柱层析法对菌株snef8有效成分的初步分离

采用硅胶柱层析的方法,对snef8菌株中的杀线虫活性物质进行了初步的分离,生物法测定各组份的杀线虫活性,减压浓缩有活性的组份制备粗提液.结果表明:菌液中的杀线虫活性物质主要集中于第8～第12号管中,其杀线虫活性接近于100%.此法不但可以除去部分杂质,提高粗提液的纯度,还可以应用制备型硅胶柱,大量制备富集粗提液,将其作...     

黑蜣(Bess beetle)纤维素酶的组分及其特性分析

以30日龄的黑蜣幼虫为材料,对其整体匀浆后组织液中的纤维素酶组成及活性进行了分析。结果表明:黑蜣体内存在内切β-1,4-葡聚糖酶(Cx酶)、β-葡萄糖苷酶;β-葡萄糖苷酶和Cx酶的最适宜温度为分别为40℃和50℃;Cx酶和β-葡萄糖苷酶分别在pH值为6.4和5.2时具有最大的活力;β-葡萄糖苷酶和Cx酶的最适宜反应时间为80 min。Cx酶具有比β-葡萄糖苷酶更强的温度适应性,在30～50℃温度范围内可维持最大酶活力的66.20%。     

Aggregate-related changes in network patterns of nematodes and ammonia oxidizers in an acidic soil

Nitrification plays a central role in global nitrogen cycle, which is affected by biological interaction between soil microfauna and microorganisms. However, the complexity of soil biotic communities made it difficult to reveal organizational principles of the community and the interactions among species. Here, we used the network analysis to decipher the interactions between nematodes and ammonia oxidizers within aggregate fractions under 10-year manure application, and examine their associations with soil variables and potential nitrification activity (PNA). Three aggregate fractions included large macroaggregates (>2000 μm, LA), small macroaggregates (250–2000 μm, SA), and inter-aggregate soil and space (<250 μm, IA). Aggregate factions showed a remarkable effect on association networks of nematodes and ammonia oxidizers. The average connectivity (avgK) and the number of edges in overall networks increased with increasing aggregate sizes, while the average geodesic distance (GD) followed the opposite trend. The LA network could be viewed as a better organized or a better operational soil food web with more functional interrelated members than the SA and IA networks. The modules related to PNA were significantly correlated and clustered together as meta-modules in networks of aggregate fractions. The role-shifts prevailed among the network members such as significant module memberships (MMs) and generalist/specialist operational taxonomic units (OTUs). A half of shared nodes were further identified as shared MMs, dominated by ammonia-oxidizing bacteria (AOB) especially for Nitrosospira cluster 3a and 10. Soil pH could explain partly the shift of module hubs in different networks, while grazing by bacterivores might account for three exclusively connecters related to Nitrososphaera clusters 1.1. The strongly coupled modules correlated positively to pH and total carbon (TC), regardless of aggregate fractions. The network analysis approach provided new insights into potential importance of network interactions between nematodes and ammonia oxidizers in soil nitrogen cycling.     

四种柱花草种子萌发中参与氮代谢的酶活性变化

研究了4种柱花草种子萌发过程中参与氮代谢的蛋白酶、肽酶和硝酸还原酶活性的动态变化。结果表明,蛋白酶的活性随着柱花草种子萌发的深入而不断上升,在种子即将露白胚根突破种皮的时候蛋白酶活性达到最大值。此后,随着胚芽的生长蛋白酶活性逐渐降低,其中热研2号柱花草的蛋白酶活性高于其他3种;肽酶活性的变化总体呈上升趋势;硝酸还原酶呈S型变化。了解柱花草种子萌发期各种酶活性的变化规律,为深入研究柱花草种子萌发时贮藏物质的动员和代谢机理提供数据。     

Soil aggregation: Influence on microbial biomass and implications for biological processes

Our 1988 paper, describing the effects of cultivation on microbial biomass and activity in different aggregate size classes, brought together the ‘aggregate hierarchy theory’ and the ‘microbial biomass concept’. This enabled us to identify the relationships between microbial and microhabitat (aggregate) properties and organic matter distribution and explain some of their responses to disturbance. By combining biochemical and direct microscopy based quantification of microbial abundance with enzyme activities and process measurements, this study provided evidence for the role of microbial biomass (especially fungi) in macroaggregate dynamics and carbon and nutrient flush following cultivation. In the last ten years environmental genomic techniques have provided much new knowledge on bacterial composition in aggregate size fractions yet detailed information about other microbial groups (e.g. fungi, archaea and protozoa) is lacking.We now know that soil aggregates are dynamic entities – constantly changing with regard to their biological, chemical and physical properties and, in particular, their influences on plant nutrition and health. As a consequence, elucidation of the many mechanisms regulating soil C and nutrient dynamics demands a better understanding of the role of specific members of microbial communities and their metabolic capabilities as well as their location within the soil matrix (e.g. aggregates, pore spaces) and their reciprocal relationship with plant roots. In addition, the impacts of environment and soil type needs to be quantified at the microscale using, wherever possible, non-destructive ‘in situ’ techniques to predict and quantify the impacts of anthropogenic activities on soil microbial diversity and ecosystem level functions.     

