Development of a fluorescent probe hydrolysis-insulated isothermal PCR for rapid and sensitive on-site detection of African swine fever virus

Highlights
1. A probe-based insulated isothermal PCR (iiPCR) assay was developed for rapid and onsite detection of ASFV.
2. The developed iiPCR showed similar sensitivity and specificity with OIE recommended real-time PCR.
3. Blood samples could be directly applied as PCR template in iiPCR without DNA extraction.     

小鼠心脏发育中组蛋白乙酰化酶GCN5和PCAF的时空表达特征

目的研究组蛋白乙酰化酶氨合成通用控制蛋白5（general control nonderepressible-5 GCN5）和p300/CREB结合蛋白相关因子（p300/CREB binding protein-associated factor PCAF）在小鼠心脏发育过程中的时空表达规律。方法选取胎龄7.5-18d、出生1d和3月成年昆明小鼠正常心脏,利用免疫组织化学法和RT-PCR或Western blot技术半定量检测GCN5和PCAF在小鼠心脏发育过程中的时空表达变化。结果1.GCN5在E7.5-E9.5心脏原基中不表达;E10.5后心内膜垫及室间隔膜部和房室瓣相对强表达,心肌组织弱表达。GCN5 mRNA在E10.5-E11.5出现高峰表达,E12.5-E15.5表达水平下降,E16.5至成年鼠期表达极低。2.PCAF在E7.5-E9.5心脏原基中广泛弱表达;E10.5后心肌膜较强表达,心内膜、房室瓣和小梁网较弱表达,室间隔肌部弱表达,室间隔膜部极弱表达;PCAF蛋白在E10.5已有相对高表达,E11.5达到表达高峰,此后表达逐渐降低,在E13.5-E15.5和E16.5-生后1d仔鼠期出现两个表达平台期,成年鼠期心脏中表达最低。结论GCN5和PCAF在发育心脏中存在不同表达分布和变化规律,表明二者均参与了心脏的发生发育过程,且二者可能与心脏的特定结构发育密切相关。     

有害赤潮藻对浮游动物影响的研究进展

有害赤潮藻对浮游动物的影响在赤潮危害的研究中越来越受到关注,本文综述了有害赤潮藻对浮游动物的毒害作用,主要表现为影响浮游动物的存活和生长,降低母体的产卵力、卵的受精率或受精卵的孵化率以及减慢胚胎发育或造成畸形,影响幼体的正常发育,抑制浮游动物的摄食和改变其正常行为等;这些毒害作用主要是由赤潮藻产生的藻毒素或其它一些胞外渗出物通过降低浮游动物对食物的利用率、造成生理损伤、破坏细胞膜、降低亲体产生的配子质量、抑制有丝分裂和胚胎发育等造成的．某些赤潮藻营养成分单一如缺乏多不饱和脂肪酸,也会对浮游动物的生长繁殖产生影响．     

An intramolecular disulfide bond is required for the thermostability of methyl parathion hydrolase, OPHC2

OPHC2, a methyl parathion hydrolase (MPH) from Pseudomonas pseudoalcaligenes C2-1 (CGMCC 1150), can degrade a wide range of organophosphate pesticides. Compared with other MPHs, OPHC2 exhibits high thermostability. Its thermostability mechanism, however, remains unknown. In the present study, sequence analysis demonstrated that two cysteines (Cys110 and Cys146) exist in OPHC2, but not in other MPHs. The three-dimensional structural model of OPHC2 performed by computer-assisted homology modelling revealed a potential stacking network with residues Cys110 and Cys146, which probably formed an intramolecular disulfide bond. Furthermore, both sodium dodecyl sulphate-polyacrylamide gel electrophoresis and thiol-titration analyses indicated that OPHC2 contains a disulfide bond. Substitution of the disulfide bond-forming cysteines with alanine, leucine or methionine residues substantially decreased the thermostability of OPHC2, suggesting that disulfide bond formation affects conformational stability. These results, combined with three-dimensional structural modelling, demonstrated that the formation of a C110-C146 disulfide bond may stabilise the conformation of OPHC2, contributing to its thermostability.     

