Cadmium stress in wheat seedlings: growth,cadmium accumulation and photosynthesis

Seedlings of wheat (Triticum aestivum L.) cultivars Jing 411, Jinmai 30 and Yangmai 10 were exposed to 0, 10, 20, 30, 40 or 50 μM of CdCl₂ in a solution culture experiment. The effects of cadmium (Cd) stress on wheat growth, leaf photon energy conversion, gas exchange, and Cd accumulation in wheat seedlings were investigated. Gas exchange was monitored at 3, 9, 24 days after treatment (DAT). Growth parameters, chlorophyll content, leaf chlorophyll fluorescence, and Cd concentration in shoot and root were measured at 24 DAT. Seedling growth, gas exchange, chlorophyll content, chlorophyll fluorescence parameters were generally depressed by Cd stress, especially under the high Cd concentrations. Cd concentration and accumulation in both shoots and roots increased with increasing external Cd concentrations. Relationships between corrected parameters of growth, photosynthesis and fluorescence and corrected Cd concentrations in shoots and roots could be explained by the regression model Y = K/(1 + exp(a + bX)). Jing 411 was found to be Cd tolerant considering parameters of chlorophyll content, photosynthesis and chlorophyll fluorescence in which less Cd translocation was from roots into shoots. The high Cd concentrations were in shoots and roots in Yangmai 10 which has been found to be a relative Cd tolerant cultivar in terms of most growth parameters.     

HSPC117 deficiency in cloned embryos causes placental abnormality and fetal death

Somatic cell nuclear transfer (SCNT) has been successfully used in many species to produce live cloned offspring, albeit with low efficiency. The low frequency of successful development has usually been ascribed to incomplete or inappropriate reprogramming of the transferred nuclear genome. Elucidating the genetic differences between normal fertilized and cloned embryos is key to understand the low efficiency of SCNT. Here, we show that expression of HSPC117, which encodes a hypothetical protein of unknown function, was absent or very low in cloned mouse blastocysts. To investigate the role of HSPC117 in embryo development, we knocked-down this gene in normal fertilized embryos using RNA interference. We assessed the post-implantation survival of HSPC117 knock-down embryos at 3 stages: E9 (prior to placenta formation); E12 (after the placenta was fully functional) and E19 (post-natal). Our results show that, although siRNA-treated in vivo fertilized/produced (IVP) embryos could develop to the blastocyst stage and implanted without any difference from control embryos, the knock-down embryos showed substantial fetal death, accompanied by placental blood clotting, at E12. Furthermore, comparison of HSPC117 expression in placentas of nuclear transfer (NT), intracytoplasmic sperm injection (ICSI) and IVP embryos confirmed that HSPC117 deficiency correlates well with failures in embryo development: all NT embryos with a fetus, as well as IVP and ICSI embryos, had normal placental HSPC117 expression while those NT embryos showing reduced or no expression of HSPC117 failed to form a fetus. In conclusion, we show that HSPC117 is an important gene for post-implantation development of embryos, and that HSPC117 deficiency leads to fetal abnormalities after implantation, especially following placental formation. We suggest that defects in HSPC117 expression may be an important contributing factor to loss of cloned NT embryos in vivo.     

A recombinant mimetics of the HIV-1 gp41 prehairpin fusion intermediate fused with human IgG Fc fragment elicits neutralizing antibody response in the vaccinated mice

HIV-1 gp41 prehairpin fusion intermediate (PFI) composed of three N-terminal heptad repeats (NHR) plays a crucial role in viral fusion and entry and represents an attractive target for anti-HIV therapeutics (e.g., enfuvirtide) and vaccines. In present study, we constructed and expressed two recombinant gp41 PFI mimetics, designated N46Fd and N46FdFc. N46Fd consists of N46 (residues 536-581) in gp41 NHR and foldon (Fd), a trimerization motif. N46FdFc is composed of N46Fd fused with human IgG Fc fragment as an immunoenhancer. We immunized mice with N46 peptide, N46Fd and N46FdFc, respectively, and found that only N46FdFc elicited neutralizing antibody response in mice against infection by HIV-1 strains IIIB (clade B, X4), 92US657 (clade B, R5), and 94UG103 (clade A, X4R5). Anti-N46FdFc antibodies inhibited PIE7 binding to PFI, blocked gp41 six-helix bundle formation, and suppressed HIV-1 mediated cell-cell fusion. These findings provide an important clue for developing recombinant gp41 PFI mimetics-based HIV vaccines.     

Epigenetic repression of microRNA-129-2 leads to overexpression of SOX4 in gastric cancer

High levels of SOX4 expression have been found in a variety of human cancers, such as lung, brain and breast cancers. However, the expression of SOX4 in gastric tissues remains unknown. The SOX4 expression was detected using immunohistochemical staining and semi-quantitative RT-PCR, and our results showed that SOX4 was up-regulated in gastric cancer compared to benign gastric tissues. To further elucidate the molecular mechanisms underlying up-regulation of SOX4 in gastric cancers, we analyzed the expression of microRNA-129-2 (miR-129-2) gene, the epigenetic repression of which leads to overexpression of SOX4 in endometrial cancer. We found that up-regulation of SOX4 was inversely associated with the epigenetic silencing of miR-129-2 in gastric cancer, and restoration of miR-129-2 down-regulated SOX4 expression. We also found that inactivation of SOX4 by siRNA and restoration of miR-129-2 induced apoptosis in gastric cancer cells.     

