Molecular character of a phosphatase 2C (PP2C) gene relation to stress tolerance in Arabidopsis thaliana

Protein phosphatases type 2C (PP2Cs) from group A, which includes the ABI1/HAB1 and PP2CA branches, are key negative regulators of ABA signaling. HAI-1 gene had been shown to affect both seed and vegetative responses to ABA, which is one of PP2Cs clade A in Arabidopsis thaliana. Transgenic plants containing pHAI-1::GUS (β-glucuronidase) displayed GUS activity existing in the vascular system of leave veins, stems and petioles. Green fluorescent protein fused HAI-1 (HAI-1-GFP) was found in the nucleus through transient transformation assays with onion epidermal cells. The water-loss assays indicated the loss-of-function mutants did not show symptoms of wilting and they had still turgid green rosette leaves. The assays of seed germination by exogenous ABA and NaCl manifested that the loss-of-function mutants displayed higher insensitivity than wild-type plants. Taken together, the final results suggest that the HAI-1 (AT5G59220) encoded a nuclear protein and it can be highly induced by ABA and wound in Arabidposis, the stress-tolerance phenotype showed a slightly improvement when HAI-1 gene was disrupted.     

Function of wheat phosphate transporter gene TaPHT2;1 in Pi translocation and plant growth regulation under replete and limited Pi supply conditions

Several phosphate transporters (PTs) that belong to the Pht2 family have been released in bioinformatics databases, but only a few members of this family have been functionally characterized. In this study, we found that wheat TaPHT2;1 shared high identity with a subset of Pht2 in diverse plants. Expression analysis revealed that TaPHT2;1 was strongly expressed in the leaves, was up-regulated by low Pi stress, and exhibited a circadian rhythmic expression pattern. TaPHT2;1–green fluorescent protein fusions in the leaves of tobacco and wheat were specifically detected in the chloroplast envelop. TaPHT2;1 complemented the Pi transporter activities in a yeast mutant with a defect in Pi uptake. Knockdown expression of TaPHT2;1 significantly reduced Pi concentration in the chloroplast under sufficient (2 mM Pi) and deficient Pi (100 μM Pi) conditions, suggesting that TaPHT2;1 is crucial in the mediation of Pi translocation from the cytosol to the chloroplast. The down-regulated expression of TaPHT2;1 resulted in reduced photosynthetic capacities, total P contents, and accumulated P amounts in plants under sufficient and deficient Pi conditions, eventually leading to worse plant growth phenotypes. The TaPHT2;1 knockdown plants exhibited pronounced decrease in accumulated phosphorus in sufficient and deficient Pi conditions, suggesting that TaPHT2;1 is an important factor to associate with a distinct P signaling that up-regulates other PT members to control Pi acquisition and translocation within plants. Therefore, TaPHT2;1 is a key member of the Pht2 family involved in Pi translocation, and that it can function in the improvement of phosphorus usage efficiency in wheat.     

竹叶菜的组织培养研究

竹叶菜为百合科鹿药属一种多年生草本植物,因做菜口感好,且营养丰富、药用价值高而成为深受人们喜爱的一种野生蔬菜.有关竹叶菜及其同属的组织培养研究均未见报道.本文以竹叶菜的顶芽为外植体,通过器官发生途径,初步探索了竹叶菜的组织培养技术,结果表明:芽诱导的合适培养基为:MS +6-BA 1.5 +NAA 0.05+0.6％琼脂+3％蔗糖;芽继代培养的合适培养基为:MS+6-BA 1.2 +NAA 0.05 +0.6％琼脂+3％蔗糖;无菌小苗生根的合适培养基为:MS+ IAA 1.5+ NAA 0.2+ 0.6％琼脂+3％蔗糖.为竹叶菜今后较大规模地栽培或生产提供种苗及技术贮备,为实现竹叶菜资源的可持续性开发利用奠定了基础.     

Overlapping functions of argonaute proteins in patterning and morphogenesis of Drosophila embryos

Argonaute proteins are essential components of the molecular machinery that drives RNA silencing. In Drosophila, different members of the Argonaute family of proteins have been assigned to distinct RNA silencing pathways. While Ago1 is required for microRNA function, Ago2 is a crucial component of the RNA-induced silencing complex in siRNA-triggered RNA interference. Drosophila Ago2 contains an unusual amino-terminus with two types of imperfect glutamine-rich repeats (GRRs) of unknown function. Here we show that the GRRs of Ago2 are essential for the normal function of the protein. Alleles with reduced numbers of GRRs cause specific disruptions in two morphogenetic processes associated with the midblastula transition: membrane growth and microtubule-based organelle transport. These defects do not appear to result from disruption of siRNA-dependent processes but rather suggest an interference of the mutant Ago2 proteins in an Ago1-dependent pathway. Using loss-of-function alleles, we further demonstrate that Ago1 and Ago2 act in a partially redundant manner to control the expression of the segment-polarity gene wingless in the early embryo. Our findings argue against a strict separation of Ago1 and Ago2 functions and suggest that these proteins act in concert to control key steps of the midblastula transition and of segmental patterning.     

Elicitation of immunity in mice after immunization with the S2 subunit of the severe acute respiratory syndrome coronavirus

The S2 domain of the severe acute respiratory syndrome coronavirus (SARS-CoV) spike (S) protein is responsible for fusion between virus and target cell membranes, and is expected to be immungenic. In this study, we investigated the immune responses against the S2 subunit in BALB/c mice, which were vaccinated either with plasmid DNA encoding the S2 domain (residues 681-1120), the recombinant S2 fragment (residues 681-980) in incomplete Freund's adjuvant, or with inactivated SARS-CoV. The increased number of specific cytotoxic cells (CTLs) and the high titer of specific antibody showed stimulation of both arms of the immune system in these groups. The shift in cytokines suggested that Th1-polarized immune response was induced by plasmid pCoVS2, meanwhile the Th2-dominant response was induced by recombinant S2 fragment and inactivated vaccine. However, the titer of neutralizing antibodies was only detectable in mice immunized with inactivated virus, but not with pCoVS2 plasmid. Taken together, the S2 domain could induce specific cellular immune response and a high level of total IgG but little neutralizing antibodies against infection by SARSCoV.     

