Catabolic activity of two phosphoric diester hydrolases in wheat leaves inoculated with brown rust,Puccinia recondita

Changes in total phospholipids and activities of two phosphoric diester hydrolases,i.e. phospholipase C and phospholipase D have been studied in inoculated wheat leaves of 25-day old plants, interacting differentially with three races of brown rust,Puccinia recondita, during early stages of pathogenesis. With progressive rust iafection the lipids and phospholipids were found to decrease in all the three interactions except at later stages (48 h and 72 h) in race 104-interaction. However, total phospholipids at various stages were found to be higher in resistant interaction (race 63) as compared to the other two interactions. An initial increase in activities of phospholipase C and D in all the three interactions was followed by a decline at the intermediate stages which continued in case of phospholipase C activity while a reverse trend was observed in phospholipase D activity towards later stages of infection. Further the enzymatic activities at various stages were found to be higher in the susceptible and intermediate interactions (with races 77 and 104 respectively) as compared to the resistant interaction.     

Fatty acids compositions of polar and non-polar lipid fractions of wheat leaves inoculated with three brown rust races during progressive pathogenesis

The fatty acids composition of the polar and non-polar lipid fractions of wheat leaves was affected due to progressive brown rust infection during early stages of pathogenesis,i.e. 0, 12, 24, 36, 48 and 72 h after inoculation. The three races ofPuccinia recondita differentially affected the composition of saturated and unsaturated fatty acids and their relative occurrence in wheat leaves. The infection of wheat by race 77 resulted in a relative decrease in fatty acid chain length as measured through C₁₆∶C₁₈ fatty acid ratio. An increase in the relative degree of unsaturation (18∶2/18∶3 acids ratio) was recorded in both lipid fractions. Such changes may be taken as one of the earliest characteristics of disease development.     

Pathogen race determines the type of resistance response in the stripe rust –Triticum dicoccoides pathosystem

Wild relatives of crop plants may serve as a promising source for screening for new disease resistance genes that can be utilized in breeding programs. Triticum dicoccoides, the wild progenitor of most cultivated wheats, was shown to harbor many resistance genes against the major diseases attacking cultivated wheat. Stripe rust is a devastating fungal disease that attacks wheat in many regions of the world. New races of Puccinia striiformis Westend. f. sp. tritici, the causative agent of stripe rust, have overcome most of the known Yr resistance genes in wheat. Therefore, there is a need to search for new resistance genes in the T. dicoccoides gene pool. A set of 120 T. dicoccoides accessions, collected from 13 populations representing different habitats in Israel and vicinity, was tested for resistance to three prevalent stripe rust races (38E134, 6E16 and 6E0). Of these 120 accessions, 14, 8 and 12% were resistant to races 38E134, 6E16 and 6E0, respectively, while 57, 2 and 4% were moderately resistant to these races, respectively. A unique resistance was found in the population of Mt Hermon where >80% of the accessions showed resistance to all races. Distribution of infection types (ITs) of race 38E134 showed a normal distribution that can fit a quantitative pattern of response, while the distributions of ITs of races 6E16 and 6E0 had excess of extreme values and therefore showing a qualitative pattern of response. anova testing the main factor effects and interaction showed significant effects of population, race and their interaction on IT. Significant positive correlations were obtained between the resistance to races 6E16 and 6E0 and humidity variables of the collections sites, while resistance to race 38E134 was positively correlated with temperature variables. These results show that the pathogen race can determine the type of resistance response, qualitative or quantitative, in the stripe rust—T. dicoccoides pathosystem. The obtained results also reveal that the distribution of resistance to different pathogen races can be affected by different climatic factors.     

