鸡Ii结合Rab5a和Rab7b分子的分析

本研究旨在探明鸡恒定链（invariant chain，Ii）与内吞体转运蛋白Rab5a和Rab7b结合的结构域和在细胞内共定位的特征。首先，用PCR和基因突变技术将Ii胞浆区与跨膜区[Ii_{（Cyt-Tra）}]、Ii CLIP （class Ⅱ-associated invariant chain peptide）-三聚体区[Ii_{（CLIP-TRIM）}]和Ii突变体[Ii_{（M81-87aa）}、Ii_{（M91-99aa）}和Ii_{（M81-99aa）}]分别插入pET-32a和pEGFP-C1构建相应的原核和真核重组质粒。其次，将构建的含有绿色荧光蛋白的重组质粒与实验室保存的含有红色荧光Rab5a和Rab7b的重组质粒共转染至人胚胎肾细胞系293 T，观察它们的共定位。将构建的原核重组质粒进行表达和纯化，最后用拉下法和免疫印迹检测Ii与Rab5a和Rab7b的结合域。结果表明，成功构建Ii结构域及Ii突变体的重组质粒。Ii_{（Cyt-Tra）}及Ii突变体均能与Rab5a和Rab7b在细胞内共定位，而Ii_{（CLIP-TRIM）}与空载体却不能。Ii的胞浆区和跨膜区是与Rab5a和Rab7b结合的功能结构域，而不是CLIP与三聚体区。综上所述，鸡Ii与Rab5a和Rab7b共定位和结合的区域是其胞浆区和跨膜区，而不是内质网腔区。这些结果提示Rab分子参与了Ii在胞内细胞器的转运机制，为进一步研究Ii及其载体在细胞内的转运机制和功能提供了新的途径。     

牛星状病毒EvaGreen实时荧光定量PCR检测方法的建立及应用

牛星状病毒(BAstV)是我国新发的犊牛腹泻病原,本试验的目的是建立检测BAstV的Real-time PCR方法。根据BAstV流行株的ORF1a基因序列设计引物,通过优化反应条件和体系,成功建立基于EvaGreen检测BAstV的Real-time PCR方法。结果表明,该检测方法的Ct值与标准品模板在1.36×10¹~1.36×10⁸拷贝/μL线性关系良好,相关系数R²=0.999,扩增效率为93.79%;该方法可特异性检出BAstV,对犊牛腹泻其他相关病原呈阴性;最低检测下限为13.6拷贝/μL;批间和批内的变异系数均小于2%,重复性好。对2017年9月至2019年5月采自河南省的221份犊牛腹泻样本进行检测,BAstV的检出率为18.1%(40/221),采样场阳性率为100.0%(14/14)。本试验所建方法灵敏度高、特异性强、稳定性好,为BAstV的检测和流行病学调查提供了有力手段。     

Site‐specific management is crucial to managing Mikania micrantha

Increasingly, weeds have been taking on global distributions. With the proliferation of invasive weeds has come the challenge of managing these species over broad geographical regions, with diverse habitats and political jurisdictions. Here, we review the management of Mikania micrantha Kunth (Asteraceae; mile‐a‐minute) throughout its invaded range, extending through most of the Pacific islands and southern and south‐east Asia. Context matters when determining the best course of action for managing M. micrantha, as it has invaded a large variety of agricultural and natural systems. In Queensland, Australia and Florida, USA, M. micrantha has been targeted in relatively successful eradication campaigns, highlighting the importance of early detection and rapid response methods, while elsewhere in its invaded range, populations are either still increasing or showing limited signs of decline. An inter‐regional approach to research and management should incorporate successful management strategies employed throughout the invaded range including, but not limited to, chemical and cultural control practices, manual and mechanical control, classical biological control using the rust fungus Puccinia spegazzinii, plant–plant competition and integrated approaches utilising two or more control methods concurrently. Additional knowledge of M. micrantha genetics is required to determine if management approaches could be fine‐tuned for particular populations. Countries bordering the Mekong River formed a network in 2011 to co‐ordinate the management of invasive species such as M. micrantha. Expanding such a collaborative approach to other regions could further reduce populations of M. micrantha and limit its spread.     

