芽胞杆菌高产纤维素酶菌株的诱变选育与培养基优化

为获得高产纤维素酶菌株,采用紫外(UV)诱变和紫外-亚硝基胍(UV-NTG)复合诱变方法对新疆吐鲁番地区土壤中分离得到的产纤维素酶菌株芽胞杆菌C-8进行诱变筛选,并通过二次正交旋转组合试验和响应面试验对诱变筛选得到的菌株进行培养基的优化。结果表明,UV诱变菌株UV-5和UV-NTG比出发菌株C-8纤维素酶活分别提高了1.15倍和3.23倍;当发酵培养基中碳源浓度为3%、氮源浓度为1.5%、吐温-80的浓度为0.15%时,纤维素酶活达到453.20 U·m L-1,是出发菌株C-8的5.49倍和复合诱变菌株UV-NTG-10的1.7倍。本研究结果为纤维素酶的扩大发酵以及后期工业化生产提供了理论依据。     

E.coli谷氨酸脱羧酶高产菌株选育及发酵条件研究

以普通大肠杆菌(E. coli)为出发菌株,以L-谷氨酸、溴甲酚绿(pH3.8~5.4)为筛选及指示剂,经亚硝基胍(NTG)、UV诱变处理,得到了L-谷氨酸脱羧酶活性变异菌株,其L-谷氨酸脱羧酶（GAD）活性大幅度提高,比出发菌株酶活性提高了2.22倍。优化实验显示：pH值、发酵时间、诱导剂L-谷氨酸、硫酸镁对GAD产酶有较大影响。通过对产酶发酵培养基中几种成份单因素变动及正交优化实验,对该菌株产GAD的诱导剂L-谷氨酸、营养要求和发酵条件进行了研究,优化后的发酵培养基组成为牛肉膏5 g/L,蛋白胨10 g/L,氯化钠3 g/L,L-谷氨酸0.5 g/L,葡萄糖1.5 g/L,磷酸二氢钾2 g/L,硫酸镁0.7 g/L。发酵条件是：pH6.8,接种菌龄20h,发酵时间20h。在优化条件下突变菌株GAD活性可达6790.7U,是出发菌株的2.76倍。     

空间诱变选育万隆霉素高产菌株

为获得万隆霉素高产菌株,以菌株C2507为出发菌株连续2次进行空间诱变,正突变率分别为21.33%和16.5%,筛选得到高产突变株W3-356,万隆霉素产量提高1.83倍,连续传代试验表明其产素能力具有遗传稳定性。突变株W3-356的生长速度加快,在6种营养培养基上,其形态特征有一定改变,在摇瓶发酵过程中,突变株W3-356对碳源和氮源的利用率高于出发菌株,发酵周期缩短了12h。     

产IAA菌株的UV和DES诱变筛选及培养条件优化

为了得到高产吲哚-3-乙酸(IAA)菌株,采用紫外(UV)诱变和硫酸二乙酯(DES)诱变的方法,对从烟草根际土壤中分离得到的一株产IAA并具有溶有机磷性状的促生菌进行诱变选育,并对获得的目标菌株的发酵培养条件进行正交优化。结果表明,菌株经UV诱变和DES诱变均能有效提高其IAA产量。诱变条件为 UV(15 W,30 cm)照射1 min或2 mg·mL^-1 DES处理30 min时,得到1株高产IAA的菌株UV19,其IAA产量为33.77 mg·L^-1,是出发菌株产量的299.48%。菌株UV19经10代继代,其产IAA能力和溶有机磷性状稳定遗传。菌株UV19产IAA培养发酵优化条件为10%装瓶量、初始pH值8、培养温度25℃、接种量3%、培养时间96 h、含500 mg·L^-1色氨酸的LB培养基,优化后IAA产量高达75.47 mg·L^-1,为优化前的2.23倍。本研究结果为菌肥及生物法生产IAA提供了基础材料及技术方案。     

基于~(60)Co-γ射线辐照的木糖乙醇发酵差异性突变菌株筛选及其发酵特性

本研究利用60Co-γ辐射源对树干毕赤酵母CICC1960进行辐照诱变,获取木糖乙醇发酵优良菌株和差异性突变菌株群体,为进一步利用比较基因组学方法深入研究木糖乙醇代谢机制提供菌种材料。经筛选获得7株木糖乙醇转化差异性突变菌株,以乙醇木糖比为指标,可将各突变菌株的发酵能力由高到低排列为:a1、X4、i11、K2、X21、CICC1960、a11、c10,乙醇木糖比均值分别为0.3143、0.3037;0.2911、0.2678、0.2578、0.2311、0.2230、0.1220。其中菌株a1、X4、i11为优良菌株,菌株a1在发酵72h时乙醇产量达到顶峰15.6 g·L-1,高出出发菌株140%,乙醇木糖比高出出发菌株37.9%;菌株X4在发酵60h时乙醇产量达到顶峰15.4 g·L-1,高出出发菌株234.8%,乙醇木糖比高出出发菌株33.1%;菌株i11在发酵60h时乙醇产量达到顶峰14.8 g·L-1,高出出发菌株221.7%,乙醇木糖比高出出发菌株29%。     

