Analysis of mitochondrial DNA from somatic hybrid cell lines of Saccharum officinarum (sugarcane) and Pennisetum americanum (pearl millet)

Mitochondrial DNA (mtDNA) from cell suspension cultures of two intergeneric somatic hybrids of Pennisetum americanum (pearl millet) + Saccharum officinarum (sugarcane) was examined by restriction endonuclease digestion and hybridization with sorghum mtDNA cosmids. The mtDNA of one somatic hybrid was indistinguishable from that of pearl millet, while the second exhibited a combination of parental mtDNAs, suggesting mitochondrial fusion. Several novel, possibly recombinant, mtDNA restriction fragments were detected in this hybrid, which may have resulted from intergenmic recombination.Florida Agriculture Experiment Station Journal Series No: 8090.     

The contribution of abiotic processes to buried litter decomposition in the northern Chihuahuan desert

Summary Creosobebush (Larrea tridentata) fine litter was treated with either the general biocide HgCl₂ and CuSO₄ or water (controls) and buried 5 cm beneath the soil surface in the northern Chihuahuan Desert. The treated litter showed significantly less mass loss than controls during the three month summer-autumn field study; controls lost about 20% of the original mass while treated litter lost less than 2%. In addition, the total nitrogen content of the control litter increased from an initial concentration of about 14.08 g kg^-1 to 17.62 g kg^-1 dry weight by the end of the study, while treated litter nitrogen content decreased to 13.30 g kg^-1. Results suggest abiotic processes other than leaching have little effect on the decomposition of buried litter in this environment.     

Assembly of the barley light-harvesting chlorophyll a/b proteins in barley etiochloroplasts involves processing of the precursor on thylakoids

A barley gene encoding the major light-harvesting chlorophyll a/b-binding protein (LHCP) has been sequenced and then expressed in vitro to produce a labelled LHCP precursor (pLHCP). When barley etiochloroplasts are incubated with this pLHCP, both labelled pLHCP and LHCP are found as integral thylakoid membrane proteins, incorporated into the major pigment-protein complex of the thylakoids. The presence of pLHCP in thylakoids and its proportion with respect to labelled LHCP depends on the developmental stage of the plastids used to study the import of pLHCP. The reduced amounts of chlorophyll in a chlorophyll b-less mutant of barley does not affect the proportion of pLHCP to LHCP found in the thylakoids when import of pLHCP into plastids isolated from the mutant plants is examined. Therefore, insufficient chlorophyll during early stages of plastid development does not seem to be responsible for their relative inefficiency in assembling pLHCP. A chase of labelled pLHCP that has been incorporated into the thylakoids of intact plastids, by further incubation of the plastids with unlabelled pLHCP, reveals that the pLHCP incorporated into the thylakoids can be processed to its mature size. Our observations strongly support the hypothesis that after import into plastids, pLHCP is inserted into thylakoids and then processed to its mature size under in vivo conditions.     

Proton transfer reactions in aqueous solutions of pyridoxamine phosphate

UV-visible and ¹³C NMR measurements described in the literature and our ³¹P NMR measurements support the following mechanism of proton transfer reactions in aqueous solutions of pyridoxamine phosphate: Only the tautomeric equilibrium between neutral form, A _N, and zwitterion, A _Z, which is analogous to the tautomeric equilibrium of 3-hydroxypyridine in aqueous solution, is important, and that equilibrium does not change upon the dissociation of the second phosphate proton. With these simplifying assumption, we have simulated the relaxation spectrum of the proton transfer reactions of pyridoxamine phosphate in water using parameters from analogous reactions and compared it with our ultrasound and temperature jump measurements. We have found that the relaxation process measured by the temperature jump experiment is mainly caused by the overall reaction A _N=A _Z (or A _N ^- =A _Z ^- ) and the ultrasound absorption at the isoelectric point between pK₂ and pK₃ is mainly caused by the overall reaction .     

A berberine-aniline blue fluorescent staining procedure for suberin,lignin, and callose in plant tissue

Summary A fluorescent staining procedure to detect suberin, lignin and callose in plants has been developed. This procedure greatly improves on previous methods for visualizing Casparian bands in root exodermal and endodermal cells, and performs equally well on a variety of other plant tissues. Berberine was selected as the most suitable replacement forChelidonium majus root extract after comparing the staining properties of the extract with those of four of its constituent alkaloids. Aniline blue counterstaining efficiently quenched unwanted background fluorescence and nonspecific berberine staining, while providing a fluorochrome for callose. When used with multichambered holders which allow simultaneous processing of freehand sections, this efficient staining procedure facilitates morphological studies involving large numbers of samples.Abbreviations ISCC-NBS Inter-Society Color Council-National Bureau of Standards - UV ultraviolet light     

Enzymatic production of L-tryptophan in a reverse micelle reactor

The enzymatic production of tryptophan from indole and serine was investigated in a micellar solution of the surfactant Brij 56 in cyclohexane. An anion exchanger was employed to facilitate the transfer of tryptophan and serine between the water pool of the reverse micelle and the bulk organic phase. The influence of potassium ion, water content, pH, and co-surfactant on enzyme activity is reported. Kinetic studies indicate that the enzyme is not inhibited by indole in the micellar system and that the enzyme is more stable in reverse micelles than in bulk water. The design of a continuous reverse micelle reactor, which accommodates both product recovery and enzyme reactivation, is discussed.     

Experimental reovirus myocarditis in newborn mice. Electron microscopic observations

Reovirus is a double-stranded RNA-virus which induces myocarditis in newborn mice. Due to the large diameter of the viral particles (70-75 nm) it can be detected by electron microscopy. Subcutaneous inoculation of 0.05 ml reovirus type 3 (TCID50-titer: 10(8.5)/ml) into newborn NMRI-mice (12-18 h after birth) caused a grey-yellow mottling on the ventricular surface first seen on the 5th day after birth. At the same time muscle fiber necrosis was observed which increased with time. Electron microscopic investigations of the diseased heart muscle disclosed a marked interstitial oedema, swelling of the tubular system and sarcoplasmic reticulum, and degenerative changes in the mitochondria of individual myocardiocytes as early as the 2nd post-inoculation day. Simultaneously, an enlarged Golgi-apparatus and an increasing number of lysosomes, partially exhibiting acid phosphatase activity, was detected in the perinuclear region of ventricular myocardiocytes. On the 5th day after infection, viruses were detected either within single membrane vesicles, dispersed in cytoplasm or as aggregated clusters in the perinuclear region. These in vivo electron microscopic findings correspond with observations of virus propagation in cell-culture systems.