慢性蜜蜂麻痹病病毒核酸与蛋白质组分的研究

从自然感染的意大利麻痹病蜂(APis mellifera)头部,经二次差速离心与蔗糖梯度离心获得纯化的慢性蜜蜂麻痹病病毒(CBPV)。纯化的CBPV制备物感染正常蜜蜂,4天后出现典型的麻痹症状,接着死亡,平均死亡率分别为95％与100％。SDS-聚丙烯酰胺凝胶电泳分析,二次差速离心初步纯化的病毒制备物含有多条蛋白带,而蔗糖密度梯度纯化的病毒制备物仅含有单一的多肽带。5％、7.5％与10％SDS-聚丙烯酰胺凝胶电泳分析,均检测出一种病毒蛋白质,分子量大约为24,200道尔顿,而且不同凝胶浓度检测的蛋白质分子量相近。慢性蜜蜂麻痹病病毒核酸也用SDS-聚丙烯酰胺凝胶电泳分析,结果表明,凝胶中有5条带,对核酸酶敏感,证明该病毒含有5个单股RNA组分。对慢性蜜蜂麻痹病病毒的基因组结构进行了讨论。     

短期氮添加对东灵山三种森林土壤呼吸的影响

为了分析氮沉降对我国温带地区森林土壤碳循环的影响,以北京东灵山的阔叶林(辽东栎林)和针叶林(华北落叶松林和油松林)为研究对象,通过模拟氮沉降的方式(10g N·m-2·a-1,大约5倍于大气氮沉降速率),探讨了不同温带森林土壤呼吸对氮沉降的短期响应。结果表明:短期氮添加降低了阔叶林土壤呼吸速率,而提高了针叶林土壤呼吸速率,但其短期效应未达到显著性水平。不同森林类型间,土壤呼吸速率(P0.001)和生长季土壤呼吸释放总量(P0.001)均存在显著差异,整体表现为:辽东栎林油松林华北落叶松林,土壤温度是引起不同森林类型间土壤呼吸差异的主要因素。温度-水分双因素模型可以较好地模拟野外条件下3种森林类型土壤呼吸与温度和水分间的关系,解释率约为47%~87%。此外,氮添加可以改变土壤呼吸对温度和水分变化的响应:氮添加后在较高温度且较低水分情况下,土壤呼吸速率明显上升,此时土壤呼吸对温度变化更加敏感。实验结果揭示了氮沉降对我国温带地区不同森林类型土壤呼吸的影响,但其复杂的影响机制仍有待进一步研究。     

出芽短梗霉膨大细胞分选及产糖分析

本研究运用Percoll密度梯度离心的方法对出芽短梗霉Aureobasidium pullulans的两种细胞形态进行分选,并对两种形态的细胞进行多糖产量的分析。通过对转速、分选时间、Percoll分离液浓度的优化,确定了两种细胞形态分选效果最佳的条件是Percoll分离液浓度为60%、转速为5 000r/min、离心时间为30min。经过光学显微镜和透射电子显微镜观察发现上层为酵母状细胞（YL）、下层为膨大细胞（SC）,并发现膨大细胞外有明显的薄膜包被,且产大量多糖。也为今后在相应状态下研究出芽短梗霉膨大细胞的其他代谢机理提供了可行的方法,满足后续研究的需要。     

Effects of simulated nitrogen deposition on fine root morphology,nitrogen and phosphorus efficiency of Pinus massoniana clone under phosphorus deficiency

Aims In forest ecosystems with phosphorus (P) deficiency, the impact of atmospheric nitrogen (N) deposition on nutritional traits related to N and P uptake potentially affect plant growth and vegetation productivity. The objective of this study was to explore the effects of simulated N deposition on fine root morphological characteristics and effiency of N and P absorption in Pinus massoniana under under low P stress.
Methods Two clones of P. massoniana seedling with different P efficiency (high P efficiency 19-5 vs. low P efficiency 21-3) were used. A two-year pot experiment was applyed with treatments of two P conditions, (i.e. homogeneous low P availability vs. heterogeneous low P availability) and three N deposition levels (0, 30 and 120 kg N·hm^-2·a^-1; i.e., N0, N30, or N120, respectively) .
Important findings 1) The growth of P. massoniana seedling was interactively affected the three factors: simulated N deposition, P condition and genotypes. Simulated N deposition increased the seedling height and dry mass under heterogeneous P deficiency, but did not significantly affect those traits under homogeneous P deficiency. Under heterogeneous P deficiency and N120 treatment, the seedling height and dry mass of clone 19-5 were 1.1 times and 1.6 times higher than that of clone 21-3, respectively. 2) Fine root length and surface area decreased as root diameter increased. N deposition significantly stimulated proliferation of fine root with diameter ≤1.5 mm, while roots with diameters ranged from 1.5 to 2.0 mm and over 2.0 mm were not influenced. The length of fine root ≤1.5 mm in diameter accounted for 90.4%-92.8% of the total root length and was not affected by N deposition. 3) Under the heterogeneous low P condition, clone 19-5 was found to respond to the simulated N deposition with increased root length and surface area in fine-root diameter class of ≤1.5 mm. Additionally, in compared with control, its N and P absorption efficiency were significantly enhanced 93.3% and 148.4%, respectively under N120 treatment. However, the N and P absorption efficiency of clone 21-3 was less affected by the simulated N deposition. The N and P use efficiency had no notable variation. Finally, we found that the proliferation of fine-root ≤1.5 mm in diameter and high N (P) absorption efficiency maybe the adaptive mechanisms of P. massoniana responding to atmospheric N deposition under P deficiency.     

