高压脉冲电场对酿酒酵母蛋白质及DNA的影响

为探究高压脉冲电场（PEF）对微生物的钝化机理,本实验主要采用Lowry法、SDS-PAGE电泳、APIZYM试剂盒、流式细胞仪等方法测定PEF处理对酿酒酵母蛋白质含量、胞内酶活性和DNA等的影响。结果显示：PEF处理后,酿酒酵母胞内蛋白质含量显著降低(p<0.05),SDS-PAGE 电泳图分析发现酿酒酵母胞内蛋白谱图不清晰,且有小分子和 60 KDa 以上大分子蛋白谱带缺失,酿酒酵母细胞内 13 种酶的活性均有不同程度的下降,细胞DNA含量降低,部分DNA变性。结论：PEF钝化微生物与其胞内酶失活、蛋白质和DNA发生外泄或变性有关。     

脉冲电场对果胶酯酶的活性及构象的影响

为了采用非热杀菌技术以保持果汁的新鲜度,研究了高压脉冲电场(PEF)对果胶酯酶(PE)的活性及酶构象的影响效果。结果表明PEF可以钝化PE的活性：随脉冲电场电场强度的增强(5～25 kV/cm)和脉冲数的增加(207～1449个)PE酶活钝化效果增强;在PEF处理后立刻测定发现25 kV/cm、1449个脉冲时,PE酶活性最高降低了65.3％。PE酶活变化与PEF电场强度和脉冲数之间存在良好的线性相关,R²分别达到0.988和0.953。PEF处理(621个脉冲)后PE的荧光强度增加,整体变化趋势是随电场强度增强荧光强度先增加后下降,但荧光强度值都低于热处理(100℃处理5 min)后酶蛋白的荧光强度值。因此PEF可以有效钝化果胶酯酶的活性,并且改变果胶酯酶的构象。     

高密度CO2对虾优势腐败菌的杀菌效果及机理

为了探讨高密度CO2（dense phase carbon dioxide,DPCD）对水产品腐败菌的杀菌效果和机制,以一株凡纳滨对虾优势腐败菌（Chryseobacterium sp. LV1）为研究对象,研究了DPCD处理温度（30～55℃）、压力（5～25 MPa）、时间（5～60 min）对杀菌效果的影响,分析了DPCD处理前后该菌理化性质的变化。结果表明：DPCD对其具有较好的杀菌效果,温度升高、压力增大、延长时间都会增强杀菌效果,而且超临界CO2比亚临界CO2的杀菌效果好。当在45℃、15 MPa和55℃、15 MPa下处理菌悬液30 min时,菌落总数均能下降5个对数;菌悬液的pH值从6.97分别下降至5.58和5.56;细胞外蛋白由最初的78.69 μg/mL分别增至151.91和157.40 μg/mL,200～800 nm范围内的吸光度值增大,这说明DPCD处理改变了其细胞膜的通透性,造成胞内蛋白质和核酸泄漏;可溶性和不溶性蛋白的电泳图谱发生变化,说明DPCD处理能够诱导Chryseobacterium sp. LV1可溶性蛋白质变性,降低其溶解度;能够钝化与其新陈代谢相关的14种酶类;但不会造成其DNA的降解。因此,DPCD处理致使其理化性质的改变可能是杀菌的主要原因之一。研究结果将为DPCD技术在对虾加工中的应用提供参考。     

