A gene encoding alanine racemase is involved in spore germination in <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis>

Alanine racemase is a major component of the exosporium of Bacillus cereus spores. A gene homologous to that of alanine racemase (alrA) was cloned from Bacillus thuringiensis subsp. kurstaki, and RT-PCR showed that alrA was transcribed only in the sporulating cells. Disruption of alrA did not affect the growth and sporulation of B. thuringiensis, but promoted l-alanine-induced spore germination. When the spore germination rate was measured by monitoring DPA release, complementation of the alrA disruptant reduced the rate of l-alanine-induced spore germination below that of even wild-type spores. As previously reported for spores of other Bacillus species, d-alanine was an effective and competitive inhibitor of l-alanine-induced germination of B. thuringiensis spores. d-cycloserine alone stimulated inosine-induced germination of B. thuringiensis spores in addition to increasing l-alanine-induced germination by inhibiting alanine racemase. d-Alanine also increased the rate of inosine-induced germination of wild-type spores. However, d-alanine inhibited inosine-induced germination of the alrA disruptant spores. It is possible that AlrA converted d-alanine to l-alanine, and this in turn, stimulated spore germination in B. thuringiensis. These results suggest that alrA plays a crucial role in moderating the germination rate of B. thuringiensis spores.     

糖蜜草(Melinis minutiflora Beauv.)内生固氮菌分离鉴定

糖蜜草（Melinis minutiflora Beauv．）是热带地区的一种优良牧草。采用选择性培养基在厌氧和好氧两种培养条件下,从糖蜜草根、茎中都可分离得到具有较强固氮酶活性的菌株。通过SDS-PAGE全细胞蛋白电泳技术快速聚类分析表明,来源于糖蜜草中的菌株为同一类群。16S rDNA序列分析和总DNA的G＋c％含量进一步确定糖蜜草中所分离的菌株属于固氮螺菌属（Azospirillum）,与产脂固氮螺菌（Azospirillum lipoferum）亲缘关系较近。BIOLOG板测定结果显示,糖蜜草菌株TMCY243对多种碳源具有很强的适应性,与产脂固氮螺菌（A．lipoferum）的模式菌株DSM 1691存在着较大的差异。以上结果表明,糖蜜草内生固氮菌为固氮螺菌属的一个新类群。     

Structure and Function of sanV: A Gene Involved in Nikkomycin Biosynthesis of Streptomyces ansochromogenes

A 6.3-kb BamHI-BglII DNA fragment was cloned from cos20 by using chromosome walking strategy. It was partially sequenced with the result that there is a possible ORF of 1272 nucleotides. The ORF designated sanV was deposited in GenBank under accession no. AF469955. Database search indicated that the deduced protein of sanV shows 28% identity and 44% similarity over 405 amino acid residues to the large component (E) of glutamate mutases from Clostridium cochlearium. Gene disruption was performed to study the function of sanV. It was found that sanV disruptants exhibited much poorer inhibition to the test strain than the wild-type S. ansochromogenes 7100. Furthermore, HPLC analysis indicated that the sanV disruptants almost did not produce nikkomycins X and Z, whereas they accumulated new nikkomycins O_x and O_z, which revealed that sanV is an important gene involved in the biosynthesis of the peptidyl moiety of nikkomycins. Received: 13 June 2002 / Accepted: 27 July 2002     

