应用Illumina MiSeq高通量测序技术分析河西走廊地区盐碱土壤微生物多样性

【目的】以甘肃省河西走廊地区的9个盐碱土壤样品(原生盐碱土、次生盐碱土、农田土)为材料,研究该地区盐碱土壤中微生物群落的多样性。【方法】提取土壤微生物总DNA,应用Illumina Mi Seq高通量测序技术进行分析。【结果】从分布在河西走廊3个流域的9个盐碱土样品中共获得325 089条微生物的16S r RNA基因序列。冗余分析和热图分析表明,原生盐碱土与次生盐碱土、原生盐碱土与农田土微生物群落构成差异较大,次生盐碱土与农田土微生物群落差异较小。土壤p H对微生物群落组成的影响最显著。多样性指数和稀释性曲线分析得出,在9个土壤样品中,S6号Shannon指数最大,S1号Shannon指数最小,S1号Simpson指数最大,S6号Simpson指数最小,说明原生盐碱土的微生物群落多样性最低,次生盐碱土的微生物群落多样性最高。盐碱土壤中主要的微生物群落包括9个门,其中变形菌门占主导地位,其余依次是放线菌门、拟杆菌门、酸杆菌门、浮霉菌门、绿弯菌门、芽单胞菌门、厚壁菌门和疣微菌门。原生盐碱土和农田土中占优势的微生物群落是变形菌门,次生盐碱土中占优势的微生物群落是放线菌门。【结论】河西走廊地区盐碱土壤中微生物多样性非常丰富,存在大量的微生物类群,尤其是在次生盐碱土壤中。     

小鼠淋巴细胞抗原CTLA-4的胞外肽段表达及纯化

目的: 真核细胞表达小鼠淋巴细胞抗原CTLA-4胞外段肽,研究表达肽段与抗原呈递细胞B7分子结合后减轻小鼠淋巴细胞刺激后的增殖抑制,从而启动T淋巴细胞进一步增殖。方法:从小鼠脾脏淋巴细胞获得总RNA,通过逆转录PCR扩增出CTLA-4全长基因,克隆并测序。依据胞外段序列和真核表达载体pcDNA3.1序列,合成引物扩增胞外片段,两者经内切核酸酶处理、连接构建重组表达pcDNA3.1载体,重组质粒经测序验证后,采用lipofectamine 2000转染入小鼠肝癌细胞Hepa1-6,经G418筛选获得稳定表达细胞株。结果:获得小鼠CTLA-4胞外段真核表达载体和小鼠肝癌细胞Hepa1-6稳定表达转染细胞株,制备了CTLA-4胞外肽段,经His标签抗体和小鼠CTLA-4抗体Western blot检测表达蛋白带均呈阳性。结论:获得CTLA-4胞外段肽,为进一步研究该肽的作用打下基础。     

天童常绿阔叶树种栲树生殖个体大小及其生殖构件特征

对浙江天童木荷-栲树林内的常绿阔叶树种栲树(Castanopsis fargesii Franch.)的生殖个体大小、生殖构件的分布及其动态变化特征进行了研究。结果表明, 该地区栲树生殖个体的胸径在17~50 cm 间, 平均胸径为31. 2±8.0 cm, 平均年龄约36.3±6.6 年;林缘附近的生殖个体小于木荷-林内。相对稳定的群落和比较丰富的土壤养分条件有利于生殖枝数量和花序数量的增多。栲树生殖个体的数量在两年中变化较大, 部分栲树个体可以在连续年份中生殖。从枝系水平分析:在持续生殖的栲树个体上, 生殖枝数量有明显变化, 并非所有的生殖枝在两年中都可开花或结果, 保持连续生殖的枝系约占48.2%。栲树果序枝数量在连续年份有明显差异(p < 0.01), 而且果序枝上的幼蕾数、果实数量及结实率等都有明显差异(p < 0.05)。     

Social perspectives on the use of reference conditions in restoration of fire‐adapted forest landscapes

