Nonhost resistance of rice to rust pathogens

Rice is atypical in that it is an agricultural cereal that is immune to fungal rust diseases. This report demonstrates that several cereal rust species (Puccinia graminis f. sp tritici, P. triticina, P. striiformis, and P. hordei) can infect rice and produce all the infection structures necessary for plant colonization, including specialized feeding cells (haustoria). Some rust infection sites are remarkably large and many plant cells are colonized, suggesting that nutrient uptake occurs to support this growth. Rice responds with an active, nonhost resistance (NHR) response that prevents fungal sporulation and that involves callose deposition, production of reactive oxygen species, and, occasionally, cell death. Genetic variation for the efficacy of NHR to wheat stem rust and wheat leaf rust was observed. Unlike cereal rusts, the rust pathogen (Melampsora lini) of the dicotyledenous plant flax (Linum usitatissimum) rarely successfully infects rice due to an apparent inability to recognize host-derived signals. Morphologically abnormal infection structures are produced and appressorial-like structures often don't coincide with stomata. These data suggest that basic compatibility is an important determinate of nonhost infection outcomes of rust diseases on cereals, with cereal rusts being more capable of infecting a cereal nonhost species compared with rust species that are adapted for dicot hosts.     

Tissue culture of the alternate hosts of wheat,rye and oat rusts and their response to in vitro inoculation with the rust pathogens

Alternate host plants of cereal rust fungi are necessary for studying the rust sexual cycle and pathogenicity. These plants are usually difficult to propagate through cloning, while seed-propagated plants may have variable responses to the pathogen. To overcome these obstacles, tissue culture, under controlled and aseptic conditions, was utilized for clonal propagation and in vitro inoculation of the following species: Rhamnus palaestinus Boiss., the alternate host of oat (Avena spp.) crown rust (Puccinia coronata Corda); Thalictrum speciosissimum L., the alternate host of brown leaf rust of wheat (Puccinia recondita f. sp. tritici Eriks. & Henn.); and Lycopsis arvensis L., the alternate host of rye (Secala spp.) leaf rust (Puccinia recondita f. sp. recondita Rob. & Desm.). Shoot culture procedures for initial establishment and proliferation were developed for all three alternate host species. Shoot cultures were multiplied at rates ranging from 0.3 to 1.7 shoots/week. Successful infection following inoculation with teliospores of the corresponding rust fungi was obtained for R. palaestinus and T. speciosissimum but not for L. arvensis. The hardening and acclimatization efficiency of rooted T. speciosissimum and L. arvensis was of 80–90%. The propagation efficiency for R. palaestinus was not successful because of the low rate and poor quality of its rooting. It is concluded that the in vitro system might be used as an alternative method for inoculation and multiplication of alternate hosts of cereal rusts, although more experimentation is needed to define accurately the appropriate conditions for the proper infection response. This revised version was published online in June 2006 with corrections to the Cover Date.     

The relationship between hypersensitive tissue and resistance in wheat seedlings infected with Puccinia graminis tritici

Data obtained during the first 120 hr. of the infection process in several resistant and susceptible varieties of wheat infected with Puccinia graminis tritici did not show any consistent relationship between the growth of the rust colony and the rate of increase of host tissue showing hypersensitive necrosis. Furthermore, no apparent relationship existed between the area of the rust colony at any given time after inoculation, and the amount of hypersensitive necrotic tissue per unit area of the leaf colonized. Thus, it is suggested that hypersensitive necrosis of the host tissue is a consequence and not the cause of resistance of wheat to infection by P. graminis tritici.     

Production of teliospores by some races of Puccinia graminis f. sp. tritici in detached wheat and barley leaves

Five races of Puccinia graminis f. sp. tritici cultured on wheat and barley leaves in two nutrient solutions were studied for teliospore formation by subjecting them to varying treatments of temperature and light. The early appearance as well as a high percentage of teliospore formation occurred in 100 ppm benzimidazole solution on wheat or barley leaves kept at 30°C and 500 footcandles of light. The feasibility of maintaining and multiplying races of Puccinia graminis f. sp. tritici in continuous cultures on detached leaves depends on various factors, the most important being the onset of the teliostage of the fungus. The appearance of the teliospore denotes the culmination of the sporophytic or repeating stage of the wheat rusts. In this paper, some factors that influence the production of teliospores by certain races of Puccinia graminis tritici in detached leaves are discussed.     