人铜锌超氧化物歧化酶基因在乳酸乳球菌中的食品级表达

以lacF基因为食品级选择标记,构建了乳酸乳球菌食品级基因表达系统,并进而实现了人铜锌超氧化物歧化酶基因在乳酸乳球菌中的食品级表达。首先构建了含有lacF基因两侧同源DNA序列(0.5kb)的整合型质粒pUCEmDE,通过pUCEmDE与乳酸乳球菌MG5267染色体上单拷贝的乳糖操纵子之间的同源双交换,构建了lacF基因缺失突变的食品级受体菌WZ103 (Lac-),并经PCR及Lac表型检测所验证。然后构建了互补质粒pMG36eF,其lacF基因的表达受组成型的强启动子P32的控制。将其电转化导入WZ103后,Lac+表型得到恢复,表明WZ103中lacF基因的功能可被互补质粒pMG36eF上的lacF基因互补。随后,以互补质粒pMG36eF为基础,构建了不含任何抗生素抗性选择标记的人铜锌超氧化物歧化酶基因的食品级表达质粒pWZ104。通过非变性聚丙烯酰胺凝胶电泳和SOD活性凝胶染色分析,检测到WZ103(pWZ104)中Cu/Zn SOD的表达,并且具有生物活性。     

酵母菌絮凝的分型及其生理生化特性的研究

通过对410余株酵母菌进行絮凝测定,从中筛选到5株强絮凝菌。依据不同糖对其絮凝水平的抑制,将5株强絮凝菌分为Flo 1型和NewFlo型。对这两种絮凝型菌株的相关生理生化特性进行了研究。结果表明,Flo 1型菌絮凝只受甘露糖抑制,它对高温(70℃)、蛋白酶E、胰蛋白酶敏感,而对蛋白酶K、糜蛋白酶、Ca\+\{2+\}\,pH有一定耐受性。NewFlo型菌絮凝受甘露糖等多种糖抑制,它对高温(70℃)、各种蛋白酶、Ca\+\{2+\}、pH均较敏感。这两种类型菌株絮凝的最适Ca\+\{2+\}浓度为10mmol/L～1mol/L,最适pH为3.0～45。     

流域景观格局与河流水质的多变量相关分析

流域内的景观格局改变是人类活动的宏观表现,会对河流水质产生显著影响,因此明确影响水质变化的关键景观因子,对于深入了解景观对水质的影响机制具有重要的研究价值。选择广东省淡水河流域为研究对象,以2007年ALOS卫星影像以及水质监测数据为基础,运用空间分析和多变量分析方法,分析淡水河流域景观格局与河流水质的相关关系。用包括流域和河岸带尺度的景观组成和空间结构信息的景观指数表征景观格局,用Spearman秩相关分析、多元线性逐步回归模型和典型相关分析(CCA)研究景观指数和水质指标的相关关系。研究结果表明:林地、城镇用地和农业用地占淡水河流域总面积超过90%,其中城镇用地超过20%。多元线性逐步回归分析和CCA结果说明水质指标受到多个景观指数的综合影响,反映了景观格局对水质的复杂影响机制。流域景观格局对河流水质有显著影响,流域尺度的景观指数比河岸带尺度的景观指数对水质影响更大。城镇用地比例是影响耗氧污染物和营养盐等污染物浓度最重要的景观指数,林地和农业用地对水质的影响较小。另外,景观破碎化对pH值、溶解氧和重金属等水质指标有显著影响。CCA的第一排序轴解释了景观指数与水质指标相关性的54.0%,前两排序轴累积能解释景观指数与水质指标相关性的87.6%,前两轴分别主要表达了城市化水平和景观破碎化水平的变化梯度。淡水河流域的景观格局特征从上游到下游呈现出城市—城乡交错—农村的景观梯度,水质变化也对应了这个梯度的变化,说明人类活动引起的流域土地覆盖及土地管理措施变化会对水质变化产生显著影响。     

槐尺蛾卵期及幼虫期的预测预报

槐尺蛾是园林重要绿化树之一的国槐、龙爪槐最大食叶害虫,既食尽叶片影响观赏价值和绿化成果,又排泄大量粪便和吐丝下垂,有碍市容市貌．该虫发生世代多,准确预报各代幼虫发生期,则是防治的关键．经多代饲养研究测定卵期C＝10.39±0.84（℃）,K＝67.57±3.03（DD）;幼虫期C＝9.16±0.42（℃）,K＝241.26±5.21（DD）．其结果符合实际发生情况.