Up-regulation of miR-181a attenuates releases of CSE-induced pro-inflammatory factors and expression of collagen IV,fibronectin and α-SMA in human bronchial epithelial cells

AIM: To investigate the effect and potential mechanism of microRNA-181a (miR-181a) on cigarette smoke extract (CSE)-induced the productions of pro-inflammatory factors and the expression of collagen IV, fibronectin and α-smooth muscle actin (α-SMA) in human bronchial epithelial cells (HBECs). METHODS: CSE-induced miR-181a expression was detected by RT-qPCR in the HBECs. After tansfected with miR-181a mimic, the releases of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), IL-6 and transforming growth factor-β1 (TGF-β1) were measured by ELISA, the protein expression of collagen IV, fibronectin and α-SMA was determined by Western blot. The activation of NF-κB/TGF-β1/Smad3 pathway was also evaluated by Western blot. RESULTS: CSE increased the levels of TNF-α, IL-1β, IL-6 and TGF-β1 and the expression of collagen IV, fibronectin and α-SMA, and decreased the expression of miR-181a in the HBECs (P<0.05). However, transfected with miR-181a mimic partially prevented the releases of TNF-α, IL-1β, IL-6 and TGF-β1, and inhibited the expression of collagen IV, fibronectin and α-SMA (P<0.05). Additionally, the activation of NF-κB/TGF-β1/Smad3 evoked by CSE was attenuated after transfected with miR-181a mimic. CONCLUSION: Up-regulation of miR-181a prevents the releases of CSE-induced pro-inflammatory factors and expression of collagen IV, fibronectin and α-SMA in the HBECs, and its mechanism may be related to the inhibition of NF-κB/TGF-β1/Smad3 pathway.     

Development of a linear mixed-effects individual-tree basal area increment model for masson pine in Hunan Province,South-central China

ABSTRACT

An individual-tree basal area increment model was developed for masson pine based on 26276 observations of 13,138 trees in 987 sample plots from the 7th (2004), 8th (2009), and 9th (2014) Chinese National Forest Inventory in Hunan Province, South-central China. The model was built using a linear mixed-effects approach with sample plots included as random effects since the data have a hierarchical stochastic structure and biased estimates of the standard error of parameter estimates could be a consequence of applying ordinary least square (OLS) for regression. In addition, within-plot heteroscedasticity and autocorrelation were also considered. The final mixed-effects model was determined according to the Akaike information criterion (AIC), Bayesian information criterion (BIC), log-likelihood (Loglik), and the likelihoodratio test (LRT). The results revealed that initial diameter (DBH), the sum of the basal area (m²/ha) in trees with DBHs larger than the DBH of the subject tree (BAL), number of trees per hectare (NT), and elevation (EL) had a significant impact on individual-tree basal area increment. The mixed-effects model performed much better than the basic model produced using OLS. Additionally, the variance structure of the model errors was successfully modeled using the power function. However, the autocorrelation structures were not defined because there was no autocorrelation amongst the data. It is believed that the final model will contribute to the scientific management of the masson pine.     

环境因子对中肋骨条藻生长及叶绿素荧光特性的影响

为了解温度、光照和磷酸盐及其交互作用对中肋骨条藻(Skeletonema costatum)生长及叶绿素荧光特性的影响,每个环境因子设置3个水平[温度:17、23、29℃;光照:80、120、160μmol photons/(m~2·s);磷酸盐:0.1、1、10μmol/L],考虑环境因子间的两两交互作用,采用L18(3~7)正交实验表安排室内培养实验,研究中肋骨条藻叶绿素a浓度和光合活性的变化。结果表明,3因素3水平的实验中,中肋骨条藻在10μmol/L磷酸盐浓度下叶绿素a峰值能达到较高水平,其中最优环境因子水平组合为23℃、120μmol photons/(m~2·s)、10μmol/L。在培养期间,磷酸盐浓度对中肋骨条藻叶绿素a峰值造成极其显著的影响(P0.01),温度、光照及两两间的交互作用未对叶绿素a峰值造成显著影响(P0.05)。中肋骨条藻在10μmol/L磷酸盐浓度下光合活性更高,但光能利用效率α并未随磷酸盐浓度表现出明显差异。当中肋骨条藻处于10μmol/L和1μmol/L磷酸盐浓度时,光照对最大量子产量F_v/F_m造成显著影响(P0.05);10μmol/L磷酸盐浓度下,F_v/F_m在80和120μmol photons/(m~2·s)光强下较高;在1μmol/L磷酸盐浓度下,F_v/F_m在120μmol photons/(m~2·s)光强下最低。     

