An in Vivo Study of Substrate Specificities of Acyl-Lipid Desaturases and Acyltransferases in Lipid Synthesis in Synechocystis PCC6803

The cyanobacterium Synechocystis PCC6803 was fed heptanoic acid to study the substrate specificities of desaturases and acyltransferases in lipid synthesis. This aliphatic acid was elongated to C15, C17, and C19 fatty acids, which were incorporated into polar glycerolipids and desaturated. The double bonds were located at the [delta]6, [delta]9, [delta]12, and [omega]3 positions of the fatty acids. This suggests that the [delta]9 desaturase counts the carbon number from the carboxy terminus, whereas the so-called [delta]15 desaturase counts from the methyl terminus. The counting mechanisms of the [delta]6 and [delta]12 desaturases are not fully understood. In the distribution of fatty acids at the sn positions of the glycerol moiety, the C17, C18, and C19 fatty acids were located at the sn-1 position, whereas the C15 and C16 fatty acids were located at the sn-2 position. This suggests that glycerol-3-phosphate acyltransferase specifically transfers heptadecanoic, octadecanoic, and nonadecanoic acids, whereas 1-acylglycerol-3-phosphate acyltransferase specifically transfers pentadecanoic and hexadecanoic acids.     

Cloning of cDNAs for genes that are early-responsive to dehydration stress (ERDs) inArabidopsis thaliana L.: identification of three ERDs as HSP cognate genes

InArabidopsis thaliana L., accumulation of abscisic acid (ABA) began to increase 2 h after plants had been subjected to dehydration stress and reached maximum levels after 10h. Differential hybridization was used to isolate 26Arabidopsis cDNAs with gene expression induced by a 1 h dehydration treatment. The cDNA clones were classified into 16 groups based on Southern blot hybridization, and named ERD (early-responsive todehydration) clones. Partial sequencing of the cDNA clones revealed that three ERDs were identical to those of HSP cognates (Athsp70-1, Athsp81-2, and ubiquitin extension protein). Dehydration stress strongly induced the expression of genes for the three ERDs, while application of ABA, which is known to act as a signal transmitter in dehydration-stressed plants, did not significantly affect the ERD gene expression. This result suggests that these HSP cognates are preferentially responsive to dehydration stress inA. thaliana, and that signaling pathways for the expression of these genes under conditions of dehydration stress are not mainly mediated by ABA. We also discuss the possible functions of these three ERD gene products against dehydration stress.     

Serotypes and mating types of clinical strains ofCryptococcus neoformans isolated in Taiwan

Twenty-one strains ofCryptococcus neoformans isolated from patients in Taiwan were characterized for serotypes and mating types. Slide agglutination test was performed with 8 factor-specific sera (Iatron Company, Japan) to determine the serotypes. Wheat bran agar (WBA) and malt extract agar (MEA, Wickerham) media were used for the mating tests. Twenty of the isolates were of serotype A, and one was serotype B. Except for 2 strains of serotype A, all of the serotype A strains mated withFilobasidiella neoformans var.neoformans, mating type a. The only serotype B strain mated withF. neoformans var.bacillispora mating type a in MEA medium. These data revealed the low prevalence (1/21; 4.8%) ofC. neoformans var.gattii in Taiwan, a subtropically located island.     

Temperature dependence of the photosynthetic activities in the thylakoid membranes from the blue-green alga Anacystis nidulans.

The effects of temperature and ethidium bromide on the banding of heat-denatured DNA was studied during equilibrium centrifugation in density gradients of NaI. Centrifugation at 10 degrees C prevents the partial renaturation of Escherichia coli DNA and Clostridium perfringens DNA that occurs at 20 degrees C. A centrifugation temperature of --5 degrees C is required to prevent renaturation of T7 phage DNA. Ethidium bromide decreases renaturation of Escherichia coli DNA during centrifugation at 20 degrees C and causes a small shift in the buoyant density of both denatured and native DNA. Equilibrium centrifugation at lower temperatures prevents DNA renaturation and permits increased utilization of the large buoyant density difference between native and heat-denatured DNA in gradients of NaI.     

