高山被孢霉高产ARA的原生质体融合子筛选

利用原生质体融合技术,对两株单一优良性状差异显著的高山被孢霉菌株F24和G12进行原生质体融合,再采用双灭活或流式细胞分选法筛选优良性状相结合的高产菌株,对双亲菌株原生质体的制备、融合和再生的相关条件进行了研究,并对双灭活和流式细胞分选的具体方法进行了研究.经研究发现,采用培养16 h的高山被孢霉菌丝于混合酶液(蜗牛酶15.0 mg·m L~(-1)、纤维素酶6.0 mg·m L~(-1)、溶菌酶0.5 mg·m L~(-1)和几丁质酶0.2 mg·m L~(-1))中30℃酶解4 h为原生质体制备的最佳条件.并探究得出两种融合子筛选方法的最佳方案,成功筛选出融合子,进一步再生培养验证ARA产量,最终挑选出一株ARA产量达6.32 g·L~(-1)的高产菌株.以前尚未发现此方法在高山被孢霉上的应用,为高山被孢霉育种提供了一种有效的手段.     

新西兰青霉(Penicillium novae-zeelandiae)原生质体的制备与再生研究

利用酶对新西兰青霉菌的菌丝进行酶解获得原生质体,探究了影响原生质体制备和再生的因素,包括酶种类,酶浓度,菌丝培养时间,酶解时间和温度,pH,二硫苏糖醇(DTT)预处理等.结果显示最佳酶解条件为:菌丝体培养48h,用0.05mol·L~(-1) DTT预处理0.5h,以0.8mol·L~(-1)氯化钠作为稳定剂,31℃条件下经Lywallzyme(10mg·mL~(-1))酶解4h,原生质体数量达到4.75×107 g~(-1),再生率为18.0%.     

蒙古口蘑与双孢蘑菇原生质体融合育种研究

以蒙古口蘑和双孢蘑菇为出发菌株,研究了酶浓度、酶解时间、菌龄和渗透压稳定剂、再生培养基等对蒙古口蘑和双孢蘑菇原生质体制备和再生的影响.结果表明,蒙古口蘑原生质体制备和再生的最佳条件是,培养5 d的菌丝体用2.5%(w/w)的溶壁酶酶解4 h,渗透压稳定剂为0.6 mol/L KCl,MB为再生培养基,制备率为7.53×1010个/L,再生率为8.4×10-4;双孢蘑菇原生质体制备和再生的最佳条件是,培养5 d的菌丝体用2%(w/w)的溶壁酶酶解3 h,渗透压稳定剂为0.6 mol/L KCl,PQA为再生培养基,制备率为6.96×1010个/L,再生率为7.4×10-4.对两种菌原生质体的释放过程进行形态观察,均为顶端释放.采用双亲灭活原生质体融合技术对蒙古口蘑和双孢蘑菇原生质体融合进行研究,蒙古口蘑原生质体采用热灭活,65℃处理30 min,灭活率达100%,双孢蘑菇采用紫外灭活,在15 W紫外灯下,距离30 cm,处理20 min,灭活率达100%,两菌株按1∶1混合,以30%PEMG(6 000)为促融剂,融合10 min,融合率为5.6×10-5.经遗传稳定性检验,融合菌株的遗传稳定率为80%,从菌落特征、菌丝形态、菌丝生长量、酯酶、游离全蛋白、乳酸脱氢酶、乙醇脱氢酶和过氧化物酶等同工酶方面对融合子进行筛选,获得一株具有双亲性状的融合株.     

Blakeslea trisporaH1^-原生质体制备与再生条件研究

以β-胡萝卜素生产菌株Blakeslea trispora H1-为研究对象,对其原生质体形成及再生条件进行了研究.得知最佳形成与再生条件为:以摇瓶方式培养菌丝体,培养菌龄为56 h,酶解温度为30 ℃,酶质量分数2.0%,混合酶组成为m(蜗牛酶)∶m(纤维素酶)＝6∶4,酶解时间为120 min,渗稳剂为0.6 mol/L MgSO4·7H2O.Blakeslea trispora H1-原生质体形成数为7.9×106 个/mL,再生率为3.9%.     

