Contents of Volume 53

Plant Molecular Biology -     

INTRASPECIFIC VARIATION AND THE ROLE OF SUPERPARASITISM IN DIAPAUSE INDUCTION OF TRICHOGRAMMA EVANESCENS WEST-WOOD*

Abstract The diapause of two populations of Trichogramma evanescens Westwood (T. evanescens A and T. evanescens B), collected from different Iranian insect pests, was studied. T. evunescens A in the eggs of Corcyra cephalonica was easily induced to stable diapause with constant 8°C, 15 % and fluctuating temperature (11–23) C -11C. T. evanescens B could not be induced to diapause with the same temperatural regime and host. The experiments showed that the host of maternal generations and the ability of avoiding super-parasitism may play an important role in intraspecific variation of T. evunescens in diapause induction.     

Membrane-trafficking RabA4c involved in the effect of glycine betaine on recovery from chilling stress in Arabidopsis

Glycine betaine (GB) can confer tolerance to several types of stress at low concentrations, either after application to plants or in transgenics engineered to overproduce GB. Based on earlier studies on levels of GB in plants and evidence for effects on gene expression, we hypothesized that at least part of this effect could be ascribed to the activation of the expression of stress tolerance genes. Using a strategy based on high-throughput gene expression analysis with microarrays followed by confirmation with northern blots, we identified Arabidopsis genes upregulated in roots that reinforce intracellular processes protecting cells from oxidative damage and others that appear to be involved in reinforcing a scavenging system for reactive oxygen species (ROS) in cell walls. Upregulated genes in roots include those for the membrane-trafficking RabA4c, the root-specific NADPH-dependent ferric reductase (FRO2) localized to the plasma membrane, mitochondrial catalase 2 and the cell wall peroxidase ATP3a. Comparative studies with wild-type Arabidopsis and knockout mutants for the membrane-trafficking RabA4c gene demonstrated that the mutants respond only slightly to GB, if at all, compared with wild-type in relation to root growth recovery after chilling stress, demonstrating the role of RabA4c in relation to the GB effect. The results point toward links between oxidative stress, gene expression, membrane trafficking and scavenging of ROS such as superoxide and hydrogen peroxide in relation to GB effects on chilling tolerance in plants.     

Characterization of cDNA of lycopene beta-cyclase responsible for a high level of beta-carotene accumulation in Dunaliella salina.

Lycopene beta-cyclase (Lyc-B) is the key enzyme in the catalysis of linear lycopene to form cyclic beta-carotene, an indispensable part of the photosynthetic apparatus and an important source of vitamin A in human and animal nutrition. Studies showing that the microalga Dunaliella salina can accumulate a high level of beta-carotene are lacking. We hypothesize that D. salina is closely involved with the catalytic mechanism of Lyc-B and the molecular regulation of its gene. In this study, we used RT-PCR and RACE-PCR to isolate a 2475 bp cDNA with a 1824 bp open reading frame, encoding a putative Lyc-B, from D. salina. Homology studies showed that the deduced amino acid sequence had a significant overall similarity with sequences of other green algae and higher plants, and that it shared the highest sequence identity, up to 64%, with Lyc-B of Chlamydomonas reinhardtii. Codon analysis showed that synonymous codon usage in the enzyme has a strong bias towards codons ending with adenosine. Two motifs were found in the Lyc-B sequence, one at the N terminus, for binding the hypothetical cofactor FAD, and the other was a substrate carrier motif in oxygenic organisms shared by an earlier carotenogenesis enzyme, phytoene desaturase, and Lyc-B. A tertiary structure prediction suggested that the catalytic or binding site structure within LycB from D. salina is superior to that of both H. pluvialis and C. reinhardtii. The LycB protein from D. salina was quite removed from that of H. pluvialis and C. reinhardtii in the phylogenetic tree. Taken as a whole, this information provides insight into the regulatatory mechanism of Lyc-B at the molecular level and the high level of beta-carotene accumulation in the microalga D. salina.     

染色体辐射效应的微机检测 Ⅱ断片率 微核率与细胞畸变率间的相关性检测

本文继前文后,按照设计的线性回归程序在“IBM—PC/XT”微型计算机上,进一步检测了断片率、微核率与细胞畸变率之间的相关性,肯定了微核测定法,断片测定法可以替代染色体畸变分析法。     

Bi（Ⅲ）与金属硫蛋白作用性质研究

 Ｂｉ（Ⅲ）与金属硫蛋白作用性质研究张保林,黄辉,朱凌燕,岳晟,唐雯霞（南京大学配位化学研究所,配位化学国家重点实验室,南京２１００９３）如何降低顺铂或其它抗癌铂的毒性,一直是癌症化疗中的重要课题之一,最近研究发现预先给大鼠或肺癌病人服用铋盐,可以极大...     