Determination of Races of Pseudomonas syringae pv. glycinea Occurring in Europe

Fifty-eight strains of Pseudomonas syringae pv. glycinea were collected from France, Germany, Hungary, Italy, Poland, The Ukraine and the former Yugoslavia. The bacterial cultures were fluorescent on King's medium B, oxidase negative, produced levan, and induced a hypersensitive reaction (HR) on tobacco leaves within 24h. The race of each strain was determined by inoculating a set of seven differential soybean cultivars: Acme, Chippewa, Flambeau, Harosoy, Lindarin, Merit and Norchief.Conditions of plant cultivation, bacterial inoculation and plant incubation had to be standardized scrupulously to obtain reproducible results. Authentic strains belonging to races 1, 4, 5, and 6 produced the expected reactions on the differentials. However, strains of race 2 and race 4 induced identical responses on the differential cultivars. The differentiating critierion between these two races was the ability of race 2 to produce a brown diffusable pigment on King's medium B. The most prevalent race, occurring in every European country studied, was race 4. This is the most aggressive race of P. glycinea, since it infects all the cultivars of the set of differentials. From 58 strains tested, 42 belonged to race 4, 4 to race 6, and 6 to race 9. For two strains race identification was impossible. The remaining 4 isolates did not fit into the pattern of known races. It is proposed that these strains belong to a new race (no. 10) which is similar to race 5, but can inf, ect the soybean cultivar ‘Lindarin’. On the other hand, race 10 can not infect cv. ‘Chippewa’, in contrast to race 9.     

Genetic studies of leaf and stem rust resistance in six accessions of Triticum turgidum var. dicoccoides.

Six accessions of Triticum turgidum var. dicoccoides L. (4x, AABB) of diverse origin were tested with 10 races of leaf rust (Puccinia recondita f.sp. tritici Rob. ex Desm.) and 10 races of stem rust (P. graminis f.sp. tritici Eriks. &Henn.). Their infection type patterns were all different from those of lines carrying the Lr or Sr genes on the A or B genome chromosomes with the same races. The unique reaction patterns are probably controlled by genes for leaf rust or stem rust resistance that have not been previously identified. The six dicoccoides accessions were crossed with leaf rust susceptible RL6089 durum wheat and stem rust susceptible 'Kubanka' durum wheat to determine the inheritance of resistance. They were also crossed in diallel to see whether they carried common genes. Seedlings of F1, F2, and BC1F2 generations from the crosses of the dicoccoides accessions with RL6089 were tested with leaf rust race 15 and those from the crosses with 'Kubanka' were tested with stem rust race 15B-1. The F2 populations from the diallel crosses were tested with both races. The data from the crosses with the susceptible durum wheats showed that resistance to leaf rust race 15 and stem rust race 15B-1 in each of the six dicoccoides accessions is conferred by a single dominant or partially dominant gene. In the diallel crosses, the dominance of resistance appeared to be affected by different genetic backgrounds. With one exception, the accessions carry different resistance genes: CI7181 and PI 197483 carry a common gene for resistance to leaf rust race 15. Thus, wild emmer wheat has considerable genetic diversity for rust resistance and is a promising source of new rust resistance genes for cultivated wheats.     

Differential Accumulation of Translatable mRNA Species During a Specific Host-pathogen Interaction Between Puccinia recondita tritici and Triticum aestivum

The time-course of mRNA induction in specific cultivar-race combinations was examined using a criss-cross interaction system consisting of nearly isogenic lines of wheat (Sinvalocho M.A. and Gamma 1R) and genetically related pathogenic races of leaf rust Puccinia recondita f. sp. tritici . Infection stimulated the differential accumulation of mRNA species, identified by in vitro translation, after only three days of inoculation with rust spores. Comparisons of different host-pathogen combinations showed polypeptide pattern changes likely to be associated with gene-for-gene relationships. At least two specific mRNAs which code for polypeptide bands of 34 and 24 kDa are associated with the compatible interaction mediated by genes A₁,/A₂ from Gamma 1R wheat line and virulence gene p_t/p₂ of rust race FO₁. Comparisons made using a mutant clone of rust, which elicits an inverse criss-cross relationship with the same wheat lines, are consistent with the proposed specificity of the detected changes.
In addition, two wheat mRNAs (coding for polypeptide bands of 20.5 and 32.5 kDa) were elicited by infection with rust race FO₁ regardless the plant genotype or reaction type. A cDNA clone involved in this kind of race specific induction has been isolated.     

