Rust Fungus Research Articles

Genome-wide identification and functional characterization of wheat Brassinazole-resistant transcription factors in response to abiotic stresses and stripe rust infection.

Brassinazole-resistant (BZR) transcription factors (TFs) are key players in brassinolides (BRs) signaling pathway, which is widely involved in regulating plant growth and development, as well as in plant responding to a variety stresses. Despite their critical roles, little is known about BZR TFs in wheat. In this study, we performed genome-wide analysis of BZR gene family from wheat genome, and 20 TaBZRs were identified. Based on the phylogenetic relationships of TaBZR and BZRs from rice and Arabidopsis, all BZR genes were clustered into four groups. The intron-exon structural patterns and conserved protein motifs of TaBZRs showed high group specificity. TaBZR5, 7, and 9 were significantly induced after salt, drought treatment, and stripe rust infection. However, TaBZR16, which was significantly upregulated under NaCl application, was not expressed during wheat-stripe rust fungus interaction. These results indicated that BZR genes in wheat play different roles in response to various stresses. The results of this study will lay a foundation for further in-depth functional studies of TaBZRs and will provide information for the breeding and genetic improvement of wheat against drought and salt stresses.

Diploid Nuclei Occur throughout the Life Cycles of Pucciniales Fungi.

Within Eukaryotes, fungi are the typical representatives of haplontic life cycles. Basidiomycota fungi are dikaryotic in extensive parts of their life cycle, but diploid nuclei are known to form only in basidia. Among Basidiomycota, the Pucciniales are notorious for presenting the most complex life cycles, with high host specialization, and for their expanded genomes. Using cytogenomic (flow cytometry and cell sorting on propidium iodide-stained nuclei) and cytogenetic (FISH with rDNA probe) approaches, we report the widespread occurrence of replicating haploid and diploid nuclei (i.e., 1C, 2C and a small proportion of 4C nuclei) in diverse life cycle stages (pycnial, aecial, uredinial, and telial) of all 35 Pucciniales species analyzed, but not in sister taxa. These results suggest that the Pucciniales life cycle is distinct from any cycle known, i.e., neither haplontic, diplontic nor haplodiplontic, corroborating patchy and disregarded previous evidence. However, the biological basis and significance of this phenomenon remain undisclosed. IMPORTANCE Within Eukaryotes, fungi are the typical representatives of haplontic life cycles, contrasting with plants and animals. As such, fungi thus contain haploid nuclei throughout their life cycles, with sexual reproduction generating a single diploid cell upon karyogamy that immediately undergoes meiosis, thus resuming the haploid cycle. In this work, using cytogenetic and cytogenomic tools, we demonstrate that a vast group of fungi presents diploid nuclei throughout their life cycles, along with haploid nuclei, and that both types of nuclei replicate. Moreover, haploid nuclei are absent from urediniospores. The phenomenon appears to be transversal to the organisms in the order Pucciniales (rust fungi) and it does not occur in neighboring taxa, but a biological explanation or function for it remains elusive.

Integrative analysis of the multi-omics reveals the stripe rust fungus resistance mechanism of the TaPAL in wheat.

Wheat is one of the major food crops in the world. However, stripe rust fungus significantly decreases wheat yield and quality. In the present study, transcriptomic and metabolite analyses were conducted in R88 (resistant line) and CY12 (susceptible cultivar) during Pst-CYR34 infection due to the limited availability of information regarding the underlying mechanisms governing wheat-pathogen interactions. The results revealed that Pst infection promoted the genes and metabolites involved in phenylpropanoid biosynthesis. The key enzyme gene TaPAL to regulate lignin and phenolic synthesis has a positive resistance contribution to Pst in wheat, which was verified by the virus-induced gene silencing (VIGS) technique. The distinctive resistance of R88 is regulated by the selective expression of genes involved in the fine-tuning of wheat-Pst interactions. Furthermore, metabolome analysis suggested that lignin biosynthesis-related metabolite accumulation was significantly affected by Pst. These results help to elucidate the regulatory networks of wheat-Pst interactions and pave the way for durable resistance breeding in wheat, which may ease environmental and food crises around the world.

Insights into Diversity, Distribution, and Systematics of Rust Genus Puccinia.