利福霉素高产菌株快速筛选方法的建立及应用

利福霉素发酵过程中pH变化规律能够反映菌体对不同营养物质的利用情况及菌体生理特性的变化,并与最终放瓶效价有一定的关联。据此建立了一种以发酵前期(±50 h)pH变化为判断依据的新的快速筛选菌种的方法,即发酵前期pH下降到谷底较早的菌株高产的可能性很大(概率接近70%),并在双亲株灭活种间融合法选育利福霉素SV菌株中进行了应用,筛选到了高产融合菌株F-3及F-16,两菌株分别比出发菌株效价提高了11%和17%。同时,得到了两个亲株(利福霉素SV产生菌U-32、利福霉素B产生菌X#-1)的酶解条件(U-32:10 m g/mL、34°C、2 h;X#-1:10 m g/mL、34°C、3 h)及原生质体双灭活标记条件(U-32:UV 20 m in;X#-1:60°C、50 m in),双灭活原生质体用500 g/L PEG融合,融合频率约1.04×10-5。     

卡介苗菌株和结核分枝杆菌H37Ra菌株原生质体的制备研究

分别以卡介苗菌株(BCG)及结核分枝杆菌国际标准无毒株H37Ra菌株(H37Ra)为受体菌,制备上述两种菌株的原生质体,同时通过优化细菌菌龄、酶解浓度、酶解温度以及酶解时间等影响因素,探索出制备原生质体形成及再生的最优条件。结果显示:摸索出制备BCG和H37Ra菌株的原生质体条件:在对数生长期的两亲本菌株,经0.01 mol/L EDTA,0.01%β-巯基乙醇溶液预处理;酶解浓度为12 mg/mL,酶解温度为37℃,酶解时间为5 h,可制备出活性较高的两种菌株的原生质体。由此可知,成功制备了BCG与H37Ra菌株的原生质体,并能在高渗固体培养基上再生。本实验为进一步研究该两种菌株原生质体融合试验奠定了基础。     

利用菊粉产山梨醇酵母融合菌株的构建

介绍高产菊粉酶的克鲁维酵母(Kluyveromycessp.)Y85菌株与产山梨醇的酿酒酵母(Saccharomycescerevisiae)E15菌株,通过属间原生质体融合构建直接利用菊粉产山梨醇的酵母融合菌株.结果认为,以0.1%EDTA-巯基乙醇为脱壁预处理剂,蜗牛酶浓度2%(W/V),酶解温度30℃,E15和Y85两菌株细胞酶解时间分别为40min和30min,原生质体形成率和再生率分别达90%和25%以上.融合的适宜条件为添加30%(W/V)PEG和10mmol/LCaCl2,融合时间为30min,融合率可达2.64×10-6以上.利用两亲株自然性状差异检出融合菌株,并经DNA含量、菊粉酶活性、细胞形态与大小、菌落形态等测定加以确证.其中F27融合菌株在适宜条件下,山梨醇摇瓶发酵产量可达4.87g/100mL,并且具有遗传稳定性能.     

交替假单胞菌BYS-2产褐藻胶裂解酶条件研究

以褐藻酸钠作为褐藻胶裂解酶产生菌初筛培养基唯一碳源,从鲍鱼养殖水样中分离获得一株产褐藻胶裂解酶的微生物,形态学和分子生物学16S rDNA鉴定结果显示,该菌株为交替假单胞菌Pseudoalteromonas sp.BYS-2.对该菌株进行产酶条件研究,获得最适产酶配方为:褐藻酸钠1 g·L~(-1),葡萄糖0.5 g·L~(-1),酵母膏10g·L~(-1),黄豆饼粉3 g·L~(-1),(NH_4)_2HPO_4 3 g·L~(-1),初始培养基pH8.0;最佳产酶条件为:接种量2%(体积分数),装液量50 m L/250 m L,发酵温度20℃,摇床转速200 r·min~(-1),在此条件下发酵24 h酶活力可达5.29 U·m L~(-1),比优化前(1.57 U·m L~(-1))提高2.37倍.     

