Inhibition of selected fungi by bacterial isolates fromTripsacum dactyloides L.

Summary In an incubation study addition of green manure caused a reduction in the ammonia volatilized initially from both sodic and reclaimed soils but extensive volatilization occurred from the sodic soil, amended with green manure, after the tenth day till the conclusion of the experiment after 9 weeks. Volatilization from the reclaimed soil was much less. There was a slight build up of organic carbon and ammoniacal nitrogen in both the soils though greater in the reclaimed soil. More of nitrate and nitrite accumulated in the sodic soil.     

华西雨屏区植被恢复对紫色土酸化的影响

退耕还林是控制水土流失、恢复森林植被和改善生态环境的重要举措。植被恢复进程中会发生土壤酸化,进而影响植物生长,厘清不同还林树种对土壤酸化的影响程度及作用机制有助于更好地评估退耕还林工程的整体生态效益。以华西雨屏区紫色土坡耕地(玉米)为对照,分析了退耕20a后柳杉纯林、柳杉-光皮桦混交林、慈竹林和茶园土壤pH的变化及其垂直分异。结果表明：玉米地、慈竹林、茶园、混交林和柳杉纯林0—50 cm土壤pH平均值分别为5.66、5.55、5.12、5.03和5.00,相较于玉米地,柳杉纯林、混交林和茶园土壤pH值显著下降(P<0.01),土壤酸化严重。相较于玉米地土壤pH值的均匀分布,植被恢复不同类型人工林pH值随土壤深度的增加而增加,10—50 cm土壤pH值(4.89—5.90)显著高于0—10 cm土壤pH值(4.72—5.21)(P<0.01),表层土壤酸化最明显。土壤pH值与土壤有机质含量呈极显著负相关,而与风化指数Na/K比值呈显著正相关,有机质含量和Na/K比值共同解释了土壤pH值变化的53.9%,表明植被恢复引起的土壤有机质积累与长石类矿物风化是驱动紫色土酸化的重要过程...     

大型海藻在珊瑚礁退化过程中的作用

随着全球范围珊瑚礁的退化,大型海藻在珊瑚礁区的覆盖度呈增多的趋势。大型海藻的大量生长,妨碍了珊瑚的生长、繁殖、恢复等过程。概括起来,大型海藻对珊瑚生长、繁殖及恢复过程所产生的不利影响主要包括:(1)大型海藻通过与珊瑚竞争空间和光照而影响珊瑚生长;(2)大型海藻与珊瑚直接接触时,通过摩擦作用及释放化感物质而影响珊瑚生长;(3)大型海藻的大量生长打破了珊瑚与海藻的竞争平衡,珊瑚为应对大型海藻的入侵而把用于生长和繁殖的能量转移到组织修复与防御上,进而造成珊瑚繁殖能量的减少;(4)大型海藻通过影响珊瑚幼虫的附着及附着后的存活率,而阻碍珊瑚群落的发展;(5)海藻还能通过富集沉积物、释放病原体及扰乱珊瑚共生微生物的生长等而间接影响珊瑚生长。明确的竞争机制有利于研究海藻与珊瑚的相互作用过程。在总结前人对海藻与珊瑚的竞争机制研究的基础上,把两者的竞争机制划分成物理机制、化学机制、微生物机制三大类,物理机制是研究得比较透彻的竞争机制,而化学机制与微生物机制则需要更深入的研究,是当前研究的热点。目前,我国对珊瑚礁中底栖海藻与珊瑚的相互作用研究甚少;鉴于此,对底栖海藻功能群的划分类型以及三大类型底栖海藻对珊瑚的作用特点做了简要介绍,并对珊瑚礁退化的现状和退化珊瑚礁区内海藻的表现做了概述。在此基础上,再综述国外关于大型海藻对珊瑚的影响研究进展,指出我国应该加强对南海珊瑚礁区大型海藻的种类分布及丰富度等的调查,评价大型海藻对南海珊瑚礁的影响现状;并结合生理学、分子生物学技术和生态学研究手段,在细胞与分子水平上探索海藻对珊瑚的影响机制,以期为珊瑚礁生态系统的保护提供参考。     

土壤中棉花黄萎病菌SYBR Green Ⅰ荧光RT-PCR定量检测技术研究

为实现田间土壤棉花黄萎病菌的早期检测,建立了土壤中棉花黄萎病菌的SYBR Green I荧光定量PCR检测方法。以含342bp PCR扩增产物的阳性质粒为参考,构建了标准曲线,并对该曲线的特异性、敏感性、可重复性进行了评价。结果表明,该方法具有快速、特异性强、敏感度高等特点。检测范围在3.8×103-3.8×108copies/μL之间有良好的线性关系,相关系数R2为0.996,扩增效率为101.5%,灵敏度比常规PCR方法高102倍。     

Green fluorescent protein-tagged Edwardsiella tarda reveals portal of entry in fish

The application of green fluorescent protein (GFP) to identify the portal(s) of entry of bacterial pathogens in animal hosts was studied using the fish pathogen Edwardsiella tarda and blue gourami, Trichogaster trichopterus. An immersion challenge model was utilized to mimic natural infection conditions in fish. Gastrointestinal tract, gills and the body surface of fish were found to be the sites of entry of virulent E. tarda (PPD130/91) by histological and infection kinetics studies. On the other hand, avirulent E. tarda (PPD125/87) was mainly found in the gastrointestinal tract, and the bacterial population in tissue declined over a period of 7 days.     