Puccinia, which comprises 4000 species, is the largest genus of rust fungi and one of the destructive plant pathogenic rust genera that are reported to infect both agricultural and nonagricultural plants with severe illnesses. The presence of bi-celled teliospores is one of the major features of these rust fungi that differentiated them from Uromyces, which is another largest genus of rust fungi. In the present study, an overview of the current knowledge on the general taxonomy and ecology of the rust genus Puccinia is presented. The status of the molecular identification of this genus along with updated species numbers and their current statuses in the 21st century are also presented, in addition to their threats to both agricultural and nonagricultural plants. Furthermore, a phylogenetic analysis based on ITS and LSU DNA sequence data available in GenBank and the published literature was performed to examine the intergeneric relationships of Puccinia. The obtained results revealed the worldwide distribution of Puccinia. Compared with other nations, a reasonable increase in research publications over the current century was demonstrated in Asian countries. The plant families Asteraceae and Poaceae were observed as the most infected in the 21st century. The phylogenetic studies of the LSU and ITS sequence data revealed the polyphyletic nature of Puccinia. In addition, the presences of too short, too lengthy, and incomplete sequences in the NCBI database demonstrate the need for extensive DNA-based analyses for a better understanding of the taxonomic placement of Puccinia.

A Novel Mitovirus PsMV2 Facilitates the Virulence of Wheat Stripe Rust Fungus.

Wheat stripe rust, caused by the obligate biotrophic fungus Puccinia striiformis f. sp. tritici (Pst), seriously affects wheat production. Here, we report the complete genome sequence and biological characterization of a new mitovirus from P. striiformis strain GS-1, which was designated as "Puccinia striiformis mitovirus 2" (PsMV2). Genome sequence analysis showed that PsMV2 is 2658 nt in length with an AU-rich of 52.3% and comprises a single ORF of 2348 nt encoding an RNA-dependent RNA polymerase (RdRp). Phylogenetic analysis indicated that PsMV2 is a new member of the genus Unuamitovirus within the family Mitoviridae. In addition, PsMV2 multiplied highly during Pst infection and it suppresses programmed cell death (PCD) triggered by Bax. Silencing of PsMV2 in Pst by barley stripe mosaic virus (BSMV)-mediated Host Induced Gene Silencing (HIGS) reduced fungal growth and decreased pathogenicity of Pst. These results indicate PsMV2 promotes host pathogenicity in Pst. Interestingly, PsMV2 was detected among a wide range of field isolates of Pst and may have coevolved with Pst in earlier times. Taken together, our results characterized a novel mitovirus PsMV2 in wheat stripe rust fungus, which promotes the virulence of its fungal host and wide distribution in Pst which may offer new strategies for disease control.

Insect herbivore and fungal communities on Agathis (Araucariaceae) from the latest Cretaceous to Recent.

Agathis (Araucariaceae) is a genus of broadleaved conifers that today inhabits lowland to upper montane rainforests of Australasia and Southeast Asia. A previous report showed that the earliest known fossils of the genus, from the early Paleogene and possibly latest Cretaceous of Patagonian Argentina, host diverse assemblages of insect and fungal associations, including distinctive leaf mines. Here, we provide complete documentation of the fossilized Agathis herbivore communities from Cretaceous to Recent, describing and comparing insect and fungal damage on Agathis across four latest Cretaceous to early Paleogene time slices in Patagonia with that on 15 extant species. Notable fossil associations include various types of external foliage feeding, leaf mines, galls, and a rust fungus. In addition, enigmatic structures, possibly armored scale insect (Diaspididae) covers or galls, occur on Agathis over a 16-million-year period in the early Paleogene. The extant Agathis species, throughout the range of the genus, are associated with a diverse array of mostly undescribed damage similar to the fossils, demonstrating the importance of Agathis as a host of diverse insect herbivores and pathogens and their little-known evolutionary history.

The same boat, different storm: stress volatile emissions in response to biotrophic fungal infections in primary and alternate hosts

ABSTRACT Rust infection results in stress volatile emissions, but due to the complexity of host-pathogen interaction and variations in innate defense and capacity to induce defense, biochemical responses can vary among host species. Fungal-dependent modifications in volatile emissions have been well documented in numerous host species, but how emission responses vary among host species is poorly understood. Our recent experiments demonstrated that the obligate biotrophic crown rust fungus (P. coronata) differently activated primary and secondary metabolic pathways in its primary host Avena sativa and alternate host Rhamnus frangula. In A. sativa, emissions of methyl jasmonate, short-chained lipoxygenase products, long-chained saturated fatty acid derivatives, mono- and sesquiterpenes, carotenoid breakdown products, and benzenoids were initially elicited in an infection severity-dependent manner, but the emissions decreased under severe infection and photosynthesis was almost completely inhibited. In R. frangula, infection resulted in low-level induction of stress volatile emissions, but surprisingly, in enhanced constitutive isoprene emissions, and even severely-infected leaves maintained a certain photosynthesis rate. Thus, the same pathogen elicited a much stronger response in the primary than in the alternate host. We argue that future work should focus on resolving mechanisms of different fungal tolerance and resilience among primary and secondary hosts.

