Cladogenesis and endemism in Tanzanian mole‐rats,genus Fukomys: (Rodentia Bathyergidae): a role for tectonics?

African mole‐rats of the family Bathyergidae are subterranean hystricomorph rodents found throughout sub‐Saharan Africa, where the distributional ranges of the most speciose taxa are divided by the African Rift Valley. In particular, mole‐rats of the genera Heliophobius and Fukomys are distributed widely, and their adaptive radiation appears to have been strongly influenced by the geological process of rifting. As a result, virtually all members of the genus Fukomys occur in locations west of the Rift Valley. However, a small number of isolated populations occur east of the Rift Valley in Tanzania, where Heliophobius is widespread and is the predominant bathyergid rodent. Phylogenetic analysis of mitochondrial cytochrome b sequences of previously unstudied Tanzanian mole‐rats (genus Fukomys) and geographically adjacent populations strongly suggests that vicariance in the Western Rift Valley has subdivided populations of mole‐rats and, together with climatic changes, played a role in the isolation of extralimital populations of Fukomys in Tanzania. Together with molecular clock‐based estimates of divergence times, these results offer strong support for the hypothesis that the observed patterns of cladogenesis are consistent with tectonic activity in the ‘Mbeya triple junction’ and Rungwe volcanic province between Lakes Rukwa and Nyasa. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 100 , 337–352.     

Prevalence of white spot syndrome virus (WSSV) in wild shrimp Penaeus monodon in the Philippines

Prevalence of white spot syndrome virus (WSSV) was determined using polymerase chain reaction (PCR) methodology on DNA extracted from the gills of wild black tiger shrimp Penaeus monodon collected from 7 sampling sites in the Philippines. These 7 sampling sites are the primary sources of spawners and broodstock for hatchery use. During the dry season, WSSV was detected in shrimp from all sites except Bohol, but during the wet season it was not detected in any site except Palawan. None of the WSSV-PCR positive shrimp showed signs of white spots in the cuticle. Prevalence of WSSV showed seasonal variations, i.e. prevalence in dry season (April to May) was higher than in the wet season (August to October). These results suggest that WSSV has already become established in the local marine environment and in wild populations of P. monodon. Thus, broodstock collected during the dry season could serve as the main source of WSSV contamination in shrimp farms due to vertical transmission of the virus in hatcheries.     

Paraoxonases are associated with intestinal inflammatory diseases and intracellularly localized to the endoplasmic reticulum

We demonstrated previously that the paraoxonase (PON1/2/3) genes and proteins are expressed in human intestinal biopsies and in Caco-2 cells. The current study aims were to explore whether PON1/2/3 expression is different in inflammatory bowel diseases (IBD) or celiac disease compared to healthy controls, and to explore the intracellular localization of PON1/2/3. Our results showed that significantly fewer biopsies expressed PON1 and PON3 in the duodenum of celiac patients (PON1, P<0.0001; PON3, P=0.03), in the terminal ileum of Crohn's patients (PON1, P=0.001; PON3, P=0.008), and in the colon of UC patients (PON1, P=0.02; PON3, P=0.06) compared to controls. Since all three disorders share markedly elevated inflammatory mediators we explored the PON1/2/3 mRNA expression on cytokine stimulation. No changes were observed in Caco-2 and HT29 cells. Immunofluorescence experiments localized PON1/2/3 exclusively to the endoplasmic reticulum (ER) in both CaCo-2 and HT29 cells. These results demonstrate for the first time a novel relationship between PON1 and PON3 expression and several inflammatory gastrointestinal disorders. Together with the localization of PON1/2/3 enzymes to the ER, it may be suggested that PON1/2/3 may have extracellular functions as part of the host response in IBD and celiac disease.     

