Mycosporine glutamine and related mycosporines in the fungus Pyronema omphalodes

The structure of mycosporine glutamine, a new compound, has been established and its presence demonstrated in two fungi Pyronema omphalodes and Glomerella cingulata. Mycosporine glutamic acid has been isolated from Helvella leucomelaneae. Co-occurrence of normycosporine glutamine, mycosporine glutamine and glucosylmycosporine glutaminol has been demonstrated in the fungus P. omphalodes. A biosynthetic pathway is proposed. Mycosporines have been compared by HPLC.     

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.     

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.     

New diterpenes from Chamaecyparis pisifera

Three new diterpenes, pisiferin, 15-hydroxyferruginol, and O-methyl pisiferic acid have been isolated and the presence of 12-hydroxyabieta-8,11,13-trien-20-al has been confirmed in studies of the leaves of Chamaecyparis pisifera. The structure of pisiferin, a phenolic diterpene with a new carbon skeleton, has been determined by chemical degradation. The presence of diterpenes of the ferruginol type in the leaves of four varieties of C. pisifera has been examined.     

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.     

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.     

The medicinal fungus Cordyceps militaris: research and development

Ophiocordyceps sinensis (syn. Cordyceps sinensis) is a highly valued medicinal fungus. This entomopathogen has a limited distribution, has been overharvested in the wild, and its stromata have not been artificially cultivated. Another entomopathogenic fungus, Cordyceps militaris (commonly known as orange caterpillar fungus), has chemical capacities similar to those of O. sinensis, but unlike O. sinensis, its stromata can be easily cultivated. Consequently, C. militaris is being studied as an alternative to O. sinensis, and the large-scale production of stromata is receiving substantial attention. Significant research has been conducted on the genetic resources, nutritional and environmental requirements, mating behavior, and biochemical and pharmacological properties of C. militaris. The complete genome of C. militaris has recently been sequenced. This fungus has been the subject of many reviews, but few have focused on its biology. The current paper reviews the biological aspects of the fungus including host range, mating system, cytology and genetics, insect- and non-insect nutritional requirements, environmental influence on stroma development, and commercial development.     

Antimicrobial Agent-Associated Colitis and Diarrhea

Although antimicrobial agent-associated colitis has been recognized as a clinicopathologic entity for years, the cause of this disease has been determined only recently. Virtually all cases of pseudomembranous colitis and some cases of antimicrobial agent-associated nonspecific colitis or diarrhea have been shown to be caused by a toxin of Clostridium difficile. Methods for cultivating C difficile from feces and for detecting the toxin have been developed. Oral administration of vancomycin has proved to be effective for the treatment of C difficile-induced colitis, although isolated instances of relapse after treatment have been documented.The discovery of C difficile as a human intestinal pathogen has provided an explanation for some, but not all cases of antimicrobial agent-associated diarrhea. The epidemiology, pathogenesis and means of prevention of C difficile toxin-induced diarrhea remain to be determined.     

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.     

Predation on endemic Hawaiian land snails by the invasive snail Oxychilus alliarius

The native land snails of the Hawaiian Islands have suffered numerous extinctions and continue to decline rapidly. A leading cause of these extinctions has been the impact of non-native predators. Oxychilus alliarius is a European predatory snail that has been in Hawaii since at least 1937 and has become one of the most abundant invasive snails in the islands. It is a facultative carnivore and will also consume non-animal food. However, this species has been mostly ignored as a threat and the majority of conservation attention has been focused on Euglandina rosea, the Rosy Wolf Snail. Here we report predation by O. alliarius on two rare and endemic Hawaiian snail species: Kaala subrutila (Helicarionidae) and Pleuropoma cf. sandwichiensis (Helicinidae). The wide distribution of O. alliarius in Hawaii, its high abundance, and now compelling evidence that it preys on native (and probably endangered) snails, indicate that it could be a much greater threat than previously thought.     

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.     

(+)-catechin 3-rhamnoside from Erythroxylum novogranatense

A new flavanol glycoside has been isolated from stems of Erythroxylum novogranatense and its structure has been elucidated on the basis of MS and ¹H NMR spectroscopy and hydrolytic studies as (+)-catechin 3-O-α-L-rhamnopyranoside. On a similar basis of chemical and spectroscopic evidence, the presence of ombuin 3-O-rutinoside has been established. Furthermore, the occurrence of procyanidin biflavanoids has been demonstrated by the characterization of B₁ and B₃ as the first representatives of B-type proanthocyanidins in the genus Erythroxylum.     

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.     

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.     

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.     

Construction of a deletion strain and expression vector for the Escherichia coli NADH:ubiquinone oxidoreductase (Complex I)

Complex I of Escherichia coli is encoded by 13 consecutive genes, called the nuo operon. A chromosomal deletion of all nuo genes has been achieved by homologous recombination. A vector that encodes all of the nuo genes has been constructed, and it expresses a functional enzyme.     

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.     

Identification of a chlorosis-inducing toxin of Pseudomonas glycinea as coronatine

A toxin causing chlorosis in bean and soybean leaves has been isolated from liquid cultures of Pseudomonas glycinea, and purified. It has been identified as coronatine, a toxin produced also by Pseudomonas coronafaciens var. atropurpurea.     

Isolation and characterization of thioredoxin h from poplar xylem

Redox-dependent regulation based on disulphide/dithiol exchange reactions has been extensively studied in herbaceous plants, but up to now, there is no information concerning these systems in trees. Based on existing ESTs, a cDNA coding for a thioredoxin h has been isolated from a xylem poplar cDNA library. The nucleotidic sequence of poplar thioredoxin h displays significant homology to other thioredoxins h isolated from plants. It shows a variation in the active site with the sequence WCPPC instead of the more canonical WCGPC sequence found in most thioredoxins. The cDNA sequence has been introduced in an expression plasmid (pET3d) in order to express the corresponding recombinant polypeptide. The protein has been expressed to a high level and purified from Escherichia coli cells with a very high yield. Several of the physical and kinetic characteristics of this redox protein are described and found to be similar to other thioredoxin h. On the other hand, its stability to heat denaturation, is very different from those of other thioredoxins h characterized so far.     

The involucrin gene of the tree shrew: recent repeat additions and the relocation of cysteine codons

The coding region of the involucrin gene of Tupaia glis has been cloned and sequenced. It resembles the involucrin coding region of other non-anthropoid mammals in possessing a segment of related, short tandem repeats at a defined location, but in Tupaia, there has been recent serial duplication of a repeat into which a cysteine codon had earlier been introduced. As a result of the duplication, there is a total of as many as six cysteine codons in the segment of repeats, a number larger than for any other species yet examined. In Rattus there has been a comparable but independent addition of cysteine codons, and both Tupaia and Rattus have eliminated an otherwise conserved cysteine codon 75 located close to but outside the segment of repeats. In Tupaia, this elimination probably occurred by gene conversion. Also independently, the gene of Canis has added cysteine codons to the segment of repeats but has not yet lost cysteine 75. It is proposed that the gain and the loss of cysteine codons are parts of a multi-stage program of cysteine relocation.