免疫金试纸法快速检测盐酸克伦特罗

试验选取自身蛋白制备克盐酸伦特罗克伦特罗载体抗原,免疫动物后产生高效价的特异性抗体,纯化抗体与胶体金结合成免疫金,制备成简易的检测试纸条。通过检测试纸条敏感性、保存期、金标抗体工作浓度选择、操作方法对比、实验检测及与单抗金标试纸条平行检测等试验,证明建立的免疫金试纸法快速检测盐酸克伦特罗具有实用性、可靠性和稳定性。自身全血清作载体可使抗盐酸克伦特罗的ELISA效价高达4万以上。免疫金试纸法对盐酸克伦特罗的最小检测量为40ng·mL-1,试纸条在室温保存150d不影响检测结果。实验检测86份ELISA试剂盒阴性样品和42份阳性样品,其符合率分别为100%和95.2%。市售单抗金标试纸条检测结果与ELISA法的阳性符合率为95%(19/20)。     

Microbial immobilisation of C rhizodeposits in rhizosphere and root-free soil under continuous C labelling of oats

A greenhouse experiment was conducted by growing oats (Avenasativa L.) in a continuously ¹³CO₂ labeled atmosphere. The allocation of ¹³C-labeled photosynthates in plants, microbial biomass in rhizosphere and root-free soil, pools of soil organic C, and CO₂ emissions were examined over the plant's life cycle. To isolate rhizosphere from root-free soil, plant seedlings were placed into bags made of nylon monofilament screen tissue (16 μm mesh) filled with soil. Two peaks of ¹³C in rhizosphere pools of microbial biomass and dissolved organic carbon (DOC), as well as in CO₂ emissions at the earing and ripeness stages were revealed. These ¹³C maxima corresponded to: (i) the end of rapid root growth and (ii) beginning of root decomposition, respectively. The δ¹³C values of microbial biomass were higher than those of DOC and of soil organic matter (SOM). The microbial biomass C accounted for up to 56 and 39% of ¹³C recovered in the rhizosphere and root-free soil, respectively. Between 4 and 28% of ¹³C assimilated was recovered in the root-free soil. Depending on the phenological stage, the contribution of root-derived C to total CO₂ emission from soil varied from 61 to 92% of total CO₂ evolved, including 4-23% attributed to rhizomicrobial respiration. While 81-91% of C substrates used for microbial growth in the root-free soil and rhizosphere came from SOM, the remaining 9-19% of C substrates utilized by the microbial biomass was attributable to rhizodeposition. The use of continuous isotopic labelling and physical separation of root-free and rhizosphere soil, combined with natural ¹³C abundance were effective in gaining new insight on soil and rhizosphere C-cycling.     

Ca2+浓度对罗氏沼虾Gq蛋白α亚基亚细胞定位的影响

采用免疫胶体金电镜技术研究光暗适应条件下不同Ca2+浓度对罗氏沼虾(Macrobrachium rosenbergi)感光细胞中Gq蛋白α亚基亚细胞定位的影响.结果表明,对于光适应组,在高Ca2+溶液、生理溶液和低Ca2+溶液中细胞质与感杆束中胶体金密度的比值是3.51、2.13和0.93.对于暗适应组,在高Ca2+溶液、生理溶液和低Ca2+溶液中细胞质与感杆束中胶体金密度的比值是3.01、1.08和0.47.这些表明了罗氏沼虾感光细胞中Gq蛋白α亚基在暗适应条件下同时分配在感杆束和细胞质中.在光照和细胞质内Ca2+浓度适度升高的条件下,膜结合的Gq蛋白α亚基进而转化为可溶性的Gq蛋白α亚基.     