Adaptive divergence for a fitness-related trait among invasive Ambrosia artemisiifolia populations in France

The impact of natural selection on the adaptive divergence of invasive populations can be assessed by testing the null hypothesis that the extent of quantitative genetic differentiation (Q(ST) ) would be similar to that of neutral molecular differentiation (F(ST) ). Using eight microsatellite loci and a common garden approach, we compared Q(ST) and F(ST) among ten populations of an invasive species Ambrosia artemisiifolia (common ragweed) in France. In a common garden study with varying water and nutrient levels, we measured Q(ST) for five traits (height, total biomass, reproductive allocation, above- to belowground biomass ratio, and days to flowering). Although low F(ST) indicated weak genetic structure and strong gene flow among populations, we found significant diversifying selection (Q(ST) > F(ST) ) for reproductive allocation that may be closely related to fitness. It suggests that abiotic conditions may have exerted selection pressure on A. artemisiifolia populations to differentiate adaptively, such that populations at higher altitude or latitude evolved greater reproductive allocation. As previous studies indicate multiple introductions from various source populations of A. artemisiifolia in North America, our results suggest that the admixture of introduced populations may have increased genetic diversity and additive genetic variance, and in turn, promoted the rapid evolution and adaptation of this invasive species.     

葡寡糖对LPS诱导小鼠氧化应激与炎症反应调节作用的研究

本文研究了低、中、高三种不同聚合度葡寡糖(LGOS、MGOS、HGOS)对小鼠氧化应激与炎症反应的调节,并探讨其可能的作用机制。在采食正常日粮基础上,各组小鼠每日分别灌胃生理盐水、低聚果糖(FOS)、LGOS、MGOS、HGOS。21 d后,腹腔注射脂多糖(LPS)诱导小鼠建立氧化应激与炎症反应模型,分别测定LPS刺激后4 h和18 h时血清和肝脏中活性氧(reactive oxygen species,ROS)水平、总抗氧化能力(total antioxidant ca-pacity,T-AOC)、过氧化氢酶(catalase,CAT)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)活性、丙二醛(Malondialdehyde,MDA)含量、肝脏中一氧化氮(NO)含量、一氧化氮合酶(nitric oxide synthase,NOS)活性、血清中白细胞介素1(Interleukin-1,IL-1)和肿瘤坏死因子(tumor necrosis factor,TNF-a)含量。结果表明:三种聚合度葡寡糖均能显著降低LPS刺激小鼠血清、肝脏中ROS水平(P<0.05),降低肝脏中NO含量、T-NOS、iNOS活性、血清中炎性细胞因子(IL-1、TNF-a)含量(P<0.05),显著提高总抗氧化能力(T-AOC)和抗氧化酶(CAT、GSH-Px)活性(P<0.05)。葡寡糖具有保护机体免受氧化损伤,减缓炎症反应的作用,并随着平均聚合度(degree ofpolymerization,DP)的增加,其调节功能逐渐增强。     

微生物絮凝剂在养殖废水处理中的应用

微生物絮凝剂作为一种新型的絮凝剂,因其安全、高效等特性,正逐渐成为目前水产养殖废水处理研究的热点。主要从微生物絮凝剂的概念、絮凝机理、特点、研究现状、应用实例等方面,分析了微生物絮凝剂作为水质改良剂在水产养殖中的应用前景,并就今后的研究趋势作了展望。     

The Botrytis cinerea phytotoxin botcinic acid requires two polyketide synthases for production and has a redundant role in virulence with botrydial

The grey mould fungus Botrytis cinerea produces two major phytotoxins, the sesquiterpene botrydial, for which the biosynthesis gene cluster has been characterized previously, and the polyketide botcinic acid. We have identified two polyketide synthase (PKS) encoding genes, BcPKS6 and BcPKS9, that are up-regulated during tomato leaf infection. Gene inactivation and analysis of the secondary metabolite spectra of several independent mutants demonstrated that both BcPKS6 and BcPKS9 are key enzymes for botcinic acid biosynthesis. We showed that BcPKS6 and BcPKS9 genes, renamed BcBOA6 and BcBO9 (for B. cinerea botcinic acid biosynthesis), are located at different genomic loci, each being adjacent to other putative botcinic acid biosynthetic genes, named BcBOA1 to BcBOA17. Putative orthologues of BcBOA genes are present in the closely related fungus Sclerotinia sclerotiorum, but the cluster organization is not conserved between the two species. As for the botrydial biosynthesis genes, the expression of BcBOA genes is co-regulated by the Gα subunit BCG1 during both in vitro and in planta growth. The loss of botcinic acid production does not affect virulence on bean and tomato leaves. However, double mutants that do not produce botcinic acid or botrydial (bcpks6Δbcbot2Δ) exhibit markedly reduced virulence. Hence, a redundant role of botrydial and botcinic acid in the virulence of B. cinerea has been demonstrated.     