菜田种养结合模式下施肥方式对土壤编码碱性磷酸酶基因微生物群落的影响

为明确菜田种养结合模式下施肥方式对土壤编码碱性磷酸酶基因phoD的微生物群落结构和多样性的影响机制,通过Illumina MiSeq高通量测序手段,系统分析了4种不同施肥方式,即不施肥（CK）、施常规化肥（CF）、施有机肥（OF）、有机无机肥混施（MF）对花菜收获时0−20cm土层土壤理化性质、碱性磷酸酶（ALP）、微生物量磷（MBP）以及phoD微生物群落的影响。结果表明：（1）与CK相比,OF处理可显著提高土壤有机质、总氮、速效磷和Ca含量47.83%、38.46%、104.81%和69.21%（P<0.05）;OF和MF处理均显著提高ALP活性;CF和OF分别显著增加MBP含量56.12%和195.16%,OF处理中MBP含量最高（105.40mg·kg⁻¹）;（2）菜田种养结合模式不同施肥处理中假单胞菌属（Pseudomonas）为优势属,CF和MF较CK显著降低了假单胞菌属相对丰度33.39%和45.52%;施肥降低土壤phoD微生物Chao1指数,MF提高其多样性（Simpson）和均匀度（Simpsoneven）;（3）影响phoD微生物群落结构的关键环境因子为MBP、AP、ALP;phoD微生物α多样性指数与土壤性状指标无显著相关性。因此,菜田种养结合模式下,不同施肥处理改变了土壤理化和生物性质,从而驱动了土壤phoD微生物群落组成、结构和多样性变化。     

鱼源嗜水气单胞菌外膜蛋白基因ompTS在枯草芽孢杆菌中的表达及检测

用PCR方法从枯草芽胞杆菌（Bacillus subtilis）、鱼源嗜水气单胞菌（Aeromonas hydrophila）的基因组DNA和质粒pGFP-2中,分别扩增P43启动子、不含信号肽的外膜蛋白基因ompTS和不含启动子的绿色荧光蛋白基因。各PCR产物经测序、酶切、连接到大肠杆菌（Escherichia coli）-枯草芽胞杆菌穿梭载体pNW33N相应位点分别构建了穿梭表达载体pNWP43gfp和pN-WP43omp。电转枯草芽胞杆菌构建菌株BS01（pNWP43gfp）,在荧光显微镜下发出明亮的绿色荧光;电转枯草芽胞杆菌构建菌株BS01（pNWP43omp）,表达的外膜蛋白经SDS-PAGE和Western blot检测,结果表明,穿梭表达载体pNWP43gfp和pNWP43omp构建成功。构建的枯草芽胞杆菌在P43启动子的调控下,分别实现了绿色荧光蛋白基因和外膜蛋白基因ompTS的表达。     

高压脉冲电场对酵母活性变化与致死双重效应研究

为探究高压脉冲处理后酿酒酵母在工业生产领域的实际应用,以酿酒酵母为材料,研究高压脉冲电场处理后酿酒酵母活性变化和致死双重效应。在控制电场强度为唯一变量,其他影响因素不变(脉宽、频率、作用时间、脉冲个数等)的前提下对酵母悬浮液进行PEF处理,测定PEF处理后酿酒酵母的存活率和浓度及酵母活性的变化。结果表明,在电场强度1~30 k V·cm-1范围内,酵母细胞死亡比例随着电场强度的升高而增加,非特异性内酯酶活性增强的酵母细胞比例随着电场强度的升高而减少。当电场强度较低时,非特异性内酯酶活性增加的酵母比例高于死亡酵母细胞比例;电场强度大于10 k V·cm-1时,死亡酵母细胞比例高于非特异性内酯酶活性增加的酵母细胞比例;PEF处理过程中出现的亚致死酵母细胞比例在10 k V·cm-1时达到最大值。低电场PEF处理可以提高酵母细胞活性和发酵能力且酵母死亡比例较低;高电场PEF处理对酿酒酵母具有灭活效果。本研究结果对酿酒酵母的工业应用具有指导意义。     

接种单细胞微生物对土壤团聚体形成及其稳定性的影响

将根瘤菌（Rhizobium sp.）G-01、肠杆菌（Enterobacter sp.）San8及粘红酵母（Rhodotorula mucilaginosa）R1等3株产胞外多聚物（Extracellular polymeric substances,EPS）的单细胞微生物接种于供试土壤,进行土培试验和盆栽黄瓜试验,研究接种微生物及作物根系对土壤团聚体形成、团聚体组成变化的影响。结果表明,3株单细胞微生物分泌EPS的量分别为2.12 mg（1010 cells）?1、0.56 mg（1010 cells）?1、172.71 mg（1010 cells）?1,其主要组分是多糖、其次是蛋白质,且含有羟基、羧基、羰基等官能团;土培试验后,接种产EPS单细胞微生物处理土壤的大团聚体（> 0.25 mm）含量比接种丝状微生物处理提高了71.31%,而盆栽试验后各处理土壤的大团聚体含量（> 0.25 mm）比土培试验后土壤显著增加,这可能是由于植株根系挤压及其分泌物作用促进了土壤大团聚体形成并提高了团聚体稳定性所致。     