Changes in proteins and isoenzymes in leaves of wheat when infected by stem rust

Summary Biuret assay, gel electrophoresis and immunochemistry were used to study concentrations, forms and activities of proteins of uredospores of Puccinia graminis Pers. f. sp. tritici, in healthy wheat leaves, wheat leaves that had been inoculated with incompatible races of stem rust and leaves which had become rusted.The soluble proteins of primary leaves increased by 25–117% following infection by compatible races of stem rust. There was a corresponding decrease of proteins in uninfected younger leaves. Infection by an incompatible strain of rust led to a temporary 29% increase in soluble proteins.Immunoelectrophoresis and gel electrophoresis of infected leaves showed the presence in them of the forms of malate dehydrogenase, glucose-6-phosphate dehydrogenase, catalase and -amylase characteristic of the rust fungus. In the infected leaves, the activity of certain bands of host glucose-6-phosphate dehydrogenase and catalase changed with the development of the pathogen; the malate dehydrogenase and -amylase of the host were unaffected. In leaves inoculated with an incompatible race there were no obvious changes of any of these enzymes.     

HOST-PATHOGEN INTERACTIONS IN NATURAL POPULATIONS OF LINUM MARGINALE AND MELAMPSORA LINI: I. PATTERNS OF RESISTANCE AND RACIAL VARIATION IN A LARGE HOST POPULATION

Populations of wild flax, Linum marginale and its associated rust fungus Melampsora lini growing at Kiandra, New South Wales, Australia, were sampled during the 1986–1987 growing season. Thirteen different races of M. lini were detected in a sample of 96 isolates. The distribution of isolates was uneven: race A comprised 73% of the samples; race N, 8%; and race H, 5%; while the remaining races were represented by only one or two samples. The dominance of race A increased over the course of the growing season, comprising 67% of the early season samples and increasing to 78% for those collected late in the season. The overall diversity of the pathogen population decreased late in the growing season, but this trend was not statistically significant. The average virulence of individual isolates of the pathogen population increased during the growing season. This trend was most pronounced among the minor races, where the mean number of differential hosts infected increased from 4.58 for early season samples to 5.12 and 5.08 for mid and late season samples, respectively. In contrast to the virulence pattern in the pathogen, the L. marginale population displayed a more even distribution of resistance. In a sample of 67 plants 10 resistance phenotypes were detected from their pattern of resistance/susceptibility to seven pathogen isolates. No phenotype had a frequency that exceeded 30%. Resistance phenotypes were randomly distributed on both a population level and on a fine scale.     

Induced chitinase activity in resistant wheat leaves inoculated with an incompatible race of Puccinia striiformis f. sp. tritici,the causal agent of yellow rust disease

Chitinase specific activity was measured spectrophotometrically in wheat leaf tissues during the compatible and incompatible interactions with Puccinia striiformis f. sp. tritici, the causal agent of yellow rust disease. The wheat cultivar, Federation^* 4/Kavkaz, was inoculated with virulent (134E134A+) or avirulent (4EOA+) races of P. striiformis f. sp. tritici in the first leaf stage. The results showed that chitinase activity pattern was similar in both compatible and incompatible interactions up to 72 hrs after inoculation. However, the specific activity increased rapidly in the incompatible reaction thereafter. In susceptible reaction, chitinase activity gradually declined after 72 hrs post-inoculation reaching a level similar to that in the control plants two weeks after inoculation. Chitinase specific activity in resistance response was at least three times greater than that in the susceptible reaction two weeks following the inoculation. Electrophoresis of native polyacrylamide gel impregnated with 0.1% (w/v) glycol chitinas the substrate revealed the presence of eight chitinase isoforms with relative electrophoretic mobility (R_m) values ranging from 0.11 to 0.64 in the resolving gel. This revised version was published online in June 2006 with corrections to the Cover Date.     

