate that UV-A has an important role in the germination and morphogenesis of the species, but on the other hand, sporelings of P. haitanensis are more sensitive to UV-B radiation than juveniles.

Control of cocklebur seed germination by nitrogenous compounds : Nitrite, nitrate, hydroxylamine, thiourea, azide and cyanide

Germination of non-dormant upper cocklebur (Xanthium pinsylvanicum Wallr.) seeds was stimulated by not only CS(NH2)2 but also NH2OH, KCN and NaN3. This stimulation was not via the enhancement of aerobic C2H4 production. NH2OH, KCN and NaN3 in certain concentrations promoted the initial growth of axial and/or cotyledonary parts, but the degree of growth promotion by NH2OH, NaN3 and KCN was slight compared with that by CS(NH2)2. As in the case of CS(NH2)2, however, the germinationstimulating effect of NH2OH disappeared rapidly as the preceding imbibition period was prolonged. In contrast, KCN and NaN3 were still effective in stimulating the germination of aged seeds maintained on a water substratum, as previously seen with anaerobiosis. Anaerobic induction was enhanced not only by NaN3 and KCN but also by NH2OH, KNO3, KNO2 CO(NH2)2 and CS(NH2)2 applied during the anaerobic treatment, but without causing an increase in anaerobic production of C2H4. Furthermore, KCN and NaN3, given prior to the anaerobic treatment acted additively with anaerobic induction. The germination-stimulating actions of nitrogenous compounds are discussed in comparison with those of C2H4 and anaerobiosis.

Fruit anatomy,seed germination and seedling development in the Japanese seagrass phyllospadix

Phyllospadix iwatensis Makino and phyllospadix japonicus Makino have similar frunt morphology and anatomy.The rhomboid fruit of Japanese phyllospadix is dark brown in colour and is characterized by two arms bearing stiff inflected bristles which can act as an anchoring system. The fruit covering consists of a thin cuticular seed coat and pericarp remains mainly fibrous endocarp. In the groove region of the fruit.the cuticular seed coat and endocarp are replaced by nucellus cells with wall in growths and crushed pigment strands with lignified walls.these tissues appera to control the transfer of nutrients to developing seed.the seed is oval with a small embryo and a large hypocotyl. the embryo is straight and simple,with the plumule containing three leaf primordia and a pair of root primordia surrounded by a cotyledon.the hypocotyl has large vontral lobe containing central provascular tissue and two small dorsal lobes.the hypocotyl contains starch.lipid and protein.and acts as a nutrient store.the seed of P.iwatensis has a dormancy period of 2-6 weeks and germination eventually reaches-65%.but is not synchronized.during germination the leaves emerge first.and then after at least three young leaves have formed and abseised.the roots emerge,usually?6 months after the commencement of germination.Utilizaton of the nutrient reserves is initially from the perihpery of the hypocotyl and then progressively towards its centre.

Seed germination and seedling morphology of the seagrass, Halophila engelmannii (Hydrocharitaceae)

Seeds of Halophila engelmannii Aschers., that were collected in Redfish Bay, Texas, at weekly intervals from mid-May to mid-June 1986, began to germinate 3–4 weeks after collection. Most of the collections subsequently showed an increase in the rate of germination under increased light intensity and all had a stoppage of germination after transfer to darkness, indicating a light requirement to break endogenous seed dormancy. During the 5 weeks after seeds germinated, seedlings in soil culture produced a rosette of six leaves before the appearance of a rhizome bud in the axil of the third leaf. The first node of the rhizome produced a root and an upright shoot with a pseudowhorl of three to five leaves.