小麦TCTP基因的克隆及白粉菌诱导下的表达

翻译控制肿瘤蛋白(translationally controlled tumor protein,TCTP)最初在鼠肿瘤细胞中被发现,研究表明TCTP广泛存在于动植物细胞中,并具有多种生物学功能。本研究用RT PCR和RACE技术在抗白粉病栽培小麦Brock中克隆了一个TCTP基因,该基因全长766 bp,推测编码一个168个氨基酸的多肽。ScanProsite分析表明,该多肽链具有2个TCTP特征结构区（TCTP1和TCTP2）和7个可能的功能位点。表达半定量分析发现,该基因受华北地区流行的优势小种15号白粉菌诱导,且随着诱导时间的增加其表达量增加。本研究将可能在小麦白粉病抗性研究领域开辟新的研究思路。     

Peroxidase Activity in Leaves of Wheat Cultivars Differing in Resistance to Erysiphe graminis DC.

The peroxidase activities in leaves from resistant and susceptible cultivars of wheat infected and non-infected by Erysiphe graminis DC were studied. In non-infected wheat, soluble and ionic bound peroxidase activity level was found to be higher in the resistant cultivar than that in the one susceptible to Erysiphe graminis DC. After infecting wheat leaves with Erysiphe graminis DC a remarkable increase in the activity of soluble and ionic bound peroxidases was detected 5 days after inoculation only in the resistant cultivar. In the susceptible cultivar a high increase in the activity of the soluble and ionic bound peroxidases occurred only 15 days after inoculation. Using ion exchange chromatography four peroxidase fractions were obtained from infected susceptible and resistant cultivars as from non-infected ones. The fraction II in non-inoculated resistant cultivars was much higher than that in the susceptible one. This fraction increased after inoculation in both cases reaching a higher level in resistant cultivars. Fraction I was higher in the susceptible cultivar. Electrofocusing profiles of peroxidase from the susceptible and resistant cultivar differed from one another. New peroxidase bands after inoculation appeared only in the resistant cultivar.     

Deposition of Erysiphe graminis Conidia on a Barley Crop

Naturally released Erysiphe graminis conidia were trapped (on horizontal slides, on vertical sticky cylinders and in suction traps) in a barley crop infected with powdery mildew and the numbers of single spores and of clumps of different sizes deposited on the traps were counted. The efficiencies of impaction calculated from deposits and wind speed measurements were higher than expected from mean wind speed measurements. The values were consistent with the hypothesis that spores were removedpredominantly in gusts. More than half the conidia were removed in clumps of two or more spores. The measurements suggest that clumps were more effectively deposited than single spores. The measurements demonstrate that spore release mechanisms can influence spore deposition significantly, especially close to the source.     

Ethirimol-sensitivity in populations of Erysiphe graminis from plots of spring barley

Barley plots, cv. Proctor were sown with either ethirimol-treated or untreated seed. At seedling emergence, plots were inoculated with isolates of Erysiphe graminis either sensitive or insensitive to ethirimol, or left uninoculated. Mildew samples taken from the coleoptile leaves of seedlings, and later from leaf 6 (next but one to the flag leaf) were compared for ethirimol sensitivity using probit analysis. Probit lines were derived using ‘Wadley's analysis’. This analysis uses the number of survivors to estimate the total number of individuals originally present. In the test for ethirimol sensitivity a count of colonies on inoculated test leaves was taken to represent survivors. Microscopic counting of the original inoculum was thus avoided. The analyses snowed that the initial leaf inoculation treatments established mildew populations in the plots, which differed considerably in ethirimol sensitivity. Some 6 wk after inoculation the levels of sensitivity in these populations remained unchanged even though inoculum coming into the experiment from external sources was often much more insensitive than the mildew in some plots. Probit lines of response to ethirimol had shallow slopes, showing that large variability exists in the pathogen for this character.     

Dispersal of Erysiphe graminis and Lycopodium clavatum spores near to the source in a barley crop

In experiments to study dispersal of spores in a crop of barley, a 4-m wide strip of the cultivar Zephyr (a mildew-susceptible variety) was a source of mildew (Eyrsiphe graminis) conidia. Small suction traps, previously calibrated in a wind tunnel, were used to measure spore concentration within and above the crop. Large concentrations of conidia were measured in the crop at the downwind edge of the Zephyr strip but these decreased rapidly with distance downwind. At 1 m concentrations were halved and by 4 m they were no greater than the background concentration measured in the crop upwind of the source. Next to the source, concentrations were much greater within than above the crop and net spore movement (flux) was upwards out of the crop; by 4 m downwind concentrations were greater above the crop and spore flux was reversed. Lycopodium clavatum spores were released in the same crop from a line of point sources. Concentrations also decreased rapidly downwind but, with no background of spores, numbers remained greater within than above the crop further from the source than for E. graminis. Even so by 7 m downwind concentrations in the crop had declined to less than those above. Deposition of L. clavatum spores onto horizontal glass slides in the crop agreed with that expected by settling. However, impaction onto vertical cylinders among plants was much greater than predicted. The reason is not known although turbulent air-flow around plants may, in some way, enhance impaction. Many E. graminis conidia near the source were deposited in clumps. This prevented any accurate prediction of deposition rates as fall speeds of clumps (necessary for prediction) were not known. Not surprisingly, deposition on horizontal slides often exceeded that expected from settling of single spores although it was not always greatest where clumps predominated. The proportion of spores deposited on vertical cylinders and horizontal slides located among plants ranged from 0–02 to 0–27 and from 0–019 to 0–127 of the area dose, respectively. Although these may seem to be small trapping efficiencies, the same deposition rates in a crop with many leaves and stems would rapidly filter most spores from air in the crop and can explain why concentrations were observed to decline so rapidly.     

