Hyperspectral signal decomposition and symptom detection of wheat rust disease at the leaf scale using pure fungal spore spectra as reference

This study establishes a method to detect and distinguish between brown rust and yellow rust on wheat leaves based on hyperspectral imaging at the leaf scale under controlled laboratory conditions. A major problem at this scale is the generation of representative and correctly labelled training data, as only mixed spectra comprising plant and fungal material are observed. For this purpose, the pure spectra of rust spores of Puccinia triticina and P. striiformis, causal agents of brown and yellow rust, respectively, were used to serve as a spectral fingerprint for the detection of a specific leaf rust disease. A least-squares factorization was used on hyperspectral images to unveil the presence of the spectral signal of rust spores in mixed spectra on wheat leaves. A quantification of yellow and brown rust, chlorosis and healthy tissue was verified in time series experiments on inoculated plants. The detection of fungal crop diseases by hyperspectral imaging was enabled without pixel-wise labelling at the leaf scale by using reference spectra from spore-scale observations. For the first time, this study shows an interpretable decomposition of the spectral reflectance mixture during pathogenesis. This novel approach will support a more sophisticated and precise detection of foliar diseases of wheat by hyperspectral imaging.     

Histological responses in Hordeum chilense to brown and yellow rust fungi

The histological reaction of 16 Hordeum chilense accessions to Puccinia recondila, P. hordei and P striiformis was investigated. It is not possible to generalize about the reaction of H. chilense to these rust fungi. Several lines showed a susceptible infection type to a brown rust harvested on H. juhatum. One of these lines had an intermediate infection type to the wheat brown rust fungus. The H. chilense reaction to the yellow rust fungi ranged from susceptible to resistant. The lines differed in their response to the rust fungi, as expressed by the level of early abortion of sporelings, and the frequency of plant-cell necrosis associated with the infection sites. All the lines caused complete early abortion associated with infrequent necrosis of the rye brown rust fungus. The responses were to some extent rust non-specific. The differences in response between the lines may reflect differences in general defence mechanisms.     

Dry-dispersal and rain-splash of brown (Puccinia recondita f.sp. tritici) and yellow (P. striiformis) rust spores from infected wheat leaves exposed to simulated raindrops

The dispersal of spores from lesions of brown ( Puccinia recondita f.sp. tritici ) or yellow ( P. striiformis ) rusts of wheat by impacting drops was studied. Using a generator of uniform-size drops, drops of 2.5, 3.4, 4.2 and 4.9 mm in diameter were released from rest at heights of 5, 50 and 100 cm above horizontal and primary leaves uniformly covered with sporulating lesions. Dry-dispersal and rain-splash occurred simultaneously in response to drop impaction. A coloration technique allowed separate counting of dry-dispersed and rain-splashed spores caught on slides. More spores were rain-splashed than dry-dispersed. Neither removal mechanism affected in-vitro germination of spores, which was higher in brown than in yellow rust. For both rusts, the number of both dry-dispersed and rain-splashed spores, as well as their travel distance, increased with drop diameter and fall height. The fall speed of incident drops in relation to diameter and fall height was obtained by solving numerically the equation of vertical drop motion. The number of spores removed by a given impacting drop was found to be a power function of the calculated kinetic energy of the impacting drop. Based on this experimental relationship, a simulation study showed the relevance of rain type in the removal of spores.     

Life-history of Phragmidium rubi-idaei on red raspberry in the United Kingdom

Phragmidium rubi-idaei produced five spore stages on red raspberry and overwintered as teliospores adhering to canes. In the field spermagonia appeared in a single flush in early summer on leaves of the primocanes (first-year canes) and on leaves of the lateral shoots of fruiting canes, and were followed by aecia, uredinia and telia. Aecia and uredinia occasionally appeared on petioles and sepals and uredinia on drupelets of mature fruit. Lesions caused by aecia and uredinia on the bases of primocanes did not become cankerous in the following fruiting year. Necroses associated with large aecia produced a shot-hole effect and leaflets heavily affected by either aecia, uredinia or telia abscissed prematurely.
The sequence of spore stages on pot-grown plants inoculated with mature overwintered teliospores took 60 days to the reappearance of telia. The rate of development of uredinia and associated necroses was faster on younger than on older leaves but the cumulative spore yield was not significantly different.
Optimum temperature for germination of urediniospores in vitro (18.4–20.9°C) was higher than for mature teliospores (14.6–18.4°C). Light inhibited germination of both types of spores. Teliospore germination was inhibited by wavelengths of 590–750 nm, but this was partially reversible under other light conditions. A reversible inhibition of basidiospore formation was also observed under shorter wavelengths.     