RNAi Technology: A New Path for the Research and Management of Obligate Biotrophic Phytopathogenic Fungi.

Powdery mildew and rust fungi are major agricultural problems affecting many economically important crops and causing significant yield losses. These fungi are obligate biotrophic parasites that are completely dependent on their hosts for growth and reproduction. Biotrophy in these fungi is determined by the presence of haustoria, specialized fungal cells that are responsible for nutrient uptake and molecular dialogue with the host, a fact that undoubtedly complicates their study under laboratory conditions, especially in terms of genetic manipulation. RNA interference (RNAi) is the biological process of suppressing the expression of a target gene through double-stranded RNA that induces mRNA degradation. RNAi technology has revolutionized the study of these obligate biotrophic fungi by enabling the analysis of gene function in these fungal. More importantly, RNAi technology has opened new perspectives for the management of powdery mildew and rust diseases, first through the stable expression of RNAi constructs in transgenic plants and, more recently, through the non-transgenic approach called spray-induced gene silencing (SIGS). In this review, the impact of RNAi technology on the research and management of powdery mildew and rust fungi will be addressed.

Phylogeny and taxonomy of three species of Blastospora (Pucciniales) from East Asia

ABSTRACT Three species of the rust fungus genus Blastospora, Bl. betulae, Bl. itoana, and Bl. smilacis, have been reported in East Asia. Although their morphological characteristics and life cycles have been investigated, their phylogenetic positions have not been clarified sufficiently. Phylogenetic analysis showed that these three species were included into Zaghouaniaceae of Pucciniales. However, Bl. betulae was phylogenetically distinct from Bl. itoana and Bl. smilacis and different from other genera. Based on this result, and applying recent International Code of Nomenclature decisions/recommendations/requirements, Botryosorus, gen. nov., and Bo. deformans,, comb. nov., were applied for Bl. betulae. Two new combinations, Bl. radiata for Bl. itoana and Bl. makinoi for Bl. smilacis, were also applied. Their host plants and distribution were described based on literature records. Zaghouania yunnanensis, comb. nov., was proposed for Cystopsora yunnanensis as a result of this analysis.

Phenolic acids and defense-related enzymes in host-pathogen interaction in Senecio aegyptius and the rust fungus, Puccinia lagenophorae

The objective of this study was to investigate the effect of the rust fungus Puccinia lagenophorae on total phenols, phenolic acids, and defense-related enzymes in the leaf tissues of the host, Senecio aegyptius. The infection increased the total phenol and defense-related enzymes in the host leaf tissues when compared to healthy tissues. The analysis using HPLC revealed the existence of 10 phenolic acids in healthy and infected host leaves. The infection increased the amounts of caffeic, chlorogenic, ferulic, p-coumaric, and salicylic acids. The examination by TEM revealed the presence of peroxidase activity on the host microbodies, Golgi bodies, endoplasmic reticulum, and the thickening of xylem vessels. After the infection, the activity was localized at the penetration sites of fungal haustoria and in amorphous materials found between the intercellular hyphae and the host cell wall. The heaviest peroxidase activity was detected in heavily infected vascular tissue, due to the monokaryotic infection.

Promotion of the growth and yield of Zea mays by synthetic microbial communities from Jala maize.