水稻对硅元素吸收特性研究

[目的]研究水稻对硅元素的吸收特性。[方法]以水稻为材料,采用水培试验,设置0.085、0.850 mmol/L 2个硅水平,明确水稻对硅的吸收模式。[结果]在2个硅处理下水稻在12 h的总吸硅量明显高于蒸腾吸硅量,二者都随着时间的延长逐渐升高,并且蒸腾吸硅量占总吸硅量的比值表现为高硅浓度处理高于低硅浓度处理;2个处理在6 h内的硅吸收都受到了代谢抑制剂和低温的抑制,并且表现为高硅浓度的抑制程度低于低硅浓度的抑制程度;2个处理在12 h内木质部汁液硅浓度随时间的延长逐步升高,而外部溶液硅浓度随时间的延长逐步降低,并且木质部汁液硅浓度比外部溶液的硅浓度高数倍。[结论]水稻对硅的吸收存在主动吸收和被动吸收2个过程,尤其在高硅浓度处理下,被动吸收更不能忽视。     

Replication of neurovirulent equine herpesvirus type 1 (EHV-1) in CD172a+ monocytic cells

Equine herpesvirus type 1 (EHV-1) is responsible for respiratory disorders, abortion and myeloencephalopathy (EHM) in horses. Two pathotypes of EHV-1 strains are circulating in the field: neurovirulent (N) and non-neurovirulent (NN). For both strains, CD172a⁺ monocytic cells are one of the main carrier cells of EHV-1 during primary infection, allowing the virus to invade the horse’s body. Recently, we showed that EHV-1 NN strains showed a restricted and delayed replication in CD172a⁺ cells. Here we characterize the in vitro replication kinetics of two EHV-1 N strains in CD172a⁺ cells and investigate if the replication of these strains is similarly silenced as shown for EHV-1 NN strains. We found that EHV-1 N replication was restricted to 7–8% in CD172a⁺ cells compared to 100% in control RK-13 cells. EHV-1 N replication was not delayed in CD172a⁺ cells but virus production was significant lower (10^3.0 TCID₅₀/10⁵ inoculated cells) than in RK-13 cells (10^8.5 TCID₅₀/10⁵ inoculated cells). Approximately 0.04% of CD172a⁺ cells produced and transmitted infectious EHV-1 to neighbour cells compared to 65% of RK-13 cells. Unlike what we observed for the NN strain, pretreatment of CD172a⁺ cells with histone deacetylases inhibitors (HDACi) did not influence the replication of EHV-1 N strains in these cells. Overall, these results show that the EHV-1 replication of N strains in CD172a⁺ cells differs from that observed for NN strains, which may contribute to their different pathogeneses in vivo.     

新形势下农业科研单位廉政建设工作的现状与对策研究

该文阐述了当下农业科研单位党风廉政建设的新形势,从硬件建设、科研经费使用、人事任免和学术管理等几个廉政风险点详细分析了目前农业科研事业单位反腐败工作所面临的新局面和新挑战,并根据近年来涌现出的主要问题,从思想教育、制度建设等方面提出了具有针对性的对策和措施。     

基于转录组测序的半滑舌鳎长链非编码RNA的初步鉴定与分析

以抗病家系与易感家系半滑舌鳎为材料,进行哈维弧菌感染实验,并对易感家系感染前(CsSU)、易感家系感染后(CsSC)、抗病家系感染前(CsRU)、抗病家系感染后(CsRC)4组进行转录组测序,根据RNA-seq数据挖掘半滑舌鳎长链非编码RNA信息;通过生物信息学分析,筛选出与抗哈维弧菌病相关的差异长链非编码RNA(long non-coding RNA, lncRNA)。结果显示,共识别出4 584个lncRNA座位,包含5 714个转录本;其基本特征与编码基因的比较分析,lncRNA的GC含量低于编码基因,单外显子基因数多于编码基因,转录本的平均长度长于编码基因,基因表达量低于编码基因。对4组样品进行两两比较(CsRU vs CsSU、CsRC vs CsSC、CsRC vs CsRU、CsSC vs CsSU)分别筛选出818、813、261、140个差异表达lncRNA,其中CsRU与CsSU之间、CsRC与CsSC之间lncRNA数目差异最多,通过聚类分析确定了各实验组的表达模式之间的联系,CsRU与CsSU之间的表达模式最为相近。通过共表达分析,预测出lncRNA和274个编码基因可能存在14 539种相互关系,并进行了功能注释,进而筛选出7个关键lncRNA。qRT-PCR结果显示,差异表达lncRNAs的表达模式和转录组数据得到的基本一致。研究结果为揭示lncRNA在半滑舌鳎抗哈维弧菌免疫调控反应中的作用提供重要的参考数据。