复合诱变选育高产赤藓糖醇解脂亚罗酵母及其发酵工艺优化

为获得赤藓糖醇高产菌株，以解脂亚罗酵母WT5为出发菌株，采用⁶⁰Co-γ射线和常压室温等离子体(ARTP)对其进行复合诱变处理。为进一步提高赤藓糖醇产量，采用单因素筛选结合Plackett-Burman(PB)试验方法优化培养基组成，同时对培养模式进行改进。结果表明，复合诱变能有效提升解脂亚罗酵母性能，其中⁶⁰Co-γ射线最佳辐射剂量和ARTP最优处理时间为1 200 Gy、60 s。诱变处理后获得优良菌株CA20，其赤藓糖醇产量为60.80 g·L^-1，是出发菌株WT5的2.41倍。此外，PB拟合结果确认发酵最优培养基组分为：43 g·L^-1葡萄糖、1.92 g·L^-1酵母浸出粉、2.98 g·L^-1蛋白胨、4.70 mg·L^-1硫酸铵、6.85 g·L^-1氯化钠、3.30 mg·L^-1磷酸二氢钾、0.65 mg·L^-1维生素B1、49 mg·L^-1     

微波诱变选育高产生长素及耐药性根瘤菌株研究

为获得高效分泌IAA(吲哚乙酸Indoleacetic acid),同时具有卡那霉素(Kanamycin)和氨苄青霉素(Ampicillin)双抗药性的根瘤菌突变株,以红豆草根瘤菌(Rhizobium Onobrychis vicilfolia)RS-1为原始菌株进行微波诱变处理,对微波诱变参数进行了优化,并以含抗生素平板筛选出卡那霉素和氨苄青霉素双抗药突变菌株,以多次传代的方式考察了突变菌株的遗传稳定性和生长素产量。结果显示,最优微波筛选参数为800W、6s,该参数下诱变获得5株高产生长素及双耐药突变株RSW-14、RSW-55、RSW-62、RSW-96和RSW-107。传代试验表明RSW-96是稳定的高产生长素、耐药突变株,表现在RSW-96传代6次后对80mg/L卡那霉素和300mg/L青霉素具有抗性,培养4d和24d的IAA产量比原始菌株提高了40.09%和50.15%,对单株生物量增加效果高于原始菌株25.58%,具有良好的促生效应。     

酸性β-甘露聚糖酶高产菌株的诱变育种

摘要：以一株实验室保藏的酸性β-甘露聚糖酶产生菌黑曲霉（Aspergillus niger）LW-1作为原始出发菌株,首先经自然分离筛选出一株产酶较稳定的N-9作为诱变出发菌株,再经真空微波和甲基磺酸乙酯（EMS）逐级诱变处理,采用基础发酵培养基固态发酵筛选以及斜面传代培养,获得了一株高产、稳产酸性β-甘露聚糖酶的E-30菌株。其产β-甘露聚糖酶活性达36 675 U/g,是原始出发菌株（17 048 U/g）的2.15倍。E-30菌株三角瓶麸曲种子保藏两个月,产酶活性稳定。     

产γ-氨基丁酸的乳酸菌株筛选及诱变

γ-氨基丁酸(-γaminobutyric acid,GABA)是中枢神经系统一种重要的抑制性神经递质。本研究从鲜奶中分离得到1株高产GABA的乳酸菌株hjxj-01,经初步鉴定为短乳杆菌。实验结果表明,在含5%的L-谷氨酸钠的GYP培养基中,此乳酸菌产GABA最大积累浓度为7 g/L。在此基础上,又先后使用了紫外线和γ射线对出发菌株进行了诱变处理。以正突变率为标准确定诱变条件。30W的紫外灯下,距离45cm照射及照射时间50 s为紫外线诱变的较佳条件;60Co射线诱变的适宜剂量为300 Gy。诱变后得到1株突变菌株hjxj-08119,经连续传代12次,遗传性状稳定,平均GABA产量达到17 g/L,较出发菌株hjxj-01提高142.9%。     

~(60)Coγ射线诱变选育热凝胶多糖高产菌株的研究

利用60 Coγ射线对产生热凝胶多糖 (Curdlan)出发菌株GM 2 4的菌体细胞与原生质体分别进行辐照处理 ,发现60 Coγ射线对GM 2 4原生质体的诱变效应明显优于对其菌体细胞的作用。对最终诱变筛选获得的 1株高产稳定的Curdlan生产突变株A81研究表明 :与GM 2 4相比 ,其Curdlan产量提高了 5 0 4%,发酵周期缩短了 1 8%,发酵终止后底物残糖由 1 5 3g L降至7 4g L。糖的转化率从 38 7%提高至 5 8 2 %。这说明在Curdlan生产菌株的筛选中60 Coγ辐照是一种有效的方法     

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,     

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.     

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.