桓仁林蛙和桓仁产东北林蛙的染色体组型

用骨髓细胞制片法对桓仁林蛙（Rana huanrenesis）和桓仁产东北林蛙（R．dybowskii）的染色体组型进行了报道,两种林蛙的染色体数均为2n=24,都可配成12对,按照相对长度可分为3组,A组包括第1～5对,为大型染色体（相对长度〉9．0）;B组是第6对,为中型染色体（相对长度在7．0～9．0之间）;C组包括第7～12对,这一组为小型染色体（相对长度〈7．0）。在两种林蛙的染色体组型中未发现有异型性染色体。桓仁林蛙的第1、3、4、5对为中部着丝点染色体,第9对为端部着丝点染色体,其余各对为亚中部着丝点染色体。东北林蛙的第1、2、3、4、5、6、8对为中部着丝点染色体,第9对为端部着丝点染色体,其余各对为业中部着丝点染色体。两种林蛙的第11对染色体长臂有明显的次缢痕。     

Mutagenesis breeding of high echinocandin B producing strain and further titer improvement with culture medium optimization

Bioprocess and Biosystems Engineering - A combination of microbial strain improvement and statistical optimization is investigated to maximize echinocandin B (ECB) production from Aspergillus...     

梨果实愈伤组织褐腐病菌侵染过程cDNA-SRAP差异分析

梨果实褐腐病危害果实,可造成果实运输和储藏过程中严重的经济损失。本研究构建了褐腐病菌侵染梨果实愈伤组织离体系统,对梨褐腐病菌侵染其果实愈伤组织不同时期进行细胞学观察分析,基于cDNA-SRAP技术,分析该过程中差异基因表达,以期分离克隆与梨果实抗病反应过程相关的防卫基因。结果表明：与未侵染的梨果实愈伤组织相比,侵染12～60h过程中梨果实褐腐病菌从表面逐渐深入到内部细胞;30个SRAP引物组合共扩增出457条带,其中差异回收条带数为16条,差异比率为3.5%。最终获得5条差异基因表达条带。核酸序列同源性分析表明,其中1条差异基因片段未搜索到任何同源蛋白,2条差异基因片段经序列比对,序列相似度一致,与苹果属的肉桂醇乙酰脱氢酶（CAD）同源性为96%;其他2条差异基因片段分别与DNA结合蛋白（DBP）和寡肽转运蛋白（OPT）基因序列同源,其同源性为85%和78%,因此暂将这3个基因命名为PbCAD、PbDBP和PbOPT。荧光定量PCR结果表明,PbCAD基因在褐腐病菌侵染梨果实愈伤组织12和24h时相对表达量最高,为对照的2.94和2.66倍;PbOPT基因在褐腐病菌侵染梨果实愈伤组织12～36h时相对表达量明显升高,为对照的2.17～2.46倍,而其他时期表达量均与对照接近;PbDBP基因表达量在整个侵染时期均与对照接近。因此我们推测PbCAD和PbOPT基因可能为梨褐腐病菌侵染梨果实愈伤组织响应的相关防卫基因。     

Improvement of 1,3-dihydroxyacetone production from Gluconobacter oxydans by ion beam implantation

Improvement of dihydroxyacetone (DHA) production by mutagenesis of ion beam implantation and medium optimization using response-surface methodology (RSM) were investigated in this work. More than 1000 mutant strains were selected through a mutagenesis method using N(+) ions implantation with a dose of 60?×?(2.6?×?10(13)) ions/cm(2) and energy of 10?keV. Several high-yield mutant strains were showed the potent application for DHA production and the genetically stable mutant strain G. oxydans ZJB09113 was selected for optimization of cultivation condition by RSM. The optimal medium for DHA fermentation is composed (in g/L) of yeast extract 4.88, CaCO(3) 2.00, and glycerol 52.86?mL/L (initial pH 4.89). The maximal DHA concentration of 40.0?g/L was achieved after 24?hr of shaken flask fermentation at 30°C with 150?rpm, and 196.3% increase in DHA production in comparison with unoptimized conditions.     

物分子系统发育与进化研究的基因选择、引物设计与应用策略

引物选择、设计与应用策略是植物分子系统发育与进化研究的关键环节。本文综述了基因选择的原则、引物设计的技巧以及如何有效地利用所涉及的片段获取相应的PCR片段的方法。     

Plant drought tolerance assessment for re-vegetation in heterogeneous karst landscapes of southwestern China