马尾松幼苗光合产物的运输与分配特征

【目的】解析光合产物在马尾松幼苗植株中的运输和分配规律,揭示马尾松生产力形成过程,为探讨不同环境干扰下马尾松光合产物分配过程的变化提供参考依据。【方法】采用 13 C同位素脉冲标记法,对1.5年生马尾松幼苗进行标记,标记结束后第0、2、5、17、24、72、120、168、216和 360 h ,按不同部位对马尾松幼苗进行全收获取样,测定 13 C含量,以监测近期合成的光合产物在马尾松幼苗中的运输和分配规律,同时测定光合产物全碳、非结构性碳水化合物(NSC)在各个器官的积累量。【结果】1)标记的光合产物在针叶中合成后,向各库器官的运输量随着时间延长由多逐渐减少,具体表现为标记结束后0~24 h内最多,24~216 h逐渐减少,216 h之后运输完成,且59%以上标记的光合产物在0~24 h内运输到各库器官。2)光合产物运输趋于稳定后,在各器官的分配大小依次为1年生叶>当年生叶>根>茎干>1年生枝>当年生枝,与生物量的分配大小一致,但与库活力大小不同,其库活力依次为当年生叶>当年生枝>1年生叶>根>1年生枝>茎干。3)各器官全碳和NSC积累量的分配与近期合成光合产物的分配大小一致,依次为1年生叶>当年生叶>根>茎干>1年生枝>当年生枝。【结论】马尾松幼苗光合产物运输速率大于0.1 m ·h -1 ,59%以上标记的光合产物在合成后的0~ 24 h 内完成向各个库器官输出。新合成的光合产物在各器官中的积累量表现为功能器官(叶和根)居多,这一分配规律有利于马尾松幼苗阶段的生长。     

Earthworm (<Emphasis Type="Italic">Metaphire guillelmi</Emphasis>) effects on rice photosynthates distribution in the plant–soil system

An experiment was conducted to investigate the effects of earthworm (Metaphire guillelmi) activities on rice photosynthates distribution in plant–soil system through ¹⁴C pulse-labelling method. Rice was planted in pots, and maize straw was mulched on the surface with or without earthworms. Rice plants at tillering stage or heading stage were labelled with ¹⁴CO₂. Plant and soil were sampled 15 days after labelling at the tillering or heading stage and at harvest. Rice growth was inhibited by earthworms (M. guillelmi) at early stage, but the inhibition disappeared at later stage. Earthworms significantly (P < 0.01) increased the ¹⁴C percentage in root at day 15 after tillering stage labelling, but the effect disappeared at harvest. Earthworms (M. guillelmi) significantly (P < 0.01) increased the ¹⁴C percentage in root at day 15 after heading stage labelling and increased ¹⁴C percentage in soil at harvest. Earthworms decreased the percentages of total organic ¹⁴C (TO¹⁴C) present as microbial biomass ¹⁴C (MB¹⁴C) and increased the percentages of total organic ¹⁴C present as dissolved organic ¹⁴C (DO¹⁴C) at all sampling times. It is suggested that earthworms might alter the transfer of plant photosynthates from the aboveground to the belowground, and thus, soil active C pool. However, these data should be also confirmed in the field.     

Estimating symbiotic N2 fixation in Robinia pseudoacacia

Estimating symbiotic di‐nitrogen (N₂) fixation is challenging, especially when working with woody N₂ fixers in field trials. Fortunately, isotope methods based on ¹⁵N natural abundance or on ¹⁵N artificial enrichment (dilution method) make it possible to estimate the proportion of nitrogen derived from the atmosphere (Ndfa) in N₂‐fixing species. These methods have been extensively used in the field for herbaceous species, much less for tree species such as alder and acacia, and rarely for black locust (Robinia pseudoacacia). The objectives of this study were to characterize the fixation potential of black locust in a plantation by using the two ¹⁵N isotope methods in the field, and to document values of isotope fractionation occurring during N₂ fixation (the B value). B values were estimated both by growing trees on an N‐free medium in controlled conditions (B_lab) and by making Ndfa calculated with the natural abundance method converge with Ndfa calculated with the ¹⁵N dilution method in the field (B_field). The two methods gave consistent estimates of the B value. B values ranging between –1.4 and –3.2‰ were found, varying with the age of the plant material. Up to 76% of the N in the black locust trees came from the atmosphere, representing more than 45 kg N ha^?1 over five years and confirming that black locust may be well adapted to N‐poor soils.     