模拟氮沉降和干旱对准噶尔盆地两种一年生荒漠植物生长和光合生理的影响

氮素和水分是荒漠生态系统的两个主要限制因子, 研究两者对荒漠植物的效应有助于深入了解荒漠生态系统对全球变化的响应。该文选择准噶尔盆地荒漠地区两种常见的一年生植物涩荠(Malcolmia africana)和钩刺雾冰藜(Bassia hyssopifolia), 设置0、0.18和0.72 g N·m ^-2·week ^-13个施氮浓度和湿润与干旱两个土壤水分处理, 研究模拟氮沉降增加和干旱对其生长和光合生理的影响。结果表明: (1)两种植物的根长、根重、叶片数、叶面积、总生物量和冠根比均随着施氮浓度的增加而增加, 干旱能够抑制氮对植物生长的促进作用, 但是, 氮的增加同时也能部分缓解干旱对植物生长的影响。与钩刺雾冰藜相比, 涩荠的根长、生物量和冠根比更易受氮增加和干旱的影响。(2)两种植物的最大净光合速率、叶绿素含量、可溶性蛋白含量随着氮浓度增加而增加, 但涩荠和钩刺雾冰藜对氮增加和干旱的生理响应也有所不同, 涩荠的响应更加敏感。两种植物对氮沉降和干旱胁迫响应的差异可能是其生活型等生物学特性差异所引起。通过对两种一年生植物的生长和光合生理分析表明, 在古尔班通古特沙漠, 春季丰富的降水和氮素增加将有利于涩荠和钩刺雾冰藜的生长和生产力的增加, 相对地下生长, 地上部分增加更显著。当干旱季节来临时, 氮的增加又能够在一定程度上降低干旱对这两种植物的负效应, 说明其对干旱具有一定的生态补偿作用。     

Responses of nitrogen and phosphorus resorption from leaves and branches to long-term nitrogen deposition in a Chinese fir plantation

为了解森林养分内循环对全球变化的响应, 基于长期模拟氮沉降试验, 研究了杉木(Cunninghamia lanceolata)人工林不同龄级(一年生、二年生和衰老)叶和枝的氮(N)、磷(P)养分分配及其再吸收特征, 并分析了不同模拟N沉降处理时间(7年和14年)杉木叶N、P养分再吸收差异。在12年生杉木中开展模拟N沉降试验, 以尿素(CO(NH₂)₂)为N源, 设N0、N1、N2和N3 4个处理水平, 施氮量分别为0、60、120和240 kg·hm ^-2·a ^-1, 每个处理重复3次。结果表明: (1)叶和枝在衰老过程中碳(C)、N和P含量逐渐降低, 且叶的C、N和P含量比枝高; N含量大小依次为一年生叶>二年生叶>衰老叶>一年生枝>二年生枝>衰老枝, 且N3 > N2 > N1 > N0, 而C:N则呈现相反的趋势; 衰老器官的C:N、C:P、N:P比新鲜器官高; N沉降增加了不同龄级叶和枝(除二年生叶外)的N、N:P和C:P, 但降低了P和C:N。(2)叶和枝的N、P养分再吸收率(RE_N、RE_P)随龄级的增加至衰老有规律地递减, 且RE_P > RE_N; 受长期N沉降的影响, RE_N叶(28.12%) <枝(30.00%), 而RE_P则为叶(45.82%) >枝(30.42%); 杉木叶和枝N:P与RE_N:RE_P之间存在极显著的线性相关关系。(3)随N沉降处理时间的增加, 叶RE_N呈降低态势, 各处理(N1、N2和N3)分别降低了9.85%、3.17%和11.71%; 而RE_P则明显上升, 分别增加了71.98%、42.25%和9.60%。研究结果表明: 不同器官、不同龄级的养分再吸收率随氮沉降处理的水平、处理时间而所有不同; RE_N:RE_P与N:P之间存在紧密关系。     

Effects of nitrogen addition on leaf traits of common species in natural Pinus tabuliformis forests in Taiyue Mountain,Shanxi Province,China

