Ability of Pollen to Germinate prior to Anthesis and Effect of Desiccation on Germination

The ability of pollen to germinate prior to anthesis was tested using Easter lily (Lilium longiflorum L.) and corn (Zea mays L.). Lily pollen normally dries to a low moisture content between anthesis and pollination while corn does not. The corn pollen germinated well (about 73%) when removed from anthers 1 day before anthesis and placed on culture medium. The lily pollen germinated poorly (0 to 5%) when harvested one to six days before anthesis. However, the lily pollen harvested one or two days before anthesis gave greatly improved germination (about 55%) after it was dried to a low moisture content. The results indicate that an internal control prevents premature germination of lily pollen and that drying is the final stage of pollen maturation. A different sort of regulatory mechanism must operate to prevent premature germination of corn pollen.     

Pollen tube growth following compatible and incompatible intraspecific pollinations in Petunia hybrida

The observation that both compatible and incompatible pollen tubes grow at identical speeds on the stigma in many plants with gametophytically controlled self-incompatibility (SI) systems has, in Petunia, been extended to cover all other facets of pollen behaviour on this tissue. On entry into the stylar transmitting tissue both types of tubes accelerate, but the compatible achieve a higher terminal velocity than do the incompatible, which eventually slow and stop. Grafting experiments show that the top 1 mm of the stylar tissue can play an important rôle in determining the future development of the pollen tube. Following mixed pollinations, proportionally too many compatible pollen tubes reach the ovary than would be expected from the results of pure compatible and incompatible pollinations indicating that incompatible pollen in some way helps prime the style for growth of compatible pollen tubes. This data is considered in terms of recent structural studies of these tissues, and related to the pollination conditions pertaining to Petunia populations in the field.Abbreviation SI self-incompatibility     

In vivo isolation of a pPJ3a-λ cosmid mediated by the Tn2307 transposon

From an E. coli cell harboring plasmid pPJ3b (= pPJ3a::Tn2301) and infected with phage λ, we have isolated two defective phages having inserted pPJ3a DNA and Tn2301 in their genomes. One of them has been extensively characterized: it behaves like a cosmid, i.e., upon injection into the cell, its DNA circularizes and replicates as a plasmid (pPJ10); it can be packaged again in λ heads, provided the presence of a phage helper. Furthermore, heteroduplex analysis has shown that in pPJ10, the transposon Tn2301 is inverted compared to its direction in pPJ3b. We give evidence suggesting that this type of inversion is in part mediated by Tn2301.     

Opiate receptor antagonism by l-prolyl-l-leucyl-glycinamide, MIF-I

Chronic administration of l-prolyl-l-leucyl-glycinamide (MIF-I) to mice slightly reduced morphine's antinociceptive activity in the hot-plate test and modified the biphasic motor activity response to morphine. MIF-I antagonized the initial depression of activity and potentiated the increased motor activity phase. Chronic treatment of rats with MIF-I prevented morphine's antinociceptive activity in the tail flick test. MIF-I partly antagonized the inhibition by morphine of the coaxially stimulated guinea-pig ileum preparation. The inhibition of the ileum produced by ethylketocyclazocine was weakly antagonized by MIF-I. In contrast, MIF-I had no effect on the inhibition of the stimulated mouse vas deferens produced by Leu-enkephalin. The findings show that MIF-I weakly and selectively inhibits μ-type opiate receptors which suggests that MIF-I could be an endogenous inhibitor of opiate receptors.     

Nonsense mutations in the amylomaltase gene and other loci of Streptococcus pneumoniae

Molecular Genetics and Genomics - Maltose-negative mutations in the amylomaltase gene of Streptococcus pneumoniae were examined for the presence of nonsense mutations. Out of 28 single-site mutants...     

