The origin and genetic diversity of the causal agent of Asian soybean rust,Phakopsora pachyrhizi,in South America

A sequence‐based approach was used to investigate molecular genetic variations in Phakopsora pachyrhizi, an obligate biotrophic pathogen that causes Asian soybean rust. In Argentina, the samples came from uredinium‐bearing leaves taken from 11 soybean fields; in Brazil, the samples comprised urediniospores from leaves of 10 soybean genotypes that had been grown in three experimental stations during two growing seasons. PCR‐based cloning techniques were used to generate DNA sequences for two gene regions and alignments were supplemented with data from GenBank. A total of 575 sequences for the internal transcribed spacer region (18 ribotypes) and 160 partial sequences for a housekeeping gene encoding ADP‐ribosylation factor (10 haplotypes) were obtained. Ribotype accumulation curves predicted that about 20 bacterial clones would recover 5–6 ribotypes (c. 70–80% of the total molecular variation) per locality. The samples from the three experimental stations in Brazil displayed most (14 out of 16) ribotypes found worldwide; the lack of genetic structure and differentiation at a diverse geographic scale suggests that both local and distant sources provide airborne inoculum during disease establishment. Soybean genotypes with resistance genes for the Asian soybean rust did not decrease the molecular genetic variation of fungal populations.     

Diversity and distribution of pathotypes of the soybean rust fungus Phakopsora pachyrhizi in East Africa

Phakopsora pachyrhizi is a biotrophic fungus that causes rust on soybean, leading to devastating yield losses. Development of resistant cultivars for deployment in different geographic regions requires a comprehensive understanding of the prevalent P. pachyrhizi pathotypes. To determine the pathotypes existing in four East African countries, 65 isolates were tested on 11 soybean host differentials. In addition, the virulence spectrum of isolates collected from the same region over multiple years was compared. The majority of the isolates (54%) belonged to pathotype 1000, which was found in all countries. The pathotypes with the most complex virulence spectrum, which comprised isolates from Kenya and Malawi, were virulent on four differentials. All pathotypes were virulent on soybean genotypes carrying the Rpp1 resistance gene to P. pachyrhizi, but they were avirulent on cultivars carrying the Rpp1b, Rpp2, or Rpp3 gene, as well as on cultivar No6-12-1 that carries Rpp2, Rpp4, and Rpp5. Two of the pathotypes were virulent on cultivar UG 5 that carries Rpp1 and Rpp3 and on Hyuuga that carries Rpp3 and Rpp5. The isolates collected from different countries differed in their virulence spectrum across the years. Shannon's index (H) and Simpson's index (S) of diversity indicated that the isolates from Malawi were more diverse (H = 1.55, S = 0.90) while those from Uganda had lower diversity (H = 0.78, S = 0.46 ). The Rpp genes that were found to provide resistance to all pathotypes of P. pachyrhizi can be employed for soybean breeding aimed at durable rust resistance.     

Evaluation of soybean genotypes for resistance against the rust-causing fungus Phakopsora pachyrhizi in East Africa

Soybean rust, caused by the biotrophic fungus Phakopsora pachyrhizi, is the most important foliar disease of soybean (Glycine max) worldwide. Deployment of resistant soybean cultivars is the best option for managing this disease. Genes conferring resistance to P. pachyrhizi have been identified, but pathotypes of the rust fungus overcoming these resistance genes have also been found. To identify novel resistance genes, soybean genotypes from both local and international sources were screened at multiple locations in Tanzania and Uganda in 2016 and 2017. The results from this screening revealed that infection types, disease severities, and sporulation levels varied among the genotypes and locations. The majority of the genotypes had tan-coloured (TAN) lesions and developed moderate sporulation, implying susceptibility, while only seven of the 71 lines had reddish-brown (RB) lesions and showed low disease severities in all of the screening environments. We identified seven genotypes that were the most resistant to rust in the most locations over the two years. These genotypes will be useful for further studies and, ultimately, for rust management, as they show broad resistance to various pathotypes of the rust fungus.     