Aims Our objectives were to explore the effects of simulated nitrogen deposition on leaf traits and ecological stoichiometry characteristics of common species in natural Pinus tabuliformis forests.
Methods We conducted the experiment of nitrogen (N) addition from 2009 to 2013 in the natural Pinus tabuliformis forests in Taiyue Mountain, Shanxi, China. The levels of N addition were 0 (control), 50 (low-N), 100 (medium-N) and 150 (high-N) kg·hm^-2·a^-1, respectively. Eleven common plant species in 12 20 m × 20 m plots were selected, including Pinus tabuliformis, Quercus mongolica, Acer ginnala, Corylus mandshurica, Cornus bretschneideri, Spiraea salicifolia, Lonicera maackii, Carex callitrichos, Diarrhena mandshurica, Anemone tomentosa, and Polygonatum odoratum. Nine leaf traits were measured, including leaf thickness (LT), specific leaf area (SLA), leaf dry matter content (LDMC), leaf nitrogen content (LNC), leaf phosphorus content (LPC), and other four.
Important findings We found that: 1) LT and SLA of Polygonatum odoratum significantly differed among four levels of N addition. Leaf area (LA) and LDMC of several species, such as Spiraea salicifolia, had significant difference among the N addition concentration. LNC of all species, chlorophyll content (CC) and LPC of most species increased significantly with the addition of N. Leaf N:P of 9 species varied significantly, and leaves with different types and ages showed different responses to N addition. 2) Leaf traits were significantly correlated with each other. For instance, SLA was significantly positively correlated with LNC and LPC. In contrast LT was negatively connected with LNC and LPC. In addition, the degree of correlation changed with the level of N addition. 3) The pattern of species distribution in leaf trait space was consistent with the prediction from the theory of Leaf Economic Spectrum (LES). N addition drove species moving along axis 1 in the trait space, and propelled them towards different directions along axis 2, which indicated that these species tended to take the “fast investment-return” strategy. These results suggested that with the change of environmental conditions, plants changed their survival strategy and adjusted resource allocation to maintain the stability of communities. This is the inherent characteristic of plants, thus the formation of LES did not depend on the environment change.     

N添加对西南亚高山红桦林根系分泌物及其介导的养分转化过程的影响

目前缺乏对根系分泌物通量以及相关生态后果对不同氮(N)沉降水平响应方向和幅度的深入理解,该研究以西南亚高山典型的红桦(Betula albosinensis)林为研究对象,通过野外原位N添加试验模拟不同氮沉降水平(对照组,0 kg·hm^-2·a(-1);低氮处理,25 kg·hm^-2·a(-1);高氮处理,50 kg·hm^-2·a(-1)),分析了红桦林根系分泌物C输入通量及其介导的根际土壤养分循环过程对不同N添加水平的差异化响应,试图揭示不同N添加处理对红桦根系分泌物C输入通量及其介导的土壤养分转化过程的影响。结果表明：(1)N添加显著抑制了红桦林根系分泌物C输入速率(其中低氮(N25)条件下单位根生物量根系分泌速率均值降低约14.87%)和年C输入通量(低氮条件下降低了约45.01%)(P<0.05),其高氮处理的抑制效应更强。(2)N添加显著抑制了红桦林N矿化速率及其相关的微生物胞外酶活性(P<0.05),并显著降低了其根际效应;N沉降显著抑制了根系分泌物C输入通量及其介导的土壤养分转化过程,并且这种抑...     

土壤微生物对气候变暖和大气N沉降的响应

气候变暖和大气N沉降是近一、二十年来人们非常关注的全球变化现象,它们所带来的一系列生态问题已成为全球变化研究的重要议题。它们不仅影响地上植被生长和群落组成,还直接或间接地影响土壤微生物过程,而土壤微生物对此做出的响应正是生态系统反馈过程中非常重要的环节。该文分别从气候变化对土壤微生物的影响(土壤微生物量、微生物活动和微生物群落结构)和土壤微生物对气候变化的响应(凋落物分解、养分利用与循环以及养分的固持与流失)两个角度,综述近期土壤微生物对气候变暖和大气N沉降响应与适应的研究进展。气候变暖和大气N沉降对土壤微生物的影响更多地反映在微生物群落的结构和功能上,而土壤微生物量、微生物活动和群落结构的变化又会通过改变凋落物分解、养分利用和C、N循环等重要的土壤生态系统功能和过程做出响应,形成正向或负向反馈,加强或削弱气候变化给整个陆地生态系统带来的影响。然而,到目前为止土壤微生物的响应对陆地生态系统产生的最终结果仍是未决的关键性问题。     

狂犬病疫苗蔗糖密度梯度离心纯化工艺开发

目的:开发冻干人用狂犬病疫苗(鸡胚成纤维细胞)的连续流蔗糖密度梯度离心纯化工艺。方法:分别比较两种不同初始蔗糖浓度和不同上样速度对纯化效果的影响,初步确定纯化工艺;通过多批次实验确定样品收集范围;比较不同浓缩倍数条件下杂质去除和抗原回收情况,确定合适的收获液浓缩比例;比较不同批次样品纯化后的杂质去除率和重复性,判定本纯化工艺的稳定性。结果:选取60%作为初始蔗糖浓度,在上样速度为150～200ml/min时,可以有效地对10倍浓缩的病毒收获液进行纯化;卵清蛋白、牛血清白蛋白和庆大霉素去除率分别达到99%,95%和95%,且工艺具有极好的稳定性。结论:开发的连续流蔗糖密度梯度离心技术可以作为冻干人用狂犬病疫苗(鸡胚成纤维细胞)的产业化纯化工艺。