Effect of temperature, light and host on prepenetration development of Puccinia graminis avenae and Puccinia coronata avenae

The influence of temperature and light on prepenetration development of single and mixed isolates of Puccinia graminis avenae and Puccinia coronata avenae was studied on 0–2% water agar and on leaves of three oat cultivars and on three non-cultivated species of Avena. Germination of uredospores of P. graminis avenae and P. coronata avenae occurred best at 10–30^oC and at 20^oC respectively. The optimum temperature for germ-tube growth and for appressorial formation was 20^oC for both rusts. An inverse relationship was observed between light intensity and prepenetration development with maximal germination of uredospores, germ-tube growth and appressorial formation occurring in darkness. Under optimum conditions maximum percentage germination and appressorium formation of both rusts was attained within 4 and 12 h after inoculation respectively. The proportion of germinated uredospores of crown rust which gave rise to appressoria was about twice that observed for stem rust. No significant differences were observed in prepenetration development between the single and mixed race inocula of the two rusts. Although germination of uredospores was significantly greater on water agar than on oat leaves, there were no significant differences in prepenetration development of the rusts on the various oat cultivars and species examined. Consequently, the data failed to indicate the presence of resistance mechanisms operating during the prepenetration phase of the infection process on the cultivars and species examined.     

Puccinia chunjii, a close relative of the cereal stem rusts revealed by molecular phylogeny and morphological study

A rust specimen with macroscopic similarities to the cereal stem rusts was collected on Elymus sp. from Gansu province, China. Phylogenetic analyses of ITS and COI DNA sequences indicated the fungus was closely related but distinct as a strongly supported sister taxon to the Puccinia graminis species complex. Microscopic examination revealed diagnostic teliospore characteristics, differentiating it from P. graminis and other morphologically similar rusts. Herein, we designate a name for this new lineage, Puccinia chunjii sp. nov.     

Effect of chronic irradiation on plant resistance to biotic stress in 30-km chernobyl nuclear power plant exclusion zone

It was established in greenhouse experiments that infection with powdery moldew (Erysiphe graminis DC. f. sp. tritici Em. Marchal) and brown rust (Puccinia triticana Erikss. & Henn.) of three wheat (Triticum aestivum L.) cultivars (Mironovskaya 808, Polesskay 70, and Kiyanka) grown from seeds, collected in the Chernobyl exclusion zone, was 1.5–2.0 times higher than of plants grown from control seeds. On field trials in the Chernobyl zone, wheat plant resistance to biotic stress was reduced. At artificial infection with brown rust, the disease development was enhanced on plots with increased radiation background. One of the mechanisms of the declined phytoimmunity potential under the action of low doses of chronic irradiation is evidently a reduced activity of plant proteinase inhibitors. Thus, in wheat and rye (Secale cereale L., cv. Saratovskaya) grains, their activity reduced by 35–60% as compared to control. Active form and race formation in the population of the cereal stem rust causal agent (Puccinia graminis Pers.) was observed in the Chernobyl zone. A “new” population of this fungus with high frequency of more virulent clones than in other Ukraine regions was distinguished. The results obtained independently in greenhouse and field trials performed in the Chernobyl zone demonstrated radiation stress influence on the pathogen-plant interactions. They indicate a necessity of monitoring the microevolutionary processes occurring in both plants and their pathogens under conditions of technogenic stresses.     

Hyperparasitism of Aphanocladium album on Aecidiospores and Teliospores of Puccinia graminis f. sp. tritici

Aphanocladium album invaded rapidly 100% of aecidiospores of Puccinia graminis f. sp. tritici resulting in total collapse of the cells. Only 2% of teliospores were invaded. A. album penetrated through the germ pores. The precocious formation of teliospores which occurs immediately following uredial infection may be regarded as defence mechanism of the rust against its hyperparasite.     