Plant growth-promoting bacteria (PGPB) are a source of nutrient supply, stimulate plant growth, and even act in the biocontrol of phytopathogens. However, these phenotypic traits have rarely been explored in culturable bacteria from native maize landraces. In this study, synthetic microbial communities (SynCom) were assembled with a set of PGPB isolated from the Jala maize landrace, some of them with additional abilities for the biocontrol of phytopathogenic fungi and the stimulation of plant-induced systemic resistance (ISR). Three SynCom were designed considering the phenotypic traits of bacterial strains, including Achromobacter xylosoxidans Z2K8, Burkholderia sp. Z1AL11, Klebsiella variicola R3J3HD7, Kosakonia pseudosacchari Z2WD1, Pantoea ananatis E2HD8, Pantoea sp. E2AD2, Phytobacter diazotrophicus Z2WL1, Pseudomonas protegens E1BL2, and P. protegens E2HL9. Plant growth promotion in gnotobiotic and greenhouse seedlings assays was performed with Conejo landrace; meanwhile, open field tests were carried out on hybrid CPL9105W maize. In all experimental models, a significant promotion of plant growth was observed. In gnotobiotic assays, the roots and shoot length of the maize seedlings increased 4.2 and 3.0 times, respectively, compared to the untreated control. Similarly, the sizes and weights of the roots and shoots of the plants increased significantly in the greenhouse assays. In the open field assay performed with hybrid CPL9105W maize, the yield increased from 11 tons/ha for the control to 16 tons/ha inoculated with SynCom 3. In addition, the incidence of rust fungal infections decreased significantly from 12.5% in the control to 8% in the treatment with SynCom 3. All SynCom designs promoted the growth of maize in all assays. However, SynCom 3 formulated with A. xylosoxidans Z2K8, Burkholderia sp. Z1AL11, K. variicola R3J3HD7, P. ananatis E2HD8, P. diazotrophicus Z2WL1, and P. protegens E1BL2 displayed the best results for promoting plant growth, their yield, and the inhibition of fungal rust. This study demonstrated the biotechnological eco-friendly plant growth-promoting potential of SynCom assemblies with culturable bacteria from native maize landraces for more sustainable and economic agriculture.

Major proliferation of transposable elements shaped the genome of the soybean rust pathogen Phakopsora pachyrhizi

With >7000 species the order of rust fungi has a disproportionately large impact on agriculture, horticulture, forestry and foreign ecosystems. The infectious spores are typically dikaryotic, a feature unique to fungi in which two haploid nuclei reside in the same cell. A key example is Phakopsora pachyrhizi, the causal agent of Asian soybean rust disease, one of the world’s most economically damaging agricultural diseases. Despite P. pachyrhizi’s impact, the exceptional size and complexity of its genome prevented generation of an accurate genome assembly. Here, we sequence three independent P. pachyrhizi genomes and uncover a genome up to 1.25 Gb comprising two haplotypes with a transposable element (TE) content of ~93%. We study the incursion and dominant impact of these TEs on the genome and show how they have a key impact on various processes such as host range adaptation, stress responses and genetic plasticity.

Intrinsically Disordered Kiwellin Protein-Like Effectors Target Plant Chloroplasts and are Extensively Present in Rust Fungi.

The effector proteins produced by plant pathogens are one of the essential components of host-pathogen interaction. Despite being important, most of the effector proteins remain unexplored due to the diversity in their primary sequence generated by the high selection pressure of the host immune system. However to maintain the primary function in the infection process, these effectors may tend to maintain their native protein fold to perform the corresponding biological function. In the present study, unannotated candidate secretory effector proteins of sixteen major plant fungal pathogens were analyzed to find the conserved known protein folds using homology, ab initio, and AlphaFold/Rosetta Foldprotein dimensional (3D) structure approaches. Several unannotated candidate effector proteins were found to match various known conserved protein families potentially involved in host defense manipulation in different plant pathogens. Surprisingly a large number of plant Kiwellin proteins fold like secretory proteins (> 100) were found in studied rust fungal pathogens. Many of them were predicted as potential effector proteins. Furthermore, template independent modelling usingAlphaFold/Rosetta Fold analysis and structural comparison of these candidates also predicted them to match with plant Kiwellin proteins. We also found plant Kiwellin matching proteinsoutside rusts including several non-pathogenic fungi suggesting the broad function of these proteins. One of the highest confidently modeled Kiwellin matching candidates effectors, Pstr_13960 (97.8%), from the Indian P. striiformis race Yr9 was characterized using overexpression, localization, and deletion studies in Nicotiana benthamiana. The Pstr_13960 suppressed the BAX-induced cell death and localized in the chloroplast. Furthermore, the expression of the Kiwellin matching region (Pst_13960_kiwi) alone suppressed the BAX-induced cell death in N. benthamiana despite the change of location to the cytoplasm and nucleus, suggesting the novel function of the Kiwellin corefold in rust fungi. Molecular docking showed that Pstr_13960 can interact with plant Chorismate mutases (CMs) using three loops conserved in plant and rust Kiwellins. Further analysis of Pstr_13960 showed to contain Intrinsically disordered regions (IDRs) in place of the N-terminal β1/β2 region found in plant Kiwellins suggesting the evolution of rust Kiwellins-like effectors (KLEs). Overall, this study reports the presence of a Kiwellin protein-like fold containing a novel effector protein family in rust fungi depicting a classical example of the evolution of effectors at the structure level as Kiwellin effectors show very low significant similarity to plant Kiwellin at the sequence level.

