N-isobutyl-trans-2-trans-4-eicosadienamide and other alkaloids of fruits of Piper guineense

A novel alkaloid, N-isobutyl-trans-2-trans-4-eicosadienamide, has been isolated from the fruits of Piper guineense and fully characterized. The structure of the compound has been confirmed by an unambiguous synthesis of the tetrahydro derivative. The known alkaloid, δαβ-dihydropiperine has also been isolated and the position of the double bond in this compound confirmed by the use of an NMR shift reagent.     

A new intracellular phytase of enterobacteria: Isolation and characterization

A phytase (myo-inositol-1,2,3,4,5,6-hexakisphosphate phosphohydrolase) has been isolated for the first time from a cell lysate of Pantoea vagans 3.2 enterobacteria and studied. The enzyme has been assigned to the class of histidine acid phosphatases (EC 3.1.3.26). It has been purified to homogeneity, and its primary structure has been determined. The molecular weight of the protein is 46 kDa, and K _m is 0.28 mM. Some physicochemical properties of the enzyme have been examined.     

Mapping of BamHI and SmaI DNA restriction sites on the genome of the nuclear polyhedrosis virus of the alfalfa looper, Autographa californica

The DNA of the nuclear polyhedrosis virus of the alfalfa looper, Autographa californica (AcNPV), has been analyzed with restriction endonucleases BamHI and SmaI. The molecular weight of the BamHI fragments, SmaI fragments, and BamHI + SmaI fragments has been determined. The molecular weight of AcNPV DNA is calculated to be about 82 million. A presumptive physical map of the BamHI and SmaI restriction sites on the AcNPV genome has been constructed.     

A CIS-Dominant Mutation in ASPERGILLUS NIDULANS Affecting the Expression of the amdS Gene in the Presence of Mutations in the Unlinked Gene, amdA

A mutant producing very high levels of the acetamidase enzyme encoded by the amdS gene has been isolated in a strain containing the amdA7 mutation, which itself causes high levels of this enzyme. Genetic analysis has shown that this mutation, designated amdI66, is adjacent to the amdS gene and is cis-dominant in its effect. The amdI66 mutation has little effect on amdS expression when present in strains not containing the amdA7 mutation. Two other amdA mutations investigated also interact with the amdI66 mutation to result in high acetamidase levels. No interaction between amdI66 and any of the other putative regulatory genes affecting amdS expression has been observed. The amdI66 mutation has been located by fine structure mapping at the extreme end of the controlling region, which has previously been defined by genetic mapping (Hynes 1979). Analysis of this region has been extended by mapping new mutations resulting in loss of amdS expression. One of these defines the most extreme site capable of mutation to loss of gene function found so far.     

Isolation and characterization of a new bacillary phytase

From cell lysates of a recombinant Escherichia coli strain, Bacillus ginsengihumi phytase has been isolated and studied for the first time. The enzyme has been purified to homogeneity, and its primary structure has been determined. It has been concluded that phytase belongs to the class of β-propeller phosphatases. The molecular weight of the protein is 41 kDa, and pI is 4.8. Some physicochemical properties of the enzyme have been investigated.     

Identification in various chlorate-resistant mutants of a protein involved in the activation of nitrate reductase in the soluble fraction of a chlA mutant of Escherichia coli K-12

We report some properties of Protein PA which has been isolated from the soluble fraction of a chlB mutant after anaerobic growth in the presence of KNO₃. This protein has been identified by its capacity to reactivate nitrate reductase present in the soluble fraction of a chlA mutant by the complementation process. The presence of active Protein PA in the chlB mutant is independent of the presence of oxygen or of nitrate during growth. In contrast, the addition of sodium tungstate to the growth medium leads to the formation of inactive Protein PA which is not able to activate nitrate reductase in the chlA-soluble extract by complementation. Inactive Protein PA has been quantitated immunologically. The partial purification of Protein PA has been achieved from various chlorate-resistant mutants (chlA?chlG). The establishment of particular complementation systems comprising the soluble extracts of chlA or chlB mutants and partially purified Protein PA from soluble fractions of different chlorate-resistant mutants, has allowed the quantitative estimation of this protein. The analysis by ‘rocket immunoelectrophoresis’ using an antiserum specific for Protein PA has shown that inactive Protein PA is present in approximately equivalent amounts in the chlA, chlE, chlG and chlD mutants     

