Bacterial Endophytic Communities in the Grapevine Depend on Pest Management

Microbial plant endophytes are receiving ever-increasing attention as a result of compelling evidence regarding functional interaction with the host plant. Microbial communities in plants were recently reported to be influenced by numerous environmental and anthropogenic factors, including soil and pest management. In this study we used automated ribosomal intergenic spacer analysis (ARISA) fingerprinting and pyrosequencing of 16S rDNA to assess the effect of organic production and integrated pest management (IPM) on bacterial endophytic communities in two widespread grapevines cultivars (Merlot and Chardonnay). High levels of the dominant Ralstonia, Burkholderia and Pseudomonas genera were detected in all the samples We found differences in the composition of endophytic communities in grapevines cultivated using organic production and IPM. Operational taxonomic units (OTUs) assigned to the Mesorhizobium, Caulobacter and Staphylococcus genera were relatively more abundant in plants from organic vineyards, while Ralstonia, Burkholderia and Stenotrophomonas were more abundant in grapevines from IPM vineyards. Minor differences in bacterial endophytic communities were also found in the grapevines of the two cultivars.     

Molecular characterization of SLC24A5 variants and evaluation of Nitisinone treatment efficacy in a zebrafish model of OCA6

Skin pigmentation is a highly heterogeneous trait with diverse consequences worldwide. SLC24A5, encoding a potent K⁺‐dependent Na⁺/Ca²⁺ exchanger, is among the known color‐coding genes that participate in melanogenesis by maintaining pH in melanosomes. Deficient SLC24A5 activity results in oculocutaneous albinism (OCA) type 6 in humans. In this study, by utilizing a exome sequencing (ES) approach, we identified two new variants [p. (Gly110Arg) and p. (IIe189Ilefs*1)] of SLC24A5 cosegregating with the OCA phenotype, including nystagmus, strabismus, foveal hypoplasia, albinotic fundus, and vision impairment, in three large consanguineous Pakistani families. Both of these variants failed to rescue the pigmentation in zebrafish slc24a5 morphants, confirming the pathogenic effects of the variants. We also phenotypically characterized a commercially available zebrafish mutant line (slc24a5^ko) that harbors a nonsense (p.Tyr208*) allele of slc24a5. Similar to morphants, homozygous slc24a5^ko mutants had significantly reduced melanin content and pigmentation. Next, we used these slc24a5^ko zebrafish mutants to test the efficacy of nitisinone, a compound known to increase ocular and fur pigmentation in OCA1 (TYR) mutant mice. Treatment of slc24a5^ko mutant zebrafish embryos with varying doses of nitisinone did not improve melanin production and pigmentation, suggesting that treatment with nitisinone is unlikely to be therapeutic in OCA6 patients.     

综述——新型纳米生物材料的研发：磁性纳米材料：化学合成、功能化与生物医学应用

磁性纳米材料,由于其独特的磁学性能、小尺寸效应,被广泛应用于生物医学领域．本文总结了磁性纳米材料的化学设计与合成、表面功能化方法,及其在核磁共振成像、磁控治疗、磁热疗和生物分离等生物医学领域的应用进展．     

Functional Null Mutations of MSRB3 Encoding Methionine Sulfoxide Reductase Are Associated with Human Deafness DFNB74

The DFNB74 locus for autosomal-recessive, nonsyndromic deafness segregating in three families was previously mapped to a 5.36 Mb interval on chromosome 12q14.2-q15. Subsequently, we ascertained five additional consanguineous families in which deafness segregated with markers at this locus and refined the critical interval to 2.31 Mb. We then sequenced the protein-coding exons of 18 genes in this interval. The affected individuals of six apparently unrelated families were homozygous for the same transversion (c.265T>G) in MSRB3, which encodes a zinc-containing methionine sulfoxide reductase B3. c.265T>G results in a substitution of glycine for cysteine (p.Cys89Gly), and this substitution cosegregates with deafness in the six DFNB74 families. This cysteine residue of MSRB3 is conserved in orthologs from yeast to humans and is involved in binding structural zinc. In vitro, p.Cys89Gly abolished zinc binding and MSRB3 enzymatic activity, indicating that p.Cys89Gly is a loss-of-function allele. The affected individuals in two other families were homozygous for a transition mutation (c.55T>C), which results in a nonsense mutation (p.Arg19X) in alternatively spliced exon 3, encoding a mitochondrial localization signal. This finding suggests that DFNB74 deafness is due to a mitochondrial dysfunction. In a cohort of 1,040 individuals (aged 53–67 years) of European ancestry, we found no association between 17 tagSNPs for MSRB3 and age-related hearing loss. Mouse Msrb3 is expressed widely. In the inner ear, it is found in the sensory epithelium of the organ of Corti and vestibular end organs as well as in cells of the spiral ganglion. Taken together, MSRB3-catalyzed reduction of methionine sulfoxides to methionine is essential for hearing.     

Resistance to Chilo infuscatellus (Lepidoptera: Pyraloidea) in transgenic lines of sugarcane expressing Bacillus thuringiensis derived Vip3A protein

Molecular Biology Reports - Sustainable agriculture requires management of insect pests through resistance development. The biological potential of Cry toxins and Vip protein, derived from Bacillus...     