Reaction of some wheat cultivars and breeding lines to Puccinia striiformis f. sp. tritici hot races in Iran

Many physiological races of Puccinia striiformis f. sp. tritici which cause stripe rust in wheat can be determined in different parts of the world. The emergence of new races with different pathogenicity which happens very quickly breaks cultivars resistant and cause disease. Therefore, breeding cultivar for resistance to different pathogenic races should be continued. In this research, pathogenicity of two isolates collected from two regions of Iran were determined by using wheat yellow rust differential lines, which indicated race 70E50A+ and 6E18A+ The responses of 30 wheat genotypes were separately evaluated in the forms of randomized complete block design with three replicates in the seedling stage under greenhouse condition. The components of resistance including latent period and infection type were recorded. Results indicated genotypes were evaluated in terms of both traits and were significant at 1% level. Also, the results from pathogenicity study indicated of effective gene/s included Yr1, Yr2+, Yr3, Yr4, Yr5, Yr10, Yr15, Yr24, Yr26, YrSP, YrND, YrSD and YrSU. From the genotypes studied in the greenhouse condition, 39% of the genotypes showed complete resistance to both races. Probably, resistance genes, Yr32 and YrCV, or the other unknown genes which are types of seedling resistance are either alone or in combination of one another cause strength in resistant genotypes.     

紫外线诱导小麦条锈菌毒性突变及突变体的RAPD分析

以夏孢子相对致死率90%左右作为紫外线处理最适时间,发现8min为小麦条锈菌条中29号单孢菌系(CY29-3)最适处理时间,其夏孢子相对致死率达88.97%。CY29-3的夏孢子经紫外线处理、扩繁、筛选品种筛选及4代的稳定选择后,获得了两个毒性变异的突变菌株。毒性突变产生的Jubi菌株对尤皮Ⅱ号的毒性增强,反应型由野生菌系的0型变为4型;非毒性突变产生的Funo菌株在阿夫上的反应型由野生菌系的4型变为2型。研究结果表明两突变菌株在鉴别寄主上的反应型和毒性范围也明显不同于野生菌系,说明紫外线诱导的小麦条锈菌变异是不定向和复杂的。对野生菌系和两个突变菌株进行RAPD分析后发现,两突变菌株与野生菌系之间的DNA多态性存在显著差异,多态率分别为10.58%和11.57%,说明紫外线可使小麦条锈菌基因组DNA发生较大的变化且突变位点比较复杂。     

Histological Observations on Uromyces phaseoli and Puccinia recondita Infection in Allopurinol-treated Susceptible Plants

The development of rust after administering allopurinol, a specific inhibitor of xanthine oxidoreductase, via roots was studied at the histological level in leaves of susceptible‘Pinto 111’bean plants inoculated with Uromyces phaseoli and‘Thatcher',‘Mentana’and‘Leopardo’wheat plants challenged with Puccinia recondita. A marked reduction and delay in fungal growth was observed in allopurinol-treated plants starting between 24 h and 48 h post-inoculation, i.e. after differentiation of the first haustoria (onset of the biotrophic plant-parasite relationship). Infection hyphae often grew twisted and convoluted in treated hosts, sometimes producing small, irregularly shaped colonies. Differentiation of subepidermal stromata in fungal colonies was delayed and restricted by the treatment and uredospore yield severely reduced. Allopurinol administration also tended to increase the proportion of haustoria which became embedded in thick translucent sheaths during the late stages of infection. These results support the view that plant xanthine oxidoreductase activity is necessary for biotrophic development of rust fungi and suggest that the inhibition of this enzyme, which impairs the pathogen metabolism, may favour some natural host responses to attack such as haustorial sheath formation.     