Nitrogen budget of a shortgrass prairie ecosystem

Summary A N budget is presented for a shortgrass prairie ecosystem. The grassland was ungrazed by domestic herbivores. The quantities of N in various plant, animal, microorganism, and soil components of the ecosystem are estimated for the date when aboveground living biomass was at its maximum for the growing season of 1973. Annual transfers of N between the various compartments were also estimated.Of the total N, 99.5% was in organic forms. The relatively inert heteropolycondensate fraction of the organic matter in the soils contained 88.8% of the N. Living organisms contained 4.2% and dead but recognizable organisms or part thereof contained 6.5% of the total N. Belowground animals contained more than 10 times as much N as abovegroud animals, but combined, animals contained less than 0.1% of the total in the system. Living plant material contained 2.5% of the total N. Seventy-two percent of the living plant N was below ground. Microorganisms contained 1.4% of the total N.Total N uptake by plants from soil solution was 2.9 g·m^-2·yr^-1. Aerial portions of plants were allocated 1.9 N·m^-2·yr^-1 although apparently 26% of this amount came from internally recycled sources. The heteropolycondensate fraction of the soil contributed 0.7 g N·m^-2·yr^-1 to mineral forms, but these components of the system were assumed to be in steady state; thus an equal amount of mineral N was allocated back to the source. Mineralization of N from plant residues was sufficient to account for all of the N taken up by plants from soil solution. Soil animals immobilized about 0.4 g N·m^-2·yr^-1 while the amount shunted to aboveground animals was trivial.     

Inhibition by ethanol, acetaldehyde and trifluoroethanol of reactions catalysed by yeast and horse liver alcohol dehydrogenases.

1. Produced inhibition by ethanol of the acetaldehyde-NADH reaction, catalysed by the alcohol dehydrogenases from yeast and horse liver, was studied at 25 degrees C and pH 6-9. 2. The results with yeast alcohol dehydrogenase are generally consistent with the preferred-pathway mechanism proposed previously [Dickenson & Dickinson (1975) Biochem. J. 147, 303-311]. The observed hyperbolic inhibition by ethanol of the maximum rate of acetaldehyde reduction confirms the existence of the alternative pathway involving an enzyme-ethanol complex. 3. The maximum rate of acetaldehyde reduction with horse liver alcohol dehydrogenase is also subject to hyperbolic inhibition by ethanol. 4. The measured inhibition constants for ethanol provide some of the information required in the determination of the dissociation constant for ethanol from the active ternary complex. 5. Product inhibition by acetaldehyde of the ethanol-NAD+ reaction with yeast alcohol dehydrogenase was examined briefly. The results are consistent with the proposed mechanism. However, the nature of the inhibition of the maximum rate cannot be determined within the accessible range of experimental conditions. 6. Inhibition of yeast alcohol dehydrogenase by trifluoroethanol was studied at 25 degrees C and pH 6-10. The inhibition was competitive with respect to ethanol in the ethanol-NAD+ reaction. Estimates were made of the dissociation constant for trifluoroethanol from the enzyme-NAD+-trifluoroethanol complex in the range pH6-10.     

Estimation of rate dissociation constants involving ternary complexes in reactions catalysed by yeast alcohol dehydrogenase

Stopped-flow studies of oxidation of butan-1-ol and propan-2-ol by NAD(+) in the presence of Phenol Red and large concentrations of yeast alcohol dehydrogenase give no evidence for the participation of a group of pK(a) approx. 7.6 in alcohol binding. Such a group has been implicated in ethanol binding to horse liver alcohol dehydrogenase [Shore, Gutfreund, Brooks, Santiago & Santiago (1974) Biochemistry13, 4185-4190]. The present result supports previous findings based on steady-state kinetic studies with the yeast enzyme. Stopped-flow studies of the yeast alcohol dehydrogenase-catalysed reduction of acetaldehyde by NADH in the presence of ethanol as product inhibitor indicate that the rate-limiting step is NAD(+) release from the enzyme-NAD(+)-ethanol product complex. This finding permits calculation of K(3), the dissociation constant for ethanol from the enzyme-NAD(+)-ethanol complex, by using the product-inhibition data of Dickenson & Dickinson (1978) (Biochem. J.171, 613-627). The calculations show that K(3) varies very little with pH in the range 5.95-8.9, and this agrees with the findings of the stopped-flow experiments described above. Absorption and fluorescence measurements on mixtures of substrates and coenzymes in the presence of high concentrations of alcohol dehydrogenase have been used to estimate values for the ratio [enzyme-NADH-acetaldehyde]/ [enzyme-NAD(+)-ethanol] at equilibrium. The values obtained were in the range 0.11+/-0.04, and this value together with estimates of K(3) was used to provide estimates of values for rate constants and dissociation constants for steps within the catalytic mechanism.     

