影响棉铃虫增殖马尾松毛虫质型多角体病毒的条件因子的初步探讨

棉铃虫为马尾松毛虫多角体病毒 (DPCPV)理想的替代寄主。试验结果表明 ,不同接毒方法、接种浓度、接种温度及接种虫龄对棉铃虫增殖病毒有影响 ,以病毒涂在人工饲料表面的接毒方法、2 0～ 2 3℃温度及 3龄虫为最佳组合条件。由于利用棉铃虫生产出的病毒 (Ha DPCPV)容易诱发棉铃虫本身的多角体病毒 (NPV) ,尤其对那些带有NPV源的虫种 ,Ha DPCPV不宜作毒种 ,因此为慎重起见 ,最好用马尾松毛虫复制出的病毒作为生产毒种。     

基于扎根理论的社区参与国家公园建设与管理的机制研究

大量研究表明自然保护地周边社区居民对保护地的态度是影响社区参与保护,实现保护地管理目标的主要因素,由此建立了"认知-态度-参与"的行为逻辑。中国国家公园体制建设希望进一步推进社区与保护地的关系,全面提升自然保护管理效率,也建立在对这一行为逻辑的认同之上。为验证社区认知与行为的关系,并为促进不同利益相关者参与国家公园建设与管理提供科学支持,研究试图分析武夷山国家公园体制试点区社区居民的国家公园的"认知-态度-参与"的行为逻辑以揭示社区参与国家公园建设与管理的深层机制。与一般研究不同在于,本研究是针对正在建设中的国家公园,使得本地利益相关方的态度有机会影响国家公园建设进程。为获得社区"认知-态度-参与"核心观念,探索社区参与的过程和方式,提出促进社区参与的方法,研究采用扎根理论这一质性研究方法,基于对本地利益相关者采用半结构化开放问题的深度访谈,归纳出概念与范畴,形成社区参与国家公园建设与管理的理论。研究首先形成45个概念与13个范畴,通过范式模型研究发现,稳定资源使用权利、引导社区参与决策、保障社区发展权利、创新社区产业模式、提高个体适应能力、降低政策风险影响等六个主范畴能够解释社区居民对国家公园的认知-态度-参与这一行为逻辑。通过归纳主范畴,研究进一步提炼出社区参与国家公园建设与管理的理论核心在于社区自然资源管理的自主性、创新性与适应性,其内在机制在于社区参与是一个自主性提高的动态过程,需要通过保障社区生计资源禀赋来实现长期的价值转化,最终使其成为社会-生态系统适应性治理的组成部分。在理论阐释基础上,研究提出从改善社区认知、提升社区参与角度推进国家公园建设的路径。研究也指出,所提出的社区参与机制理论能够提炼量化指标评价社区参与,追踪武夷山国家公园体制试点进程。     

Four New Diterpenoid Alkaloids from The Roots of Aconitum coreanum

Four new hetisine‐type C₂₀‐diterpenoid alkaloids, named as coreanines A–D ( 1 – 4 ), were isolated from the roots of Aconitum coreanum, together with thirteen known alkaloids ( 5 – 17 ). Their structures were elucidated by extensive spectroscopic methods including IR, HR‐ESI‐MS and NMR techniques. All the isolated compounds were screened for the acetylcholinesterase (AChE) inhibitory effects, and none of them showed considerable inhibitory activity.     