Light and scanning electron microscopy of aecia and aeciospores of Cronartium ribicola on Pinus koraiensis branch tissues

Morphological characteristics of aecia and aeciospores of Cronartium ribicola on Pinus koraiensis branch tissues were investigated using light and field emission scanning electron microscopy (FESEM). Mature P. koraiensis trees in Jeongseon, Korea, showed yellowish aecia on stems and branches. Aecia and surrounding tissues were excised from the lesions and vapor-fixed for FESEM imaging, which revealed morphology including intact blister-shaped, flattened, and burst forms. Light microscopy revealed yellowish aeciospores having surface projections. Aeciospores were mostly ovoid and measured approximately 20 µm long. The FESEM showed irregularly shaped cracks on the aecia that had erupted through the bark of P. koraiensis. Some aeciospores had germinated, producing two germ tubes from a spore in a burst aecium. Aeciospores had both smooth and verrucose regions on the surface, and some had concave or convex regions. Aeciospore layers and underlying fungal matrices including aecial columns were obvious in the cross-sections of aecia. Approximately 1 µm-high wart-like surface projections could be resolved and comprised less than 10 angular platelets stacked in vertical rows. Remains of the primary spore wall were present between surface projections. These results provide insights into the morphology of the heteroecious rust fungus with the help of vapor fixation and high-resolution surface imaging.

TaUAM3, a UDP‐Ara mutases protein, positively regulates wheat resistance to the stripe rust fungus

AbstractPlant cell walls are composed of polysaccharides such as cellulose, hemicelluloses, and pectins, whose location and function differ depending on plant type. UDP‐Ara mutases (UAMs) have been reported to play important roles in plant development and response to various plant stresses (abiotic and biotic). However, little work has been reported on UAM in wheat. In this study, we dissected the role of the UAM family member, UAM3, during the interaction between wheat and the stripe rust fungus, Puccinia striiformis f. sp. tritici (Pst), and in response to treatment with salicylic acid (SA). RNA interference (RNAi)‐based stable silencing of TaUAM3 resulted in decreased resistance to Pst fungus. In addition, CRISPR‐mediated genome editing (GE) of TaUAM3 enhanced the susceptibility of wheat to Pst or compromised disease resistance accompanied by increased fungal growth and decreased H2O2 production in plant tissues. Moreover, the transcript levels of pathogenesis‐related (PR) genes and reactive oxygen species (ROS)‐generating genes were down‐regulated in both the RNAi‐silenced and CRISPR‐edited plants, while the ROS‐scavenging gene, TaCAT3, was up‐regulated. Therefore, TaUAM3 positively regulates the resistance of wheat to Pst.

Three‐way fungal interactions affect the potential biological control of Himalayan balsam, Impatiens glandulifera

AbstractHimalayan balsam (Impatiens glandulifera) is one of the most invasive weeds across Europe. The rust fungus, Puccinia komarovii var. glanduliferae has been introduced as a biological control agent, but success has been patchy. Here, we investigated whether mycorrhizal and endophytic fungi can affect rust efficacy and plant growth. Over three experiments, we found that AM fungi and the rust alone or together consistently reduced plant growth, but this depended on the identity of species in the AM inoculum. Meanwhile, AM fungi increased infection frequency of the endophyte Colletotrichum acutatum. Rust inoculation had no detrimental effects on mycorrhizal colonisation or C. acutatum infection, but the latter two fungi reduced rust sporulation. However, plant size was reduced when all three fungal types were present, suggesting that a combined fungal inoculum offers a promising approach for the control of this weed.

Raveneliopsis, a new genus of ravenelioid rust fungi on Cenostigma (Caesalpinioideae) from the Brazilian Cerrado and Caatinga