Nickel potentiates soybean resistance against infection by Phakopsora pachyrhizi

Asian soybean rust (ASR), caused by the fungus Phakopsora pachyrhizi, causes significant yield losses worldwide. Nickel (Ni) plays a key role in the metabolism of some profitable crops, such as soybeans, because it is a constituent of several biomolecules and is required for the catalytic process of several enzymes. This study investigated the effect of foliar Ni treatment on the potentiation of soybean cultivar TMG 135 resistance to P. pachyrhizi infection at the microscopic, biochemical, and molecular levels. The severity of ASR decreased by 35% in plants treated with Ni. The malondialdehyde concentration, an indicator of cellular oxidative damage, was high in the leaves of plants that were not treated with Ni and was linked to ASR severity and the extensive colonization of the palisade and spongy parenchyma cells by fungal hyphae. The lignin concentration, β-1,3-glucanase activity, and expression of the URE gene and the defence-related genes PAL1.1, PAL2.1, CHI1B1, and PR-1A were up-regulated in Ni-treated plants infected with P. pachyrhizi. The information provided by this study shows the great potential of Ni to increase the basal level of soybean resistance to ASR and to complement other control methods within the context of sustainable agriculture.     

Pathogenic diversity of soybean rust in Argentina, Brazil, and Paraguay

Phakopsora pachyrhizi, the cause of soybean rust, is an economically important pathogen of soybean in South America. Understanding the pathogenicity of indigenous fungal populations is useful for identifying resistant plant genotypes and targeting effective cultivars against certain populations. Fifty-nine rust populations from Argentina, Brazil, and Paraguay were evaluated for pathogenicity in three cropping seasons, 2007/2008–2009/2010, using 16 soybean differentials. Only two pairs of P. pachyrhizi populations displayed identical pathogenicity profiles, indicating substantial pathogenic variation in the rust populations. Comparative analysis of 59 South American and five Japanese samples revealed that pathogenic differences were not only detected within South America but also distinct between the P. pachyrhizi populations from South America and Japan. In addition, seasonal changes in rust pathogenicity were detected during the sampling period. The differentials containing resistance genes (Rpp: resistance to P. p achyrhizi) Rpp1, Rpp2, Rpp3, and Rpp4, except for Plant Introduction (PI) 587880A, displayed a resistant reaction to only 1.8–14, 24–28, 22, and 36 % of South American P. pachyrhizi populations, respectively. In contrast, PI 587880A (Rpp1), Shiranui (Rpp5), and 3 Rpp-unknown differentials (PI 587855, PI 587905, and PI 594767A) showed a resistant reaction to 78–96 % of all populations. This study demonstrated that P. pachyrhizi populations from South America vary geographically and temporally in pathogenicity and that the known Rpp genes other than Rpp1 in PI 587880A and Rpp5 have been less effective against recent pathogen populations in the countries studied.     

A major variation in the virulence of the Asian soybean rust pathogen (Phakopsora pachyrhizi) in Bangladesh

Asian soybean rust (ASR) caused by Phakopsora pachyrhizi is a major threat to soybean production in Bangladesh. Understanding the yearly changes and the current status of pathogenic structures is essential for developing appropriate breeding strategies for obtaining ASR-resistant soybean lines. Thirty-four P. pachyrhizi samples were collected from ASR hotspot areas (Chandpur, Lakshmipur, Noakhali, Barisal and Bhola districts) of Bangladesh in 2018 and 2019 and evaluated for pathogenicity on 12 soybean differential lines. The tested samples showed similar and dissimilar pathogenicity patterns on the differentials, yielding 21 distinct pathotypes. The cluster analysis, principal coordinate analysis and principal component analysis of the disease phenotypes of 47 samples collected in 2016, 2018 and 2019 indicated a higher pathogenic diversity and virulence variation in the P. pachyrhizi samples of 2018 and 2019 compared to that of 2016. The pathogenicity profiles of the Bangladeshi P. pachyrhizi samples appeared distinct from those of Argentinian and Brazilian samples, but showed slight similarities with Japanese, Mexican and Paraguayan samples. Furthermore, none of the resistance genes for P. pachyrhizi (Rpp genes) was solely effective against all the tested samples from 2018 and 2019, while samples (BdRP-48, BdRP-56 and BdRP-58) virulent to all Rpp1–Rpp6 genes were detected. The Rpp-pyramided line No6–12–1, carrying Rpp2, Rpp4 and Rpp5, was capable of conferring robust resistance to these virulent samples. Altogether, these results indicate an increase in the virulence of the current ASR pathogen in Bangladesh, which can be resolved by pyramiding different resistance genes in soybean cultivars.     