Phenolic compounds of the subfamily pomoideae: A chemotaxonomic survey

The subfamily Pomoideae has been surveyed for leaf phenolics and it has been shown that flavone glycosides are present in the genera Sorbus, Aronia, Chaenomeles and Hesperomeles in addition to the previously reported occurrences in Crataegus, Malus and Pyrus. The dihydrochalcone phloridzin, a typical constituent of Malus, has also been found in Docynia. Arbutin and phenolic acid-calleryanin esters are apparently restricted to Pyrus. Naringenin and eriodictyol glucosides have been detected in Pyracantha, Sorbus, Photinia, Chaenomeles and Hesperomeles. A number of Pomoideae phenolics have been found in two Spiraeoideae genera; luteolin 7-glucoside,] luteolin 7-diglucoside, luteolin 7-rhamnosylglucoside and apigenin 7-glucoside in Exochorda and the dihydrochalcone trilobatin in Sorbaria. The chemotaxonomic evidence is consistent with the hypothesis that the Pomoideae evolved through a process of allopolyploidy from primitive members of the Spiracoideae and Prunoideae.     

Bioinformatic analysis of maize granule-bound starch synthase gene

Alignment of the Wx-gene and its homolog sequences has been conducted. A dendrogram displaying phylogenetic relationship between Poaceae family members has been built. Transfer of ancient Wx-gene from Zea mays to Dimeria lawsonii has been assumed. Primers for the exons 8–10 of polymorphic region have been designed. In silico PCR analysis has been conducted.     

Conformational basis of N-glycosylation of proteins: conformational analysis of Ac-Asn-Ala-Thr-NH2

The structural requirements of the Asn-X-Thr(Ser) sequence for the N-glycosylation of proteins has been traced to a local conformation acting as a signal for the enzymatic process. The conformational space of the smallest in vivoN-glycosylation substrate, Ac-Asn-Ala-Thr-NH₂, has been thoroughly explored using energy calculations. All the lowest energy conformers have been characterized as bended structures.     

Human nucleotide sequences related to the transforming gene of a murine sarcoma virus: studies with cloned viral and cellular DNAs

A recombinant plasmid, pI26, has been constructed by cloning into pBR322 a transforming gene of murine sarcoma virus (a Moloney strain, clone 124, MSV) synthesized by detergent-treated virions. From this plasmid a XbaI-HindIII fragment has been isolated which contains only mos-specific sequences. This mos-specific probe has been used for screening a human gene library cloned in bacteriophage λ Charon 4A. Of these, 19 clones have been isolated containing mos-related sequences. By physical mapping and molecular hybridization it has been shown that these sequences are neighboured by DNA regions related to Moloney murine leukemia virus. Recombinant phages have also been found containing human inserts related to MLV, not to the mos gene. The possible existence of murine-like endogenous retroviruses in the normal human genome, including that of a sarcoma type, is discussed. By Northern blotting, expression of the cellular c-mos gene has been detected in mouse liver treated with a hepatocarcinogen. The general significance of the suggested model for evaluating the relationship between chemical carcinogenesis and oncogene expression is discussed.     

The systematic status of the Italian wolfCanis lupus

In the past, the gray wolfCanis lupus Linnaeus, 1758, has been recognized in Italy as either the subspecieslupus oritalicus. It has also been postulated that this population has undergone introgression from the domestic dogCanis familiaris. In order to clarify these issues, multistatistical analyses were made of 10 skull measurements of 34 full grown male wolves from the Italian Peninsula, 91 other male Eurasian wolves, and 20 domestic dogs. The analyses, together with other morphological evidence and prior genetic research, support recognition of the Italian wolf as a separate subspecies,Canis lupus italicus. The same evidence indicates that the subspecies has not been affected through hybridization with the domestic dog.     