Spontaneous Genetic Variation for Leaf Rust Reaction in Sinvalocho MA Wheat

Spontaneous off-type plants for leaf rust reaction to Argentine races 66 and 77 of Puccinia recondita tritici were found at rates of 3.4 per 1000 and 0.44 per 1000, respectively, in the progeny of self-pollinated (bagged) spikes of Sinvalocho, MA wheat. Leaf rust reaction changes were associated to aneuploidy in most cases. The variation to race 66 was associated to monosomy of chromosome 6B or probably deletions on chromosome arm 6BL. Some off-type plants however showed, in subsequent generations. changes for reaction to race 77 or simultaneous changes to races 66 and 77 as well as changes in leucine aminopeptidase isozyme expression, which cannot be explained on grounds of allelic dosage. The origin of these genetic and chromosomal changesis not clear in some cases, however, a common origin could be suggested by the presence of transposonlike elements.     

Identification of a New Race of Sporisorium reilianum and Characterization of the Reaction of Sorghum Lines to Four Races of the Head Smut Pathogen

Sporisorium reilianum is the causal agent of head smut on sorghum and maize. In order to effectively utilize host resistance to control this important disease in crops, it is necessary to monitor changes in disease dynamics and virulence of the pathogen. An outbreak of head smut was recently observed in a sorghum field, near Gaoping, Shanxi, China, and research was undertaken to characterize a putative new race of S. reilianum. A set of differential sorghum lines with resistance to several conventional races was used to characterize the newly collected isolate of S. reilianum. The reactions of differential cultivars/germplasm lines to the new isolate indicate that it is a new physiological race of S. reilianum. The new race is highly virulent on sorghum line A₂V4 and its hybrid, Jinza 12, that are known as resistant to all existing Chinese races of S. reilianum, including races 1, 2, and 3. The new isolate of S. reilianum is different from all of the described races of the pathogen; thus, it is designated as race 4 of S. reilianum. Furthermore, a collection of 34 sorghum genotypes including commercial cultivars and germplasm lines was evaluated for disease reaction to the newly described race and the three known races of the pathogen.     

Differentiation-related proteins of the broad bean rust fungus Uromyces viciae-fabae,as revealed by high resolution two-dimensional polyacrylamide gel electrophoresis

On artificial polyethylene membranes providing a thigmotropic signal, uredospores of the broad bean rust fungus Uromyces viciae-fabae differentiated a series of infection structures which in nature are necessary to invade the host tissue through the stomata. Within 24 h germ tubes, appressoria, substomatal vesicles, infection hyphae and haustorial mother cells were developed successively. Alterations in protein metabolism during infection structure differentiation of this obligate plant pathogen were analyzed in the absence of the host plant by high resolution two-dimensional polyacrylamide gel electrophoresis (2-DE) and silver staining. The norm pattern representing the 2-DE protein patterns of the whole developmental sequence of infection structures of U. viciae-fabae showed 733 spots. During infection structure differentiation 55 proteins were newly formed, altered in quantity, or disappeared. Major alterations in the protein pattern occurred during uredospore germination and when infection hyphae were formed. Uredospore germination was characterized by a decrease of acidic proteins and an increase mainly of proteins with isoelectric points ranging from weakly acidic to basic.Abbreviations 2-DE two-dimensional polyacrylamide gel electrophoresis - DAPI 4,6-diamino-phenylindol - kDa kilo Dalton - pl isoelectric point - PMSF phenylmethylsulfonyl fluoride - SDS-PAGE sodium dodecyl sulfate polyacrylamide gel electrophoresis     

Cavity spot of carrots: II. Cell-wall-degrading enzymes secreted by Pythium and pathogen-related proteins produced by the root cells

