The Streptomyces plasmid SCP2*: its functional analysis and development into useful cloning vectors

Detailed restriction maps of the plasmid SCP2* and its deletion derivative pSCP103 were constructed. DNA fragments carrying hygromycin (Hyg), thiostrepton (Thio) or viomycin-resistance (VioR) determinants were inserted into pSCP103, and various segments were deleted from the resulting plasmids. Changes in plasmid phenotypes associated with these insertions and deletions allowed the localisation and characterisation of plasmid replication, stability, transfer and fertility functions. Several useful cloning vectors were constructed. They are able to maintain large (greater than 30 kb) DNA inserts, with stable inheritance at a low copy number (1-2 per chromosome) and without structural rearrangements, in Streptomyces hosts. The vectors have a broad host range in the genus Streptomyces. One of them (pIJ903) is a shuttle vector for Streptomyces and Escherichia coli.     

Upper limits of temperature for growth inChlorella (Chlorophyceae)

The upper limit of temperature for growth is a species-specific character in the genusChlorella. The limits of 14Chlorella species range from 26–30°C (C. saccharophila) to 38–42°C (C. sorokiniana), withC. fusca var.vacuolata (34°C) andC. kessleri (34–36°C) assuming an intermediate position. Thus, there is no wide gap in the temperature limits between the normal (“low-temperature”) species ofChlorella and the “high-temperature” species,C. sorokiniana.     

Sulphide tolerance in coastal halophytes

The effect of sulphide on the growth of several species of salt-marsh plants was investigated. Relative growth rates were significantly reduced in two upper-marsh species, Festuca rubra and Atriplex patula, and in the lower-marsh species Puccinellia maritima. However the growth of Salicornia europaea, a species frequently associated with sulphide-containing sediments, was unaffected. In a separate experiment the wide ranging halophyte Aster tripolium, also appeared to be tolerant of sulphide at a concentration frequently encountered in salt marshes. Sulphide pretreatment inhibited the activity of two metallo-enzymes, polyphenol oxidase and external phosphatase, in plants from the upper marsh, but had no effect on enzymes from P. maritima or S. europaea. The rate of respiration by root tissue was significantly reduced in all of the species investigated but whereas the uptake of ⁸⁶rubidium was markedly inhibited in the other three species, uptake by S. europaea showed a significant stimulation. Similarly, whereas sulphide-grown plants of F. rubra, A. patula and P. maritima had a considerably reduced tissue iron content, the total iron concentration in S. europaea tissues was comparable to that of the controls. When the sulphide-tolerant species A. tripolium was grown in sulphide-containing media there was no significant effect on the tissue concentration of any of the elements investigated. These results are discussed in relation to possible mechanisms of sulphide toxicity and resistance.     

Effects of monovalent cations on red cell shape and size

Human erythrocytes were incubated in isotonic solutions of different monovalent cations. The apparent size of the red cells measured on scanning electron microscopic pictures decreases in the order Li⁺>Na⁺=K⁺>Rb⁺. These differences in size are abolished after pretreatment with trypsin, which removes a large part of the charges associated with membrane glycoproteins. Shape alterations are also observed. Normal biconcave shapes are visible after Na⁺ or K⁺ incubation, whereas Li⁺ leads to flabby, flattened cells with a certain tendency to crenation, and Rb⁺ causes more pronounced biconcavity with a certain tendency to cupping. The overall effects of pretreatment with trypsin are similar to those of Li⁺. Our results provide evidence that the electrostatic repulsion of glycoproteins and other charged membrane components may play an essential role in maintaining red cell shape.     

低温对不同耐寒力的黄瓜幼苗子叶的各细胞器中NAD~+-苹果酸脱氢酶的影响

NAD~+-MDH在黄瓜子叶中的定位是细胞溶质中占总活性的55～59％,线粒体为38～35％,叶绿体为7％。其同工酶谱亦为细胞溶质中带数最多,全青为5条,粤早3号为4条,线粒体和叶绿体均为1条,品种间无明显差异。黄瓜幼苗随低温胁迫的加剧,伤害逐步加重,子叶电解质渗出率明显增加,NAD~+-MDH活性亦不断下降,其中叶绿体的NAD~+-MDH对低温最敏感,1±1℃处理就能反映品种间耐寒力的差异。叶绿体和线粒体的NAD~+-MDH同工酶对低温的反应与活性变化一致,谱带数没有差异,只是活性降低。细胞溶质部分酶带较多,各条酶带对低温的反应不同。     

菠萝叶膜脂脂肪酸组成的温度效应及其与抗寒性的关系

生长在不同季节的菠萝叶膜脂脂肪酸的配比存在着明显差异;随着大气温度的下降,18:1含量显著减少,18:2和18:3含量增加。不同品种均表现出一致的变化趋势。致害低温破坏了膜脂,使较不抗寒品种的16:0含量增加,18:2和18:3含量减少;较抗寒品种这种变化则较不显著。适当低温锻炼能改变膜脂脂肪酸的代谢过程,16:0和18:1含量减少,18:3含量增加。当处于更低温度时,除了16:0和18:1继续减少外,有一部分18:2也脱饱和而转变为18:3。因之明显地增加了膜脂中18:3的含量和脂肪酸的不饱和度,从而有利于抗寒性的提高。而品种间的抗寒性差异亦是在此低温期间表现出来。