应用酶标仪动力学方法监测棉蚜的抗药性

用酶标仪动力学测定法对3个抗性水平不同的棉蚜品系（R1、R2和R3）和1个敏感品系（S）的羧酸酯酶进行了研究,S、R1、R2和R3品系对α-乙酸萘酯（α-NA）的平均比活力分别为57．10、1171．69、1236．14和3293．00μmol·mgpro－1·min-1（分光光度计终点测定法）和38．24、85．27、198．14和762．25mOD·min－1·aphid-1（酶标仪动力学法）。终点测定法结果显示出不同品系间最大相差达60倍;酶标仪动力学测定法研究表明,4个棉蚜品系羧酸酶活性与其抗药性程度显著相关。通过对这两种方法的比较,酶动力学方法的测定结果更可靠。     

富营养化对白洋淀底栖-浮游耦合食物网结构和功能的影响

国际湖沼学的长期研究发现,一个完整的湖泊生态系统应包括底栖食物网和浮游食物网,而营养条件变化会显著改变浅水湖泊中底栖-浮游食物网的结构和功能。为了明晰富营养化对浅水湖泊底栖-浮游耦合食物网结构和功能的影响,以浅水草型湖泊——白洋淀为研究区,运用野外监测和ECOSIM与ECOPATH(Ew E)模型相结合方法,构建白洋淀底栖-浮游耦合食物网的概念模型,模拟1982—2011年间富营养化对白洋淀底栖路径和浮游路径的结构和功能影响:(1)野外监测的结果表明,从1999年至今白洋淀一直处于富营养化状态;(2)Ew E模型模拟结果表明1982—2006年,总生物量呈下降趋势,下降比例达66.38%;能流路径从以底栖路径为主转变为以浮游路径为主;(3)运用Pearson相关分析,结果表明:浮游植物与TN(r=0.67,P0.01)和TP(r=0.37,P0.05)呈显著正相关,而底栖藻类和大型沉水植物与TN(r=0.77,P0.01;r=0.67,P0.01)和TP(r=0.54,P0.01;r=0.36,P0.05)呈显著负相关。富营养化是白洋淀底栖初级和次级生产力向浮游初级和次级生产力转变的主要驱动力。采用科学的方法准确评估富营养化对湖泊底栖-浮游耦合食物网结构和功能的影响,可为湖泊生态系统管理提供技术和方法支持。     

艾比湖湿地芦苇根际土壤氨氧化古菌的多样性和群落结构

【目的】旨在揭示耐盐植物芦苇根际与非根际土壤AOA群落结构间的差异,为深入研究盐生植物根际土壤微生物与耐盐性之间的关系提供理论基础。【方法】应用高通量测序技术以氨单加氧酶基因(amoA)为分子标记,对新疆艾比湖湿地荒漠生态系统不同季节(春、夏、秋)芦苇根际与非根际土壤氨氧化古菌(AOA)的多样性和群落结构进行研究。【结果】结果表明,不同季节芦苇根际土壤AOA多样性和丰富度存在差异,相比非根际土壤,夏季和秋季芦苇根际土壤AOA多样性较低丰富度较高,春季多样性较高丰富度较低。芦苇根际土壤中AOA的多样性为春季夏季秋季。AOA群落组成分析表明,土壤样品中AOA群落主要集中在泉古菌门(Crenarchaeota)和奇古菌门(Thaumarchaeota),其中泉古菌门为主要优势菌门。RDA分析表明,含水量(SM)、有机质(SOM)、总氮(TN)和pH是影响芦苇根际土壤AOA群落多样性和丰富度的主要环境因子。【结论】不同季节芦苇根际土壤AOA多样性及丰富度存在差异,相比非根际土壤,芦苇根际土壤AOA更丰富。     

一个新型耐热普鲁兰酶的结构与功能

新型普鲁兰酶的研究对于普鲁兰酶制剂的国产化、打破国外垄断具有非常重要的意义。从我国云南腾冲地区轮马热泉的淤泥中分离获得了一株耐热普鲁兰酶产生菌LM 18-11,经16S rDNA序列系统进化树分析,确定该菌为厌氧芽胞杆菌Anoxybacillus属种,并从中克隆获得了耐热普鲁兰酶的编码基因,该酶在55℃-60℃、pH 5.6-6.4的范围内具有最大的反应活性。此外,该酶具有较好的热稳定性,在60℃下处理48 h,仍可保持50%以上的活力;动力学分析该酶的Vmax和Km分别为750 U/mg和1.47 mg/mL,是目前文献报道中比活力最高的耐热普鲁兰酶。同时还对该酶进行了晶体结构分析,结果显示该酶具有?-淀粉酶家族中典型的结构,在N端具有一个特殊的底物结合域,该结构域的缺失导致比活力和底物结合力都有相应降低,Vmax和Km分别为324 U/mg和1.95 mg/mL。同时,将该普鲁兰酶编码基因导入枯草芽胞杆菌中,在P43启动子的控制下,普鲁兰酶基因获得了高效表达,胞外酶活可达42 U/mL,相比初始菌种,表达活力提高40倍以上。研究表明该普鲁兰酶具有很好的应用前景。     

