Water uptake by barley roots as affected by the osmotic and matric potential in the rhizosphere

Summary The water uptake rates of roots in saline soils are depressed by the simultaneously decreasing matric and osmotic water potentials in the soil surrounding the roots (rhizospheric soil). Unfortunately there are no reliable tools available for direct measurements of the effect of decreasing water potentials in the rhizospheric soil on the uptake rate of soil water by roots. This paper presents some results of a vegetation technique for studying the effect of different combinations of osmotic and matric water potentials in the rhizospheric soil on the water uptake rates of barley roots. Water uptake rates were reduced to a greater extent by decreasing soil matric water potentials than by decreasing soil osmotic water potentials. According to the results of this experiment, there was no relationship between the total soil water potential of a sandy soil and the water uptake rates when the roots were exposed to different combinations of and .     

Analysis of differential gene expression in the central nervous system of Schistocerca gregaria by differential display PCR

Within the optic lobes, the mushroom bodies or other parts of the insect brain, information is processed in an area-specific manner. To study the molecular basis of the abilities of the respective areas, the central nervous system of the desert locust Schistocerca gregaria was dissected into different parts, the optic lobes, the “midbrain”, and the thoracic ganglia. Using a simple electrophoretic approach we were able to show area-specific expression of proteins exclusively present in the optic lobes. To study brain area-specific gene expression in more detail, we adapted the differential display polymerase chain reaction to the specific needs of this project. A number of differentially expressed amplicons were identified. The majority of them could be reamplified and their differential expression verified by northern blot analysis. To demonstrate the efficiency of the approach two amplicons with complementary expression patterns were further analysed. Accepted: 31 August 1997     

Subtractive differential display: a modified differential display technique for isolating differentially expressed genes

Differential display (DD) is a novel PCR-based technique, very commonly used to study differentially expressed genes at the mRNA level. In this paper we report a modified version of this technique that we have used to study the differences between the mRNA population from brain tissue of adult and old rats. We have modified the technique to enhance reproducibility and reduce false positives and redundancy. It is fast and does not require any expensive or uncommon reagent. We choose to call it as subtractive differential display as it is a differential display performed over subtracted mRNA population. We have used this protocol successfully to clone a number of age-related differentially expressed sequences from rat brain that need to be sequenced to establish the gene identity.     

Isolation of genes differentially expressed during the fruit body development of Pleurotus ostreatus by differential display of RAPD

To analyze genes involved in fruit body development of Pleurotus ostreatus, mRNAs from three different developmental stages: i.e., vegetative mycelium, primordium, and mature fruit body, were isolated and reverse-transcribed to cDNAs. One hundred and twenty random PCR amplifications were performed with the cDNAs, which generated 382, 394, 393 cDNA fragments from each developmental stage. From these fragments, four cDNA clones specifically expressed in primordium or mature fruit body were detected. Sequence analysis and database searches revealed significant similarity with triacylglycerol lipase, cytochrome P450 sterol 14 alpha-demethylase and developmentally regulated genes of other fungi. Northern blot analyses confirmed that all of the four cDNAs were unexpressed in mycelium, thus stage-specific genes for fruit body formation of P. ostreatus were successfully isolated.     

Global analysis of gene expression by differential display

Differential display (DD) is one of the most commonly used approaches for identifying differentially expressed genes. However, there has been lack of an accurate guidance on how many DD polymerase chain reaction (PCR) primer combinations are needed to display most of the genes expressed in a eukaryotic cell. This study critically evaluated the gene coverage by DD as a function of the number of arbitrary primers, the number of 3′ bases of an arbitrary primer required to completely match an mRNA target sequence, the additional 5′ base match(s) of arbitrary primers in first-strand cDNA recognition, and the length of mRNA tails being analyzed. The resulting new DD mathematical model predicts that 80 to 160 arbitrary 13mers, when used in combinations with 3 one-base anchored oligo-dT primers, would allow any given mRNA within a eukaryotic cell to be detected with a 74% to 93% probability, respectively. The prediction was supported by both computer simulation of the DD process and experimental data from a comprehensive fluorescent DD screening for target genes of tumor-suppressor p53. Thus, this work provides a theoretical foundation upon which global analysis of gene expression by DD can be pursued.     

优异大麦种质资源的抗性鉴定和评价

1998～1999年,将1997年筛选出的191个农艺性状表现优良的大麦品种进行大麦黄花叶病、大麦白粉病、大麦赤霉病、大麦条纹病及耐盐等抗性的鉴定评价,结果表明,抗及高抗大麦黄花叶病品种60份,抗大麦赤霉病品种59份,高抗大麦条纹病品种89份,抗大麦白粉病品种19份,芽期耐盐品种17份,苗期耐盐品种2份.     

New perspective on the mechanism of alleviating salt stress by spermidine in barley seedlings

Salt stress is considered to be a major limiting factor for plant growth and crop productivity. Salt injuries in plants are mostly due to excess Na⁺ entry. A possible survival strategy of plants under saline environments is the effective compartmentation of excess Na⁺ by sequestering Na⁺ in roots and inhibiting transport of Na⁺ from roots to shoots. Our previous study showed that exogenous application of polyamines (PAs) could attenuate salt injuries in barley plants. In order to further understand such protective roles of PAs against salt stress, the effects of spermidine (Spd) on sodium and potassium distribution in barley (Hordeum vulgare L.) seedlings under saline conditions were investigated. The results showed that exogenous application of Spd induced reductions in Na⁺ levels in roots and shoots with comparison of NaCl-treated plants, while no significant changes in K⁺ levels were observed. Correspondingly, the plants treated with Spd exogenously maintained high values of [K⁺]/[Na⁺] as compared with salt-stressed plants. Moreover, it was shown by X-ray microanalysis that K⁺ and Na⁺ accumulated mainly in the exodermal intercellular space and cortical cells of roots under salinity stress, and low accumulation was observed in endodermal cells and stelar parenchyma, indicating Casparian bands possibly act as ion transport barriers. Most importantly, Spd treatment further strengthened this barrier effects, leading to inhibition of Na⁺ transport into shoots. These results suggest that, by reinforcing barrier effects of Casparian bands, exogenous Spd inhibits Na⁺ transport from roots to shoots under conditions of high salinity which are beneficial for attenuating salt injuries in barley seedlings.     

