地黄SCoT分子标记体系的建立和指纹图谱的构建

该研究采用L_(25)(5~6)正交设计和单因素两种方法,对影响地黄SCoT-PCR反应的5个因素(模板DNA浓度,引物浓度,ddH_2O和Mix的用量以及退火温度)进行了优化。结果表明:优化后的反应体系总体积为25μL,含有8μL ddH_2O,1μL模板DNA(80 ng·μL~(-1)),1μL引物(8μmol·L~(-1))和15μL Mix,退火温度为45℃。运用30份地黄种质材料,对优化的SCoT-PCR正交体系进行多次重复验证,获得了多态性丰富、条带清晰的扩增图谱,证明该反应体系稳定可靠。利用该体系对32条SCoT引物进行两次筛选,得到14个扩增产物清晰、重复性好且多态性条带相对较高的引物。利用SCoT_4等5条引物构建了上述地黄2个种共30份种质的SCoT指纹图谱。利用这5个SCoT引物指纹图谱可将7个地黄常用栽培品种区分开。这表明SCoT分子标记体系适用于地黄主要品种亲缘关系及遗传多样性的研究,所构建的指纹图谱也为地黄常见的7个栽培品种的区分提供参考依据。     

协同分布式实验2.0 (CDE 2.0):生态学野外研究新方法

全球野外实验网络是近年来兴起的生态学实验方法,产生了“协同分布式实验”(CDE)、“分布式协作实验”(DCE)的野外实验网络和英国生物学记录中心(BRC)的野外观察网络等具体方法,但均存在尺度小、周期短或数据偏差问题.野外实验网络的协议设计应秉持实验优于观察测量、数据数量优于数据质量和尺度优于操作的原则.综合上述方法,本研究提出利用公众科学(citizen science)获得大范围、多尺度和长周期的数据,采用环境因子作为协变量对公众参与者的实验数据集进行验证后,将其作为后验概率与作为主观概率的生态学家实验数据集进行比对,以完成数据有效性验证,克服公众参与者数据质量瑕疵,并提出了相应的统计学模型.讨论了生态学实验数据中应用先验概率、先验与后验概率的逻辑关系以及后验概率发现演化过程新关系的可能性.这种方法更符合统计学采样规范,并增加了大尺度时空的样本量.该方法有望为生态学研究所寻找的一般性理论提供方法支持,新方法可称之为“协同分布式实验2.0”(CDE 2.0).     

黑果枸杞提取物泡腾片的制备及其质量评价

为了解决黑果枸杞中花青素稳定性问题,本文采用Box-Behnken设计对黑果枸杞提取物泡腾片配方进行优化,并对其进行质量评价。采用酸碱混合制粒压片法,通过单因素实验,筛选出片剂所需的辅料:崩解剂、填充剂、润滑剂以及甜味剂。采用响应面试验,结合感官评价进行处方优化,从而确定最优配方:柠檬酸32%、黑果枸杞提取物25%、碳酸氢钠24%、乳糖15%、甜蜜素3%、聚乙二醇6000 1%;对最优配方进行质量评价,各项指标均符合规定,其中花青素含量为8.06 mg/g。该泡腾片表面光滑,泡腾效果好,具有黑果枸杞香气,为实际生产提供理论依据。     

一株高产Monacolin K紫红曲霉菌株筛选、鉴定及发酵条件优化

为从天然发酵红曲米中分离的30株红曲霉菌株中筛选高产MonacolinK的菌株,并对其产MonacolinK的发酵条件进行优化。实验采用高效液相色谱法(HPLC)筛选到9株具有产MonacolinK能力的红曲霉菌株,其中以编号ZX26的菌株产MonacolinK能力最高,发酵液中Monacolin K产量达到107.6mg/L,并且产MonacolinK能力具有良好的稳定性。微生物形态学结合ITS基因同源性分析结果表明,编号ZX26菌株为紫红曲霉。进一步采用单因素试验和正交试验法优化紫红曲霉ZX26产MonacolinK的发酵条件,结果表明在培养基组分为葡萄糖70g/L,牛肉膏15g/L,NaNO32g/L,MgSO4·7H2O0.5g/L,KH2PO41.5g/L时,其最优发酵条件为:发酵初始pH4.0,接种量为7%,培养温度30℃,发酵10天,在此条件下,紫红曲霉ZX26发酵液中MonacolinK产量达到271.36mg/L,相对于培养条件优化前MonacolinK产量提高152.19%,经验证此培养条件下MonacolinK产量最佳。     