Centric diatom algae of volcanic Verkhneavachinsk lakes (Kamchatka)

Eleven representatives of diatom algae of the class Centrophyceae from the genera Aulacoseira (1), Conticribra (1), Cyclotella (5), Discostella (1), Stephanodiscus (2), and Thallasiosira (1) have been found in the phytoplankton of 5 volcanic lakes (Bakening volcano, Kamchatka). Stephanodiscus popovskayae, a species new for science, has been discovered in all the lakes, and a species new for the flora of Russia, Cyclotella gordonensis, has been found only in three lakes. The closest similarity in the composition of centric diatoms has been revealed between the lakes of a single water system: Lake Medvezh’e and Lake Verkhneavachinskoe.     

A Rapid Protocol of Crude RNA/DNA Extraction for RT-qPCR Detection and Quantification of 'Candidatus Phytoplasma prunorum'

Many efforts have been made to develop a rapid and sensitive method for phytoplasma and virus detection. Taking our cue from previous works, different rapid sample preparation methods have been tested and applied to Candidatus Phytoplasma prunorum (‘Ca. P. prunorum’) detection by RT-qPCR. A duplex RT-qPCR has been optimized using the crude sap as a template to simultaneously amplify a fragment of 16S rRNA of the pathogen and 18S rRNA of the host plant. The specific plant 18S rRNA internal control allows comparison and relative quantification of samples. A comparison between DNA and RNA contribution to qPCR detection is provided, showing higher contribution of the latter. The method presented here has been validated on more than a hundred samples of apricot, plum and peach trees. Since 2013, this method has been successfully applied to monitor ‘Ca. P. prunorum’ infections in field and nursery. A triplex RT-qPCR assay has also been optimized to simultaneously detect ‘Ca. P. prunorum’ and Plum pox virus (PPV) in Prunus.     

Molecular biotechnology of marine algae in China

Molecular biotechnology of marine algae is referred to as the biotechnology on the identification, modification, production and utilization of marine algal molecules. It involves not only the manipulation of macromolecules such as DNA, RNA and proteins, but also deals with low molecular weight compounds such as secondary metabolites. In the last decade, molecular systematic researches to investigate the relationship and to examine the evolutionary divergence among Chinese marine algae have been carried out by Chinese scientists. For example, RAPD has been widely used in several laboratories to elucidate genetic variations of the reds, such as Porphyra, Gracilaria, Grateloupia and the greens such as Ulva and Enteromorpha. Some important data have been obtained. The study on molecular genetic markers for strain improvement is now in progress. In 1990s, genetic engineering of economic seaweeds such as Laminaria, Undaria, Porphyra, Gracilaria and Grateloupia has been studied in China. For Laminaria japonica, the successfully cultivated kelp in China, a model transformation system has been set up based on the application of plant genetic techniques and knowledge of the algal life history. Progress has been made recently in incorporating a vaccine gene into kelp genome. Evidence has been provided showing the expression of gene products as detectable vaccines. In the present paper, the progress of molecular biotechnological studies of marine algae in China, especially researches on elucidating and manipulating nucleic acids of marine algae, are reviewed.     

A comparative study of the pumping activity of some fouling animals from the White Sea

The pumping activity of the bivalve Mytulus edulus, Hiatella arctica, and solitary ascidian Styela rustica from the White Sea has been studied. The dependence of the pumping activity on the mass and size of animals has been analyzed. It was shown that the mussel is the most active filter-feeder, that the least value of the pumping activity has been registered for H. arctica, and that S. rustica occupies an intermediate position according this index. The results obtained are examined in connection with the competitive relationship of the animals in the fouling community.     