Adenosine/Adenine-Containing Nucleotides in the Leaves and Roots of Young Barley Plants after Infection of the Primary Leaf by Erysiphe graminis

The levels of adenosine, the adenine nucleotides AMP, ADP, ATP, as well as NAD, NADP, protein and chlorophyll were determined in young barley plants of which the primary leaves were infected by Erysiphe graminis DC. f. sp. hordei MARCHAL. The largest changes of these metabolite levels, compared to the non-infected control, occurred in the infected leaf and to a lesser degree also in the roots and in healthy younger leaves. The increase in the levels of most metabolites in the primary leaf revealed the sink property of this infected tissue and possible stress or defence reactions of the host, whereas the reductions in the roots showed the impaired supply of this natural sink organ due to the infection. Changes in the healthy leaves were most pronounced in the tertiary leaf and may reflect metabolic stimulation in that healthy organ. The changes of the adenosine pool, a precursor of the adenine nucleotides, were discussed in terms of translocation and its possible role as a precursor for fungal purine nucleotide synthesis.     

Analysis of the Effects of Erysiphe graminis DC. on the Growth of Barley

Three barley varieties, Plumage Archer, Proctor, and Haisa II,inoculated soon after emergence with Erysiphe graminis, thecause of powdery mildew, grew and yielded less than barley repeatedlysprayed with lime sulphur. Size of roots was affected more than tops, so the dry-weightratio of root/total plant was decreased. The smaller tops reflectedfewer shoots which, later, were also smaller than on sprayedplants. Decreases in leaf area paralleled decreases in dry weight oftops. Powdery mildew affected the balance between the absorbingsystem and the assimilating and transpiring system by decreasingthe average dry weight of roots per unit leaf area from 0.92to 0.63 mg./cm². Disease also decreased the efficiency of unitleaf area; from 12 to 68 days after inoculation, the mean netassimilation rate was 226.6 in sprayed controls and 166.0?8.6m.g./dm.²/week in the inoculated series. Powdery mildew retarded stem elongation and possibly causedstunting. Fewer and smaller ears were produced by inoculatedthan by sprayed plants.     

Deposition of Erysiphe graminis Conidia on a Barley Crop II. Consequences for Spore Dispersal

Measured deposition rates of Erysiphe graminis conidia are often greater than those calculated from spore fall speed, v^s, and mean wind speed. The predictions of two gradient transfer models of spore dispersal in crops, in which sedimentation and inertial impaction were calculated from v^s and local wind speed or estimated from measured values (Mc Cartney and Bainbridge 1987), were compared. Comparisons of the models suggest that deposition rates close to the source may be underestimated by a factor of two and deposition half distances may be doubled when deposition is calculated conventionally. The results illustrate the importance of accurately describing deposition processes when constructing spore dispersal models.     

Extrachromosomal plasmid-like DNA in the obligate parasitic fungus Erysiphe graminis f. sp. hordei

Summary The obligate parasitic fungus, Erysiphe graminis f. sp. hordei, was found to harbour plasmid-like extrachromosomal DNA. A 1.35-kb fragment of this 9kb plasmid was cloned into the pUC12 vector. No homology was detected to nuclear or mitochondrial DNA. As only about half of the 27 isolates examined contained plasmid-like DNA, this appears to be inessential for fungal survival. The plasmid is frequent in European isolates and is found in both newly collected isolates and in isolates kept under laboratory conditions for many years. No correlation between presence of plasmid and specific avirulence/virulence genes was found. The plasmid appear to be located in the mitochondria.     

The genetical control of incomplete forms of resistance to Erysiphe graminis in spring barley

Eight spring barley lines exhibiting incomplete resistance to powdery mildew in field trials were crossed and back-crossed to a common susceptible parent to produce the basic generations (P₁ P₂, F₁ F₂, B₂ B₂) required for a genetical investigation. By means of joint scaling tests, disease assessments on adult plants in a field trial were used to estimate the genetic parameters contributing to resistance. Lines with major genes, which appeared partially resistant due to the presence of corresponding virulence, were characterised by the presence of a significant dominance component in the analysis. Partially resistant lines which either lacked identifiable major genes or with major genes that were ineffective in the trial, but with good background resistance, could also be distinguished. Significant additive and epistatic effects were detected in all crosses. The prospect of enhancing partial resistance through hybridisation and selection is discussed.     

Evidence for Resistance-inducing Compounds in Conidia of Erysiphe graminis f.sp. hordei

Preparations of Erysiphe graminis f.sp. hordei conidia were spray-applied to the first leaf of barley plants which were subsequently challenge inoculated with virulent powdery mildew. The powdery mildew reducing effect of the preparations was assessed by scoring the outcome of the challenge inoculation. Homogenates of ungerminated conidia, germinated conidia, and methanol-water extracts of germinated conidia reduced the number of powdery mildew colonies. Cell wall fragments from ungerminated conidia, germinated conidia, and conidial germination fluid obtained from conidia germinated in aqueous suspension did not reduce the number of powdery mildew colonies. Microsconical analysis of the infection course following challenge inoculation indicated that the powdery mildew reducing effect is due partly to induced resistance.