辅酶Q_(10)生产菌株粟酒裂殖酵母原生质体制备和再生研究

研究对辅酶Q10生产菌株粟酒裂殖酵母原生质体制备和再生条件进行了优化,确定了原生质体制备及再生的最佳条件为:茵龄20 h,蜗牛酶浓度2 g/L,酶溶液pH值6.5,酶解温度25℃,酶解时间2 h.通过对粟酒裂殖酵母原生质体制备及再生条件的研究,建立了制备粟酒裂殖酵母原生质体的方法,以期进一步通过原生质体诱变获得辅酶Q10高产菌株.     

草菇原生质体的制备与融合研究

项目对低温草菇和高温草菇菌株进行了酶解制备原生质体及原生质体的融合进行了研究.研究结果表明:以融壁酶Novozym234对低温草菇和高温草菇酶解效果较好,酶解浓度为2.5%,稳渗剂以0.5Mol/L蔗糖为佳,pH值保持在8.5左右,以30%的助溶剂(PEG4000-6000)作为融合诱导剂进行草菇原生质体融合,并获得融...     

Characterization of Ordovician carbonate reservoirs, southeastern Saskatchewan, Canada

The discovery of the prolific Ordovician Red River reservoirs in 1995 in southeastern Saskatchewan was the catalyst for extensive exploration activity which resulted in the discovery of more than 15 new Red River pools. The best yields of Red River production to date have been from dolomite reservoirs. Understanding the processes of dolomitization is, therefore, crucial for the prediction of the connectivity, spatial distribution and heterogeneity of dolomite reservoirs.The Red River reservoirs in the Midale area consist of 3～4 thin dolomitized zones, with a total thickness of about 20 m, which occur at the top of the Yeoman Formation. Two types of replacement dolomite were recognized in the Red River reservoir: dolomitized burrow infills and dolomitized host matrix. The spatial distribution of dolomite suggests that burrowing organisms played an important role in facilitating the fluid flow in the backfilled sediments. This resulted in penecontemporaneous dolomitization of burrow infills by normal seawater. The dolomite in the host matrix is interpreted as having occurred at shallow burial by evaporitic seawater during precipitation of Lake Almar anhydrite that immediately overlies the Yeoman Formation. However, the low δ18O values of dolomited burrow infills (-5.9‰～ -7.8‰, PDB) and matrix dolomites (-6.6‰～ -8.1‰, avg. -7.4‰ PDB) compared to the estimated values for the late Ordovician marine dolomite could be attributed to modification and alteration of dolomite at higher temperatures during deeper burial, which could also be responsible for its 87Sr/86Sr ratios (0.7084～0.7088) that are higher than suggested for the late Ordovician seawaters (0.7078～0.7080). The trace amounts of saddle dolomite cement in the Red River carbonates are probably related to "cannibalization" of earlier replacement dolomite during the chemical compaction.     

A computer generator for randomly layered structures

AcomputergeneratorforrandomlylayeredstructuresYUJia shun1,2,HEZhen hua2(1.TheInstituteofGeologicalandNuclearSciences,NewZealand;2.StateKeyLaboratoryofOilandGasReservoirGeologyandExploitation,ChengduUniversityofTechnology,China)Abstract:Analgorithmisintrod…     

海南岛地体及其毗邻陆缘晚中生代—新生代古地磁研究和构造演化

本文叙述了对海南岛及其毗邻大陆边缘白垩纪到第四纪地层岩石进行古地磁研究的全部工作过程。通过分析岩石中剩余磁矢量的磁偏角及磁倾角的变化,提出海南岛白垩纪以来经历的构造演化模式如下:早期伴随顺时针旋转而向南迁移,后期伴随逆时针转动并向北运移。联系该地区及邻区的地质、地球物理资料,对海南岛上述的构造地体运动提出以下认识:北部湾内早期有一拉张作用,主要是该作用使湾内地壳显著伸长减薄,形成北部湾盆地。从而导致了海南岛的早期构造运动,而海南岛后期的构造运动则主要是受南海海底扩张的影响。海南地体运动规律的阐明对于了解北部湾油气盆地的形成演化有重要的理论和实际意义。     

Exciton Seebeck in Molecular Systems

Various applications relevant to the exciton dynamics,such as the organic solar cell,the large-area organic light-emitting diodes and the thermoelectricity,are operating under temperature gradient.The potential abnormal behavior of the exicton dynamics driven by the temperature difference may affect the efficiency and performance of the corresponding devices.In the above situations,the exciton dynamics under temperature difference is mixed with     