Specificity of Prehaustorial Resistance to Puccinia hordei and to Two Inappropriate Rust Fungi in Barley

ABSTRACT To elucidate the specificity of prehaustorial resistance to inappropriate rust fungi, we studied two populations of recombinant inbred lines of barley that segregated for partial resistance (PR) to Puccinia hordei and for the resistance to the inappropriate rust species P. recondita f. sp. tritici and P. hordei-murini. PR to P. hordei is prehaustorial and nonhypersensitive, and its level can be assessed accurately by measuring the latent period of the fungus. The resistance to the inappropriate rust species is a combination of prehaustorial (nonhypersensitive) and posthaustorial (hypersensitive) mechanisms. The amount of nonhypersensitive, early abortion of P. recondita f. sp. tritici and P. hordei-murini sporelings reflects the degree of prehaustorial defense to the two inappropriate rust species. All lines showing a long latent period of P. hordei also had a relatively high level of early abortion of the growth of P. recondita f. sp. tritici and P. hordei-murini. This indicates that genes for PR to P. hordei are also effective against these two inappropriate rust species. The reverse was not necessarily true; some lines showing a high level of early abortion of P. recondita f. sp. tritici and P. hordei-murini had a low level of PR to P. hordei. Moreover, lines with a similar level of prehaustorial resistance to P. recondita f. sp. tritici could differ considerably in their prehaustorial resistance to P. hordei-murini. This indicates that genes for prehaustorial resistance may exhibit rust species specificity.     

Histology of the infection of tritordeum and its parents by cereal brown rusts

The reactions of tritordeum lines and their Hordeum chilense and Triticum spp. parents to infection by Puccinia recondita f.sp. tritici, P. recondita f.sp. agropyri and P. hordei were studied at the seedling stage. The histological observations indicated that tritordeum behaves as the wheat parent, whatever the H. chilense parental line reaction. Tritordeum is to be considered a host of wheat brown rust where genes for hypersensitive resistance may occur; these are apparently contributed by the wheat parent. Both H. chilense and wheat are highly resistant to barley brown rust, and the reaction of tritordeum is that of the wheat parent with respect to the levels of necrosis associated with the early aborted infection units. The tritordeum reaction is also that of the wheat parent to a rust collected on H. jubatum (putatively P. recondita f.sp. agropyri ), with the susceptibility or resistance of H. chilense being overruled by the wheat parent reaction.     

Occurrence of <Emphasis Type="Italic">Puccinia</Emphasis> spp. spores in Madeira Island and their phytopathological importance

The genus Puccinia represents rust infections, which are responsible for great productivity losses in crops of commercial and ornamental plants worldwide. This work is aimed at determining the occurrence of Puccinia spp. spores in Madeira Island in order to infer the exposure risks from a phytopathological point of view. A phytopathological analysis was performed in 203 local plant samples between January 2003 and December 2012. During the same period the airborne concentrations of rust spores were monitored following well-established guidelines. Aerobiological data was compared with meteorological records. Based on macro and microscopic analyses, five species of rusts were identified: P. horiana, P. buxi, P. porri, P. pelargonii-zonalis, and P. sorghi, and they were found mostly in spring and summer. A total of 20 samples out of 203 analysed plants (9.8%), were infected with Puccinia spores, i.e., P. horiana (5.9%), P. buxi (1.47%), P. porri and P. pelargonii-zonalis (0.98%), and P. sorghi (0.5%). During the studied period Puccinia spores attained an annual average concentration of 126 spores m^?3 and most of them were recorded between March and October. Meteorological factors were determinant in fluctuations in spore concentration. Relative humidity was the parameter that favoured the biggest release and dispersal of the rust spores, whereas rainfall revealed a significant negative effect. Rusts do not represent an important plant pathogen in Madeira Island, as shown by the low infection frequencies and levels of airborne spore concentrations.     