极端耐盐碱菌株的筛选及其菌肥对盐碱条件下小麦生长和土壤环境的影响

为获得具有盐碱地改良应用潜力的耐盐碱菌株,将采集于东营盐碱地的土样稀释涂布于pH 9、盐浓度100 g·L-1的Gibbson改良培养基上,共获得18株细菌.通过提高盐浓度、pH值,最终获得在pH 12、盐浓度20%的条件下仍然可以生长的极端耐盐碱菌株N14.对N14进行形态学、生理生化特征和16S rDNA序列分析鉴定,结果表明:菌株N14为马氏芽孢杆菌.盆栽试验结果表明,与盐碱土(CK)相比,N14菌肥可以显著提高小麦的生物量,株高、鲜重、干重分别提高了21.8%、57.9%、41.7%;显著增加小麦叶绿素a、叶绿素b、叶绿素总量,增长率分别为36.4%、20.0%、31.7%;显著提高盐碱土壤中的蔗糖酶、脲酶和碱性磷酸酶的活性,增长率分别为23.2%、68.8%、106.5%;显著提高小麦根系的超氧化物歧化酶、过氧化物酶和过氧化氢酶的活性,增长率分别为109.6%、17.8%、50%;显著减少小麦根系丙二醛的含量,减少率为39.8%.本研究为极端耐盐碱菌的应用提供了一条新思路,为盐碱地的改良提供了一条新途径.     

石灰岩石器使用功效的模拟实验——以重庆巫山玉米洞遗址为例

本文以巫山玉米洞遗址为例,对石灰岩石器进行复制与使用功效的模拟实验,结果表明有些石灰岩在石器打制层面表现出较好的适用性,石灰岩石器在常规的砍砸、切割、挖掘等行为活动中表现出良好的功能实用性和效率。通过实验标本与遗址出土石器的对比分析,我们认为该遗址应存在优选天然毛坯直接使用而形成的“使用石器”,类型主要集中在砍砸器和手镐等重型工具,但二次加工的石器比“使用石器”更为有效耐用;有尖类石器在作用于硬性对象或结构不稳定、受力不均时容易产生残损,与出土有尖类石器较高的残损率相吻合;把手修理在石器使用过程中显得非常必要,为遗址出土石器普遍存在把手修理现象提供了合理解释。     

Adaptation of the toxic freshwater cyanobacterium Microcystis aeruginosa to salinity is achieved by the selection of spontaneous mutants

The cyanobacterium Microcystis aeruginosa causes most of the harmful toxic blooms in freshwater ecosystems. Some strains of M. aeruginosa tolerate low‐medium levels of salinity, and because salinization of freshwater aquatic systems is increasing worldwide it is relevant to know what adaptive mechanisms allow tolerance to salinity. The mechanisms involved in the adaptation of M. aeruginosa to salinity (acclimation vs. genetic adaptation) were tested by a fluctuation analysis design, and then the maximum capacity of adaptation to salinity was studied by a ratchet protocol experiment. Whereas a dose of 10 g NaCl L^?1 completely inhibited the growth of M. aeruginosa, salinity‐resistant genetic variants, capable of tolerating up to 14 g NaCl L^?1, were isolated in the fluctuation analysis experiment. The salinity‐resistant cells arose by spontaneous mutations at a rate of 7.3 × 10^?7 mutants per cell division. We observed with the ratchet protocol that three independent culture populations of M. aeruginosa were able to adapt to up to 15.1 g L^?1 of NaCl, suggesting that successive mutation‐selection processes can enhance the highest salinity level to which M. aeruginosa cells can initially adapt. We propose that increasing salinity in water reservoirs could lead to the selection of salinity‐resistant mutants of M. aeruginosa.     