MicroRNA‐155 prevents necrotic cell death in human cardiomyocyte progenitor cells via targeting RIP1

To improve regeneration of the injured myocardium, cardiomyocyte progenitor cells (CMPCs) have been put forward as a potential cell source for transplantation therapy. Although cell transplantation therapy displayed promising results, many issues need to be addressed before fully appreciating their impact. One of the hurdles is poor graft‐cell survival upon injection, thereby limiting potential beneficial effects. Here, we attempt to improve CMPCs survival by increasing microRNA‐155 (miR‐155) levels, potentially to improve engraftment upon transplantation. Using quantitative PCR, we observed a 4‐fold increase of miR‐155 when CMPCs were exposed to hydrogen‐peroxide stimulation. Flow cytometric analysis of cell viability, apoptosis and necrosis showed that necrosis is the main cause of cell death. Overexpressing miR‐155 in CMPCs revealed that miR‐155 attenuated necrotic cell death by 40 ± 2.3%via targeting receptor interacting protein 1 (RIP1). In addition, inhibiting RIP1, either by pre‐incubating the cells with a RIP1 specific inhibitor, Necrostatin‐1 or siRNA mediated knockdown, reduced necrosis by 38 ± 2.5% and 33 ± 1.9%, respectively. Interestingly, analysing gene expression using a PCR‐array showed that increased miR‐155 levels did not change cell survival and apoptotic related gene expression. By targeting RIP1, miR‐155 repressed necrotic cell death of CMPCs, independent of activation of Akt pro‐survival pathway. MiR‐155 provides the opportunity to block necrosis, a conventionally thought non‐regulated process, and might be a potential novel approach to improve cell engraftment for cell therapy.     

Single nucleotide polymorphism genotyping by heteroduplex analysis in sunflower (<Emphasis Type="Italic">Helianthus annuus</Emphasis> L.)

Single nucleotide polymorphisms (SNPs) and insertions/deletions (indels) are increasingly used for cultivar identification, construction of genetic maps, genetic diversity assessment, association mapping and marker-assisted breeding. Although there are several highly sensitive methods for the detection of polymorphisms, most of them are often beyond the budget of medium-throughput academic laboratories or seed companies. Heteroduplex analysis by enzymatic cleavage (CEL1CH) or denaturing high-performance liquid chromatography (dHPLC) has been successfully used to examine genetic variation in several plant and animal species. In this work, we assess and compare the performance of both methods in sunflower by genotyping SNPs from a set of 24 selected polymorphic candidate genes. The CEL1CH method allowed us to accurately detect allele differences in 10 out of 24 regions using an in-house prepared CEL1 enzyme (celery single strand endonuclease 1, Apium graveolens L.). Similarly, a total of 11 regions were successfully optimized for dHPLC analysis. As a scaling-up approach, both strategies were tested to genotype either 42 SNPs/indels in 22 sunflower accessions from the local germplasm bank or 33 SNPs/indels in 90 recombinant inbred lines (RILs) for genetic mapping purposes. Summarizing, a total of 601 genotypes were efficiently analyzed either with CEL1CH (110) or dHPCL (491). In conclusion, CEL1CH and dHPLC proved to be robust, complementary methods, allowing medium-scale laboratories to scale up the number of both SNPs and individuals to be included in genetic studies and targeted germplasm diversity characterization (EcoTILLING).