Molecular mapping of Asian soybean rust resistance in soybean landraces PI 594767A,PI 587905 and PI 416764

Asian soybean rust (ASR), caused by Phakopsora pachyrhizi, is one of the most serious diseases of soybean. The soybean landraces PI 594767A, PI 587905 and PI 416764 previously showed high levels of resistance to a wide range of ASR fungus, while the genetic basis of the resistance has yet to be understood. In this study, the ASR resistance loci were mapped using three independent mapping populations, POP‐1, POP‐2 and POP‐3 derived from crosses BRS184 × PI 594767A, BRS184 ×  PI 587905 and BRS184 × PI 416764, respectively. In each population, the resistance to ASR segregated as a single gene, but the resistance was dominant in PI 594767A and PI 587905 and incompletely dominant in PI 416764. The resistance genes from both PI 594767A and PI 587905 were mapped on chromosome 18 corresponding to the same location as known resistance locus Rpp1. Quantitative trait locus (QTL) analysis performed on POP‐3 identified the putative ASR resistance locus in PI 416764 on the defined region of chromosome 6 where Rpp3 was located. The QTLs detected by the mapping explained about 67–72% of the phenotypic variation in POP‐3. Cluster analysis based on disease reactions to 64 ASR populations demonstrated the presence of at least two types of functional resistant Rpp1 alleles: strong and weak allele(s), e.g. soybean accession PI 594767A and PI 587905 carry the strong resistant Rpp1 allele(s). Introducing or pyramiding strong Rpp1 allele(s) in elite soybean cultivars is expected to be useful against the South American rust population.     

Potentiation of soybean resistance against Phakopsora pachyrhizi infection using phosphite combined with free amino acids

Considering the importance of Asian soybean rust (ASR), caused by Phakopsora pachyrhizi, in the decrease in soybean yield, this study investigated the potential of using phosphite combined with l -α-free amino acids (referred to as induced resistance [IR] stimulus hereafter) to boost defence responses of soybean plants against P. pachyrhizi infection. Plants were sprayed with water (control), acibenzolar-S-methyl (ASM) or IR stimulus and noninoculated or inoculated with P. pachyrhizi. Urediniospore germination was not affected by the IR stimulus in vitro. Reduced ASR severity, lower malondialdehyde concentration and less colonization of leaf tissues by P. pachyrhizi (lower TEF-1α expression from 1 to 15 days after inoculation [dai]) occurred for IR stimulus-sprayed plants. The pattern of gene expression for IR stimulus-sprayed and infected plants was strikingly similar but sometimes more remarkable than that in ASM-sprayed and infected plants. Higher production of phenolics and lignin along with stronger up-regulation of PAL1.3 (5 and 10 dai), PAL2.2 (3 dai), PAL3.1 (1, 3 and 5 dai), ICS1 (5 dai), CHIA1 (1, 5 and 10 dai), CHI1B1 (5 dai), PR-1A (5 and 10 dai), NR1-2 (5 and 10 dai) and INR-2 (5 and 10 dai) for IR stimulus-sprayed plants increased their resistance against ASR. In addition, IR stimulus-sprayed and infected plants showed less impairment of the photosynthetic apparatus and maintained high concentrations of chlorophyll a + b and carotenoids. These findings highlight the potential of using this IR stimulus for developing a well-tuned and effective defensive strategy in soybean plants against P. pachyrhizi infection.     

Systemic properties of myclobutanil in soybean plants, affecting control of Asian soybean rust (Phakopsora pachyrhizi)

BACKGROUND: The demethylation inhibitor (DMI) fungicide myclobutanil can be an effective component of spray programmes designed to control the highly destructive plant pathogen Phakopsora pachyrhizi Syd. & P. Syd., causal agent of Asian soybean rust. Myclobutanil is known from previous studies in grapevines to be xylem mobile. This study investigates the mobility profile of myclobutanil in soybean as an important component of its effective field performance. RESULTS: Over a 12 day period under greenhouse conditions, a constant uptake of myclobutanil from leaflet surfaces into the leaflet tissue was observed. Once in the leaflet, myclobutanil was seen to redistribute throughout the tissue, although no movement out of leaflets occurred owing to a lack of phloem mobility. The ability of myclobutanil to redistribute over distance within the soybean plant was revealed when visualizing movement of the compound to foliage above the point of application on the plant stem. An efficacy bioassay demonstrated that the systemic properties of myclobutanil allow control of disease at a point remote from the initial site of compound application. CONCLUSION: It is suggested that the high degree of xylem systemicity displayed by myclobutanil in soybean foliage is a contributory factor towards its commercial effectiveness for control of Asian soybean rust. Copyright © 2008 Society of Chemical Industry     