Microbial response to exudates in the rhizosphere of young beech trees (Fagus sylvatica L.) after dormancy

Plants act as an important link between atmosphere and soil: CO2 is transformed into carbohydrates by photosynthesis. These assimilates are distributed within the plant and translocated via roots into the rhizosphere and soil microorganisms. In this study, 3 year old European beech trees (Fagus sylvatica L.) were exposed after the chilling period to an enriched ¹³C–CO₂ atmosphere (δ¹³C = 60‰ – 80‰) at the time point when leaves development started. Temporal dynamics of assimilated carbon distribution in different plant parts, as well as into dissolved organic carbon and microbial communities in the rhizosphere and bulk soil have been investigated for a 20 days period. Photosynthetically fixed carbon could be traced into plant tissue, dissolved organic carbon and total microbial biomass, where it was utilized by different microbial communities. Due to carbon allocation into the rhizosphere, nutrient stress decreased; exudates were preferentially used by Gram-negative bacteria and (mycorrhizal) fungi, resulting in an enhanced growth. Other microorganisms, like Gram-positive bacteria and mainly micro eucaryotes benefited from the exudates via food web development. Overall our results indicate a fast turnover of exudates and the development of initial food web structures. Additionally a transport of assimilated carbon into bulk soil by (mycrorhizal) fungi was observed.     

Spatial structure of deciduous forest stands with contrasting human influence in northwest Spain

Five contrasting deciduous forest stands were studied to characterize the spatial structural variability in human-influenced forests. These stands are representative of cultural forest types widely represented in western Europe: one plantation, two coppices, one wood-pasture forest and one high forest stand. All stems with DBH > 5 cm were measured and mapped, and stem DBH distributions, spatial structure of DBH, spatial point patterns and spatial associations were analysed. Spatial autocorrelation for DBH was calculated with Moran’s I correlograms and semivariograms. Complete spatial randomness hypothesis for spatial point patterns, and both independence and random labelling hypotheses for spatial associations were analysed using Ripley’s K function. The results showed that tree sizes were conditioned by particular former management systems, which determined unimodal symmetric, positively skewed or compound DBH distributions. Spatial structure was more complex when human influence became reduced. Coppice stands showed clumped spatial patterns and independence among size classes, as a consequence of sexual and vegetative establishment of new stems in open areas. The largest clumping intensity was observed in the wood-pasture with an intermediate disturbance frequency and low inter-tree competition. The high forest stand displayed spatial traits consistent with the gap-dynamics paradigm, such as clumping of smaller trees, random arrangement of larger trees, negative association between juveniles and adults, and high structural heterogeneity. It can be expected that after cessation of human interference, coppices and wood-pastures would evolve to a more heterogeneous structure, probably with a higher habitat and species diversity.     

Factors affecting the virulence of Ralstonia solanacearum and its colonization on tobacco roots

Tobacco bacterial wilt caused by Ralstonia solanacearum is a serious disease affecting tobacco cultivation in southwest China. The response surface methodology was employed to evaluate the optimal conditions of tobacco bacterial wilt, and green fluorescent protein gene (gfp) labelling was applied to monitor the location and survival dynamics of R. solanacearum (Rs::gfp) on tobacco roots and in soil under these optimal conditions. The results showed that the highest wilt incidence was 91.13%, which occurred when the population reached 6.6 × 10⁶ CFU/g soil, the temperature was 30.55 °C, and the humidity was >81.42%. The Rs::gfp densely colonized the root tips and root hairs, and cells of Rs::gfp were observed intermittently in the elongation zone or at the point of the emerging lateral roots. The Rs::gfp number in the rhizosphere soil was 10.75‐, 73.13‐ and 74.86‐times higher than that in the bulk soil at 10, 15 and 20 days after transplantation, respectively. Increased colonization by Rs::gfp was related to the population of the pathogen, the environmental temperature and the humidity in the soil. These three conditions determined whether R. solanacearum would induce tobacco wilt. This is the first study to investigate factors affecting the virulence of a tobacco wilt bacterial pathogen, which is important for conducting field diagnosis and biocontrol of tobacco bacterial wilt.