The loss of culturability by Escherichia coli cells in seawater depends on availability of phosphate ions and phosphate transport systems

Using strains with or without the PhoE porin or different components of the phosphate regulon, we determined that maintenance of the culturability of Escherichia coli in seawater depended significantly on the presence of structures allowing access of phosphate ions to the periplasm, then to the cytoplasm of cells. Cells totally deprived of the two main phosphate transport systems (Pit, Pst) exhibited the highest loss of culturability. Most of this effect resulted from the loss of the high-affinity Pst system, and more specifically that of the periplasmic phosphate-binding protein PhoS. Survival was enhanced in seawater supplemented with phosphate (0.5 mm), whether or not these structures were present. From an ecological point of view, it is assumed that the presence of phosphate ions, even at low concentrations, can influence the behavior of E. coli cells in seawater. Offprint requests to: M.J. Gauthier     

Characterization of Acetylcholine-induced Increases in Cytosolic Free Calcium Concentration in Individual Rat Pancreatic β-cells

The intracellular free Ca²⁺ ion concentration ([Ca²⁺]i) was measured using fura-2 microspec-trofluorimetry in individual rat pancreatic β-cells prepared by enzymatic digestion and fluorescence-activated cell sorting. The mean basal concentration of [Ca²⁺]i in β-cells in the presence of 4.4 mM glucose and 1.8 mM Ca²⁺ was 112±1.6 nM (n=207). The action of acetylcholine (ACh) was concentration-dependent, and raising the concentration resulted in [Ca²⁺]i spikes of increasing amplitude and duration in some, but not all of the β-cells. In addition, the β-cells demonstrated variable sensitivity to ACh. The increases in [Ca²⁺]i were rapid, transient and were blocked by atropine at 10^-6M. A brief exposure to 50 mM K⁺ resulted in a transient increase in [Ca²⁺]i similar to that induced by ACh, but resistant to atropine. A high concentration of ACh (100μL 10^-4M or 10^-3M) induced [Ca²⁺]i oscillations in 11 out of 57 β-cells in the presence of 4.4 mM glucose. Using calcium channel blockers and Ca²⁺ free medium, the source of the increase in [Ca²⁺]i was deduced to be from extracellular spaces. Changing the temperature from 22 to 37°C did not affect the action of ACh on [Ca²⁺]i. These data strongly suggest that ACh exerted a direct action on [Ca²⁺]i in normal rat pancreatic β-cells and support a role for Ca²⁺ as a second messenger in the action of ACh.     

α-Domain of human metallothionein IA can bind to metals in transgenic tobacco plants

We have introduced a genetically marked Dissociation transposable element (Ds ^HPT ) into tomato (Lycopersicon esculentum) by Agrobacterium tumefaciens-mediated transformation. Probes for the flanking regions of the T-DNA and transposed Ds ^HPT elements were obtained with the inverse polymerase chain reaction (IPCR) technique and used in RFLP linkage analyses. The RFLP map location of 11 T-DNAs carrying Ds ^HPT was determined. The T-DNAs are distributed on 7 of the 12 tomato chromosomes. To explore the feasibility of gene tagging strategies in tomato using Ds ^HPT , we examined the genomic distribution of Ds ^HPT receptor sites relative to the location of two different, but very closely linked, T-DNA insertion sites. After crosses with plants expressing Ac transposase, the hygromycin phosphotransferase (HPT) marker on the Ds element and the excision markers β-glucuronidase (GUS) and Basta resistance (BAR) facilitated the identification of plants bearing germinally transposed Ds ^HPT elements. RFLP mapping of 21 transposed Ds ^HPT elements originating from the two different T-DNA insertions revealed distinct patterns of reintegration sites.     

豇豆单孢锈菌中病毒样颗粒中双链RNA的存在

从北京西郊清华园附近田间豇豆上采集的豇豆单孢锈菌(Uormyces vignal Barcl)夏孢子。萌发后提取双链RNA,电泳分析可测出300—8000碱基对的三组双链RNA。从萌发的孢子中通过差迷离心提取病毒样颗粒,可获得二种类型的病毒样颗粒,一种直径为35—40nm的等轴颗粒。另一种为长短不等的棒状颗粒,用提纯物提取核酸电泳分析与直接从孢子中提取的双链RNA有相同的核酸带,从而证明这些双链RNA存在于病毒样颗粒中。     

榆耳有性结构及生活史的研究

本文通过电镜扫描、石腊切片及用苏木精染色法和DAPI荧光染色,对榆耳子实体有性结构进行观察,证实榆耳子实体菌盖结构分三层:上表层为毛层,表面着生有排列较密集顶端游离的菌丝,它们相互粘连呈菌丝束;中间层为髓部,由较疏松而相互交织在一起的薄壁菌丝组成,菌丝间充满胶质物质;下表层为子实层,表面起伏不平,呈不规则的疣状突起,上面着生担子和囊状体,担子无隔膜棍棒形,外表有不规则的网状纹饰,其顶部着生4个瓶梗状小梗,每个小梗上着生1个椭圆形或腊肠形担孢子,大小为2.5—3.0×6.0—6.5μm,担孢子表面有不规则的网状纹饰结构。在担子间的囊状体为长圆柱形或圆锥形,表面有较密的不规则的网状纹饰。 榆耳有性生殖为异宗配合。绝大多数担孢子含一个细胞核,很少数担孢子含两个细胞核。孢子萌发为一端萌发,也有少数为两端萌发。初生菌丝单核,不能形成子实体,当两种不同遗传性的交配型的初生菌丝结合后,形成具有锁状联合结构的双核菌丝,并可发育成子实体。榆耳具有典型减数分裂过程,不具有减数分裂后核分裂行为,四个子核分别进入四个担孢子内。 在初生菌丝或次生菌丝上,均可产生间生的或顶生的厚垣孢子。经过温度、光照和紫外线照射的诱发,均未发现有其它类型的无性孢子产生。因此,榆耳菌的生活史和大多数担子     

蕉类的细胞遗传学研究

本文对芭蕉属正蕉组内的一些有代表性的野生种和包括香蕉,大蕉在内的二倍体和三倍体食用栽培蕉,共１８个材料进行了核型比较分析,对其中５个重要的栽培蕉品种的花粉母细胞减数分裂进行了观察,从核型、染色体配对以及染色体 特殊分离行为等３个方面得到的证据表明,并非所有香蕉才同源三倍体,有一些香蕉品种的３个染色体组之间同源程度很低, 通过对一些特征染色体的分析,推测这种香蕉的单染色体组很可能来自ＢＢ野生蕉、大蕉     

水稻悬浮细胞的超低温保存研究

本研究分析了水稻悬浮细胞的生理状态和各种冷冻前处理等因素对超低温保存后细胞存活率的影响。结果表明,继代后培养3—5天,处于对数生长期的细胞,采用二步冷冻法,超低温保存后存活率最高。电镜超微结构观察显示,0.5mol/L山梨醇预培养,10%DMSO 0.5mol/L山梨醇复合保护剂处理,液泡显著变小,数目明显减少,从而降低了细胞内自由水含量,增强细胞的抗冷冻能力。在上述合适的前处理和冷冻-化冻条件下,超低温保存水稻悬浮细胞的恢复生长率为58%,恢复生长的细胞转移到分化培养基上,可再生健壮绿苗, 移植到盆钵,在温室中长成正常结实的植株。