Formation of satellite uredinia as an important trait related to grapevine colonization by Phakopsora meliosmae-myrianthae

Phakopsora meliosmae-myrianthae, the causal agent of Asian grapevine leaf rust, significantly reduces the photosynthetic efficiency of grapevine leaves in green symptomless tissues surrounding lesions. This study took a close look at grapevine leaf colonization kinetics by P. meliosmae-myrianthae and compared it to P. pachyrhizi–soybean and Uromyces appendiculatus–bean colonization. It is already known from the literature that soybean rust, similar to grapevine rust, has a negative effect on leaf photosynthesis greater than would be expected based on visual lesions. However, in contrast to soybean and grapevine rusts, the effect of bean rust on leaf photosynthesis is proportional to the diseased leaf area. Colonization progress was monitored by fungal biomass assessed via histological staining and quantitative polymerase chain reaction (qPCR). Individual lesions of P. meliosmae-myrianthae on grapevine, P. pachyrhizi on soybean and U. appendiculatus on common bean leaves were evaluated every 3–4 days, and the number of uredinia was counted. Staining showed that mycelial colonization did not extend beyond the lesion border. The number of P. pachyrhizi and P. meliosmae-myrianthae uredinia within the lesions increased over time (on average 14-fold), whereas the number of U. appendiculatus uredinia remained the same. These findings were corroborated by qPCR, which revealed a greater increase in fungal biomass for Phakopsora spp. than for U. appendiculatus until 12 days post-inoculation. The high number of satellite uredinia within lesions might be directly related to the impact of this pathogen in photosynthetic efficiency on symptomless areas of diseased grapevine leaves. This study identified accelerated formation of satellite uredinia as an important feature of grapevine colonization by P. meliosmae-myrianthae.     

Coinfection of soybean plants with Phytophthora sojae and soybean cyst nematode does not alter the efficacy of resistance genes

The soybean cyst nematode (SCN) Heterodera glycines and the oomycete Phytophthora sojae are among the most damaging pathogens of soybean worldwide. Resistant cultivars are commonly used to manage these diseases. As it is known that the presence of SCN can facilitate the development of other pathogens, it is important to verify if there is a synergistic activity between SCN and P. sojae. The purpose of this study was to evaluate a possible interaction on susceptible and resistant soybean lines. The plants were inoculated with one or both organisms at different stages (5 or 10 days old). Two levels of SCN inoculum (2,000 and 10,000 eggs/plant) and different timing between SCN and P. sojae inoculation (2, 5, or 8 days) were compared. The results on 5-day-old plants showed that SCN did not influence P. sojae development. The resistant cultivar to P. sojae remained effective (0% mortality) and susceptible cultivars exhibited high mortality (100%) in the presence or absence of SCN. Experiments on 10-day-old plants showed that SCN resistance was not affected by the presence of P. sojae. SCN inoculum density and timing of P. sojae infection did not affect the virulence of these pathogens and the efficacy of resistance genes. However, the number of SCN cysts was decreased by more than 50% (p < .001) when P. sojae was coinfesting the susceptible cultivar. This suggests that P. sojae might indirectly influence SCN development by reducing the root mass. This study confirmed that resistant cultivars remain a valid option for the management of P. sojae and SCN.     

Fungicide sensitivity and monocyclic parameters related to the Phakopsora pachyrhizi–soybean pathosystem from organic and conventional soybean production systems

The failure of chemical control of soybean rust has been related to the selection of less sensitive isolates, and the infection capacity of such isolates could have implications for the management of the disease. The aims of the present study were to compare the sensitivity to tebuconazole and azoxystrobin and the monocycle of soybean rust using isolates of Phakopsora pachyrhizi from two soybean fields with different production systems (organic and conventional) in 2012/13 and 2013/14 seasons, and to monitor mutations in the CYP51 gene. To assess the sensitivity to tebuconazole and azoxystrobin, detached leaf tests and in vitro germination, respectively, were used. To evaluate the monocycle, detached leaves were inoculated with a urediniospore suspension and evaluated daily by counting the number of uredia. The occurrence of the mutations in CYP51 was investigated by a pyrosequencing assay. In both 2012/13 and 2013/14 seasons, the EC₅₀ to tebuconazole was lower for the population from the organic system (0.41 and 0.10 μg mL^?1, respectively) compared to the conventional system (1.60 and 4.44 μg mL^?1, respectively), while the EC₅₀ to azoxystrobin was similar for both populations. The lower sensitivity to tebuconazole and azoxystrobin was associated with F120L + Y131H mutations in CYP51, and the F129L mutation in CYTB, respectively. The monomolecular model fitted to monocycle data and parameters related to the maximum asymptote and the AUDPC were superior for organic than the conventional system.     

