Deoxymikanolide adversely altered physiology and ultrastructure of Ralstonia solanacearum.

Deoxymikanolide (DEO) was isolated from Mikania micrantha Bunge and identified as a novel antibacterial compound previously. However, the mode of antimicrobial mechanism of DEO was not clear but hypothesized to affect the morphology and physiology of Ralstonia solanacearum cells. In this study, we confirmed our hypothesis via transmission electron microscopy (TEM) observation and comprehensive physiological analyses, including electric conductivity, glycan and phosphorus metabolism, activities of antioxidant enzymes (catalase, peroxidase, and superoxide dismutase), intrabacterial reactive oxygen species (ROS), and malondialdehyde (MDA) levels. We found that glycan and phosphorus metabolism, electric conductivity, intracellular ROS and MDA levels of R. solanacearum cells were significantly increased, while the activities of three antioxidant enzymes were significantly inhibited by DEO treatment. Moreover, TEM analysis showed that DEO treatment led to an early-stage of cell shrinkage, intermediate-stages of cytoplasmic damage, and a final-stage of cell disruption. Altogether, our data presented here indicate that DEO could adversely affect the physiology and morphology of R. solanacearum cells and be treated as an alternative antibacterial treatment in the future.

One new flavonoid from Solanum rostratum.

One new flavonoid, 5,7,8,4'-tetrahydroxy-3-methoxyflavone-8-O-ß-d-xylopyranoside (2), along with other four known flavones (1, 3-5) were isolated from the aerial parts of Solanum rostratum. 8-hydroxyflavonoid was isolated from series Androceras for the first time. The structure of the new compound 2 was determined on the basis of spectroscopic techniques, including IR, NMR and HRESI-MS. The chemotaxonomic significance of these compounds was summarised.

Genetic variation in the invasive weed Mikania micrantha (Asteraceae) suggests highways as corridors for its dispersal in southern China.

BACKGROUND AND AIMS: Roads as corridors of seed or fruit spatial dispersal have major impacts on the establishment and spread of invasive species, but their precise role in population genetic variation remains poorly understood. The South American weed Mikania micrantha has spread rapidly across southern China since its introduction to the Shenzhen area in 1984. This study investigated how its genetic diversity is distributed along highways, and whether highways have acted as corridors for the rapid expansion of M. micrantha METHODS: Twenty-seven roadside populations were sampled along four highways in southern China, and 787 samples were examined using 12 microsatellite markers. Variation in genetic diversity among populations was quantified and patterns of genetic differentiation were analysed. KEY RESULTS: A high level of genetic diversity was found at both the species and the population levels in this self-incompatible plant (expected heterozygosity = 0·497 and 0·477, respectively; allelic richness = 2·580 and 2·521, respectively). The Wright F-statistic value among populations (0·044, P < 0·01) and the analysis of molecular variance (91 % of genetic variation residing within populations, 9 % among populations within highways and 0 % among the four highways) showed a relatively low level of genetic differentiation among populations, while the principal coordinate and cluster analyses also indicated a lack of clear geographical genetic structure among populations. The calculated Nm value of 5·5 signifies strong gene flow. CONCLUSIONS: The pattern of genetic variation is consistent with facilitated dispersal along highways. The genetic admixtures among the roadside populations imply the occurrence of multiple population introductions during colonization. The long-distance dispersal of seeds associated with vehicular transportation on highways may have played important roles in shaping the genetic variation. This finding highlights the importance of highways as corridors for the spread of M. micrantha in southern China.

Recombinant Passenger Proteins Can Be Conveniently Purified by One-Step Affinity Chromatography.

Fusion tag is one of the best available tools to date for enhancement of the solubility or improvement of the expression level of recombinant proteins in Escherichia coli. Typically, two consecutive affinity purification steps are often necessitated for the purification of passenger proteins. As a fusion tag, acyl carrier protein (ACP) could greatly increase the soluble expression level of Glucokinase (GlcK), α-Amylase (Amy) and GFP. When fusion protein ACP-G2-GlcK-Histag and ACP-G2-Amy-Histag, in which a protease TEV recognition site was inserted between the fusion tag and passenger protein, were coexpressed with protease TEV respectively in E. coli, the efficient intracellular processing of fusion proteins was achieved. The resulting passenger protein GlcK-Histag and Amy-Histag accumulated predominantly in a soluble form, and could be conveniently purified by one-step Ni-chelating chromatography. However, the fusion protein ACP-GFP-Histag was processed incompletely by the protease TEV coexpressed in vivo, and a large portion of the resulting target protein GFP-Histag aggregated in insoluble form, indicating that the intracellular processing may affect the solubility of cleaved passenger protein. In this context, the soluble fusion protein ACP-GFP-Histag, contained in the supernatant of E. coli cell lysate, was directly subjected to cleavage in vitro by mixing it with the clarified cell lysate of E. coli overexpressing protease TEV. Consequently, the resulting target protein GFP-Histag could accumulate predominantly in a soluble form, and be purified conveniently by one-step Ni-chelating chromatography. The approaches presented here greatly simplify the purification process of passenger proteins, and eliminate the use of large amounts of pure site-specific proteases.

Synthesis of bifunctional molecules containing [12]aneN3 and coumarin moieties as effective DNA condensation agents and new non-viral gene vectors.

