Cellulose degradation and monitoring of viscosity decrease in cultures of Cellulomonas uda grown on printed newspaper

The rheological behavior of cultures of Cellulomonas uda with shredded printed newspaper as the carbon source was studied. The initial substrate concentrations ranged from 23 to 60 g/L. The changes in apparent viscosity were followed on-line by applying a commercially available process viscometer and discretely using a rotational viscometer with an anchor impeller. During the time of highest cellulose degradation, the broths exhibited a pseudoplastic behavior which could be explained satisfactorily by the power-law model. At the end of cultivation when cellulose degradation slowed down, the broths became Newtonian in behavior. Endo-1,4-beta-glucanase, 1,4-beta-xylanase, beta-glucosidase, and beta-xylosidase activities were also determined during cultivation as well as cellulose degradation and cell mass production. The beginning of endoglucanase formation and the start of the final viscosity decrease of the bacterial paper pulp suspensions could be correlated.     

黄芩苷对嗜水气单胞菌生物膜形成的影响及体内外的抑菌作用

[目的] 探讨中药单体黄芩苷对嗜水气单胞菌在体内外生长及生物膜形成的影响。[方法] 体外实验中,利用牛津杯法检测抑菌圈直径,结晶紫法检测生物膜的形成,通过泳动实验检测黄芩苷对嗜水气单胞菌运动性的影响,紫外吸收法检测细胞膜完整性,用透射电镜技术观察黄芩苷对细菌形态的影响。体内实验利用草鱼为对象检测黄芩苷对嗜水气单胞菌增殖的影响。[结果] 黄芩苷在体外对嗜水气单胞菌有明显的抑菌效果,通过对生物膜的研究发现黄芩苷对生物膜形成具有抑制作用,并同时抑制其运动性。同时黄芩苷可以破坏细胞结构,并增加了细胞膜通透性。体内实验结果显示黄芩苷对嗜水气单胞菌具有清除作用,且具有一定的浓度依赖性。[结论] 黄芩苷在体内外均具有抑制嗜水气单胞菌增殖的作用,有望在水产养殖病害防治工作中得到应用。     

Expression characterization and activity analysis of a cathepsin B from Pacific abalone Haliotis discus hannai

Cathepsin B (EC 3.4.22.1) is a member of lysosomal cysteine protease and has a papain-like fold. In mammals, it is involved in protein degradation and other physiological processes including immune response. However, little is known about the function of cathepsin B in mollusks. In this study, we identified and analyzed a cathepsin B homolog (HdCatB) from Pacific abalone (Haliotis discus hannai), an economically important mollusk species cultured in East Asia. HdCatB is composed of 336 amino acid residues and its mature form is predicted to start at residue 86. HdCatB possesses typical domain architecture of cathepsin B and contains a propeptide region and a cysteine protease domain, the latter containing the four active site residues (Q108, C114, H282, and N302) that are conserved in many different organisms. HdCatB shares 40–60% overall sequence identities with the cathepsin Bofa number of vertebrates and invertebrates and is phylogenetically very close to mollusk cathepsin B. Quantitative real time RT-PCR analysis revealed that HdCatB expression occurred in multiple tissues and was upregulated by bacterial infection. Recombinant HdCatB purified from Escherichia coli exhibited apparent protease activity, which was optimal at 45 °C and pH 6.0. These results indicate that HdCatB is a bioactive protease that is likely to be implicated in the immune response of abalone during bacterial infection.     

长江口不同水域浮游动物数量特征比较

为了分析长江口浮游动物与环境的关系,从2004年到2006年分别于5月份(平水期)和8月份(枯水期)在长江口水域共设22个观测站进行6次海洋调查.结果显示:长江口浮游动物丰度8月份高于5月份,北支略大于南支,长江口口外大于口内. 在大多数情况下,5月份丰度近海略大于咸淡交错水域,8月份相反.长江口浮游动物丰度和分布具有不确定性,水团的季节变化,外海水和冲淡水交汇的位置是影响长江口浮游动物分布特征的重要因素.对长江口南支水域而言,5月份中华华哲水蚤(Sinocalanus sinensis), 8月份火腿许水蚤(Schmackeria poplesia)是最主要的优势种;咸淡交错水域和北支优势种种类较多,真刺唇角水蚤(Labidocera euchaeta)在5月份和8月份都是主要优势种,其次是小拟哲水蚤(Paracalanus parvus).针刺拟哲水蚤(Paracalanus aculeatus),背针胸刺水蚤(Centropages dorsispinatus),太平洋纺锤水蚤(Acartia pacifica)和火腿许水蚤仅为8月份的主要优势种.对近海而言,中华哲水蚤(Calanus sinicus)是主要的优势种,其次是五角水母(Muggiaea atlantica)和肥胖箭虫(Sagitta enflata).     