运动性骨骼肌疲劳亚细胞机制的探讨

本实验采用持续性下坡跑运动,观察大鼠骨骼肌运动后不同时相线粒体形态、代谢、机能等指标的变化,结果表明：大鼠运动后即刻线粒体钙含量、细胞膜丙二醛（ＭＤＡ）值明显增加,ＡＴＰ含量和细胞膜Ｎａ＋,Ｋ＋－ＡＴＰ酶活性下降;运动后２４ｈ线粒体钙含量、ＭＤＡ值增加最明显,ＡＴＰ含量仍未恢复,细胞膜Ｎａ＋,Ｋ＋－ＡＴＰ酶活性基本恢复,线粒体体密度、平均体积比运动前明显增加,比表面缩小;运动后４８ｈＡＴＰ含量完全恢复,线粒体钙含量、ＭＤＡ值开始恢复。本研究结果提示,急性运动引起的细胞膜脂质过氧化加强、线粒体形态、代谢机能异常抑制线粒体氧化磷酸化过程、减少ＡＴＰ生成可能是运动性骨骼肌疲劳的亚细胞机制之一。耐力训练可以通过改善线粒体形态、代谢、机能提高机体的运动能力。     

基于变权模型的舟山群岛生态安全预警

生态安全预警是生态安全研究的重要内容,对维护区域生态安全具有重要的指示意义.本文以浙江省舟山群岛为例,基于驱动力、压力、状态、影响、响应(DPSIR)框架模型构建了生态安全预警指标体系,使用变权模型对2000-2012年舟山市生态安全的预警等级进行测度,并使用马尔科夫预测方法对2013-2018年生态安全警情进行了预测.结果表明:变权模型能够有效地满足舟山群岛生态安全动态预警研究需要;2000-2012年,舟山群岛生态安全预警指数由0.286波动上升至0.484,警度等级从“重警”演变为“中警”,指示灯由“橙灯”演化为“黄灯”;2013-2018年,舟山群岛生态安全预警等级预测结果为“中警”,指示灯为“黄灯”.研究结果可为维护舟山群岛生态安全提供参考.     

Expression of baculovirus anti-apoptotic genes p35 and op-iap in cotton (Gossypium hirsutum L.) enhances tolerance to verticillium wilt

Background

Programmed cell death plays an important role in mediating plant adaptive responses to the environment such as the invasion of pathogens. Verticillium wilt, caused by the necrotrophic pathogen Verticillium dahliae, is a serious vascular disease responsible for great economic losses to cotton, but the molecular mechanisms of verticillium disease and effective, safe methods of resistance to verticillium wilt remain unexplored.

Methodology/Principal Findings

In this study, we introduced baculovirus apoptosis inhibitor genes p35 and op-iap into the genome of cotton via Agrobacterium-mediated transformation and analyzed the response of transgenic plants to verticillium wilt. Results showed that p35 and op-iap constructs were stably integrated into the cotton genome, expressed in the transgenic lines, and inherited through the T₃ generation. The transgenic lines had significantly increased tolerance to verticillium wilt throughout the developmental stages. The disease index of T₁–T₃ generation was lower than 19, significantly (P<0.05) better than the negative control line z99668. After treatment with 250 mg/L VD-toxins for 36 hours, DNA from negative control leaves was fragmented, whereas fragmentation in the transgenic leaf DNA did not occur. The percentage of cell death in transgenic lines increased by 7.11% after 60 mg/L VD-toxin treatment, which was less than that of the negative control lines''s 21.27%. This indicates that p35 and op-iap gene expression partially protects cells from VD-toxin induced programmed cell death (PCD).

Conclusion/Significance

Verticillium dahliae can trigger plant cells to die through induction of a PCD mechanism involved in pathogenesis. This paper provides a potential strategy for engineering broad-spectrum necrotrophic disease resistance in plants.