Interactive Effects Between Reindeer and Habitat Fertility Drive Soil Nutrient Availabilities in Arctic Tundra

Herbivores impact nutrient availability and cycling, and the net effect of herbivory on soil nutrients is generally assumed to be positive in nutrient-rich environments and negative in nutrient-poor ones. This is, however, far from a uniform pattern, and there is a recognized need to investigate any interactive effects of herbivory and habitat fertility (i.e., plant C/N ratios) on soil nutrient availabilities. We determined long-term effects of reindeer on soil extractable nitrogen (N) and phosphorus (P) and their net mineralization rates along a fertility gradient of plant carbon (C) to N and P ratios in arctic tundra. Our results showed that reindeer had a positive effect on soil N in the more nutrient-poor sites and a negative effect on soil P in the more nutrient-rich sites, which contrasts from the general consensus. The increase in N availability was linked to a decrease in plant and litter C/N ratios, suggesting that a shift in vegetation composition toward more graminoids favors higher N cycling. Soil P availability was not as closely linked to the vegetation and is likely regulated more by herbivore-induced changes in soil physical and chemical properties. The changes in soil extractable N and P resulted in higher soil N/P ratios, suggesting that reindeer could drive the vegetation toward P-limitation. This research highlights the importance of including both the elements N and P and conducting studies along environmental gradients in order to better understand the interactive effects of herbivory and habitat fertility on nutrient cycling and primary production.     

Iron supplementation promotes in vitro shoot induction and multiplication of <Emphasis Type="Italic">Baptisia australis</Emphasis>

C₄ plants can efficiently accumulate CO₂ in leaves and thus reduce wasteful oxygen fixation by the RuBisCO enzyme. Three C₄ enzymes, namely carbonic anhydrase (CA), phosphoenol pyruvate (PEPC) and pyruvate orthophosphate dikinase (PPDK), were over expressed in Oryza sativa L. ssp. indica var. Khitish under the control of green tissue specific promoters PD_54o, PEPC and PPDK, respectively. Integration of these genes was confirmed by Southern hybridization. The relative expression of PEPC, CA and PPDK were, respectively, 6.75, 6.57 and 3.6-fold higher in transgenic plants compared to wild type plants (control). Photosynthetic efficiency of the transgenic plants increased significantly along with a 12?% increase in grain yield compared to wild type plants. Compared to control plants, transgenic plants also showed phenotypic changes such as increased leaf blade size, root biomass, and plant height and anatomical changes such as greater leaf vein number, bundle sheath cells, and bulliform cells. Our findings indicate that the combined over expression of these three enzymes is an efficient strategy for incorporating beneficial physiological and anatomical features that will enable subsequent yield enhancement in C₃ rice plants.     

Structure-activity relationship at the proximal phenyl group in a series of non-peptidyl N-type calcium channel antagonists.

Selective N-Type Voltage Sensitive Calcium Channel (VSCC) antagonists have shown utility in several models of pain and ischemia. We report the structure-activity relationship at the proximal phenyl group in a series of non-peptidyl VSCC blockers.     

The α-<Emphasis Type="SmallCaps">l</Emphasis>-fucosidase from <Emphasis Type="Italic">Sulfolobus solfataricus</Emphasis>

Glycoside hydrolases form hyperthermophilic archaea are interesting model systems for the study of catalysis at high temperatures and, at the moment, their detailed enzymological characterization is the only approach to define their role in vivo. Family 29 of glycoside hydrolases classification groups α-l-fucosidases involved in a variety of biological events in Bacteria and Eukarya. In Archaea the first α-l-fucosidase was identified in Sulfolobus solfataricus as interrupted gene expressed by programmed −1 frameshifting. In this review, we describe the identification of the catalytic residues of the archaeal enzyme, by means of the chemical rescue strategy. The intrinsic stability of the hyperthermophilic enzyme allowed the use of this method, which resulted of general applicability for β and α glycoside hydrolases. In addition, the presence in the active site of the archaeal enzyme of a triad of catalytic residues is a rather uncommon feature among the glycoside hydrolases and suggested that in family 29 slightly different catalytic machineries coexist.