Microscopic Observation on the Development of Puccinia striiformis in the Spike and Stem of Wheat

The majority of germ tubes of the pathotype CYR32 of Puccinia striiformis f.sp. tritici formed on the surface of spike organs of the susceptible wheat cv. Suwon 11 penetrated through the stomatal pore, only a few germ tubes formed small appressoria over the stomata. In the lemma, palea and glume, the stripe rust fungus spread between the parenchyma cells close to the inner epidermal layer, but the fungus did not develop between the thick‐walled cells near the outer epidermal layer of these organs. In the awn and stem, spread of the stripe rust was confined to the intercellular spaces of the chlorophyll parenchyma, beneath the invaded stomatal pore of the epidermis and the urediniospores to be released disrupted the epidermis. In the caryopsis, the spread of hyphae was restricted to the intercellular spaces of the pericarp cells.     

Host and environmental effects on post-penetration development of Puccinia graminis avenae and P. coronata avenae

After inoculation of Avena sterilis and the oat cultivars Algerian and Garry with Puccinia graminis avenae the time required for the eruption of pustules of the rust was markedly less at 30–35 than at 20–25 ^oC. In addition, the area of pustules and number of uredospores produced were significantly greater at 30–35 than at 20 ^oC. Peaks of uredospore production occurred between 12 and 22 days after inoculation. In comparable experiments, the time required for pustules of P. coronata avenae to erupt, and the size of pustules, were relatively insensitive to change of temperature, although weight of uredospores produced was greater at 20 than at 30 ^oC. Peaks of uredospore production occurred between 14 and 18 days after inoculation. Both rusts showed straight-line relationships between pustule area and number of uredospores produced. The percentage of infection foci that developed into pustules was similar with both rusts and on all the oat cultivars examined. Both rusts produced susceptible reaction types on all the hosts tested. Pustules of P. graminis avenae were smaller and fewer and generation time longer on cv. Garry than on cv. Algerian or Avena sterilis and the numbers of pustules per unit of inoculum of both rusts were greatest on Algerian, least on Garry. It is suggested that these quantitative differences in phases of the infection process contribute towards the ‘slow-rusting’ reaction of cv. Garry.     

Development of simple sequence repeat markers for the plant pathogenic rust fungus Puccinia triticina

Eighteen polymorphic di‐ and trinucleotide simple sequence repeat markers were developed for the phytopathogenic rust fungus Puccinia triticina. The allelic diversity varied from two to nine alleles per locus. Levels of observed heterozygosity ranged from 0.095 to 0.952. Seven of the loci deviated significantly from Hardy–Weinberg equilibrium (P < 0.002) with 70% having levels of observed heterozygosity higher than expected heterozygosity. Null allele(s) were observed for locus PtSSR76 with a frequency of 9%. A preliminary screen of other cereal rust fungi (P. coronata, P. graminis, P. recondita and P. striiformis) indicated that these primer pairs are specific to P. triticina.     

Mapping genes Lr53 and Yr35 on the short arm of chromosome 6B of common wheat with microsatellite markers and studies of their association with Lr36

The rust resistance genes Lr53 and Yr35, transferred to common wheat from Triticum dicoccoides, were reported previously to be completely linked on chromosome 6B. Four F ₃ families were produced from a cross between a line carrying Lr53 and Yr35 (98M71) and the leaf rust and stripe rust susceptible genotype Avocet “S” and were rust tested using Puccinina triticina pathotype 53-1,(6),(7),10,11 and Puccinia striiformis f. sp. tritici pathotype 110 E143 A+. The homozygous resistant lines produced infection types of “;1−” and “;N” to these pathotypes, respectively. The Chi-squared tests indicated goodness-of-fit of the data for one leaf rust gene and one stripe rust gene segregation. Linkage analysis using this population demonstrated recombination of 3% between the genes. Microsatellite markers located on the short arm of chromosome 6B were used to map the genes, with the markers cfd1 and gwm508 being mapped approximately 1.1 and 4.5 cM, respectively, proximal to Lr53. Additional studies of the relationship between Lr36, also located on the short arm of chromosome 6B, and Lr53 indicated that the two genes were independent.     