Soybean cultivar performance in the presence of soybean Asian rust,in relation to chemical control programs

Soybean rust is caused by an obligate parasite (Phakopsora pachyrhizi) which has spread in Brazil in each new season since 2001 and, despite the efforts to control the disease, losses have occurred every year. Its control demands several tactics amongst which chemical control with fungicides is the main method and remains indispensable. Control strategies such as the use of cultivars with partial resistance are desirable, but are not yet commercially available. The present study analyzed the existing differences in the reactions of short, medium and long cycle soybean cultivars against Asian rust and their responses to fungicide sprays. The experiment was conducted at Uberlandia-MG, Brazil, under field conditions from December 2007 to May 2008, in the Syngenta Seeds Experimental Station. The high pressure of the disease in the experiment simulated the natural pressure that the disease often reaches in Brazil. The studied variables were: visual severity (percentage of infected leaf area), percentage defoliation and productivity (kg ha⁻¹). Disease severity was expressed as AUDPC (area under disease progress curve). Variance analysis and comparison of means by the Tukey test (5% significance) were done for all variables studied. Significant differences were observed between cultivar effects and chemical control programs. The results obtained here indicate that the cultivars M-Soy 8199RR and Emgopa 315RR were less susceptible to disease, and that a control program termed “monitoring” (in which the appearance of new pustules of the pathogen were monitored to make the decision at each fungicide spray) was the most effective.     

Estimation of soybean rust uredospore terminal velocity,dry deposition,and the wet deposition associated with rainfall

Using models from atmospheric chemistry and physics, this study examined the wet deposition of single uredospores of soybean rust caused by Phakopsora pachyrhizi associated with rainfall and its importance compared with dry deposition. First, a measurement of the terminal velocity of freshly collected P. pachyrhizi uredospores was conducted in Nanning, China. The observed terminal velocities associated with different sizes of the uredospore clumps were fitted by negative exponential models. The average terminal velocity of single uredospores (0.0187 m s⁻¹) determined by the fitted models was used to estimate the dry deposition. The wet deposition of single uredospores associated with different rainfall rates was determined numerically using coupled models, in which raindrop capture efficiency of uredospores was based on Slinn’s semi-empirical model. The results showed that at a rainfall rate of 0.5 mm h⁻¹, wet deposition can remove 50% of the single uredospores in the air within 1 h. If the rainfall rate is 5 mm h⁻¹, 10 min is sufficient to remove 50% of the uredospores. The dry deposition of the single uredospores was estimated with simplified scenarios: i.e., assuming the uredospore cloud was continuously from 1,000 to 2,000 m in height above a field with a uniform concentration. In the first 16 h, almost no uredospores reached the ground, while the wet deposition caused by 2 mm h⁻¹ rainfall within 30 min was even much greater than dry deposition of 24 h duration. The comparisons indicated that the wet deposition of soybean rust uredospores was much more efficient than the dry deposition.     

Rhizoctonia solani: taxonomy,population biology and management of rhizoctonia seedling disease of soybean

Rhizoctonia solani, the most important species within the genus Rhizoctonia, is a soilborne plant pathogen with considerable diversity in cultural morphology, host range and aggressiveness. Despite its history as a destructive pathogen of economically important crops worldwide, our understanding of its taxonomic relationship with other Rhizoctonia‐like fungi, incompatibility systems, and population biology is rather limited. Among the host of diseases it has been associated with, seedling diseases inflicted on soybean are of significant importance, especially in the soybean growing regions of North America. Due to the dearth of resistant soybean genotypes, as well as the paucity of information on the mechanisms of host–pathogen interactions and other molecular aspects of pathogenicity, effective management options have mostly relied upon a combination of cultural and chemical control options. The first section of this review summarizes what is currently known about the taxonomy and systematics, population biology and molecular genetics of R. solani. The second section provides an overview of the pathology and management of rhizoctonia root and hypocotyl rot of soybean, a seedling disease of importance in North America.