落叶松褐锈病的研究1.夏孢子的发生及其形态鉴定

 落叶松褐锈病[Triphragmiopsis laricinum(Chou) Tai],是由落叶松拟三孢锈菌引起的一种叶锈病。现已知在黑龙江、吉林、辽宁三省的落叶松人工林内,均有不同程度的发生。发病率一般70~80%,重病区可达90%以上。病原菌能为害各种落叶松,以苗期和落叶松人工幼林较为严重。     

Genes Governing Resistance to Puccinia hordei in Thirteen Spring Barley Accessions

ABSTRACT Leaf rust, caused by Puccinia hordei, is an important disease of barley in many parts of the world. In the eastern United States, this disease was effectively controlled for over 20 years through the deployment of cultivars carrying the resistance gene Rph7. Isolates of P. hordei with virulence for Rph7 appeared in this region in the early 1990s rendering barley cultivars with this gene vulnerable to leaf rust infection. From a preliminary evaluation test, 13 accessions from diverse geographic locations possessed resistance to P. hordei isolate VA90-34, which has virulence for genes Rph1, 2, 4, 6, 7, 8, and 11. Each of these 13 accessions was crossed with susceptible cvs. Moore or Larker to characterize gene number and gene action for resistance to P. hordei. Additionally, the 13 accessions were intercrossed and crossed to host differential lines possessing genes Rph3, Rph5, and Rph9 to determine allelic relationships of resistance genes. Seedlings of F(1), F(2), and BC(1)F(1) populations were evaluated in the greenhouse for their reaction to P. hordei isolate VA90-34. Leaf rust resistance in six of the accessions including Collo sib, CR270.3.2, Deir Alla 105, Giza 119, Gloria, and Lenka is governed by a single dominant gene located at or near the Rph3 locus. All accessions for which the gene Rph3 was postulated to govern leaf rust resistance, except for Deir Alla 105, likely possess an allele different than Rph3.c found in Estate based on the differential reaction to isolates of P. hordei. The resistance gene in Grit and Donan is located at or near the Rph9 locus. Alleles at both the Rph3 and Rph9 loci confer resistance in Femina and Dorina. In addition to Rph3, Caroline and CR366.13.2 likely possess a second unknown recessive gene for leaf rust resistance. Resistance in Carre 180 is governed by a recessive gene that is different from all other genes considered in this study. Identification of both known and unique genes conferring leaf rust resistance in the barley germplasm included in this study provides breeding programs with the knowledge and opportunity to assess currently used sources of leaf rust resistance and to incorporate new sources of resistance into their programs.     

Characterization of brown planthopper damage on rice crops through hyperspectral remote sensing under field conditions

Field experiments were conducted to characterize the brown planthopper (BPH) (Nilaparvata lugens (Stål.) damage stress on rice crops through hyperspectral remote sensing. The BPH-damaged rice crop had higher reflectance in visible (VIS) and lower reflectance in near-infrared regions (NIR) of the electromagnetic spectrum compared with uninfested plants. Mean reflectance of the rice crop varied among different BPH damage levels in various wavebands, with the greatest variation in NIR (740–925 nm). Correlations between plant reflectance and BPH damage depicted four sensitive wavelengths, at 764, 961, 1201 and 1664 nm in relation to BPH stress on the rice crop. Three new brown planthopper spectral indices (BPHI) were formulated by combining two or more of these sensitive wavelengths. Some of the hyperspectral indices reported in the literature were also tested for their suitability to detect BPH stress on rice crops. Based on crop reflectance corresponding to the sensitive wavelengths, a multiple-linear regression model was developed (R²=0.71, RMSE=1.74, P<0.0001) and validated (R²=0.73, RMSE?=?0.71, P<0.0001) that would help to monitor BPH stress on a rice crop and to issue forewarnings to growers.     