Antenna organization and evidence for the function of a new antenna pigment species in the green photosynthetic bacterium Chloroflexus aurantiacus

Whole cells and isolated chlorosomes (antenna complex) of the green photosynthetic bacterium Chloroflexus aurantiacus have been studied by absorption spectroscopy (77 K and room temperature), fluorescence spectroscopy, circular dichroism, linear dichroism and electron spin resonance spectroscopy. The chlorosome absorption spectrum has maxima at 450 (contributed by carotenoids and bacteriochlorophyll (BChl) a Soret), 742 (BChl c) and 792 nm (BChl a) with intensity ratios of 20:25. The fluorescence emission spectrum has peaks at 748 and 802 nm when excitation is into either the 742 or 450 nm absorption bands, respectively. Whole cells have fluorescence peaks identical to those in chlorosomes with the addition of a major peak observed at 867 nm. The CD spectrum of isolated chlorosomes has an asymmetric-derivative-shaped CD centered at 739 nm suggestive of exciton interaction at least on the level of dimers. Linear dichroism of oriented chlorosomes shows preferential absorption at 742 nm of light polarized parallel to the long axis of the chlorosome. This implies that the transition dipoles are also oriented more or less parallel to the long axis of the chlorosome. Treatment with ferricyanide results in the appearance of a 2.3 G wide ESR spectrum at g 2.002. Whole cells grown under different light conditions exhibit different fluorescence behavior when absorption is normalized at 742 nm. Cells grown under low light conditions have higher fluorescence intensity at 748 nm and lower intensity at 802 nm than cells grown under high light conditions. These results indicate that the BChl c in chlorosomes is highly organized, and transfers energy from BChl c (742 nm) to a connector of baseplate BChl B792 (BChl a) presumably located in the chlorosome baseplate adjacent to the cytoplasmic membrane.     

Gastrin-amidating enzyme in the porcine pituitary and antrum. Characterization of molecular forms and substrate specificity

As is the case with many other peptide hormones of the brain and intestine, the formation of biologically active gastrin from a glycine-extended processing intermediate occurs via the action of a peptidylglycyl alpha-amidating monooxygenase (PAM). The observation that gastrin exists primarily as unamidated precursors in the pituitary but as amidated gastrin in the antrum prompted this study to examine whether the amidating enzymes in the two organs were different in their characteristics. Amidating activity was quantified by measuring the conversion of glycine-extended tridecagastrin (G13-Gly) to amidated tridecagastrin and glycine-extended hexapancreatic polypeptide (PP6-Gly) to amidated hexapancreatic polypeptide by radio-immunoassay. Two molecular forms of amidating activity were identified in both the porcine antrum and pituitary. The first, PAM-A, had an apparent Mr of 51,000 and a net negative charge at pH 7.0, whereas PAM-B was smaller (Mr approximately 30,000) and had a net positive charge at pH 7.0. Both molecular forms were similar in their cofactor requirements (copper, ascorbic acid, and catalase) and pH optima in the antrum and pituitary. The Km was significantly lower and the Vmax higher for PP6-Gly than for G13-Gly in the pituitary and antrum. These data suggest that although there is no difference between antral and pituitary PAM, the selective affinity of PAM for certain substrates may provide a mechanism for the differential amidation of different hormones within a given tissue or cell.