As approaches to ecological restoration become increasingly large scale and collaborative, there is a need to better understand social aspects of restoration and how they influence land management. In this article, we examine social perspectives that influence the determination of ecological reference conditions in restoration. Our analysis is based on in‐depth interviews with diverse stakeholders involved in collaborative restoration of fire‐adapted forest landscapes. We conducted interviews with 86 respondents from six forest collaboratives that are part of the U.S. Forest Service's Collaborative Forest Landscape Restoration Program. Collaboratives use a variety of approaches to develop reference conditions, including historic, contemporary, and future scenarios. Historical conditions prior to European settlement (nineteenth century or “pre‐settlement” conditions), or prior to more recent grazing, logging, and exclusion of fire, were the predominant type of reference used in all sites. Stakeholders described benefits and limitations of reference conditions. Primary benefits include (1) providing a science‐based framework for bringing stakeholders together around a common vision; (2) gaining social understanding and acceptance of the underlying need for restoration; and (3) serving to neutralize otherwise value‐laden discussions about multiple, sometimes competing, resource objectives. Limitations stem from (1) concerns over social conflict when reference conditions are perceived to contradict other stakeholder values and interests, (2) differing interpretations of reference condition science, (3) inappropriate application or over‐generalization of reference information, and (4) limited relevance of historical references for current and future conditions in some ecosystems. At the same time, collaboratives are adopting innovative strategies to address conceptual and methodological limitations of reference conditions.     

胞外ATP对AlCl3诱导的细胞死亡过程中胞内H2O2和Ca2+水平的影响及其生理机制分析

该实验以烟草悬浮细胞 BY 2 为材料,在烟草悬浮细胞中分别加入0.05、0.10、0.15、0.20 mmol·L^-1AlCl₃,以等体积去离子水处理的悬浮细胞液为对照,并依据前述实验结果选择0.15 mmol·L^-1 AlCl₃,分别添加5 mmol·L^-1 DMTU(H₂O₂ 抑制剂)、20 μmol·L^-1CaCl₂、15 μmol·L^-1 LaCl₃(Ca²⁺通道抑制剂)和50 μmol·L^-1 ATP设计多项处理,分析胞外ATP(eATP)对铝离子(Al³⁺)胁迫引起的植物细胞死亡及其胞内H₂O₂、Ca²⁺的影响,以揭示Al³⁺胁迫下植物调节细胞死亡的可能机制,进一步扩展对eATP功能的认知。结果显示：(1)随着 AlCl₃ 胁迫浓度的提高,细胞死亡水平和胞内H₂O₂水平上升,而胞内Ca²⁺和eATP水平则逐渐降低。(2)外援施加H₂O₂抑制剂 DMTU(二甲基硫脲)和Ca²⁺能够有效缓解AlCl₃诱导的细胞死亡水平的上升;而Ca²⁺通道抑制剂LaCl₃(三氯化镧)则加剧了AlCl₃胁迫下的细胞死亡。(3)在AlCl₃胁迫下对细胞添加外源ATP,能够缓解AlCl₃胁迫下胞内H₂O₂水平上升和Ca²⁺水平下降的同时,并显著降低AlCl₃胁迫导致的细胞死亡。研究表明, Al³⁺以剂量依赖的模式提升细胞死亡和细胞内H₂O₂的水平并降低胞内Ca²⁺和eATP水平,AlCl₃诱导的细胞死亡受到H₂O₂和Ca²⁺水平变化的调节,eATP可以通过调节H₂O₂与Ca²⁺水平缓解AlCl₃诱导的细胞死亡。推测Al³⁺胁迫可能通过抑制钙离子通道而破坏了细胞内H₂O₂和Ca²⁺之间的协同关系,外源ATP对Al³⁺诱导H₂O₂上升的缓解作用可能是由于其提升了细胞的抗氧化能力。     

Tropomyosin is localized in the nuclear matrix and chromosome scaffold of physarum polycephalum

The nuclei and chromosomes were isolated from plasmodia of Physarum polycephalum.The nuclear matrix and chromosome scaffold were obtained after the DNA and most of the proteins were extracted with DNase I and 2 M NaCl.SD-PAGE analyses revealed that the nuclear matrix and chromosome scaffold contained a 37 kD polypeptide which is equivalent to tropomyosin in molecular weight.Immunofluorescence observations upon slide preparations labeled with anti-tropomyosin antibody showed that the nuclear matrix and chromosome scaffold emanated bright fluorescence,suggesting the presence of the antigen in them.Immunodotting results confirmed the presence of tropomyosin in the nuclear matrix and chromosome scaffold.Immunoelectron microscopic observations further demonstrated that tropomyosin was dispersively distributed in the interphase nuclei and metaphase chromosomes.