ABSTRACT The multicellular discoid convex teliospore heads represent a prominent generic feature of the genus Ravenelia. However, recent molecular phylogenetic work has shown that this is a convergent trait, and that this genus does not represent a natural group. In 2000, a rust fungus infecting the Caesalpinioid species Cenostigma macrophyllum (= C. gardnerianum) was described as Ravenelia cenostigmatis. This species shows some rare features, such as an extra layer of sterile cells between the cysts and the fertile teliospores, spirally ornamented urediniospores, as well as strongly incurved paraphyses giving the telia and uredinia a basket-like appearance. Using freshly collected specimens of Rav. cenostigmatis and Rav. spiralis on C. macrophyllum, our phylogenetic analyses based on the nuc 28S, nuc 18S, and mt CO3 (cytochrome c oxidase subunit 3) gene sequences demonstrated that these two rust fungi belong in a lineage within the Raveneliineae that is distinct from Ravenelia s. str. Besides proposing their recombination into the new genus Raveneliopsis (type species R. cenostigmatis) and briefly discussing their potentially close phylogenetic affiliations, we suggest that five other Ravenelia species that are morphologically and ecologically close to the type species of Raveneliopsis, i.e., Rav. corbula, Rav. corbuloides, Rav. parahybana, Rav. pileolarioides, and Rav. Striatiformis, may be recombined pending new collections and confirmation through molecular phylogenetic analyses.

Ultrastructural studies of the host (Senecio aegyptius L.) responses after the infection by the pathogenic rust fungus, Puccinia lagenophorae Cooke

This study investigated the impact of the rust fungus, Puccinia lagenophorae on its host, Senecio aegyptius using both light (LM) and transmission (TEM) electron microscopy. The chloroplast numbers decreased during the aecial and telial stages of the pathogen (53.7 and 56.6%, respectively). The nucleus volumes were increased during both stages (10.9 and 14.1%, respectively). Infected chloroplasts and nuclei showed changes in their shape and ultrastructure. Instead of the M-haustorium, a collar was created around the D-haustorium. A neck encasing developed in both types of necrotic haustoria. This encasement may serve as the host cell’s protection against toxins created after the death of haustoria. The severe changes in the chloroplasts and nuclei of this host plant after infection by this pathogen may give this pathogen the possibility of controlling this serious weed. Future research in the molecular mechanism may explore the biological control of weeds for local farm agroecology.

A rust-fungus Nudix hydrolase effector decaps mRNA invitro and interferes with plant immune pathways.

To infect plants, pathogenic fungi secrete small proteins called effectors. Here, we describe the catalytic activity and potential virulence function of the Nudix hydrolase effector AvrM14 from the flax rust fungus (Melampsora lini). We completed extensive invitro assays to characterise the enzymatic activity of the AvrM14 effector. Additionally, we used in planta transient expression of wild-type and catalytically dead AvrM14 versions followed by biochemical assays, phenotypic analysis and RNA sequencing to unravel how the catalytic activity of AvrM14 impacts plant immunity. AvrM14 is an extremely selective enzyme capable of removing the protective 5' cap from mRNA transcripts invitro. Homodimerisation of AvrM14 promoted biologically relevant mRNA cap cleavage invitro and this activity was conserved in related effectors from other Melampsora spp. In planta expression of wild-type AvrM14, but not the catalytically dead version, suppressed immune-related reactive oxygen species production, altered the abundance of some circadian-rhythm-associated mRNA transcripts and reduced the hypersensitive cell-death response triggered by the flax disease resistance protein M1. To date, the decapping of host mRNA as a virulence strategy has not been described beyond viruses. Our results indicate that some fungal pathogens produce Nudix hydrolase effectors with invitro mRNA-decapping activity capable of interfering with plant immunity.

Electron microscopy of the uredinial and telial stages of the rust fungus, Puccinia imperatae

The purpose of this work was to assess the morphology and ontogeny of Puccinia imperatae urediniospores and teliospores growing on its host, Imperata cylindrica, for the first time using scanning (SEM) and transmission (TEM) electron microscopy. The first evidence of uredinial development is the aggregation of hyphae in host intercellular spaces under the epidermis to form an uredinial initial. Uredinial primordia become evident as compact masses of fungal hyphae. The mycelia are dikaryotic and originated from dikaryotic urediniospores, which infect a new host. Dikaryotic sporogenous cells begin to form from dikaryotic mycelium as the uredinium develops further. The fundamental cell in the urediniospore and teliospore formation is the sporogenous cell. The teliospores with their persistent pedicels arise from dense dikaryotic sporogenous tissue, which later becomes highly vacuolated. The sporogenous cell forms a spore bud, which later divides into a pedicel and teliospore mother cell with two nuclei that produce the mature teliospore. SEM examination revealed that the urediniospores are echinulate bearing conical spines on their surfaces, sometimes curving near their tips, and the echinulate ornaments are distributed over the entire urediniospore surface. SEM examination also indicated that the teliospore surface shows rugose ornamentations. This investigation proved that the morphology of the uredinia of different Puccinia species is not related to the host species, but to the rust species. The possibility of using this fungus as a biological control agent for the noxious weed, Imperata cylindrica, was also discussed.