Biosynthesis of furano-terpenes by sweet potato cell culture

Callus was induced from sweet potato root tissue on an agarmedium containing Heller's minerals, vitamins, 2,4-D, yeastextract and sucrose. Furano-terpenes were scarcely detectedin the callus. However, when the callus was transferred to aliquid culture medium and incubated with reciprocal shaking,furano-terpenes were rapidly produced mainly in the culturemedium. Furano-terpene production by the cell culture was suppressedby addition of Ceratocystis fimbriata spores or HgCl₂ to theculture medium. Yeast extract and sucrose in the culture mediumwere important for furano-terpene production. 3-Hydroxy-3-methylglutarylcoenzyme A (HMG-CoA) reductase activity increased in the cells,followed by the production of furano-terpenes. The TLC patternof furano-terpenes produced by the cell culture was essentiallythe same as that produced by sweet potato root tissue infectedby C. fimbriata or treated with HgCl₂, but the quantitativeproportion of the individual furano-terpenes in the former differedmarkedly from that in the latter. (Received January 11, 1979; )     

Determination of double bond positions of unsaturated fatty acids by a chemical ionization mass spectrometry computer system

After stereospecific oxidation, trimethylsilylated methyl esters of mono- and diunsaturated fatty acids were analyzed by combined gas-liquid chromatography-chemical ionization mass spectrometry. The positions of original double bonds were deduced from the fragment ions produced by the cleavage of the carbon-carbon bond between two trimethylsilyl ethers. These fragment ions were recorded at m/e 187 and 259 in the case of 16:1(n-7), at m/e 187 and 287 in the case of 18:1(n-7), at m/e 215 and 259 in the case of 18:1(n-9), and at m/e 172 and 259 in the case of 18:2(n-6), respectively. The diastereoisomers of monounsaturated fatty acids can be discriminated by comparing the intensities of the fragment ions at m/e 253 and 285 in the case of 16:1 and at m/e 281 and 313 in the case of 18:1. The diastereosiomers of diunsaturated fatty acids may also be distinguished from each other by comparing the intensities of the fragment ions formed by the loss of trimethylsilyl function from the characteristic ions. Mono- and diunsaturated fatty acids in liver may be dominantly in cis- 18:1(n-1) and cis-cis-18:2(n-6) as determined by a mass chromatographic technique.     

Synthesis of RNA in tissue discs of sweet potato roots after cutting or mercuric chloride treatment

Time course analysis of RNA contents of tissue discs after cuttingdisclosed a remarkable increase in total RNA during the first12 hr after cutting and this elevated level remained unchangedfor 48 hr. The elevated RNA level at 24 hr of incubation wasnot changed by subsequent HgCl₂ treatment. The incorporationrate of the label from ³H-uridine into RNA rapidly increasedimmediately after cutting and reached a maximum at about 9 hrof incubation, then decreased sharply until 24 hr and continuedto decrease gradually thereafter. The incorporation rate at24 hr of incubation was not changed by subsequent HgCl₂ treatment.The results of polyacrylamide gel electrophoresis indicatedthat bulk RNA was synthesized most actively at 9 hr of incubationthen the rate of RNA synthesis decreased gradually. (Received August 26, 1977; )     

Continuous production of glutathione by immobilized Saccharomyces cerevisiae cells

Summary Glutathione was continuously produced by an immobilized Saccharomyces cerevisiae IFO 2044 cell column. The production of glutathione was strongly influenced by the level of activity of the glycolytic pathway. This activity was maintained constant by the addition of NAD.Abbreviations ADP adenosine-5-diphosphate - ATP adenosine-5-triphosphate - NAD nicothinamide adenine dinucleotide     

GM1-gangliosidosis: Accumulation of ganglioside GM1 in cultured skin fibroblasts and correlation with clinical types

Summary Uptake of radioactivity from ¹⁴C-galactose into gangliosides by cultured skin fibroblasts was studied. G_M3 was the major ganglioside in control human fibroblasts. An increase of G_M1 was demonstrated in G_M1-gangliosidosis fibroblasts. The degree of G_M1 accumulation was correlated with the clinical types of this disease. The fibroblasts from an infantile-type patient showed a marked increase of G_M1. In late-onset types the amount of total gangliosides was only slightly increased, but the distribution of individual gangliosides was definitely abnormal; a relative increase of G_M1 was demonstrated in these cases. G_M1 -galactosidase activities were not detectable in either infantile or late-onset cases.