基因表达差异显示分析技术在创伤研究中的应用

基因是细胞增殖,分化,成熟等各项生命活动的调控中心,也是许多痢疾发生,发展和转归的决定性因素。基因表达的变化必然导致细胞,组织,器官乃至整个机体的各种异常。包括创伤在内的各种内外刺激,都可不同程度地引起基因表达的变化,最终妨碍机体健康。随着生物信息学的逐渐兴起和分子生物学的不断发展并向其他学科的逐渐渗透,业已建立起一系列研究基因表达变化的切实可行的技术手段（即“基因表达差异分析技术”,如DNA微阵列）,对捕获基因表达的种种变化具有重要价值。这些技术已经在肿瘤及其他疾病的研究中得到广泛应用,近几年也逐渐进入创伤研究领域,在一定程度上推动了创伤研究的发展。     

Salt stress enhances proline utilization in the apical region of barley roots

Accumulation of an osmoprotectant, proline, is enhanced in response to salinity in plants. Here, by immunohistochemical analysis, we demonstrated that proline transporter (HvProT) was highly expressed in the apical region of barley roots under salt stress. Free proline was accumulated more in the basal region than in the apical region of barley roots under salt stress, although expression level of HvProT was higher in the apical region. On the other hand, salt stress increased proline and hydroxyproline contents in the cell wall fraction of the root apical region, suggesting increment of proline utilization. Expression of the genes encoding cell wall proteins (proline rich protein and extensin) and cellulose synthase was induced in barley roots by salt stress. These findings indicated that free proline transported by HvProT presumably behaved as a component of cell wall synthesis in the apical region of barley roots under salt stress.     

Expression of defence-related peroxidases Prx7 and Prx8 during abiotic stresses in barley roots

The expression of defence-related peroxidases Prx7 and Prx8 in barley roots grown under selected abiotic stress conditions (toxic metals: Cd, Al, Co, Cu, Hg; drought, salinity, extreme temperatures: heat, cold) and compounds activating (2,4-D) or inhibiting (SHAM) POD activity as well as H₂O₂ and H₂O₂ scavenger (DTT) was characterized. Strong Cd concentration dependent expression of Prx8 peroxidase gene was observed, which correlated with root growth inhibition induced by Cd- and some other stress factors (heavy metals, heat and salinity). Application of H₂O₂ did not cause changes in expression of Prx8, but H₂O₂ scavenger (DTT) as well as the inhibitor (SHAM) and the activator (2,4-D) of PODs induced increase in Prx8 expression. Our results demonstrate that root growth inhibition during any disturbance of active oxygen species (AOS) in root tissue is correlated with up-regulation of Prx8 gene expression in barley roots.     

Potassium transport in salt-stressed barley roots

Salinization of the medium inhibits both K⁺ uptake by excised barley (Hordeum vulgare L.) roots and K⁺ release from their stele, as measured by short-term ⁸⁶Rb uptake and xylem exudation, respectively. Although inhibition was not specific to chloride, mannitol caused a different response from that of inorganic sodium salts, indicating that inhibition was at least partly the result of an ion effect. In roots previously exposed to low levels of NaCl, NaCl stress directly affected stelar K⁺ release, whereas in low-sodium roots stelar K⁺ release was much less salt-sensitive than K⁺ uptake.Abbreviation chCl choline chloride     

mRNA差别显示技术及其在生命科学中的应用

ｍＲＮＡ表达水平的变化决定细胞的功能状态,个体发育、细胞增殖分化与凋谢、生理刺激和药物治疗等过程,都会出现ｍＲＮＡ 表达水平的变化。阐明这种变化有助于揭示细胞生理过程的分子机制。该技术无需更多的背景资料,可快速有效地分离表达水平出现差别的基础,这些基因编剧编码的功能多肽在细胞生理过程中扮演着重要角色。     

用差异显示PCR法筛选与血管外膜细胞表型转化相关的基因

为筛选血管外膜成纤维细胞（adventitial fibroblast,AF）与肌成纤维细胞（myofibroblast,MF）间表型转化有关的基因,实验建立了大鼠胸主动脉AF和MF两种细胞模型,用差异显示聚合酶链反应（DD－PCR）技术获得表达差异片段,对差异片段进行克隆和测序分析,并用定量PCR和Northern blot对差别显示结果进行验证。用反义核酸转染技术观察骨桥蛋白（osteopontin,OPN）对AF迁移的影响。结果表明,两种表型细胞存在明显的基因表达差异,其中一个在MF下调的差异片段与GenBank中NADH脱氢酶亚单位5（NADH dehydrogenase subunit 5,Nd5）基因高度同源。另一个在MF上调的差异片段与OPN基因同源。上述差异表达结果被定量PCR及Northern blot证实。此外还有4个表达序列标志（expressed sequence-tag,EST）在GenBank中未查到同源序列。反义OPN寡脱氧核甘酸可抑制AF的迁移活动。结果提示,AF转化为MF可能与ND5基因下调、OPN上调及其它未知基因的表达改变有关。应用反义技术适度抑制OPN表达在防治血管重塑中具有重要作用。