小叶九里香枝干抑菌活性成分的研究

为了获得小叶九里香枝干中抑制炭疽病菌的活性成分,在活性追踪指导下,利用硅胶柱色谱、Sephadex LH-20凝胶柱色谱及半制备液相色谱等方法对小叶九里香枝干进行了研究。经追踪分离鉴定到1个活性化合物XY-01,利用质谱、核磁共振谱鉴定化合物为九里香甲碱(Koenidine),该化合物可有效抑制香蕉炭疽病菌和芒果炭疽病菌的生长,EC50值分别为34.28和45.88μg/mL,表明九里香甲碱抑菌活性较好、结构简单,具有开发为新型植物源杀菌剂的潜在价值,九里香甲碱系首次从小叶九里香中分离得到。     

Simultaneous detection of multiple mycotoxins in broiler feeds using a liquid chromatography tandem-mass spectrometry

Mycotoxins are secondary fungal metabolites that are typically present in grain and feed ingredients used for animal feeds. An analytical method using LC-ESI-MS/MS was developed to quantify nine mycotoxins, consisting of aflatoxin B₁ (AFB₁), AFB₂, AFG₁, AFG₂, T-2 toxin, deoxynivalenol (DON), nivalenol (NIV), zearalenone (ZEA) and ochratoxin A (OTA) in broiler feeds. In total, 100 samples of broiler feeds were collected from poultry farms in Central Thailand. The survey found that AFB₁ and ZEA were the most prevalent mycotoxins in the feed samples at percentages of 93% and 63%, respectively. The limit of detections (LODs) of investigated mycotoxins was 0.20–0.78 ng/g. AFB₂, DON, AFG₁, NIV and T-2 toxin were also detectable at low contamination levels with percentages of 20%, 9%, 7%, 5% and 1%, respectively, whereas OTA and AFG₂ were not detected in any of the feed samples. These results suggest that there is a very low level of risk of the exposure to mycotoxins in feeds obtained from broiler farms in Central Thailand.     

高粱光敏色素A基因的克隆、蛋白结构与长短日照诱导表达模式

光敏色素是一类红光/远红光受体,对植物光形态建成、外部形态和生理功能等方面起着重要的调节作用。为明晰光敏色素A在调控高粱光形态建成发育过程中的作用,本研究比较分析光温敏感程度不同的高粱品种Btx623、Rio、Tx430和LH645的PhyA基因序列和PHYA蛋白的保守结构域,利用qRT-PCR明确长短日照条件处理下PhyA基因的表达模式。结果表明,Btx623、Rio和Tx430之间PhyA基因序列存在非同义突变,LH645在PhyA基因的第7个外显子上插入了5 bp碱基序列;高粱PhyA蛋白中含有1个PAS-2结构域、1个GAF结构域、1个Phytochrome结构域、2个PAS结构域、1个组氨酸激酶A结构域和1个类似组氨酸激酶的ATP激酶结构;遗传进化树分析表明,单子叶和双子叶植物PhyA蛋白明显聚为2大类,高粱PhyA、谷子PhyA和玉米PhyA1、PhyA2聚为一个亚类,相互间亲缘关系较近,与水稻PhyA同源性相对较远;在短日照(SD)条件下各时期PhyA基因转录水平变化显著,Rio PhyA基因转录水平较Btx623和Tx430变化明显,LH645 PhyA基因转录变化不显著,在长日照(LD)条件下Rio PhyA基因转录水平低于Btx623和Tx430,LH645 PhyA基因转录水平变化不显著。以上结果表明,PhyA受光周期诱导,推测其在高粱光形态建成中起重要调控作用。本研究为进一步解析PhyA基因功能及其在调控高粱光形态建成发育过程中的作用提供理论基础。     

湖南永州尖吻蝮蛇蛇毒的蛋白组分及毒性分析

采用Nano–LC–ESI–MS/MS蛋白鉴定技术对湖南永州尖吻蝮蛇蛇毒蛋白组分进行质谱分析,应用常规体外试管法对尖吻蝮蛇蛇毒体外溶血值进行检测,采用改良寇式法对尖吻蝮蛇蛇毒LD_(50)进行测定。结果显示:尖吻蝮蛇蛇毒含有50种蛋白,蛋白相对分子质量集中在1.0×10~4～2.0×10~4(46.9%)、4.0×10~4～5.0×10~4(22.4%)、2.0×10~4～3.0×10~4(10.6%)和7.0×10~4～8.0×10~4(7.6%),其中代表性的高丰度蛋白有蛇毒金属蛋白酶A(11.7%)、蛇毒金属蛋白酶H1(9.8%)、蕲蛇类凝血酶–2(7.3%)、抗凝血酶A–A亚基(6.8%),低丰度蛋白(0.1%)有Ecto–5'–核苷酸酶、碱性磷脂酶A2 DAV–N6、生长分化因子11;蛇毒引起红细胞溶血的最低质量浓度为60.00μg/mL,尖吻蝮蛇蛇毒对KM小鼠的LD_(50)为7.1796mg/kg,攻毒小鼠出现呼吸急促、躁动不安、注射部位出现奇特瘙痒和大面积溃烂,至死亡。