Leaf Rust Fungus Research Articles

Wheat Leaf Rust Fungus Effector Protein Pt1641 Is Avirulent to TcLr1.

Wheat leaf rust fungus is an obligate parasitic fungus that can absorb nutrients from its host plant through haustoria and secrete effector proteins into host cells. The effector proteins are crucial factors for pathogenesis as well as targets for host disease resistance protein recognition. Exploring the role of effector proteins in the pathogenic process of Puccinia triticina Eriks. (Pt) is of great significance for unraveling its pathogenic mechanisms. We previously found that a cysteine-rich effector protein, Pt1641, is highly expressed during the interaction between wheat and Pt, but its specific role in pathogenesis remains unclear. Therefore, this study employed techniques such as heterologous expression, qRT-PCR analysis, and host-induced gene silencing (HIGS) to investigate the role of Pt1641 in the pathogenic process of Pt. The results indicate that Pt1641 is an effector protein with a secretory function and can inhibit BAX-induced programmed cell death in Nicotiana benthamiana. qRT-PCR analyses showed that expression levels of Pt1641 were different during the interaction between the high-virulence strain THTT and low-virulence strains FGD and Thatcher, respectively. The highest expression level in the low-virulence strain FGD was four times that of the high-virulence strain THTT. The overexpression of Pt1641 in wheat near-isogenic line TcLr1 induced callose deposition and H2O2 production on TcLr1. After silencing Pt1641 in the Pt low-virulence strain FGD on wheat near-isogenic line TcLr1, the pathogenic phenotype of Pt physiological race FGD on TcLr1 changed from ";" to "3", indicating that Pt1641 plays a non-toxic function in the pathogenicity of FGD to TcLr1. This study helps to reveal the pathogenic mechanism of wheat leaf rust and provides important guidance for the mining and application of Pt avirulent genes.

The nematode signaling molecule ascr#18 induces prepenetration defenses in wheat against a leaf rust fungus

AbstractLeaf rust, caused by the pathogenic biotrophic rust fungus Puccinia triticina (Pt), is one of the most destructive wheat diseases worldwide; its negative impact on crop yields is exacerbated by increasing temperatures due to climate change. Ascarosides are nematode pheromones that induce resistance to microbial pathogens and pests in a wide range of crops, making them valuable components in biocontrol scenarios. We investigated the effect on infection of various wheat (Triticum aestivum) genotypes with the virulent Pt race 77W × R by ascr#18, the major ascaroside secreted into the rhizosphere by plant-parasitic nematodes. Spraying the leaves with ascr#18 24 h before inoculation with fungal uredospores slowed disease development and resulted in a reduction of the number of rust pustules on treated compared to untreated leaves. Dose–response analysis over the nano- and micromolar range revealed a broad optimum concentration down to 0.01nM ascr#18. Microscopic analysis showed very early arrest of the fungus at the appressorial stage, with associated enhanced local accumulation of H2O2 and abortive stoma penetration. Similarly, ascr#18 also induced strong resistance to Pt race PKTTS, confirming its race-unspecific biocontrol activity. The results of this study are consistent with and extend previous research that has shown that ascr#18 activates plant immunity and thus protects plants from pathogens even at very low doses.

Evasion of wheat resistance gene Lr15 recognition by the leaf rust fungus is attributed to the coincidence of natural mutations and deletion in AvrLr15 gene.

Employing race-specific resistance genes remains an effective strategy to protect wheat from leaf rust caused by Puccinia triticina (Pt) worldwide, while the newly emerged Pt races, owing to rapid genetic evolution, frequently overcome the immune response delivered by race-specific resistance genes. The molecular mechanisms underlying the newly evolved virulence Pt pathogen remain unknown. Here, we identified an avirulence protein AvrLr15 from Pt that induced Lr15-dependent immune responses. Heterologously produced AvrLr15 triggered pronounced cell death in Lr15-isogenic wheat leaves. AvrLr15 contains a functional signal peptide, localized to the plant nucleus and cytosol and can suppress BAX-induced cell death. Evasion of Lr15-mediated resistance in wheat was associated with a deletion and point mutations of amino acids in AvrLr15 rather than AvrLr15 gene loss in the Lr15-breaking Pt races, implying that AvrLr15 is required for the virulence function of Pt. Our findings identified the first molecular determinant of wheat race-specific immunity and facilitated the identification of the first AVR/R gene pair in the Pt-wheat pathosystem, which will provide a molecular marker to monitor natural Pt populations and guide the deployment of Lr15-resistant wheat cultivars in the field.

The F-box protein encoding genes of the leaf-rust fungi Puccinia triticina: genome-wide identification, characterization and expression dynamics during pathogenesis.