Cardiac pathological changes of Atlantic salmon (Salmo salar L.) affected with heart and skeletal muscle inflammation (HSMI)

Heart and skeletal muscle inflammation (HSMI) is a disease of marine farmed Atlantic salmon where the pathological changes associated with the disease involve necrosis and an infiltration of inflammatory cells into different regions of the heart and skeletal muscle. The aim of this work was to characterize cardiac changes and inflammatory cell types associated with a clinical HSMI outbreak in Atlantic salmon using immunohistochemistry. Different immune cells and cardiac tissue responses associated with the disease were identified using different markers. The spectrum of inflammatory cells associated with the cardiac pathology consisted of mainly CD3(+) T lymphocytes, moderate numbers of macrophages and eosinophilic granulocytes. Proliferative cell nuclear antigen (PCNA) immuno-reaction identified significantly increased nuclear and cytoplasmic staining as well as identifying hypertrophic nuclei. Strong immunostaining was observed for major histocompatibility complex (MHC) class II in HSMI hearts. Although low in number, a few positive cells in diseased hearts were detected using the mature myeloid cell line granulocytes/monocytes antibody indicating more positive cells in diseased than non-diseased hearts. The recombinant tumor necrosis factor-α (TNFα) antibody identified stained macrophage-like cells and endothelial cells around lesions in addition to eosinophilic granular cells (EGCs). These findings suggested that the inflammatory response in diseased hearts comprised of mostly CD3(+) T lymphocytes and eosinophilic granular cells and hearts exhibited high cell turnover where DNA damage/repair might be the case (as identified by PCNA, caspase 3 and terminal deoxynucleotidyl transferase nick-end labeling (TUNEL) reactivity).     

A new and improved method for 3′-end labelling DNA using [α-32P]ddATP

We have synthesised dideoxyadenosine-5′-[α-³²P]triphosphate ([α-³²P]ddATP) at a specific activity of 3000 Ci/mmol and directly compared it with cordycepin-5′-[α-³²P]triphosphate ([α-³²P]KTP) as a means to 3′-end label DNA. The [α-³²P]ddATP was found to be three to five times more efficient than [α-³²P]KTP. Blunt and 3′-protruding ends were labelled more efficiently with [α-³²P]ddATP using terminal transferase than were the 5′-ends with [γ-³²P]ATP using polynucleotide kinase by standard methods. This improvement in efficiency of labelling DNA and the simplicity of the method allows 3′-end labelling of DNA to become a realistic alternative to 5′-end labelling. We have also compared [α-³²P]ddATP- and [α-³²P]KTP-labelled DNA in Maxam and Gilbert sequencing procedures and find that both give equally good results.     

AN ECOLOGICAL REVIEW OF CLADOPHORA GLOMERATA (CHLOROPHYTA) IN THE LAURENTIAN GREAT LAKES1

Cladophora glomerata (L.) Kütz. is, potentially, the most widely distributed macroalga throughout the world’s freshwater ecosystems. C. glomerata has been described throughout North America, Europe, the Atlantic Islands, the Caribbean Islands, Asia, Africa, Australia and New Zealand, and the Pacific Islands. Cladophora blooms were a common feature of the lower North American Great Lakes (Erie, Michigan, Ontario) from the 1950s through the early 1980s and were largely eradicated through the implementation of a multibillion‐dollar phosphorus (P) abatement program. The return of widespread blooms in these lakes since the mid‐1990s, however, was not associated with increases in P loading. Instead, current evidence indicates that the resurgence in blooms was directly related to ecosystem level changes in substratum availability, water clarity, and P recycling associated with the establishment of dense colonies of invasive dreissenid mussels. These results support the hypothesis that dreissenid mussel invasions may induce dramatic shifts in energy and nutrient flow from pelagic zones to the benthic zone.     

USE OF PULSE‐AMPLITUDE‐MODULATED FLUORESCENCE TO ASSESS THE PHYSIOLOGICAL STATUS OF CLADOPHORA SP. ALONG A WATER QUALITY GRADIENT1

This study investigated the application of pulse‐amplitude‐modulated (PAM) fluorometry as a rapid assessment of benthic macroalgal physiological status. Maximum quantum efficiency (F_v/F_m), dark–light induction curves, and rapid fluorescence light‐response curves (RLC) were measured on the filamentous macroalgal Cladophora sp. from Lake Ontario on 5 d at 16 sites spanning a gradient of light and nutrient supply. For Cladophora sp. growing in situ, light limitation was assessed by comparing average daily irradiance with the light utilization efficiency parameter (α) derived from RLCs. In this study, there was a nonlinear relationship between F_v/F_m and the degree of P limitation in macroalgae. However, only light‐saturated Cladophora sp. showed a significant positive linear relationship between F_v/F_m and P nutrient status. The absence of this relationship among light‐limited algae indicates that their photosynthetic rate would be stimulated by increased water clarity, and not by increased P supply. PAM fluorescence measures were successfully able to identify light‐saturated macroalgae and, among these, assess the degree to which they were nutrient limited. These results enable us to test hypotheses arising from numeric models predicting the impact of changes in light penetration and nutrient supply on benthic primary production.     

Biocatalytic and semisynthetic optimization of the anti-invasive tobacco (1S,2E,4R,6R,7E,11E)-2,7,11-cembratriene-4,6-diol

Tobacco cembranoids were reported to inhibit tumorigenesis. Biocatalysis of (1S,2E,4R,6R,7E,11E)-2,7,11-cembratriene-4,6-diol (1) using the symbiotic Bacillus sp. NC5, Bacillus sp. NK8, and Bacillus sp. NK7, isolated from the Red Sea sponge Negombata magnifica, afforded two new and four known hydroxylated metabolites 3-8. The use of symbiotic marine bacteria as biocatalysts for bioactive natural product scaffolds is very rare. Cembranoid 1 carbamate analogs 9-11 were prepared by its reaction with corresponding isocyanates. Cembranoid 1 and its bioconversion and carabamate products show anti-invasive activity against the human highly metastatic prostate PC-3M cancer cell line at 10-50 nM doses in Matrigel assay.