幽门螺杆菌致病岛CagL重组抗原的可溶性表达及其多克隆抗体的制备和分析*

目的:利用原核表达和蛋白质纯化技术获得高纯度的幽门螺杆菌致病岛CagL重组抗原(rCagL),利用其制备anti-CagL多克隆抗体,并分析抗体的特异性。方法:通过生物信息学软件分析rCagL的抗原结构;利用PCR长片段DNA合成技术合成不含有信号肽序列的幽门螺杆菌致病岛CagL基因,将其插入表达质粒pCzn1中,构建重组质粒pCzn1-rCagL。然后,将pCzn1-rCagL转入大肠杆菌Arctic Express中,经IPTG诱导表达后,通过Ni-IDA镍离子亲和层析纯化重组抗原rCagL,利用Western blot鉴定rCagL与His标签抗体和Anti-H. pylori抗体的免疫反应性;最后,通过rCagL辅以弗氏佐剂免疫BALB/c小鼠,制备anti-CagL多克隆抗血清,通过ELISA方法分析抗血清的特异性。结果:生物信息学软件表明重组抗原rCagL具有较好的抗原性质;重组质粒pCzn1-rCagL经双酶切和基因测序等技术鉴定,证实rCagL核苷酸序列与理论序列完全一致;基因工程菌株pCzn1-rCagL/Arctic Express在低温11℃条件经IPTG诱导表达。 SDS-PAGE实验结果证实:rCagL可实现相对高效地可溶性蛋白表达,可溶性蛋白约占包涵体的62.07%。经Ni-IDA亲和层析柱纯化,可获得高纯度rCagL,纯度约为96.6%。Western blot结果证实:重组抗原rCagL可特异性与His标签抗体和Anti-H. pylori抗体结合。ELISA结果证实:经rCagL免疫小鼠制备的多克隆抗体anti-CagL可特异性识别rCagL和H. pylori裂解物,具有较高的抗体特异性。结论:重组抗原rCagL在低温条件下可实现可溶性表达,经纯化可获得高纯度抗原蛋白;rCagL具有较好的抗原性,制备的多克隆抗体具有较好的免疫特异性,为发展H. pylori相关诊断试剂奠定了实验基础。     

荒漠土壤微生物量碳、氮变化对降水的响应

以腾格里沙漠东南缘的典型荒漠植被为研究对象,通过遮雨棚和滴灌系统设置5个降水梯度,即极端干旱处理、中度干旱处理、对照、增水处理I和增水处理II,研究了荒漠土壤微生物量碳(MBC)、氮(MBN)和微生物碳氮比(MBC/MBN)对季节、降水和土壤深度的响应规律,以期为极端降水事件影响干旱荒漠区土壤微生物量碳、氮及其循环规律的深入研究提供科学依据。结果表明:(1)MBC、MBN和MBC/MBN对降水处理的响应存在差异,三者的变化范围为:230.14—272.87 mg/kg,13.82—17.58 mg/kg,19.78—36.06。其中,降水处理对MBC、MBN的影响显著,对MBC/MBN的影响不显著,在极端干旱处理下,MBC、MBN均显著高于其他降水处理;(2)两年间的MBC、MBN和MBC/MBN差异显著,2017年较2016年MBC、MNB显著减少,MBC/MBN显著增加;(3)MBC、MBN和MBC/MBN变化均表现季节性差异,变化范围分别为:153.31—337.09 mg/kg,7.89—22.29 mg/kg,14.82—46.04,其中MBC、MBN为春季最高、秋季最低,M...     

Simultaneous nitrogen and carbon removal in a packed A/O reactor: effect of C/N ratio on microbial community structure

In this research, a novel packed anoxic/oxic moving bed biofilm reactor (MBBR) was established to achieve high-organic matter removal rates, despite the carbon/nitrogen (C/N) ratio of 2.7–5.1 in the influent. Simultaneous nitrification–denitrification (SND) was investigated under a long sludge retention time of 104 days. The system exhibited excellent performance in pollutant removal, with chemical oxygen demand and total nitrogen (TN) enhanced to 93.6–97.4% and 34.4–60%, respectively. Under low C/N conditions, the nitrogen removal process of A/O MBBR system was mainly achieved by anaerobic denitrification. The increase of C/N ratio enhanced SND rate of the aerobic section, where dissolved oxygen was maintained at the range of 4–6 mg/L, and resulted in higher TN removal efficiency. The microbial composition and structures were analyzed utilizing the MiSeq Illumina sequencing technique. High-throughput pyrosequencing results indicated that the dominant microorganisms were Proteobacteria and Bacteroidetes at the phylum level, which contributes to the removal of organics matters. In the aerobic section, abundances of Nitrospirae (1.12–29.33%), Burkholderiales (2.15–21.38%), and Sphingobacteriales (2.92–11.67%) rose with increasing C/N ratio in the influent, this proved that SND did occur in the aerobic zone. As the C/N ratio of influent increased, the SND phenomenon in the aerobic zone of the system is the main mechanism for greatly improving the removal rate of TN in the aerobic section. The C/N ratio in the aerobic zone is not required to be high to exhibit good TN removal performance. When C/NH₄⁺ and C/TN in the aerobic zone were higher than 2.29 and 1.77, respectively, TN removal efficiency was higher than 60%, which means that carbon sources added to the reactor could be saved. This study would be vital for a better understanding of microbial structures within a packed A/O MBBR and the development of cost-efficient strategies for the treatment of low C/N wastewater.