极低频脉冲电场与高压静电场对作物种子萌发影响的差异

为了明确基于极低频脉冲电场(pulsed electric field,PEF)的种子处理技术与基于高压静电场(highvoltage electrostatic field,HVEF)的种子处理技术的有效性及其差异,研究了极低频PEF和HVEF对绿豆生长的影响。结果表明,极低频PEF处理对绿豆幼苗的质量、芽长和根长都有明显的促进作用,相同强度的HVEF处理对绿豆幼苗的根长生长有一定促进作用,对芽长的影响不显著。机理研究显示,极低频PEF和HVEF处理均对萌发绿豆的氧化代谢和蛋白质代谢有一定影响,在种子萌发初期,极低频PEF处理促进了种子储藏蛋白的分解,在后期促进了新蛋白的合成,HVEF处理则对萌发后期的蛋白合成有一定促进作用。研究还发现,极低频PEF和HVEF均可通过诱发超氧阴离子自由基激活种子超氧化物歧化酶(SOD)活性,促进过氧化物酶(POD)的合成,但是,极低频PEF对SOD和POD的影响大于HVEF。讨论了极低频PEF和HVEF对绿豆萌发影响差异的原因。     

施磷对小麦旗叶氮代谢关键酶活性和子粒蛋白质含量的影响

在田间试验条件下,以强筋品种济麦20和弱筋品种山农1391为材料,研究了不同施磷水平对2类型品种小麦旗叶氮代谢关键酶活性和子粒蛋白质积累的效应。结果表明,在P2O5 0～135 kg /hm2范围内,施磷明显提高济麦20灌浆前期旗叶硝酸还原酶（NR）和谷氨酰胺合成酶（GS）活性;但却降低山农1391旗叶NR、GS活性。强筋品种济麦20在灌浆中、后期旗叶内肽酶（EP）、羧肽酶（CP）活性和蛋白质含量因施磷而提高;而弱筋品种山农1391的EP、CP活性和子粒蛋白质含量则降低。施磷使济麦20子粒清蛋白、醇溶蛋白和谷蛋白的含量提高,球蛋白含量有所降低;但弱筋品种山农1391子粒球蛋白、醇溶蛋白和谷蛋白的含量因施磷而降低,清蛋白含量未见显著变化。说明小麦施磷应考虑不同品质类型品种氮代谢及子粒蛋白质含量对施磷的反应。     

田间条件下不同钝化材料对玉米吸收镉的影响研究

选取4种钝化材料（赤泥、海泡石、钙镁磷肥和磷矿粉）开展田间试验,研究它们对玉米吸收镉与土壤有效态镉的影响。结果表明,除海泡石外,施用其他3种钝化材料均能促进玉米生长,增加玉米叶、茎与籽粒的重量。4种钝化材料都能降低玉米对镉的吸收,其中：高量赤泥（用量1.5%）能明显降低玉米茎、叶片及籽粒中镉的含量,较对照分别降低60.6%、33.6%与49.3%;高量钙镁磷肥（用量900kg·hm-2）明显降低玉米籽粒中镉含量,较对照降低57.4%。4种钝化剂明显降低土壤EDTA提取态镉、DGT提取态镉的含量。其中,高量海泡石处理的EDTA提取态镉含量最低,高量赤泥次之;高量赤泥处理的DGT提取态镉含量最低。本试验结果表明,1.5%用量的赤泥是最佳的功能钝化材料。     

