南极乔治王岛冰锥洞微生物培养探索

【目的】南极洲具备独特的环境和相对的生物地理隔离,南极洲各类生境中蕴藏了大量尚未培养和难培养的微生物,也是新颖微生物物种的重要来源之一。本研究以南极冰锥洞这类特殊生境为研究对象,通过培养条件的多样化提升南极微生物的培养率和多样性,揭示南极冰锥洞可培养微生物类群多样性,为该环境可培养微生物功能研究奠定基础,也为南极极端环境未培养微生物的培养方法提供借鉴。【方法】通过采用不同培养基添加复苏促进因子(resuscitation promoting factor, Rpf)的方式,提高南极柯林斯冰盖冰锥洞生境中微生物的可培养率,探究该生境中微生物的多样性。采用4种不同营养水平的培养基,平行添加Rpf进行菌株培养,经分离纯化与16S rRNA基因鉴定,分析冰锥洞可培养微生物的多样性及培养条件对多样性的影响。【结果】本研究共分离培养细菌407株,涵盖5个门、18个科、29个属,其中：放线菌门(Actinomycetota)为优势门,占72.73%;微杆菌科(Microbacteriaceae)为优势科,占69.78%;Lacisediminihabitans属为优势属,占45.70%。从培养基效果...     

Rice ubiquitin-conjugating enzyme OsUbc13 negatively regulates immunity against pathogens by enhancing the activity of OsSnRK1a

Ubc13 is required for Lys63-linked polyubiquitination and innate immune responses in mammals, but its functions in plant immunity still remain largely unknown. Here, we used molecular biological, pathological, biochemical, and genetic approaches to evaluate the roles of rice OsUbc13 in response to pathogens. The OsUbc13-RNA interference (RNAi) lines with lesion mimic phenotypes displayed a significant increase in the accumulation of flg22- and chitin-induced reactive oxygen species, and in defence-related genes expression or hormones as well as resistance to Magnaporthe oryzae and Xanthomonas oryzae pv oryzae. Strikingly, OsUbc13 directly interacts with OsSnRK1a, which is the α catalytic subunit of SnRK1 (sucrose non-fermenting-1-related protein kinase-1) and acts as a positive regulator of broad-spectrum disease resistance in rice. In the OsUbc13-RNAi plants, although the protein level of OsSnRK1a did not change, its activity and ABA sensitivity were obviously enhanced, and the K63-linked polyubiquitination was weaker than that of wild-type Dongjin (DJ). Overexpression of the deubiquitinase-encoding gene OsOTUB1.1 produced similar effects with inhibition of OsUbc13 in affecting immunity responses, M. oryzae resistance, OsSnRK1a ubiquitination, and OsSnRK1a activity. Furthermore, re-interfering with OsSnRK1a in one OsUbc13-RNAi line (Ri-3) partially restored its M. oryzae resistance to a level between those of Ri-3 and DJ. Our data demonstrate OsUbc13 negatively regulates immunity against pathogens by enhancing the activity of OsSnRK1a.     

有机肥施用对红壤原生生物与微生物互作的影响

为明确不同施肥处理对土壤原生生物群落、微生物碳代谢活性的影响,以南方典型旱地红壤为研究对象,基于中国科学院鹰潭红壤生态实验站玉米单作系统有机培肥长期定位试验,选取不施肥(M0)、低量猪粪(M1)、高量猪粪(M2)、高量猪粪+石灰(M3)4个处理,利用高通量测序技术研究不同猪粪处理下红壤原生生物多样性、群落结构的变化,揭示原生生物与微生物互作对土壤微生物碳代谢活性和玉米产量的影响。结果表明：(1)长期施用猪粪处理下,土壤pH、有机质(SOM)、全氮(TN)、全磷(TP)、速效磷(AP)和速效钾(AK)的含量显著提高;(2)与M0处理相比,施肥处理显著提高了原生生物生物量和多样性,并且显著改变了其群落结构,其中土壤TP、pH、AP、TN、SOM和AK是原生生物群落结构变化的重要驱动因子;(3)施肥处理显著提高了土壤细菌和真菌生物量,增加了微生物碳代谢活性(Average well color development, AWCD);(4)土壤pH和AP通过影响原生生物多样性和群落结构,间接提高了微生物碳代谢活性和玉米产量。本研究结果为提升旱地红壤的生物多样性,保障土壤健康和维持生态系统服务功...     