Achieving Unusual States of Matter Under High Pressure

The elongation method,originally proposed by Imamura was further developed for many years in our group.As a method towards O(N)with high efficiency and high accuracy for any dimensional systems.This treatment designed for one-dimensional(ID)polymers is now available for three-dimensional(3D)systems,but geometry optimization is now possible only for 1D-systems.As an approach toward post-Hartree-Fock,it was also extended to     

《Chinese Science Bulletin》SUBJECT INDEX TO VOLUME 59(2014)

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An on-line scaling method for improving scalability of a database cluster

The explosive growth of the Internet and database applications has driven database to be more scalable and available, and able to support on-line scaling without interrupting service. To support more client's queries without downtime and degrading the response time, more nodes have to be scaled up while the database is running. This paper presents the overview of scalable and available database that satisfies the above characteristics. And we propose a novel on-line scaling method. Our method improves the existing on-line scaling method for fast response time and higher throughputs. Our proposed method reduces unnecessary network use, i.e. , we decrease the number of data copy by reusing the backup data. Also, our on-line scaling operation can be processed parallel by selecting adequate nodes as new node. Our performance study shows that our method results in significant reduction in data copy time.     

An improved R-tree based on childnode''s probability

R-Tree is a good structure for spatial searching. But in this indexing structure,either the sequence of nodes in the same level or sequence of traveling these nodes when queries are made is random. Since the possibility that the object appears in different MBR which have the same parents node is different, if we make the subnode who has the most possibility be traveled first, the time cost will be decreased in most of the cases. In some case, the possibility of a point belong to a rectangle will shows direct proportion with the size of the rectangle. But this conclusion is based on an assumption that the objects are symmetrically distributing in the area and this assumption is not always coming into existence. Now we found a more direct parameter to scale the possibility and made a little change on the structure of R-tree, to increase the possibility of founding the satisfying answer in the front sub trees. We names this structure probability based arranged R-tree (PBAR-tree).     

An Index mechanism for multi-scale view in spatial databases

There are numerous geometric objects stored in the spatial databases. An importance function in a spatial database is that users can browse the geometric objects as a map efficiently. Thus the spatial database should display the geometric objects users concern about swiftly onto the display window. This process includes two operations:retrieve data from database and then draw them onto screen. Accordingly, to improve the efficiency, we should try to reduce time of both retrieving object and displaying them. The former can be achieved with the aid of spatial index such as R-tree, the latter require to simplify the objects. Simplification means that objects are shown with sufficient but not with unnecessary detail which depend on the scale of browse. So the major problem is how to retrieve data at different detail level efficiently. This paper introduces the implementation of a multi-scale index in the spatial database SISP (Spatial Information Shared Platform) which is generalized from R-tree. The difference between the generalization and the R-tree lies on two facets: One is that every node and geometric object in the generalization is assigned with a importance value which denote the importance of them, and every vertex in the objects are assigned with a importance value,too. The importance value can be use to decide which data should be retrieve from disk in a query. The other difference is that geometric objects in the generalization are divided into one or more sub-blocks, and vertexes are total ordered by their importance value. With the help of the generalized R-tree, one can easily retrieve data at different detail levels.Some experiments are performed on real-life data to evaluate the performance of solutions that separately use normal spatial index and multi-scale spatial index. The results show that the solution using multi-scale index in SISP is satisfying.     

Integrating GIS Web services based on mediating architecture

The geographic information service is enabled by the advancements in general Web service technology and the focused efforts of the OGC in defining XML-based Web GIS service. Based on these models, this paper addresses the issue of services chaining,the process of combining or pipelining results from several interoperable GIS Web Services to create a customized solution. This paper presents a mediated chaining architecture in which a specific service takes responsibility for performing the process that describes a service chain. We designed the Spatial Information Process Language (SIPL) for dynamic modeling and describing the service chain, also a prototype of the Spatial Information Process Execution Engine (SIPEE) is implemented for executing processes written in SIPL. Discussion of measures to improve the functionality and performance of such system will be included.