A series of bifunctional molecules with different combinations of macrocyclic polyamine [12]aneN3 and coumarin moieties, 4a/b and 5a/b, were synthesized by a two-step copper(I)-mediated alkyne­azide click reactions between 1,3,5-tris(azidomethyl)benzene and Boc-protected N-propynyl-[12]aneN3/7-propynyloxycoumarins. Agarose gel electrophoresis experiments indicated that bifunctional molecules 4b and 5b effectively induced complete plasmid DNA condensation at concentrations up to 40 µM. It was found that the structural variation had a major impact on the condensation behavior of these compounds. The electrostatic interaction involving the [12]aneN3 moiety can be compensated by the binding contribution of the coumarin units during the DNA condensation process. These two types of interaction showed different effects on the reversibility of DNA condensation. Results from studies using dynamic laser scattering, atomic force microscopy, and EB replacement assay further supported the above conclusion. Cytotoxicity assays on bifunctional compounds 4a/b and 5a/b indicated their low cytotoxicity. Results from cellular uptake and cell transfection experiments proved that bifunctional compounds 4b and 5b successfully served as non-viral gene vectors. Furthermore, methyl substituents attached to the coumarin unit (4b and 5b) greatly enhanced their DNA condensation capability and gene transfection. These bifunctional molecules, with the advantages of lower cytotoxicity, good water solubility, and potential structural modification, will have great potential for the development of new non-viral gene delivery agents.

Antimicrobial constituents of the leaves of Mikania micrantha H. B. K.

BACKGROUND: To isolate plant-derived compounds with antimicrobial activity from the leaves of Mikania micrantha, to determine the compounds configuration, and to evaluate their antimicrobial activity against eight plant pathogenic fungi (Exserohilum turcicum, Colletotrichum lagenarium, Pseudoperonispora cubensis, Botrytis cirerea, Rhizoctonia solani, Phytophthora parasitica, Fusarium solani, and Pythium aphanidermatum,) and four plant pathogenic bacteria (gram negative bacteria: Ralstonia dolaanacearum, Xanthomonas oryzae pv. Oryzae, Xanthomonas Campestris pv. Vesicatoria, and Xanthomonas campestris pv. Citri), and four bacteria (gram positive bacteria: Staphyloccocus aureus, Bacillus subtilis, Micrococcus luteus, and Bacillus cereus). METHODS AND RESULTS: Antimicrobial constituents of the leaves of M. micrantha were isolated using bioactivity- guided fractionation. The antifungal activity of the isolated compounds was evaluated by the inhibit hypha growth method and inhibit spore germination method. Characterization of antibacterial activity was carried out using the minimum inhibitory concentrations (MICs) and the minimum bactericidal concentrations (MBCs). MIC and MBC were determined by the broth microdilution method. Six compounds - deoxymikanolide, scandenolide, dihydroscandenolide, mikanolide, dihydromikanolide, and m - methoxy benzoic acid - have been isolated from leaves of Mikania micrantha H. B. K. Deoxymikanolide, scandenolide, and dihydroscandenolide were new compounds. The result of bioassay showed that all of isolated compounds were effective against tested strains and deoxymikanolide showed the strongest activity. CONCLUSIONS AND SIGNIFICANCE: The leaves of M. micrantha may be a promising source in the search for new antimicrobial drugs due to its efficacy and the broadest range. Meanwhile, adverse impact of M. micrantha will be eliminated.

Evaluation of the combination of 1,3-dichloropropene and dazomet as an efficient alternative to methyl bromide for cucumber production in China.

BACKGROUND: The combination of 1,3-dichloropropene (1,3-D) and dazomet (DZ) offers a potential alternative to methyl bromide (MB) for soil disinfection. MB is scheduled to be withdrawn from routine use by 2015 in developing countries. Combination treatments of 1,3-D + DZ were evaluated in a laboratory study and in two commercial cucumber fields. RESULTS: Laboratory studies found that nearly all of the tested combinations of 1,3-D and DZ displayed positive synergistic activity on root-knot nematodes (Meloidogyne spp.), two major soilborne fungi (Fusarium spp. and Phytophthora spp.) and the seeds of two major weed species (Digitaria sanguinalis and Abutilon theophrasti). Field trials revealed that the combination of 1,3-D and DZ (at 10 + 25 g m(-2) ) successfully suppressed Meloidogyne spp. root galling, sharply reduced Fusarium spp. and Phytophthora spp. and maintained high cucumber yields. The combination treatment of 1,3-D + DZ was more effective than 1,3-D or DZ alone and provided results similar to methyl bromide with respect to pest control, plant mortality, plant height, yield and income. All of the treatments were significantly better than the non-treated control. CONCLUSION: The results indicate that the tested combination of 1,3-D and DZ offers an efficient alternative to methyl bromide for cucumber production.

4-(4-Chloro-5-methyl-3-trifluoro-meth-yl-1H-pyrazol-1-yl)-6-(prop-2-ynyl-oxy)pyrimidine.

The molecule of the title compound, C(12)H(8)ClF(3)N(4)O, is twisted as indicated by the C-O-C-C torsion angle of 76.9 (3)°. Moreover, the trifluoro-methyl group shows rotational disorder of the F atoms, with site-occupancy factors of 0.653 (6) and 0.347 (6). The dihedral angle between the rings is 1.88 (12) Å.