Genetic analysis and molecular mapping of wheat genes conferring resistance to the wheat stripe rust and barley stripe rust pathogens

ABSTRACT Stripe rust is one of the most important diseases of wheat and barley worldwide. On wheat it is caused by Puccinia striiformis f. sp. tritici and on barley by P. striiformis f. sp. hordei Most wheat genotypes are resistant to P. striiformis f. sp. hordei and most barley genotypes are resistant to P. striiformis f. sp. tritici. To determine the genetics of resistance in wheat to P. striiformis f. sp. hordei, crosses were made between wheat genotypes Lemhi (resistant to P. striiformis f. sp. hordei) and PI 478214 (susceptible to P. striiformis f. sp. hordei). The greenhouse seedling test of 150 F(2) progeny from the Lemhi x PI 478214 cross, inoculated with race PSH-14 of P. striiformis f. sp. hordei, indicated that Lemhi has a dominant resistance gene. The single dominant gene was confirmed by testing seedlings of the F(1), BC(1) to the two parents, and 150 F(3) lines from the F(2) plants with the same race. The tests of the F(1), BC(1), and F(3) progeny with race PSH-48 of P. striiformis f. sp. hordei and PST-21 of P. striiformis f. sp. tritici also showed a dominant gene for resistance to these races. Cosegregation analyses of the F(3) data from the tests with the two races of P. striiformis f. sp. hordei and one race of P. striiformis f. sp. tritici suggested that the same gene conferred the resistance to both races of P. striiformis f. sp. hordei, and this gene was different but closely linked to Yr21, a previously reported gene in Lemhi conferring resistance to race PST-21 of P. striiformis f. sp. tritici. A linkage group consisting of 11 resistance gene analog polymorphism (RGAP) markers was established for the genes. The gene was confirmed to be on chromosome 1B by amplification of a set of nullitetrasomic Chinese Spring lines with an RGAP marker linked in repulsion with the resistance allele. The genetic information obtained from this study is useful in understanding interactions between inappropriate hosts and pathogens. The gene identified in Lemhi for resistance to P. striiformis f. sp. hordei should provide resistance to barley stripe rust when introgressed into barley cultivars.     

Variation for isozymes and double-stranded RNA among isolates of Puccinia striiformis and two other cereal rusts

Gel electrophoresis was used to examine the variation in isozymes and dsRNA within and between the rust species Puccinia striiformis, P. recondita and P. hordei. No differences in isozyme phenotype were found among 29 diverse isolates of the wheat-attacking form of P. striiformis (WYR). Smaller numbers of isolates of the barley-attacking form (BYR) of P. recondita and of P. hordei showed similar intra-group uniformity. There were major differences in isozyme phenotypes between the three species, while WYR and BYR differed for two of 12 enzymes. Double-stranded RNA was detected in each species and in all 26 isolates examined. For WYR and BYR, all isolates within each group had the same or a similar phenotype. In contrast, each isolate of P. recondita and P. hordei had a unique phenotype. There were differences in dsRNA phenotypes both between the three species and between WYR and BYR.
The uniformity of these rust populations and species for isozyme phenotype is contrasted with their variability in pathogenicity and with the variability in isozymes encountered in higher organisms. Uniformity may result from a feature of the biology of the rust species and populations examined, or from the relative homogeneity of the environment of biotrophic pathogens. Consistent differences in isozyme and dsRNA phenotype between the WYR and BYR isolates of P. striiformis indicate that these are two discrete populations, supporting the view that they should be recognized as f.sp. tritici and f.sp. hordei , respectively.     

Differential effects of phenylalanine ammonia lyase, cinnamyl alcohol dehydrogenase, and energetic metabolism inhibition on resistance of appropriate host and nonhost cereal-rust interactions

ABSTRACT Effects of phenylpropanoid and energetic metabolism inhibition on resistance were studied during appropriate host and nonhost cereal-rust interactions. In the appropriate barley-Puccinia hordei interaction, phenylalanine ammonia lyase (PAL) and cinnamyl alcohol dehydrogenase (CAD) inhibition reduced penetration resistance in two genotypes, suggesting a role for phenolics and lignins in resistance. Interestingly, penetration resistance of the barley genotype 17.5.16 was not affected by phenylpropanoid biosynthesis but penetration resistance was almost completely inhibited by D-mannose, which reduces the energy available in plant host cells. This suggests a parallel in the cellular basis of penetration resistance between 17.5.16 rust and mlo barleys powdery mildew interaction. Results revealed differing patterns of programmed cell death (PCD) in appropriate versus nonhost rust interactions. PAL and CAD inhibitors reduced PCD (hypersensitivity) in appropriate interactions. Conversely, they had no effect in PCD of wheat to P. hordei; whereas D-mannose dramatically reduced nonhost resistance and allowed colony establishment. The differential effects of inhibitors in the expression of the different resistances and the commonalities with the cereal-powdery mildew interaction is analyzed and discussed.     