The F-box proteins in fungi perform diverse functions including regulation of cell cycle, circadian clock, development, signal transduction and nutrient sensing. Genome-wide analysis revealed 10 F-box genes in Puccinia triticina, the causal organism for the leaf rust disease in wheat and were characterized using in silico approaches for revealing phylogenetic relationships, gene structures, gene ontology, protein properties, sequence analysis and gene expression studies. Domain analysis predicted functional domains like WD40 and LRR at C-terminus along with the obvious presence of F-box motif in N-terminus. MSA showed amino acid replacements, which might be due to nucleotide substitution during replication. Phylogenetic analysis revealed the F-box proteins with similar domains to be clustered together while some sequences were spread out in different clades, which might be due to functional diversity. The clustering of Puccinia triticina GG705409 with Triticum aestivum TaAFB4/TaAFB5 in a single clade suggested the possibilities of horizontal gene transfer during the coevolution of P. triticina and wheat. Gene ontological annotation categorized them into three classes and were functionally involved in protein degradation through the protein ubiquitination pathway. Protein-protein interaction network revealed F-box proteins to interact with other components of the SCF complex involved in protein ubiquitination. Relative expression analysis of five F-box genes in a time course experiment denoted their involvement in leaf rust susceptible wheat plants. This study provides information on structure elucidation of F-box proteins of a basidiomycetes plant pathogenic fungi and their role during pathogenesis.

How to Differentiate between Resistant and Susceptible Wheat Cultivars for Leaf Rust Fungi Using Antioxidant Enzymes and Histological and Molecular Studies?

Eight wheat cultivars, Sakha-94, Giza-171, Sids-1, Sids-12, Sids-13, Shandweel-1, Misr-1, and Misr-2, were evaluated for leaf rust at the seedling and adult stages in the 2021 and 2022 seasons. Biochemical, histological, and genetic analyses were performed to determine the link between cultivars that were either sensitive or resistant to the disease. Misr-2 and Giza-171 cultivars had the highest levels of resistance to leaf rust races in 2021 (LTCGT, STSJT, and TTTST) and 2022 (MBGJT, TTTKS, and TTTTT) at the seedling stage. However, at the adult stage, Sakha-94, Giza-171, Misr-1, and Misr-2 cultivars had the highest levels of resistance; consequently, they had the lowest final disease severity and the lowest values of AUDPC. The correlation between the seedling reaction and adult reaction was non-significant, with values of 0.4401 and 0.4793 in the 2021 and 2022 seasons, respectively. Throughout the biochemical, histological, and genetic analyses, it was observed that catalase, peroxidase, and polyphenol oxidase activities significantly increased in the resistant cultivars. The discoloration of superoxide (O2-) and hydrogen peroxide (H2O2) significantly decreased in resistant and moderately resistant wheat cultivars (Sakha-94, Giza-171, Misr-1, and Misr-2); higher hydrogen peroxide (H2O2) and superoxide (O2-) levels were recorded for the susceptible cultivars compared to the resistant cultivars. Molecular markers proved that the Lr50 gene was detected in the resistant cultivars. Puccinia triticina infections negatively affected most histological characteristics of flag leaves, especially in susceptible cultivars. The thickness of the blade (µ), the thickness of the upper and lower epidermis (UE and LE), the thickness of mesophyll tissue (MT), and bundle length and width in the midrib were decreased in susceptible cultivars such as Sids-1, Sids-13, and Shandwel-1 compared with resistant cultivars.

The Expression of Triticum aestivum Cysteine-Rich Receptor-like Protein Kinase Genes during Leaf Rust Fungal Infection.

Understanding the role of cysteine-rich receptor-like kinases (CRKs) in plant defense mechanisms is crucial for enhancing wheat resistance to leaf rust fungus infection. Here, we identified and verified 164 members of the CRK gene family using the Triticum aestivum reference version 2 collected from the international wheat genome sequencing consortium (IWGSC). The proteins exhibited characteristic features of CRKs, including the presence of signal peptides, cysteine-rich/stress antifungal/DUF26 domains, transmembrane domains, and Pkinase domains. Phylogenetic analysis revealed extensive diversification within the wheat CRK gene family, indicating the development of distinct specific functional roles to wheat plants. When studying the expression of the CRK gene family in near-isogenic lines (NILs) carrying Lr57- and Lr14a-resistant genes, Puccinia triticina, the causal agent of leaf rust fungus, triggered temporal gene expression dynamics. The upregulation of specific CRK genes in the resistant interaction indicated their potential role in enhancing wheat resistance to leaf rust, while contrasting gene expression patterns in the susceptible interaction highlighted potential susceptibility associated CRK genes. The study uncovered certain CRK genes that exhibited expression upregulation upon leaf rust infection and the Lr14a-resistant gene. The findings suggest that targeting CRKs may present a promising strategy for improving wheat resistance to rust diseases.

Wheat leaf rust fungus effector Pt13024 is avirulent to TcLr30.