低氮和干旱胁迫对富士和秦冠生长及氮素利用的影响

【目的】以富士（Fuji）、 秦冠（Qinguan）嫁接在平邑甜茶（Malus hupehensis Rehd.）上的当年生盆栽苗为试验材料,采用砂培方法,研究了缺氮胁迫和干旱对富士和秦冠生长情况、 光合参数、 植株各部位氮磷钾含量及氮素利用效率的影响,分析比较了低氮干旱条件下富士和秦冠生长及氮素利用的差异,以期为果树生产高效肥水利用提供理论指导。【方法】试验共设四个处理: 正常氮正常水（ZZ）、 低氮正常水（DZ）、 正常氮干旱（ZG）、 低氮干旱（DG）。氮素和水分均设置两个水平,分别为正常氮（6 mmol/L NO-3-N）、 低氮（0.3 mmol/LNO-3-N）、 正常供水（保持盆中砂子相对含水量为饱和含水量的80%~85%）、 干旱处理（保持盆中砂子相对含水量为饱和含水量的60%~65%）。【结果】富士和秦冠的生物量（茎和叶）、 株高茎粗等生长指标以及光合速率、 气孔导度、 蒸腾速率均为正常氮正常水（ZZ）＞低氮正常水（DZ）＞正常氮干旱（ZG）＞低氮干旱（DG）,并且相对应处理下秦冠的以上指标均高于富士;正常供水下,缺氮处理使富士、 秦冠的根冠比比正常氮处理均有所增加,富士提高了2.05%,秦冠提高了22.40%。富士和秦冠的氮、 磷、 钾含量均表现出正常氮正常水（ZZ）＞低氮正常水（DZ）＞正常氮干旱（ZG）＞低氮干旱（DG）; 氮、 钾元素含量在植株各部位的分布顺序依次是叶＞根＞茎,磷元素则是根＞叶＞茎;光合氮素利用效率（PNUE）和氮素利用效率表现为秦冠处理之间差异极显著,富士处理之间差异不显著;秦冠的PNUE和NUE明显高于富士,在低氮正常水（DZ）处理下,秦冠氮肥利用率比富士高42.07%,在低氮干旱（DG）处理下高64.14%;低氮胁迫下富士和秦冠的NUE显著提高,并且秦冠提高的幅度高于富士。【结论】施用氮肥能够显著提高富士与秦冠的干物质量,同等水肥条件下,秦冠生长优于富士;水分亏缺会减少叶片对氮的吸收,干旱条件下适度增施氮肥,可提高果树的抗旱能力;低氮干旱胁迫下秦冠的生长指标、 光合指标及氮素利用效率指标均优于富士,表现出较强的抗低氮干旱胁迫的能力。     

Biodiversity and ecogeography of wild Lactuca spp. in some European countries

Staff members of the Department of Botany of Palacký University in Olomouc and Gene Bank Department – Workplace Olomouc, Research Institute of Crop Production in Prague, Czech Republic, conducted an expedition in seven European countries (Austria, Czech Republic, France, Germany, Italy, the Netherlands, Switzerland) in August/September 1999 to collect wild Lactuca spp. germplasm and study its geographic distribution, ecology and biodiversity. During the mission, more than 600 locations were visited resulting in the collection of 602 seed samples (accessions) of wild Lactuca species and 13 seed samples of related genera (Chondrilla and Mycelis). Lactuca serriola f. serriola, L. serriola f. integrifolia, L. saligna and L. viminea subsp. chondrilliflora were prevalent in southern Europe (Italy, France), however, only L. serriola was common in central and western Europe (Austria, Czech Republic, Germany, Netherlands, Switzerland). The greatest diversity of Lactuca species was found in France, where also the most seed samples (165) were collected. The most characteristic habitats with a high density of Lactuca spp. populations were observed along roads and highways, grassy ditches, ruderal communities, and dust-heaps. Natural infections by powdery mildew (Erysiphe cichoracearum) and downy mildew (Bremia lactucae) on some wild Lactuca spp. were observed. Recent observations concerning the geographic distribution, population structure, habitats, and natural occurrence of diseases of Lactuca spp. are discussed. This assemblage of genetic resources of Lactuca spp. can serve as the basis of future studies of species diversification, spatial population structure, plant microevolution, domestication processes, and genetic variability of host-parasite interactions.     

