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.     

新疆春小麦育成品种遗传演变分析

【目的】研究新疆春小麦育成品种遗传演变规律,提高小麦资源的利用效率,为新疆春小麦品种选育和改良提供参考依据。【方法】以新疆春小麦育成品种为材料,对其主要性状进行综合评价分析,以春小麦90K芯片开展新疆春小麦育成种亲缘关系分析。【结果】新疆春小麦育成种遗传多样性丰富,平均遗传多样性指数2.005,变幅为1.902~2.181。相关性分析结果表明,蛋白质含量、湿面筋含量、穗粒数与育种年代呈极显著正相关。新疆春小麦品种选育主要是通过常规杂交育种、杂交辐射诱变育种、引种3种方式。利用小麦90K芯片将新疆春小麦育成种划分为3个类群,显示了各品种之间的亲缘关系。【结论】新疆春小麦育种遗传基础薄弱、遗传多样性逐渐散失,新疆小麦育种应加强资源收集与利用,扩大育种亲本选择,提高品种变异的丰度和广度,以多抗、高产稳产、优质高效的聚合育种,加快新品种选育进程,提高育种效率。     

Development of a method for live octopus (Octopus vulgaris) transportation for long distance at high densities

The development of new octopus‐based products with a growing economic value ensures the commercial interest of this species, making live octopus export an activity of great interest. The aim of this study was to develop a method for long‐distance transportation of live octopus at high densities. The system was composed by 220‐L tanks with cooling and aeration, where the animals were kept separated from each others. The water temperature was maintained at 10°C, after a decreasing of 1°C/hr. Live octopus transportation was tested for 48 hr at two densities: 50 kg/m³ and 100 kg/m³. During this period, water parameters were monitored. Stress response was evaluated through the analysis of haemolymph, muscle and brain tissues. No mortality was registered after 48 hr for both treatments. In all trials, water quality remained within the normal limits in both densities; there was, however, a significative increase of ammonia levels in the water. Ammonia, dopamine and Hsp70 levels were analysed in the beginning and at the end of the experiment for both densities; however, no significant differences were found among them. In general, this system seems to be a viable solution for live octopus 48‐hr transportation at a density of 100 kg/m³.     

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。     

文心兰HSP70基因的克隆及表达分析

为研究文心兰热激蛋白70基因（命名为OnHSP70）的分子特性及表达特点,以文心兰‘柠檬绿’（Oncidium hybridum ‘Honey Angel’）为材料,采用RT-PCR技术克隆OnHSP70,利用生物信息学方法分析其分子特性,通过qRT-PCR技术分析其在不同组织及不同非生物胁迫处理下的表达特点。生物信息学分析表明：该基因开放阅读框为1944 bp,编码647个氨基酸,翻译的蛋白为稳定蛋白,属于HSP70超家族,其蛋白二级结构由41.11%的α-螺旋、17.77%的延伸链、7.73%的β-转角和33.38%的无规卷曲组成,具有2个高度保守的功能域。聚类分析表明：该蛋白与铁皮石斛和深圳拟兰的HSP70亲缘关系较近。qRT-PCR分析表明：文心兰HSP70在4种不同组织器官中具有表达差异性,在花中的表达量最高;高温处理后基因的表达量明显上升;40 ℃高温胁迫4 h时表达量达峰值;茉莉酸甲酯及水杨酸处理时表达量呈现下调响应。本研究为后期该基因功能研究及提高文心兰对高温的适应性提供理论基础。