A better understanding of plant water relations is needed for evaluating the suitability of plant species to site-specific reforestation programs in the heterogeneous karst landscapes in southwestern China that are characterized by temporary water deficit. During both wet and dry periods, leaf water potentials of 65 plant species from five different growth forms were studied at three representative sites (forest, shrubland and grassland), to compare their adaptive strategies against water stress and assess their suitability for reforestation programs. Herbs showed the highest predawn and midday water potentials and smallest diurnal ranges of water potential values at all the three sites, indicating that they follow water stress avoidance strategies. During the dry period, evergreen shrubs showed low water potentials, the largest diurnal ranges and highest soluble sugar contents. This indicates that they have a tolerance strategy responding to water stress. Deciduous shrubs and trees still showed relatively large diurnal ranges of water potential values and high soluble sugar contents, and did not shed leaves when experiencing the lowest midday water potentials during the dry period. They shed leaves only later in the dry winter period when even more serious drought was experienced. Their strategies seem to include both tolerance and avoidance mechanisms. Evergreen trees revealed relatively low water potentials with smallest diurnal range water potentials at the shrubland site, especially during the dry period, which indicated their weak ability to tolerate severe water stress. Increasing degradation of the vegetation clearly impacts negatively plant water relations. Using the ranges of leaf water potentials, the relative suitability of the plants for reforestation could be evaluated.     

Effects of ulinastatin, a urinary trypsin inhibitor, on synaptic plasticity and spatial memory in a rat model of cerebral ischemia/reperfusion injury

Established therapies for cerebral ischemia-reperfusion injury are currently limited. The urinary trypsin inhibitor ulinastatin (UTI) is considered cytoprotective against ischemia-reperfusion injury in internal organs through its anti-inflammatory activity. We aimed to investigate the neuroprotective effects of UTI on learning and memory of rats after cerebral ischemia-reperfusion injury. Rats were treated with UTI at 10,000 U/kg body weight, then underwent ischemia and reperfusion by the middle cerebral arterial occlusion (MCAO) method. At various times after the onset of reperfusion, we evaluated neurologic impairment scores. Brain sections underwent immunohistochemical staining for synaptophysin and calcium-binding protein S100β. Other rats underwent the Morris water maze test to determine the effects of UTI on learning and memory. Spatial reference learning and memory were improved with UTI treatment by down-regulating S100β-positive cells and preventing the loss of neural cells. Thus, UTI has a neuroprotective role on synaptic plasticity and spatial memory with cerebral ischemia-reperfusion injury in rats.     

Use of glycerol for producing 1,3-dihydroxyacetone by Gluconobacter oxydans in an airlift bioreactor

1,3-Dihydroxyacetone can be produced by biotransformation of glycerol with glycerol dehydrogenase from Gluconobacter oxydans cells. Firstly, improvement the activity of glycerol dehydrogenase was carried out by medium optimization. The optimal medium for cell cultivation was composed of 5.6 g/l yeast extract, 4.7 g/l glycerol, 42.1 g/l mannitol, 0.5 g/l K₂HPO₄, 0.5 g/l KH₂PO₄, 0.1 g/l MgSO₄·7H₂O, and 2.0 g/l CaCO₃ with the initial pH of 4.9. Secondly, an internal loop airlift bioreactor was applied for DHA production from glycerol by resting cells of G. oxydans ZJB09113. Furthermore, the effects of pH, aeration rate and cell content on DHA production and glycerol feeding strategy were investigated. 156.3 ± 7.8 g/l of maximal DHA concentration with 89.8 ± 2.4% of conversion rate of glycerol to DHA was achieved after 72 h of biotransformation using 10 g/l resting cells at 30 °C, pH 5.0 and 1.5 vvm of aeration rate.     

Enantioselective biocatalytic hydrolysis of (<Emphasis Type="Italic">R</Emphasis>,<Emphasis Type="Italic">S</Emphasis>)-mandelonitrile for production of (<Emphasis Type="Italic">R</Emphasis>)-(−)-mandelic acid by a newly isolated mutant strain

(R)-(−)-Mandelic acid (R-MA) is an important intermediate with broad uses. Recently, R-MA production using nitrilase has been gaining more and more attention due to its higher productivity and enantioselectivity. In this work, a new bacterium WT10, which exhibited favorable nitrilase activity and excellent enantioselectivity for production of R-MA by enantioselective biocatalytic hydrolysis of (R,S)-mandelonitrile, was isolated and identified as a strain of Alcaligenes faecalis. In order to improve its nitrilase activity for industrial application, the wild-type strain WT10 was further subjected to mutagenesis using a combined LiCl–ultraviolet irradiation and low energy N⁺ ion beams implantation technique. A valuable mutant strain A. faecalis ZJUTB10 was obtained. The nitrilase specific activity of the mutant strain was greatly improved up to 350.8 U g⁻¹, in comparison with wild-type strain WT10 of 53.09 U g⁻¹. The reaction conditions for R-MA production by mutant strain A. faecalis ZJUTB10 were also optimized. Nitrilase activity in mutant strain showed a broad pH optimum at pH 7.7–8.5. The optimal temperature was 35°C. The highest production rate reached 9.3 mmol h⁻¹ g⁻¹. The results showed that mutant strain A. faecalis ZJUTB10 was a new candidate for efficient R-MA production from (R,S)-mandelonitrile and could potentially be used in industrial production.