Pasture Vegetation Elemental Analysis by Laser-Induced Breakdown Spectroscopy

Laser-induced breakdown spectroscopy (LIBS) is a new technique for the analysis of plant material. This study investigates the application of LIBS to pasture-based plant samples. The LIBS measurements were obtained from pelletized pasture samples (100 samples) that had been also analyzed by inductively coupled plasma–optical emission spectroscopy (ICP-OES) following microwave digestion for calibration and comparison purposes. Comparisons for elements sodium (Na), potassium (K), magnesium (Mg), calcium (Ca), manganese (Mn), iron (Fe), copper (Cu), zinc (Zn), boron (B), phosphorus (P), and sulfur (S) showed that LIBS could be used for almost all the standard profile total elements with concentrations down to low mg/kg levels (observed error of Na: 0.024 percent, K: 0.18 percent, Mg: 0.016 percent, Ca: 0.073 percent, P: 0.017 percent, Mn: 31 mg/kg, Fe: 150 mg/kg, Zn: 6.6 mg/kg, and B: 1.1 mg/kg). Elemental analysis at less than mg/kg levels was not possible using LIBS. The elements S and Cu were particularly difficult to analyze with reliability using LIBS at the concentration levels found in the plant samples. Replacing microwave digestion and subsequent ICP analysis with a direct analysis of dried plant samples using LIBS has the potential to improve the productivity and reduce the cost of testing.     

Properties of anaerobically digested and composted municipal solid waste assessed by linking soil mesofauna dynamics and nitrogen modelling

We have studied the effect of anaerobically digested (ADMSW) and composted municipal solid waste (CMSW) on mineralization and foodweb dynamics to verify the hypothesis that ADMSW would immobilize N right after addition to soil in contrast to addition with CMSW. Another hypothesis was that the mesofauna (enchytraeids and microarthropods) would stimulate N release from the decomposer community. We measured excretion of -N and urea-N from the mesofauna and hypothesized that enchytraeids would release urea. ADMSW and CMSW were amended to pots with sandy loam and barley. The pots were divided into treatments with or without mesofauna. Mesofauna, plant N and biomass, soil N and ergosterol (fungal biomass) were measured over a 113-day period of four equidistant samplings. Soil respiration, N mineralization and N release by the mesofauna were modelled from concurrent studies. ADMSW- and CMSW-treated soils initially (<20 days) immobilized N. The amendments did not increase plant growth substantially, and this was probably due to N-limitation in the early stages of plant growth. Enchytraeid abundance was about three times higher in ADMSW- than CMSW-treated soils, indicating that ADMSW contained more labile compounds, bacteria, and microfauna. The mesofauna did not affect N-content, but the cumulated -N excreted by the mesofauna was estimated to be substantial and about one-fifth of total plant N in ADMSW. An explanation for this discrepancy might be that in the absence of the mesofauna, other members of the detrivore and microbivore community performed the mesofauna’s function. Neither enchytraeids nor microarthropods excreted urea.     

Faculty positions: Center for Agricultural Resources Research,Chinese Academy of Sciences

正The Center for Agricultural Resources Research(CARR),the Institute of Genetics and Developmental Biology(IGDB),Chinese Academy of Sciences,invites applicants for several research group leader positions.CARR is one of the research organizations in Chinese Academy of Sciences(CAS).We seek nominations and applications from individuals who have expertise and a record of accomplishment in research areas related to ecology,agro-hydrology,     

Faculty positions: Center for Agricultural Resources Research,Chinese Academy of Sciences

正The Center for Agricultural Resources Research(CARR),the Institute of Genetics and Developmental Biology(IGDB),Chinese Academy of Sciences,invites applicants for several research group leader positions.CARR is one of the research organizations in Chinese Academy of Sciences(CAS).We seek nominations and applications from individuals who have expertise and a record of accomplishment in research areas related to ecology,agro-hydrology,     

Faculty positions:Center for Agricultural Resources Research,Chinese Academy of Sciences

正The Center for Agricultural Resources Research(CARR),the Institute of Genetics and Developmental Biology(IGDB),Chinese Academy of Sciences,invites applicants for several research group leader positions.CARR is one of the research organizations in Chinese Academy of Sciences(CAS).We seek nominations and applications from individuals who have expertise and a record of accomplishment in research areas related to ecology,agro-hydrology,     

Assessing the effectiveness of conservation measures: Resolving the “wicked” problem of the Steller sea lion