Host and environmental effects on the penetration of oats by Puccinia graminis avenae and Puccinia coronata avenae

The frequency of penetration from appressoria of Puccinia graminis avenae and P. coronata avenae varied among Avena species and between oat cultivars, although both rusts produced susceptible infection type pustules on the cultivars tested. Penetration on cv. Garry was significantly less than that on the Avena species (A. barbata, A.fatua and A. sterilis) studied and penetration of these Avena species was significantly less than on the cvs Algerian and Fulmark. When the rusts were allowed to develop into pustules on seedlings which had been inoculated with fixed amounts of inoculum, there was a direct relationship between number of pustules produced and penetration frequency. The effects of temperature, light and dew period on penetration from appressoria of ‘single race’ and ‘mixed race’ inocula was also studied on these cultivars and species. Penetration by P. graminis avenae was greatest at 30–35 °C and at light intensities of 5625 lux and above, whereas that by P. coronata avenae was greatest at 20 °C and was unaffected by artificial light intensities up to n 250 lux. Maximal penetration by P. graminis avenae and P. coronata avenae was observed after inoculated plants had been exposed to dew periods of 16 and 12 h respectively. Some penetration was observed after a dew period of 8 h. The time taken for each rust to attain maximum penetration varied from 36 to 52 h after inoculation, depending on the environment, and was usually less for P. coronata avenae than for P. graminis avenae.     

Powdery mildew resistance and Lr34/Yr18 genes for durable resistance to leaf and stripe rust cosegregate at a locus on the short arm of chromosome 7D of wheat

The incorporation of effective and durable disease resistance is an important breeding objective for wheat improvement. The leaf rust resistance gene Lr34 and stripe rust resistance gene Yr18 are effective at the adult plant stage and have provided moderate levels of durable resistance to leaf rust caused by Puccinia triticina Eriks. and to stripe rust caused by Puccinia striiformis Westend. f. sp. tritici. These genes have not been separated by recombination and map to chromosome 7DS in wheat. In a population of 110 F₇ lines derived from a Thatcher × Thatcher isogenic line with Lr34/Yr18, field resistance to leaf rust conferred by Lr34 and to stripe rust resistance conferred by Yr18 cosegregated with adult plant resistance to powdery mildew caused by Blumeria graminis (DC) EO Speer f. sp. tritici. Lr34 and Yr18 were previously shown to be associated with enhanced stem rust resistance and tolerance to barley yellow dwarf virus infection. This chromosomal region in wheat has now been linked with resistance to five different pathogens. The Lr34/Yr18 phenotypes and associated powdery mildew resistance were mapped to a single locus flanked by microsatellite loci Xgwm1220 and Xgwm295 on chromosome 7DS.     

Molecular mapping of stem and leaf rust resistance in wheat

Stem rust caused by Puccinia graminis f. sp. tritici Eriks and Henn and leaf rust caused by Puccinia triticina Rob. ex Desm. are major constraints to wheat production worldwide. In the present study, F₄-derived SSD population, developed from a cross between Australian cultivars ‘Schomburgk’ and ‘Yarralinka’, was used to identify molecular markers linked to rust resistance genes Lr3a and Sr22. A total of 1,330 RAPD and 100 ISSR primers and 33 SSR primer pairs selected ob the basis of chromosomal locations of these genes were used. The ISSR marker UBC 840₅₄₀ was found to be linked with Lr3a in repulsion at a distance of 6.0 cM. Markers cfa2019 and cfa2123 flanked Sr22 at a distance of 5.9 cM (distal) and 6.0 cM (proximal), respectively. The use of these markers in combination would predict the presence or absence of Sr22 in breeding populations. A previously identified PCR-based diagnostic marker STS638 linked to Lr20 was validated in this population. This marker showed a recombination value of 7.1 cM with Lr20.     

Notes on some rust fungi in Vietnam

Sixteen rust species and two unconnected Peridermium collected in Vietnam are enumerated. For all the rust species except Hemileia vastarix, Olivea tectonae, and Puccinia thwaitesii, these are the first records of their occurrence in Vietnam. Some taxonomic and pathological considerations are noted for two economically important pine stem rusts.     