Wheat leaf rust uredospore production and carbon and nitrogen export in relation to lesion size and density

ABSTRACT To develop mechanistic yield loss models for biotrophic fungi, we need better account for the export of dry matter, carbon, and nitrogen from the leaf into the spores. Three experiments in controlled environment chambers were performed to study the dynamics of uredospores production of Puccinia triticina on seedling leaves of wheat in relation to time, lesion density, and sporulating surface area. The detrimental effect of lesion density on the sporulation capacity of brown rust lesions was confirmed. When lesion density increased, spores production per lesion strongly decreased. However, our results showed that increasing lesion density also greatly reduces lesion size. A model was developed to summarize these relationships. Our main conclusion is that the density effect on spore production per lesion is accounted for by lesion size. When sporulation was related to the sporulating surface area, it became independent of density. As well, carbon and nitrogen contents of the spores were independent of lesion density. Our data suggest that when nitrogen available in the host is limiting, spore production is reduced but nitrogen content of spores tend to remain stable.     

Attempted somatic hybridization of Puccinia striiformis f. sp. tritici and P. striiformis f. sp. hordei

Attempts were made to produce somatic hybrids between isolates of Puccinia striiformis f. sp. tritici and f. sp. hordei. A mixed infection was produced on a common susceptible barley host, Fong Tien, using white-spored isolates of P. striiformis f. sp. tritici and yellow-spored isolates off. sp. hordei. Selection was made for non-parental spore colour on selective wheat and barley hosts, and variants thus isolated were analysed for virulence markers, and for isozyme and double-stranded RNA (dsRNA) markers, all of which clearly differentiated the parental isolates. Two white-spored (non-parental) isolates were found on the selective barley host which otherwise resembled the parental f. sp. hordei isolate in virulence, isozyme and dsRNA markers. The most likely explanation of the origin of these isolates is mutation to white spore colour in the f. sp. hordei isolate.     

Detection of airborne inoculum of Leptosphaeria maculans and Pyrenopeziza brassicae in oilseed rape crops by polymerase chain reaction (PCR) assays

The potential use of DNA-based methods for detecting airborne inoculum of Leptosphaeria maculans and Pyrenopeziza brassicae , both damaging pathogens of oilseed rape, was investigated. A method for purifying DNA from spores collected using Hirst-type spore samplers and detecting it using polymerase chain reaction (PCR) assays is described. For both pathogens, the sensitivities of the DNA assays were similar for spore-trap samples and pure spore suspensions. As few as 10 spores of L. maculans or P. brassicae could be detected by PCR and spores of both species could be detected against a background of spores of six other species. The method successfully detected spores of P. brassicae collected using spore traps in oilseed rape crops that were infected with P. brassicae. Leptosphaeria maculans spores were detected using spore traps on open ground close to L. maculans -infected oilseed rape stems. The potential use of PCR detection of airborne inoculum in forecasting the diseases caused by these pathogens is discussed.     

Removal of urediniospores of brown (Puccinia recondita f.sp. tritici) and yellow (P. striiformis) rusts of wheat from infected leaves submitted to a mechanical stress

The passive spore removal from colonies due to mechanical stress was compared in the brown (Puccinia recondita f.sp. tritici) and yellow (P. striiformis) rusts of wheat. Mechanical stress was applied using either a miniaturized wind tunnel or a centrifuge. In wind-tunnel experiments, a wind of minimum velocity of 1.3 and 1.8 m s^-1 for P. recondita f.sp. tritici and P. striiformis, respectively, applied for at least 10 seconds, was necessary to remove spores. The interaction between wind velocity and cumulated duration was significant for both rusts. At low wind velocity, a longer duration was required to remove the spores than at high wind velocity, and vice versa. In centrifugation experiments, the maximum spore removal occurred for angular velocities of 10³ and 2 10³ rotations min^-1, for P. recondita f.sp. tritici and P. striiformis, respectively, applied for 5 min. Calculation of the aerodynamic and centrifugal forces showed that the forces necessary to remove spores are greater for P. striiformis than for P. recondita f.sp. tritici. This difference can be related to the size of the dispersal unit, which is larger in P. striiformis than in P. recondita f.sp. tritici due to spore clustering. These observations are consistent with the differences in the mean spore dispersal distance, which is usually smaller in P. striiformis than in P. recondita f.sp. tritici.     