法尼醇X受体激动剂细胞筛选体系的建立与优化

本研究旨在建立一种基于双荧光素酶报告基因检测体系的法尼醇X受体(farnesoid X receptor, FXR)激动剂细胞筛选体系,以满足对FXR激动剂先导化合物的高通量筛选。通过在报告基因质粒pGL4-luc2P-Hygro中的萤火虫荧光素酶(firefly luciferase,Luc)基因上游克隆并插入来自FXR靶基因的FXR反应元件(FXR response element,FXRE)片段,构建用于筛选FXR激动剂的报告基因质粒,并结合海肾荧光素酶内参质粒,建立能够有效反映药物对FXR激动效应的双荧光素酶报告基因细胞检测体系。通过一系列优化实验,比较了过表达RXR、鼠源和人源FXR、不同的FXRE片段、FXR过表达质粒与报告基因质粒的混合比对筛选体系诱导效率和灵敏度的影响。根据上述结果,最终确定了优化条件,优化后体系Z因子达到0.83。本研究建立了一种用于FXR激动剂筛选的改良的基于双荧光素酶报告基因检测体系的细胞筛选体系,其主要特征在于,使用多段FXR靶基因上的FXRE片段叠加组成一种新型的增强型FXRE元件,而非传统的反向重复序列-1 (inverted repeats...     

洞庭平原褐家鼠年龄分组及种群年龄动态分析

本文采用1986年10月—1989年12月在洞庭平原收集的褐家鼠(Rattus norvegicus)标本,以胴体重作指标,参考繁殖特征,将褐家鼠划分5个年龄组:Ⅰ.幼体组,雌鼠<60克,雄鼠<70克;Ⅱ.亚成体组,雌鼠60—99克,雄鼠70—119克;Ⅲ.成体Ⅰ组,雌鼠100—139克,雄鼠120—169克;Ⅳ.成体Ⅱ组,雌鼠140—189克,雄鼠170—219克;Ⅴ.老体组,雌鼠>190克,雄鼠>220克。种群年龄结构的季节动态特征是:开春时以Ⅲ、Ⅳ组占优势,初夏时Ⅰ、Ⅱ组明显增加,7、8、9月各年龄组比例较均匀,冬季以Ⅱ、Ⅲ组为主。还探讨了年龄与体重、体长、尾长及繁殖率的相互关系。     

Responses of the desert green algae,Chlorella sp. to drought stress

Desert algae are important components of the desert soil crust and play an essential role in desert soil ecosystem development. Owing to their special habitat, desert algae are often exposed to harsh environments, among which drought represents the most common stress. Green algae are considered to have drought tolerance potential; however, only a few studies have investigated this. In this study, we selected the green alga Chlorella sp., which was isolated from desert soil, and studied its physiological response to polyethylene glycol (PEG) 6000-induced drought stress. The results showed that drought stress can affect the photosynthetic efficiency of Chlorella sp., reduce its water retention ability, and destroy its ultrastructure. However, Chlorella sp. can cope with drought stress through a series of physiological regulatory strategies. Protective strategies include quick recovery of photosynthetic efficiency and increased chlorophyll content. In addition, induced synthesis of soluble proteins, lipids, and extracellular polysaccharide (EPS), and accumulation of osmotic regulatory substances, such as sucrose and trehalose, also contribute to improving drought tolerance in Chlorella sp. This study provides insights into the physiological responses of Chlorella sp. to drought stress, which may be valuable for understanding the underlying drought adaptation mechanisms of desert green algae.     

Substitution of transferrin by FeCl3 in the development of a low foetal calf serum concentration medium for KB-26.5 hybridoma cell line

KB-26.5, a murine hybridoma cell line producing an IgG₃ monoclonal antibody used in blood type determination, primarily adapted to grow at 5% foetal calf serum (FCS) concentration has been adapted to grow at 0.5% FCS, maintaining its ability to produce antibodies at the same level. In the final step of adaptation, the addition of insulin, transferrin, ethanolamine and selenium to the media formulation was studied, using factorial assay techniques to check the effect of the different compounds and to optimize their required level for satisfactory growth and antibody secretion. KB-26.5 cells required only 20 g/ml of transferrin to adapt to 0.5% FCS medium. Furthermore, transferrin could be substituted by FeCl₃, at a relatively low level of 2 g/ml. Maximum cell density decreased by 31.5% in spinner flask test, but the antibody titer was maintained, thus the specific productivity increased. However, inoculum size had to be increased three-fold with 0.5% FCS medium in order to assure cell growth.