     

去N端信号肽的水痘-带状疱疹病毒糖蛋白E的原核可溶性表达及其免疫原性评价

水痘-带状疱疹病毒(varicella zoster virus,VZV)糖蛋白E(glycoprotein E,gE)是VZV亚单位疫苗的主要候选蛋白,但目前原核表达系统制备的gE蛋白以包涵体形式为主,可溶性差。本研究采用去除第1～30氨基酸序列的VZV gE胞外域基因,将其与原核表达载体pET32a连接,并转化至感受态细胞BL21(DE3)中。使用异丙基-β-D-硫代半乳糖苷(Isopropylβ-D-thiogalactoside,IPTG)诱导表达,His-tag柱纯化重组gE蛋白,蛋白质印迹法(Western blot,WB)检测其特异性。用该重组gE蛋白免疫BALB/c小鼠制备多克隆抗体,酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)和间接免疫荧光法检测多克隆抗体效价及特异性。结果显示,BL21/pET32a-VZV gE工程菌可以表达可溶性重组gE蛋白,纯化后纯度约为90%。WB鉴定该重组蛋白具有良好的免疫反应性。ELISA检测显示小鼠抗VZV gE多克隆抗体效价＞1∶10 000,间接免疫荧光实验结果显示该抗体特异性较高。结果表明,本研究在原核表达系统中成功表达可溶性重组VZV gE蛋白,同时该蛋白具有较强的免疫原性,这为VZV gE亚单位疫苗的研制和大规模生产奠定了基础。     

Fungal Diversity in Deep Sea Sediments from East Yap Trench and Their Denitrification Potential

Abstract

In recent years, the hadal trenches have been recognized as biological hot spots for deep sea researchers. Due to high hydrostatic pressure, low temperatures, high salinity and low nutrients, the microorganisms in hadal trenches may have unique community structure with potential for biotechnical application. Compared with bacteria and archaea, the diversity and ecological roles of fungi in hadal trenches remain largely unknown. The purpose of this study was to explore fungal diversity in deep-sea sediments of the Yap trench and their denitrification potential. In the present study, a total of 106 fungal strains were isolated from six sediment samples collected in the East Yap Trench. These fungi belonged to five classes (Dothideomycetes, Eurotiomycetes, Sordariomycetes, Cystobasidiomycetes, and Microbotryomycetes), thirteen genera (Acremonium, Alternaria, Aureobasidium, Aspergillus, Cladosporium, Cystobasidium, Engyodontium, Gliomastix, Lecanicillium, Penicillium, Phoma, Rhodotorula and Trichoderma) and eighteen species, based on morphological identification and ITS-rDNA sequence analysis. Among them, the dominant genus is Cladosporium, which accounting for 42.45% of the total fungal strains. Meanwhile, the denitrification potential of the fungal strains was also examined with two different denitrifying media (nitrate and nitrite as sole substrate, respectively). Two fungal strains (Acremonium sp. and Aspergillus versicolor), were found to be able to produce N₂O ex situ in the presence of nitrite. No fungus was found to produce N₂O by using nitrate. Our results suggest that fungi in hadal sediments, play important roles in nitrogen cycles.