Structural and catalytic properties of oxidized and reduced chloroplast NADP-malate dehydrogenase upon denaturation and renaturation

Chloroplast NADP-dependent malate dehydrogenase exists in two interconvertible forms: the inactive disulfide-containing form and the active dithiol form. No major difference in secondary structure or conformation was found between the oxidized and the reduced enzyme as determined by circular dichroism and intrinsic protein fluorescence. The guanidine/HCl-dependent unfolding of the enzyme is characterized by two transition midpoints: those of the reduced enzyme are lower by about 0.2 M guanidine/HCl compared to the oxidized enzyme. As shown by analytical ultracentrifugation, there was no effect of guanidine/HCl concentrations up to 0.25 M on the quaternary structure of the enzyme in its oxidized and reduced forms: both sedimentation coefficient (S20,w = 4.9 +/- 0.1 S) and sedimentation equilibrium (75 +/- 3 kDa) yield the dimer. In the oxidized state the enzyme undergoes guanidine-dependent dissociation to the monomer with a midpoint of transition at 0.5 M. The kinetics of unfolding were found to be significantly faster for the reduced than for the oxidized enzyme. Renaturation and reactivation of reduced enzyme was more rapid and occurred with higher yields (100%) than for the oxidized enzyme (60-80% yield). Furthermore, the effect of denaturants on catalytic activity, and reductive activation of the oxidized form, were studied. Both increase in protein fluorescence and a stimulatory effect on the activities at low guanidine/HCl concentrations were observed for the oxidized and the reduced form of the enzyme. Denaturants increase the rate of reductive activation of NADP-malate dehydrogenase.     

锡林河流域羊草草原群落土壤呼吸及其影响因子的研究

采用碱液吸收法对内蒙古锡林河流域羊草 (L eymus chinensis)草原群落土壤呼吸速率进行了连续 2年的野外测定 ,并通过多元回归手段对其影响因子进行了分析。结果表明 :1) 1998～ 1999年两个生长季羊草群落土壤呼吸速率的季节动态均为单峰型曲线 ,最大值出现在 7月下旬 ;2 ) CO2 释放速率、气温和土壤含水量之间有如下关系 :L n Res=5 .85 96 0 .0 12 5 M 0 .0 394T 0 .0 0 49M× T,R2 =0 .70 96 ,表明约有 70 %的土壤呼吸量变化是由气温和土壤含水量共同决定的 ,其中以温度为主 (Res=2 5 6 .75 T0 .6 5 5 7,R2 =0 .5 779) ,Q1 0 在 2 .0～ 3.0之间 ;3)羊草无放牧群落生长季的土壤呼吸总量在 2 49.4～ 32 0 .7g C.m- 2 .a- 1之间 ,放牧群落在 2 37.0～ 30 5 .6 g C.m- 2 .a- 1之间 ,放牧对土壤呼吸影响不大。     

Genetic diversity and differentiation in the critically endangered orchid (Amitostigma hemipilioides): implications for conservation

Amitostigma hemipilioides is an endangered terrestrial orchid endemic to China. In the Southwestern China, the species is found restricted growing in the karst limestone or rock in the edge of farmland, moist hillsides or river. In the present study, the genetic diversity and differentiation was estimated within and among habitats, populations and groups of this species by ISSR markers. Using 13 polymorphic primers, an intermediate level of genetic diversity was found at the species level and population level with the percentage of polymorphic bands (P) of 64.7 and 50.9 %, Shannon index of diversity (I) of 0.3873 and 0.2949, respectively. The analysis of molecular variance (AMOVA) showed that the high level of population differentiation was presented with 45.63 % relative to the total genetic variation residing among eight populations. It was noteworthy that as much as 69.17 % of the total diversity was most likely attributed to the difference among the populations in fragmentation habitat conditions, while 16.32 % of the total diversity could be attributed to the difference among the populations in stable and favorable habitat conditions. The in situ conservation is a top strategy, thus the mycorrhizal fungi and pollinators are protected by protecting habitat and avoiding fragmentation which is helpful to the cycle of this endangered orchid species and recovery of its wild populations. In addition, to maintaining the germplasm bank of this species, the ex situ conservation by habitat simulation and reintroduction has been considered in the living collection at Kaili University ecological garden and Yuntai Mountain scenic spot using the plants collected in this study.     

马铃薯Y病毒CP基因RNAi载体构建与干涉效果测定

RNA沉默(RNAi)介导的植物抗病毒研究是近年来引起广泛关注的一项植物抗病毒基因工程策略.马铃薯Y病毒(PVY)在世界范围内引起马铃薯、烟草等重要经济作物的病害,并且造成严重的经济损失.本文以PW的外壳蛋白(CP)基因为模板扩增300 bp和354 bp两个片段,正向和反向分别插入植物表达载体pROKⅡ的35S启动子下游,从而构建了以PVY的CP基因为靶标的RNAi植物表达载体pROKY300,转入农杆菌EHA105中,以农杆菌渗入法在本氏烟中瞬时表达与PVY CP同源的hairpin RNA.结果表明,瞬时表达的hairpinRNA有效干涉了PVY侵染.