The growth stimulating effect of 5-bromouracil and uracil on chlorococcal algae

The growth stimulating effect of 5-bromouracil and uracil on two strains of chlorococcal algae has been found in cell colonies grown from single cells, which were inoculated onto an agar medium. Analyses of the effect recorded in the cell cycles after treatment have revealed that the growth stimulating effect required four, or more, cell cycles to become evident. This has been proved by the number of autospores released from the treated cells and by the length of the lag phase after inoculation. The differences between the control and the treated population in some experiments with 5-bromouracil inChlorella and with uracil inScenedesmus obliquus have been visible by the unaided eye, whereas in some other experiments, they have been proved statistically. Growth stimulation has not been found only in a small number of experiments. The inhibition of growth induced by 5-bromouracil has been recorded in one experiment withScenedesmus quadricauda.     

Glucosinolates in seeds of Neslia paniculata

The glucosinolate fraction in seeds of Neslia paniculata (L.) Desv., subsp. thracica (Velen.) Bornmüller, (Cruciferae), has been isolated and subjected to enzymic hydrolysis. The isothiocyanates thus produced have been separated and identified as cheirolin (II), (R)-9-methylsulfinylnonyl (III), (R)-10-methylsulfinyldecyl (IV), and (R)-11-methylsulfinylundecyl isothiocyanate (V), the last of which has not been previously encountered as a natural compound.     

Antiviral potential of Bulgarian medicinal plants

Medicinal plants have been widely used to treat a variety of infectious and non-infectious diseases. Bulgarian flora includes 4,300 plant species, over 500 of which are rare or endemic to the country or the Balkan region. The aim of the present work is to summarize comprehensively the investigations on the antiviral activity of Bulgarian medicinal plants from the past three decades. The effect of different extracts derived from in vitro propagated plants has been examined as well. The phytochemical composition and its influence on specific steps of the viral life cycle have been discussed in this paper. The review includes the following families: Amaryllidaceae, Fabaceae, Geraniaceae, Lamiaceae, Onagraceae, Ranunculaceae, Rosaceae, Scrophulariaceae and Rhodophyta. Special attention has been paid to viruses as important human pathogens.     

Hydroperoxide isomerase: a new enzyme of lipid metabolism

An enzyme has been isolated from flaxseed (Linum usitatissimum) which utilizes the product of lipoxidase for its substrate. The enzyme, termed hydroperoxide isomerase, converts the conjugated diene hydroperoxide of linoleic acid to the corresponding monoenoic ketohydroxy fatty acid. The structure of the latter has been determined by ultraviolet, infrared, and nuclear magnetic resonance spectroscopy; periodate and permangate oxidation; gas chromatography; and thin layer chromatography. Hydroperoxide isomerase activity has also been demonstrated in crude extracts from barley (Hordeum vulgare), wheat germ (Triticum aestivum), mung beans (Phaseolus aureus), and corn (Zea mays) and from partially purified extracts of soybean (Glycine max).     

Increased Drought Tolerance through the Suppression of ESKMO1 Gene and Overexpression of CBF-Related Genes in Arabidopsis

Improved drought tolerance is always a highly desired trait for agricultural plants. Significantly increased drought tolerance in Arabidopsis thaliana (Columbia-0) has been achieved in our work through the suppression of ESKMO1 (ESK1) gene expression with small-interfering RNA (siRNA) and overexpression of CBF genes with constitutive gene expression. ESK1 has been identified as a gene linked to normal development of the plant vascular system, which is assumed directly related to plant drought response. By using siRNA that specifically targets ESK1, the gene expression has been reduced and drought tolerance of the plant has been enhanced dramatically in the work. However, the plant response to external abscisic acid application has not been changed. ICE1, CBF1, and CBF3 are genes involved in a well-characterized plant stress response pathway, overexpression of them in the plant has demonstrated capable to increase drought tolerance. By overexpression of these genes combining together with suppression of ESK1 gene, the significant increase of plant drought tolerance has been achieved in comparison to single gene manipulation, although the effect is not in an additive way. Accompanying the increase of drought tolerance via suppression of ESK1 gene expression, the negative effect has been observed in seeds yield of transgenic plants in normal watering conditions comparing with wide type plant.