Occurrence of Diploid Lines of <Emphasis Type="Italic">Puccinia graminis tritici</Emphasis> in Axenic Culture

THE dikaryophase of the wheat stem rust fungus, Puccinia graminis (Pers.) f. sp. tritici(Erikss. and E. Henn.), can now be grown on artificial medium^1,2; but cultures of this organism initiated from heavy inocula of urediospores usually sporulate or stale after 2-4 weeks and the growth period cannot be appreciably prolonged by subculture onto fresh medium^2,3. Scott and Maclean³, however, referred to the discovery of continuously subculturable isolates of P. graminis tritici. These were obtained by incubating urediospores on liquid nutrient medium, but Scott and Maclean offered no information which might explain the distinctive behaviour of the isolates. We have obtained lines of P. graminis tritici from race 126-Anz 6,7 (culture 70165) which will grow continuously on artificial medium. This communication describes how these lines were obtained and presents evidence that their mycelia are composed of diploid cells.     

Patterns of resistance of Israeli wild emmer wheat to pathogens I. Predictive method by ecology and allozyme genotypes for powdery mildew and leaf rust

The association of ecological factors and allozyme markers with genotypes of tetraploid wild emmer wheat,Triticum dicoccoides, varying in resistance to four cultures of the pathogenErysiphe graminis tritici, and to one culture ofPuccinia recondita tritici, which incite the diseases powdery mildew and leaf rust respectively, were explored theoretically and practically. The study involved 233 accessions comprising 10 populations representing the ecological range ofT. diococcoides in Israel. Our results indicate that genetic polymorphism for resistance to both pathogens is structured geographically, and is predictable by climatic as well as allozymmc markers. Three variable combinations of water factors and temperature differentials significantly explain 0.27 and 0.14 of the spatial variance for resistance to powdery mildew and leaf rust, respectively, suggesting the involvement of natural selection. Several allozyme genotypes, singly, or in combination, are significantly associated with disease resistance. We conclude thatT. dicoccoides populations in Israel, which grow in the center of diversity of the species, contain large amounts of unexploited disease resistant genotypes. The populations could be effectively screened and utilized for producing resistant cultivars by means of ecological factors and allozyme markers as predictive guidelines.     

Carotenoid biosynthesis and the evolution of carotenogenesis genes in rust fungi

Diseases caused by rust fungi pose a significant threat to global plant production. Although carotenoid pigments are produced in spores of nearly all rust species, the corresponding biosynthesis pathway(s) have not been investigated. Here, candidate genes for carotenoid biosynthesis in Puccinia graminis f. sp. tritici (Pgt) were identified, cloned and functionally complemented using specifically engineered strains of Escherichia coli. A part of the carotenoid biosynthesis pathway in rust fungi was elucidated, with only two genes, CrtYB and CrtI, catalysing the reactions from geranyl–geranyl diphosphate (GGPP) to γ-carotene. The CrtYB gene encodes a bi-functional lycopene cyclase/phytoene synthase, which catalyses the condensation of two GGPP into phytoene, as well as the cyclisation of the ψ-end of lycopene to form γ-carotene. The CrtI gene encodes a phytoene desaturase that carries out four successive desaturations of phytoene, through the intermediates phytofluene and neurosporene to lycopene. The evolution of carotenoid pigmentation in rust fungi, including Pgt, P. graminis avenae, P. graminis secalis (Pgs), P. graminis lolli, P. striiformis f. sp. tritici, P. striiformis f. sp. pseudohordei, P. striiformis f. sp. hordei, the “scabrum” rust (putative hybrids between Pgt and Pgs), P. triticina, and P. hordei, was investigated by phylogenetic analysis. Both CrtYB and CrtI were found to be closely related among rust fungi, other pathogenic fungi, and some aphids. Our results provide a springboard to increase the understanding of the physiological role(s) of carotenoid pigments in rust fungi, to better understand evolution within the Pucciniales, and to develop robust molecular diagnostics for rust fungi.