This study investigates the biochemical relationships between carrot roots and Pythium violae, the pathogen responsible for cavity spot (CS) disease. P. violae isolates obtained from CS lesions, cultured in Petri dishes on agar were used for inoculation of uninfected mature carrots. The fungus secreted a wide spectrum of enzymes that degraded the cellulose and pectic substances of the carrot cell walls. Cellulase and polygalacuronase (pg) showed the highest activity during the first day post-inoculation, subsequently declining. Pectin lyase (PnL), pectate lyase (PeL) and pectin methylesterase (PME) gradually increased to their highest levels of activity 14 to 30 days post-inoculation. This pattern of activity enables the penetration of the fungus through the walls of the host cells and the establishment of the hyphae. Several plant pathogen-related substances such as peroxidase, chitinase, glucanase and polyphenol oxidase were produced in the infected tissue. Peroxidase activity rose in the inoculated roots from day 1 post-inoculation. Chitinase, glucanase and polyphenol oxidase activities first appeared 3–4 days post-inoculation. At this time, two bands corresponding to chitinase at about 26 and 33 KDa and one band corresponding to glucanase at about 24 KDa could be resolved by SDS-PAGE.     

Development of a Differential Set for the Race analysis of Puccinia hordei Otth.

Investigations for the race analysis of P. hordei (location Cologne, W.-Germany) showed, that most of the resistant genotypes in the International differential set with Pa₁ to Pa₈ were ineffective for that purpose. Out of 80 selected barley genotypes with different degree of resistance against the field-collection RG 1975 hosts with a strongly differentiating effect were chosen. This was done using three parameters: mean resistance x?w, its standard deviation x?w and the range z. They were calculated from evaluating the reaction types of 80 barley genotypes infected with 116 single-pustule lines of barley brown rust. Eight hosts with strongly differentiating reactions towards the 116 rust lines were composed into a differential set, which could characterize the heterogeneity of the Cologne P. hordei-population essentially better and more precisely than the International differential set.     

Changes in phenol and peroxidase in the leaves ofJava citronella infected withCurvularia andropogonis

The contet of phenols, o—dihydroxyphenols and peroxidase activity in healthy andCurvularia andropogonis (Zimm.) Boedijn infected leaves ofJava citronella were determined. As a result of infection, the content of phenols and peroxidase increased two- and four-fold, respectively in necrotic lesions compared to healthy leaves. In surrounding tissue of lesions, their increase was one-and half fold only. The peroxidase activity decreased with the maturity of the necrotic lesions. Necrotic lesions produced in response to infection appear to be the consequence of higher accumulation of phenols and their oxidation by peroxidase.     

Pathogenic Diversity in and Sources of Resistance to Uromyces appendiculatus in Southern Africa

The reaction of the first (1983) common bean international differential set and other germplasm to 248 single pustule isolates of the rust fungus Uromyces appendiculatus, collected from various southern African countries, was evaluated. Eleven of the most important isolates were re‐purified and re‐inoculated, this time also on the second (2002) revised and smaller international differential set. The 248 isolates could be grouped into 44 race‐groups. These were subjected to principal coordinates analysis (PCoA). A second PCoA was carried out using 25 of the most important of the 44, together with 34 African races reported by previous authors. Isolates were generally avirulent on accessions with the resistance genes Ur‐3+, ‐5 or ‐11, as well as Compuesto Negro Chimaltenango (CNC) and A 286, all small seeded, and the most useful sources were accessions carrying both Ur‐3 and Ur‐11, for instance BelMiNeb‐RMR‐7, BelDakMi‐RMR‐14 and ‐18. Isolates were generally virulent on large seeded accessions (with, among others Ur‐4, ‐6 or ‐9), reflecting the preference for large seeded beans in southern Africa and co‐evolution of host and pathogen. No large seeded accessions showed broad resistance. The least susceptible was Plant Introduction 260418, which rated resistant to moderately susceptible to the 11 races. These observations were confirmed by field ratings on the same accessions over multiple seasons. According to the PCoA, which proved useful for the identification of differentiating accessions, southern African isolates fell into three main clusters, for which Redlands Pioneer and the South Africa cultivar Teebus were the most discriminating differentials. Other accessions that showed particularly useful differentiating ability were Olathe and 51051. Of these, only Redlands Pioneer has been included in the 2002 differential set. The PCoA grouping of the African races was similar to that of the southern African race‐groups.