Virulence spectrum of Puccinia hordei in barley production systems in Ethiopia

Barley leaf rust caused by Puccinia hordei is an important disease of barley in Ethiopia. In the 2003 and 2004 cropping seasons, surveys of P. hordei were conducted on fields in the main rainy, residual moisture and short rainy season-barley production systems. A total of 381 isolates were analysed on 12 barley differential hosts carrying different Rph resistance genes ( Rph1 – Rph12 ). Based on infection phenotypes on leaf rust ( Rph ) resistance genes, seven pathotypes were identified, namely ETPh7611, ETPh7631, ETPh6611, ETPh7651, ETPh7671, ETPh7653 and ETPh7633, with frequencies of 63·0, 21·5, 6·8, 2·9, 2·6, 2·1 and 1·2%, respectively. ETPh7611 and ETPh7631 were the most common pathotypes found in all the surveyed areas of the three production systems. ETPh7653 was found in the small rainy season production system only. ETPh7671, ETPh7653 and ETPh7633 were the most virulent, but the least frequent, pathotypes. All isolates were virulent on resistance genes Rph1 , Rph4 , Rph8 , Rph9 , Rph11 and Rph12 . Virulence against Rph3 and Rph7 was absent. Genes Rph2 , Rph6+2 , Rph5 and Rph10 were effective against 96·3, 88·9, 65 and 2·4% of the rust isolates tested, respectively.     

Wheat Leaf Rust Uredospore Production on Adult Plants: Influence of Leaf Nitrogen Content and Septoria tritici Blotch

ABSTRACT Leaf rust uredospore production and lesion size were measured on flag leaves of adult wheat plants in a glasshouse for different lesion densities. We estimated the spore weight produced per square centimeter of infected leaf, per lesion, and per unit of sporulating area. Three levels of fertilization were applied to the plants to obtain different nitrogen content for the inoculated leaves. In a fourth treatment, we evaluated the effect of Septoria tritici blotch on leaf rust uredospore production. The nitrogen and carbon content of the spores was unaffected or marginally affected by lesion density, host leaf nitrogen content, or the presence of Mycosphaerella graminicola on the same leaf. In leaves with a low-nitrogen content, spore production per lesion was reduced, but lesion size was unaffected. A threshold effect of leaf nitrogen content in spore production was however, evident, since production was similar in the medium- and high-fertilizer treatments. In leaves inoculated with M. graminicola and Puccinia triticina, the rust lesions were smaller and produced fewer spores. The relationships among rust lesion density, lesion size, and uredospore production were fitted to a model. We determined that the density effect on spore production resulted mainly from a reduction in lesion size, the spore production per unit of sporulating surface being largely independent of lesion density. These results are consistent with those obtained previously on wheat seedlings. The main difference was that the sporulation period lasted longer in adult leaves.     

Variation in the pine stem rust fungus Peridermium pini in the United Kingdom

Peridermium pini aeciospores from lesions on Scots pine ( Pinus sylvestris ) in Scotland and in Thetford Forest, East Anglia, differed in terms of germ tube morphology and cytology. The former produced long aseptate germ tubes containing a pair of nuclei. The latter produced short septate germ tubes, each cell containing a single nucleus. One third of the 'Scottish' germ tubes ended in a vesicle. Nuclei could be counted in only a small proportion of ungerminated aeciospores. On average 83% of Scottish spores had two nuclei and 4% had one. The equivalent figures for Thetford spores were 66 and 20%. A small number of aeciospore samples from other parts of England resembled Scottish spores rather than Thetford spores in germ tube morphology and cytology.
Both Scottish and Thetford spores proved capable of causing infection of wounded Scots pine shoots. In addition, some Thetford spores caused infection on leaf discs taken from certain Paeonia cultivars. From these results it is concluded that two distinct forms of Peridermium pini exist in the UK. The implications of this finding are discussed in relation to the history of pine stem rust in the country and the genetics and taxonomy of P. pini.