Isolation and characterization of polymorphic microsatellite loci in Camellia nitidissima Chi (Theaceae)

? Premise of the study: Microsatellite markers were developed in Camellia nitidissima for further population genetic studies. ? Methods and Results: Eight microsatellite markers were newly developed from C. nitidissima and 7 were transferred from other Camellia species. Two to 13 alleles per locus were identified for these microsatellites. Observed and expected heterozygosities ranged from 0.040 to 0.909, and 0.184 to 0.916, respectively. Four loci showed a significant deviation from Hardy-Weinberg equilibrium and five locus pairs displayed linkage disequilibrium. ? Conclusions: These microsatellite markers will be useful to assess the genetic variation and genetic structure of C. nitidissima.     

Genetic Control of Individual Differences in Gene-Specific Methylation in Human Brain

We have observed extensive interindividual differences in DNA methylation of 8590 CpG sites of 6229 genes in 153 human adult cerebellum samples, enriched in CpG island “shores” and at further distances from CpG islands. To search for genetic factors that regulate this variation, we performed a genome-wide association study (GWAS) mapping of methylation quantitative trait loci (mQTLs) for the 8590 testable CpG sites. cis association refers to correlation of methylation with SNPs within 1 Mb of a CpG site. 736 CpG sites showed phenotype-wide significant cis association with 2878 SNPs (after permutation correction for all tested markers and methylation phenotypes). In trans analysis of methylation, which tests for distant regulation effects, associations of 12 CpG sites and 38 SNPs remained significant after phenotype-wide correction. To examine the functional effects of mQTLs, we analyzed 85 genes that were with genetically regulated methylation we observed and for which we had quality gene expression data. Ten genes showed SNP-methylation-expression three-way associations—the same SNP simultaneously showed significant association with both DNA methylation and gene expression, while DNA methylation was significantly correlated with gene expression. Thus, we demonstrated that DNA methylation is frequently a heritable continuous quantitatively variable trait in human brain. Unlike allele-specific methylation, genetic polymorphisms mark both cis- and trans-regulatory genetic sites at measurable distances from their CpG sites. Some of the genetically regulated DNA methylation is directly connected with genetically regulated gene expression variation.     

Bt群体信号应答因子nprR基因的缺失对cry1Ac基因表达的影响

摘要：【目的】研究群体信号应答蛋白编码基因nprR在苏云金芽胞杆菌（Bacillus thuringiensis,Bt）HD-73菌株晶体蛋白形成过程中的作用。【方法】通过同源重组,构建了HD-73 nprR基因缺失突变菌株HD73（ΔnprR ）。利用启动子-lacZ融合、SDS-PAGE方法,测定不同培养基中nprR基因转录活性及nprR基因缺失对cry1Ac转录及表达的影响。【结果】启动子转录活性分析表明,在LB和SSM培养基中nprR基因从对数期结束（T0）开始表达,稳定期持续表达。在LB培养基中,nprR基因的缺失使cry1Ac基因在生长过渡期和稳定期前期转录活性显著提高,同时HD73（ΔnprR ）菌株Cry蛋白生成量也明显高于出发菌株HD-73,但是在芽胞形成释放后,Cry蛋白的表达没有明显的区别。【结论】在丰富培养基中苏云金芽胞杆菌nprR基因的缺失在生长过渡期和稳定期前期能够提高cry1Ac基因转录和表达,从而缩短了cry基因表达时间,并且Cry蛋白总产量与出发菌株相当。     

Fruits and foliage of Pueraria (Leguminosae, Papilionoideae) from the Neogene of Eurasia and their biogeographic implications

? Premise of the study: Pueraria (Leguminosae, Papilionoideae) is native in East Asia, South Asia, Southeast Asia, and Oceania and is well known as a rampant invasive weed in the southeastern United States (P. montana; better known as kudzu), but relatively little is known about its early evolution and biogeographic origin. ? Methods: On the basis of comparative analyses of the fruit and leaflet architecture of closely related extant and fossil taxa, we studied the fossil history and biogeography of Pueraria. ? Key results: Fossil Pueraria is recognized on the basis of distinctive fruit and foliage from the Mio-Pliocene of middle latitudes in China, Japan, Abkhazia, and Croatia. Recognition of P. miothunbergiana from the Mio-Pliocene of China and Japan is reinforced by a trifoliolate leaf as well as isolated lateral and terminal leaflets. Pueraria shanwangensis sp. nov. represents the first recognition of fossil Pueraria fruits. This fruit species co-occurs with P. miothunbergiana in the Middle Miocene Shanwang flora and possibly represents the same population. Pueraria maxima (Unger) comb. nov., previously named as Dolichites maximus or Desmodium maximum, is recognized on the basis of leaflets from the Miocene of Croatia and Abkhazia. Other prior fossil reports of Pueraria and Dolichites are reevaluated. ? Conclusions: Pueraria had begun to diversify by at least the Middle Miocene and had spread into the Mio-Pliocene subtropical and temperate floras of the Balkan Peninsula, the Caucasus, and eastern Asia, which suggests the present diversity of this genus in tropical Asia and Oceania might have originated from the mid-latitudes of Eurasia.