“Wicked” problems are those that are complex and that change when solutions are applied. Many conflicts in conservation fall into this category. The study approached the problem of how to constrain the apparent wickedness of a problem in the conservation management of a species by using simple empirical indicators to carry out iterative assessment of the risk to a population and to document how this risk evolves in relation to the addition of new data and the implementation of management actions. Effects of high levels of uncertainty within data and also concerning population structure were examined through stochastic simulation and by exploration of scenarios. Historical trends in the example used, the Steller sea lion, showed rapid declines in abundance in some regions during the 1980s. The current total population is 130,000-150,000 Steller sea lions through Alaska and British Columbia and this number has been stable since about 1990 in spite of regional differences in population dynamics. Regional differences in the sequence of changes in the number of pups and non-pups, suggested that an internal re-distribution of juveniles could have happened between 1980 and 1990. Current productivity also appears close to the long-term mean. Stochastic population projection using various scenarios showed that, based upon this history, the risk of extinction for the population has declined and is below reasonable thresholds for considering the population to be endangered. The trends in risk suggest that management actions taken since 1990 have probably been effective. Consequently, the conservation management objectives for the Steller sea lion are probably being met. The approach provides a mechanism, based upon experience and scenario analysis, for exploring future policy options and may help to constrain the debate amongst stakeholders about the cost-benefit trade-offs associated with different options.     

Soil-profile distribution of primary and secondary plant-available nutrients under conventional and no tillage

Nutrient distributions under no tillage (NT) compared with conventional disk-and-bed tillage (CT) management in the warm, humid region of the southeastern USA need to be assessed so that future placement, quantity, and type of fertilizers can be altered, if necessary, to efficiently match crop demands. We determined soil-profile distributions of pH, N, P, S, K, Ca, Mg, Na, Zn, Fe, Mn, and Cu to a depth of 0.9 m at the end of 8.5 years of continuous CT and NT management on a Weswood silty clay loam (fine, mixed, thermic Fluventic Ustochrept) in southcentral Texas. Most dramatic changes occurred within the 0–0.05 m depth, where soil under NT had lower pH, Fe, and Cu than under CT, but greater P, K, Zn, and Mn. Greater P and K under NT than under CT also occurred below the till-zone (0.15–0.3 m). At a depth of 0–0.3 m, soil under NT contained greater amounts of extractable P, K, Zn, Fe, Mn, and Cu than under CT. Nitrogen fertilization had little effect on nutrient distributions, except resulting in greater extractable K at 0–0.05 m and greater nitrate at 0–0.15 m. Few changes in soil-profile distributions were observed for extractable S, Ca, Mg, and Na. Long-term continuous use of NT on this fine-textured, high-fertility (except for N) soil had no apparent adverse effects on nutrient distributions relative to CT, but enhanced conservation and availability of P, K, Zn, Fe, Mn, and Cu near the soil surface where crop roots proliferate.     

Macroinvertebrates in North American tallgrass prairie soils: effects of fire, mowing, and fertilization on density and biomass

The responses of tallgrass prairie plant communities and ecosystem processes to fire and grazing are well characterized. However, responses of invertebrate consumer groups, and particularly soil-dwelling organisms, to these disturbances are not well known. At Konza Prairie Biological Station, we sampled soil macroinvertebrates in 1994 and 1999 as part of a long-term experiment designed to examine the effects and interactions of annual fire, mowing, and fertilization (N and P) on prairie soil communities and processes. For nearly all taxa, in both years, responses were characterized by significant treatment interactions, but some general patterns were evident. Introduced European earthworms (Aporrectodea spp. and Octolasion spp.) were most abundant in plots where fire was excluded, and the proportion of the total earthworm community consisting of introduced earthworms was greater in unburned, unmowed, and fertilized plots. Nymphs of two Cicada genera were collected (Cicadetta spp. and Tibicen spp.). Cicadetta nymphs were more abundant in burned plots, but mowing reduced their abundance. Tibicen nymphs were collected almost exclusively from unburned plots. Treatment effects on herbivorous beetle larvae (Scarabaeidae, Elateridae, and Curculionidae) were variable, but nutrient additions (N or P) usually resulted in greater densities, whereas mowing usually resulted in lower densities. Our results suggest that departures from historical disturbance regimes (i.e. frequent fire and grazing) may render soils more susceptible to increased numbers of European earthworms, and that interactions between fire, aboveground biomass removal, and vegetation responses affect the structure and composition of invertebrate communities in tallgrass prairie soils.