Superior neovascularization and muscle regeneration in ischemic skeletal muscles following VEGF gene transfer by rAAV1 pseudotyped vectors

Recombinant adeno-associated virus serotype 2 (rAAV2) vector has been widely employed for gene therapy. Recent progress suggests that the new serotypes of AAV showed a better performance than did AAV2 in normal tissues. Here, we evaluate the potential role of human vascular endothelial growth factor (VEGF) gene transfer using rAAV vector pseudotyped with serotype 1 capsid proteins (rAAV1) in the treatment of muscle ischemia. In ischemic skeletal muscles, the rAAV1-LacZ vector allowed higher level, broader distribution, and long-lasting gene expression compared with the rAAV2-LacZ vector. Muscle VEGF165 production following the rAAV1-VEGF165 vector injection was 5-10 times higher than that following the rAAV2-VEGF165 vector injection. VEGF165 production mediated by the rAAV1-VEGF165 vector stimulated a large set of neovascularization with relatively mature vascular structures and enhanced muscle regeneration in the ischemic skeletal muscles. Thus, the rAAV1-VEGF165 vector mediated gene transfer may be a therapeutic approach to peripheral vascular diseases.     

大肠杆菌青霉素G酰化酶基因及其邻近区域的核苷酸全序列

Some of microorganisms have been known to possess penicillin G acylase activity. The E. coli derived penicillin G acylase (PGA) can catalyze the conversion of penicillin G into phenylacetic acid and 6-amino-penicillanic acid, the latter is used as the starting compound for the industrial formation of semi-synthetic penicillins. Apart from its industrial importance, the enzyme PGA displays a number of interesting properties. Catalytically active enzyme is localized in the periplasmic space of E. coli cells and composed of two dissimilar subunits. The two subunits are apparently produced from a precursor protein, via a processing pathway hitherto unique in its features for a prokaryotic enzyme. The studies on processing of the precursor and on the relationship between structure and function of the mature enzyme are important theoretically. Previously we cloned a 3.5 kb DNA fragment from a strain (E. coli AS 1.76), which displays PGA activity. In this paper, we report a nucleotide sequence of the 3.5 kb DNA fragment containing PGA gene. After insertion of the DNA fragment into EcoR I and Hind III sites in pWR 13, pPGA 20 had been obtained. We subcloned the Hind III and Bg1 II treated fragment of 1.6 kb in length from pPGA 20 into Hind III and BamH I sites of pWR 13 to get a pPGA 1.6, and Bg1 II and EcoR I treated fragment of 1.9 kb in length into BamH I and EcoR I sites of pWR 13 to get a pPGA 1.9. The linearized pPGA 1.9 which were digested with appropriate restriction enzymes were progressively shortened from both ends respectively by digestion with Bal 31 nuclease, followed by cleavage of shortened target DNA off vector DNA molecules with appropriate restriction enzymes. The series of the DNA fragments shortened from EcoR I end were then cloned into plasmid pWR 13 which had previously digested with Hind III and Sma I enzymes (Fig. 1). The DNA fragment cloned in pWR 13 were directly sequenced on the resulted plasmids by using primer I and primer II. Thus we have obtained the complete nucleotide sequence of the 3.5 kb DNA fragment. The 3.5 kb fragment contains an intact PGA gene which is 2.6 kb.(ABSTRACT TRUNCATED AT 400 WORDS)     

Spatial Confinement of Sn/TiO2 Nanoparticles in Hollow Mesoporous Carbon Spheres Opal for Stable Zn Metal Anodes

The sluggish Zn²⁺ diffusion and high nucleation energy barrier induce uncontrolled growth of Zn dendrites and detrimental parasitic reactions, severely hampering the commercialization of Zn metal anodes (ZMAs). Herein, hollow mesoporous carbon sphere opal with confined Sn/TiO₂ clusters (HMCSST) is designed as the host to spatially regulate the Zn deposition. Owing to the capillary effect of the ordered hierarchical porous structure, the mass diffusion can be dramatically accelerated to promote a fast deposition kinetics at the interface between ZMA and electrolyte. Besides, the encapsulated ultrafine Sn/TiO₂ clusters serve as zincophilic sites to achieve both the uniform Zn deposition on the hierarchical porous opal host and the high thermodynamic stability of ZMAs. Benefiting from the structural and componential merits, the HMCSST host effectively reduces the activation energy to enable a temporal-spatial ordering of Zn nucleation and growth. As expected, the stable HMCSST-Zn electrode guarantees steady Zn platting/stripping with long-term stability over 1300 h in a symmetrical cell at a depth of discharge of 37.5%. As a proof-of-concept demonstration, an HMCSST-Zn||ammonium vanadate full cell shows a long lifespan over 5000 cycles at 10.0 A g⁻¹ with low polarization.     

醋酸棉酚诱导人粘液表皮样癌MEC-1细胞DNA双链断裂

本文研究醋酸棉酚(gossypol acetic acid,GAA)对人粘液表皮样癌细胞MEC-1体外增殖的影响,并初步探讨其抑制肿瘤细胞增殖的机制.体外培养人粘液表皮样癌细胞系MEC-1细胞,用MTT法检测GAA对MEC-1细胞增殖的影响;用中性彗星实验检测GAA对MEC-1细胞的DNA双链断裂;用免疫荧光染色法检测...