Divergent responses of Atlantic cod to ocean acidification and food limitation

In order to understand the effect of global change on marine fishes, it is imperative to quantify the effects on fundamental parameters such as survival and growth. Larval survival and recruitment of the Atlantic cod (Gadus morhua) were found to be heavily impaired by end‐of‐century levels of ocean acidification. Here, we analysed larval growth among 35–36 days old surviving larvae, along with organ development and ossification of the skeleton. We combined CO₂ treatments (ambient: 503 µatm, elevated: 1,179 µatm) with food availability in order to evaluate the effect of energy limitation in addition to the ocean acidification stressor. As expected, larval size (as a proxy for growth) and skeletogenesis were positively affected by high food availability. We found significant interactions between acidification and food availability. Larvae fed ad libitum showed little difference in growth and skeletogenesis due to the CO₂ treatment. Larvae under energy limitation were significantly larger and had further developed skeletal structures in the elevated CO₂ treatment compared to the ambient CO₂ treatment. However, the elevated CO₂ group revealed impairments in critically important organs, such as the liver, and had comparatively smaller functional gills indicating a mismatch between size and function. It is therefore likely that individual larvae that had survived acidification treatments will suffer from impairments later during ontogeny. Our study highlights important allocation trade‐off between growth and organ development, which is critically important to interpret acidification effects on early life stages of fish.     

The response of cerambycid beetles (Coleoptera: Cerambycidae) to long‐term fire frequency regimes in subtropical eucalypt forest

Fire has a varied influence on plant and animal species through direct (e.g. fire‐induced mortality) and indirect (e.g. modification of habitat) effects. Our understanding of the influence of fire regime on invertebrates and their response to fire‐induced modifications to habitat is poor. We aimed to determine the response of a beetle family (Coleoptera: Cerambycidae) to varying fire treatments and hypothesised that the abundance of cerambycid beetles is influenced by fire frequency due to modifications in habitat associated with the fire treatments. Arthropods were sampled across 3 months in annually and triennially burnt areas (treatments starting in 1952 and 1973 respectively), an area unburnt since 1946, and a former unburnt treatment, burnt by wildfire in 2006. Eleven different cerambycid taxa were collected using flight intercept panel traps, dominated by three species (Ipomoria tillides, Adrium sp. and Bethelium signiferum) which made up 99% of individuals collected. Over the sampling period the long unburnt treatment had significantly lower species richness than the triennial and wildfire treatments. Cerambycid abundance was significantly higher in the triennially burnt treatment than in all other fire treatments. Ipomoria tillides was more abundant in both frequently burnt treatments, Adrium sp. was more common in triennially burnt areas, whereas B. signiferum, was more common in the wildfire affected treatment. Some, but not all, cerambycid beetles were more common in areas with a more open understorey (i.e. resulting from frequent burning), and lower tree basal area, as this likely influences their ability to fly easily between food sources. Cerambycid abundance was positively related to the volume of coarse woody debris and healthy tree crowns. Cerambycid beetles were clearly influenced by historic fire regime, suggesting that changes in fire regime can potentially have a profound influence on arthropod assemblages, and subsequent influences on ecosystem processes, which are currently poorly understood.     

Chelate-assisted phytoextraction of lead using Fagopyrum esculentum: laboratory vs. field experiments

Abstract

The development of more sustainable remediation techniques has been receiving greater attention, as an alternative to soil excavation plan in urban gardens. An in situ phytoextraction experiment with buckwheat (Fagopyrum esculentum) was performed with a 5?mmol kg^?1 citric acid (CA) application. Joint experiments under laboratory conditions were conducted using various cultivars of F. esculentum in two soils with a Pb contamination of either geogenic or anthropogenic origin and various chelate concentrations. Results show that a minimum dose of 50?mmol kg^?1 of CA is required to lower soil pH and raise the concentration of mobile Pb–CaCl₂ for both soils. Consequently, Pb shoot uptake is increased from 6.3 to 8.9 times depending on soil type. Phytoextraction efficiency is found to be 1.3 to 2.0 times higher in the anthropogenic contaminated soil than in the soil with geogenic Pb. A scale effect has also been identified since Pb root accumulation under laboratory conditions was 2.4 times higher than in the field experiment. Despite an increase in the Pb extraction rate with CA, buckwheat appears to lack the efficiency needed to remove Pb in moderately contaminated soils. The calculated remediation period would last 166?years to remove the mobile Pb fraction.