人血红细胞酪氨酸蛋白磷酸酶（PTPP）的研究Ⅱ·胞浆PTPP的分离纯化及其主要性质

人血红细胞胞浆部分经（ＮＨ_４）_２ＳＯ_４沉淀,ＤＥＡＥ－纤维素（ＤＥ５２）柱层析,磷酸纤维素柱层析（Ｐ１１）得到部分纯化的ＰＴＰＰ,产率：５．７％,提纯１０７５倍。以（３２）￣Ｐ－Ｔｙｒ－Ｐｏｌｙ（Ｇ_４：Ｔ）作底物,测得其表征Ｋｍ约为０．５－０．８μｍｏｌ／Ｌ,该酶的最适ｐＨ和最适温度分别为７．０－７．８及３７－４０℃。Ｚｎ￣（２＋）等二价金属离子及Ｎａ_３ＶＯ_４等酸根基团对其活性有明显的抑制作用;ＥＤＴＡ、甘油及ＤＴＴ、巯基乙醇等则对其有强烈激活作用;而氟化物、酒石酸等对ＰＴＰＰ活性基本无影响。此外,某些蛋白质、氨基酸、核苷酸及抗肿瘤药物等对ＰＴＰＰ活性也都有不同程度的影响。特别是一些ＰＫｓ及ＰＰｓ在体外对ＰＴＰＰ活性也具有不同的作用。     

人血红细胞蛋白质酪氨酸磷酸酶（PTPP）的研究Ⅰ·PTPP在红细胞胞浆和膜中的分布及其影响因素

通过测定红细胞胞浆及膜中的ＰＴＰＰ活性,发现人正常血红细胞中胞浆ＰＴＰＰ活性约占红细胞ＰＴＰＰ总活性的７０－８０％,膜中只有约２０－３０％的ＰＴＰＰ活性。很多因素诸如：病变、ＰＨ值、温度、离子强度、细胞贮存时间以及药物等,对ＰＴＰＰ在胞浆与膜中的分布有影响。可以推测：膜上的ＰＴＰＰ能通过某种机制解离下来,进入胞浆;相反的过程,胞浆中的ＰＴＰＰ也能通过某种机制与膜结合。这种偶联与去偶联的具体机制及其生理功能还有待进一步探索。     

Detection of Salmonella typhi by polymerase chain reaction

A rapid and sensitive method for detection of Salmonella typhi would help in preventing the spread of outbreaks and in clinical diagnosis. In order to develop unique PCR primers to detect Salm. typhi , ribosomal RNA genes from Salm. typhi (Rawlings) were cloned in pUC18. The resulting clone was confirmed by sequencing. The cloned DNA fragment contained the 5S, part of the 23S rRNA genes and the 5S-23S spacer region (EMBL/GenBank accession No. U04734).
It was expected that the 5S-23S spacer region is divergent unlike the highly conserved 23S+5S genes. This was confirmed by comparison with the rRNA gene sequences in the EMBL/GenBank database. A pair of PCR primers specific for Salm. typhi was obtained, based on this spacer region sequence. The specificity of this pair of primers was tested with 54 Salm. typhi strains (of 27 different phage types). All these Salm. typhi strains showed the positive 300 bp PCR product with this pair of primers. Six other Salmonella species as well as six other non- Salmonella bacteria were tested and none showed the 300 bp PCR product. The sensitivity of the detection level was 0·1 pg of pure Salm. typhi genomic DNA, or approximately 40 Salm. typhi cells in a spiked food sample. This pair of primers therefore has the potential for development into a diagnostic tool for the rapid diagnosis of typhoid fever.     

肿瘤坏死因子－α（TNF－α）及白细胞介素1β（IL1－β）对胶质增生的影响及机理

本文利用小鼠大脑机械损伤模型及体外培养的胶质细胞,采用同位素渗入法观察了细胞介素及其抗体对胶质细胞增生的影响。结果表明：体外培养时ＴＮＦ－α在浓度为１０～２００ｕ／ｍｌ培养液时均能促进胶质细胞增生（Ｐ＜０．０５）,ＩＬ１－β在浓度为５～２００ｕ／ｍｌ培养液时能促进胶质细胞增生,ＴＮＦ－α＋ＩＬ１－β其促进胶质细胞增生作用更强烈,ＴＮＦ－α抗体能完全阻断ＴＮＦ－α的促增生作用,部分阻断ＴＮＦ－α＋ＩＬ１－β的促增生作用。在体实验时,ＩＬ１－β及ＴＮＦ－α的作用与离体时相似,ＩＬ１－β及ＴＮＦ－α亦能促进胶质细胞增生,二者共同作用时促细胞增生作用更强。以上结果提示,外源性ＴＮＦ－α及ＩＬ１－β能促进中枢神经损伤后胶质细胞增生且具有协同作用。