Shared patterns of species turnover between seaweeds and seed plants break down at increasing distances from the sea

We tested for correlations in the degree of spatial similarity between algal and terrestrial plants communities along 5500 km of temperate Australian coastline and whether the strength of correlation weakens with increasing distance from the coast. We identified strong correlations between macroalgal and terrestrial plant communities within the first 100 km from shore, where the strength of these marine–terrestrial correlations indeed weakens with increasing distance inland. As such, our results suggest that marine‐driven community homogenization processes decompose with increasing distance from the shore toward inland. We speculate that the proximity to the marine environment produces lower levels of community turnover on land, and this effect decreases progressively farther inland. Our analysis suggests underlying ecological and evolutionary processes that give rise to continental‐scale biogeographic influence from sea to land.     

The impact of CO2 emission scenarios and nutrient enrichment on a common coral reef macroalga is modified by temporal effects

Future coral reefs are expected to be subject to higher pCO₂ and temperature due to anthropogenic greenhouse gas emissions. Such global stressors are often paired with local stressors thereby potentially modifying the response of organisms. Benthic macroalgae are strong competitors to corals and are assumed to do well under future conditions. The present study aimed to assess the impact of past and future CO₂ emission scenarios as well as nutrient enrichment on the growth, productivity, pigment, and tissue nutrient content of the common tropical brown alga Chnoospora implexa. Two experiments were conducted to assess the differential impacts of the manipulated conditions in winter and spring. Chnoospora implexa's growth rate averaged over winter and spring declined with increasing pCO₂ and temperature. Furthermore, nutrient enrichment did not affect growth. Highest growth was observed under spring pre‐industrial (PI) conditions, while slightly reduced growth was observed under winter A1FI (“business‐as‐usual”) scenarios. Productivity was not a good proxy for growth, as net O₂ flux increased under A1FI conditions. Nutrient enrichment, whilst not affecting growth, led to luxury nutrient uptake that was greater in winter than in spring. The findings suggest that in contrast with previous work, C. implexa is not likely to show enhanced growth under future conditions in isolation or in conjunction with nutrient enrichment. Instead, the results suggest that greatest growth rates for this species appear to be a feature of the PI past, with A1FI winter conditions leading to potential decreases in the abundance of this species from present day levels.     

慢病毒介导的GFP-Hsp90αE47A基因在HepG2细胞表达及对细胞增殖性影响的研究

目的：建立真核细胞表达的GFP-Hsp90αE47A基因重组慢病毒载体三质粒包装细胞系统,并检测其对细胞增殖性的影响,为进一步研究HSP90分子伴侣功能奠定基础。方法：制备完整的重组慢病毒载体三质粒系统：转移质粒（Hsp90αE47A/psin-GFP）,包装质粒（ΔNRF）及包膜蛋白质粒（VSV-G）。磷酸钙法将三质粒共转染293T包装细胞,48h后收集病毒上清。将制备好的慢病毒颗粒感染HepG2细胞,在荧光显微镜下观察报告基因GFP的表达情况,Westernblot检测HepG2细胞GFP-Hsp90α表达。MTT法检测细胞增殖情况。结果：转染后的293T和感染后的HepG2细胞能观察到较强的绿色荧光,培养液上清病毒滴度约为3.0×103ifu/μl,HepG2细胞中有GFP-Hsp90α蛋白的表达。内源性Hsp90α表达无明显上升（为对照组的1.05±0.15倍,P〈0.05,t检验）,有明显外源性GFP-Hsp90αE47A蛋白的表达,为对照组内源性Hsp90α的0.68±0.12倍。外源性GFP-Hsp90αE47A蛋白的表达HepG2细胞增殖活性于第4d有明显抑制。（1.051±0.03vs1.349±0.05,P〈0.05,t检验）。结论：成功建立重组慢病毒载体的三质粒包装细胞系统,并将GFP-Hsp90αE47A基因在HepG2细胞中稳定表达,且并未引起细胞明显的热休克反应而导致的内源性Hsp90α增高;且能明显抑制细胞增殖,为后期Hsp90α分子伴侣功能进行研究奠定基础。     

Microwave-assisted Nile red method for in vivo quantification of neutral lipids in microalgae

In vivo determination of neutral lipids with Nile red fluorescence has been used as a rapid screening method for certain types of microalgae, but has been unsuccessful in others, particularly those with thick, rigid cell walls that prevent penetration of the fluorescence dye into the cell. To solve the problem, a microwave-assisted Nile red staining method for microalgal lipid determination was developed. In a two-step staining protocol, 50 and 60 s were selected as the optimal microwave times for the pretreatment and staining process, respectively. Moreover, several calibration methods for quantitative analysis of neutral lipids in microalgae were investigated and compared with conventional gravimetric methods. Factors that affected the in vivo quantification of cellular neutral lipids were also investigated. Application of the new method for detection and quantification of neutral lipids in a number of green microalgae was demonstrated.     

Kras突变基因真核表达载体的构建及在不同肝细胞株中的表达

目的：构建携带突变Kras基因,以增强型绿色荧光蛋白（EGFP）为报告基因的重组真核表达载体,并导入两种不同的肝细胞株中表达。方法：PCR扩增突变Kras目的基因,将该全长基因定向克隆至真核表达载体pEGFP-N1上,构建重组质粒载体。并利用脂质体转染人肝癌细胞株Huh7．5和鸡肝癌细胞株LMH,在活细胞状态下用荧光显微镜直接观察Kras-EGFP融合蛋白在细胞中的表达;用WesternBlotting方法验证Kras蛋白水平的表达。结果：酶切和测序证实pEGFP—N1-Kras重组质粒构建正确,将EGFP报告基因融合在突变的Kras基因的3’端;在Huh7．5和LMH中均观察到了绿色荧光,转染率分别为19％和53％;WesternBlott—ing也检测到融合蛋白的表达。结论：通过基因克隆方法成功构建了pEGFP—N1-Kras重组质粒载体,并且在Huh7．5和LMH中均稳定表达,为下一步筛选针对突变Kras基因的靶向药物奠定了基础。