Wheat leaf rust, caused by Puccinia triticina Eriks. (Pt), is a global wheat disease threatening wheat production. Dissecting how Pt effector proteins interact with wheat has great significance in understanding the pathogenicity mechanisms of Pt. In the study, the cDNA of Pt 13-5-72 interacting with susceptible cultivar Thatcher was used as template to amplify Pt13024 gene. The expression pattern and structure of Pt13024 were analyzed by qRT-PCR and online softwares. The secretion function of Pt13024 signal peptide was verified by the yeast system. Subcellular localization of Pt13024 was analyzed using transient expression on Nicotiana benthamiana. The verification that Pt13024 inhibited programmed cell death (PCD) was conducted on N. benthamiana and wheat. The deletion mutation of Pt13024 was used to identify the virulence function motif. The transient transformation of wheat mediated by the type III secretion system (TTSS) was used to analyze the activity of regulating the host defense response of Pt13024. Pt13024 gene silencing was performed by host-induced gene silencing (HIGS). The results showed that Pt13024 was identified as an effector and localized in the cytoplasm and nucleus on the N. benthamiana. It can inhibit PCD induced by the Bcl-2-associated X protein (BAX) from mice and infestans 1 (INF1) from Phytophthora infestans on N. benthamiana, and it can also inhibit PCD induced by DC3000 on wheat. The amino acids 22 to 41 at N-terminal of the Pt13024 are essential for the inhibition of programmed cell death (PCD) induced by BAX. The accumulation of reactive oxygen species and deposition of callose in near-isogenic line TcLr30, which is in Thatcher background with Lr30, induced by Pt13024 was higher than that in 41 wheat leaf rust-resistant near-isogenic lines (monogenic lines) with different resistance genes and Thatcher. Silencing of Pt13024 reduced the leaf rust resistance of Lr30 during the interaction between Pt and TcLr30. We can conclude that Pt13024 is avirulent to TcLr30 when Pt interacts with TcLr30. These findings lay the foundation for further investigations into the role of Pt effector proteins in pathogenesis and their regulatory mechanisms.

Simplifying Barley Leaf Rust Research: An Easy and Reproducible Infection Protocol for Puccinia hordei on a Small Laboratory Scale.

Barley (Hordeum vulgare) is one of the most important agricultural crops in the world, but pathogen infections regularly limit its annual yield. A major threat is the infection with the biotrophic leaf rust fungus, Puccinia hordei. Rust fungi have a complex life cycle, and existing resistances can be easily overcome. To address this problem, it is crucial to develop barley varieties with improved and durable resistance mechanisms. An essential step towards this goal is a simple and reproducible infection protocol to evaluate potential resistance phenotypes in the lab. However, available protocols sometimes lack detailed procedure or equipment information, use spore application methods that are not suitable for uniform spore dispersion, or require special mineral oils or engineered fluids. In addition, they are often optimized for pathogen-dedicated greenhouses or phytochambers, which may not be available to every research institute. Here, we describe an easy and user-friendly procedure to infect barley with Puccinia hordei on a small laboratory scale. This procedure utilizes inexpensive and simple tools to evenly split and apply spores to barley leaves. The treated plants are incubated in affordable and small phytocabinets. Our protocol enables a quick and reproducible infection of barley with leaf rust, a method that can easily be transferred to other rust fungi, including stripe rust, or to other plant species. Key features Step-by-step infection protocol established for barley cv. Golden Promise, the gold standard genotype for genetic transformation Plant age-independent protocol Precise spore application by using inexpensive pipe cleaners for uniform symptom formation and increased reproducibility No specialized equipment needed Includes simple spore harvesting method Protocol is applicable to other biotrophic pathogens (stripe rust or powdery mildew) and other plants (e.g., wheat) Protocol is also applicable for a detached leaf assay Graphical overview.

Dynamics of cell wall components and histochemical profile of a rust fungi gall (Basidiomycota: Pucciniales) on Byrsonima variabilis A.Juss. (Malpighiaceae)

Context An obligate biotrophic parasitism with a rust fungus led to gall formation on Byrsonima variabilis. Aims The hypothesis that the host leaf–rust fungi interaction alters the dynamics of plant cell walls and the histochemical profile toward favouring the plant cell-to-fungi cell translocation of metabolites is tested. Methods Gall samples were sectioned and submitted to anatomical, histometric, histochemical, and immunocytochemical techniques to evaluate structural alterations and the detection of primary and secondary metabolites, as well as the epitopes of glycoproteins, pectins, and hemicelluloses. Key results Fungi gall development results in the hypertrophy of the stomatal chamber and the hyperplasia of epidermis and spongy parenchyma. The cell-to-cell translocation of metabolites from plant mesophyll cells toward the rust fungi gall is favoured by the epitopes of homogalacturonans (HGs) and (1 → 5) α-l-arabinans detected in the hyphae passage sites in the pycnial and aecial stages. The arabinogalactan-proteins (AGPs) may favour mycelial nutrition and differentiation, and cell wall adhesion. HGs and arabinans confer porosity to mesophyll cell walls, which favours the traffic of molecules toward the rust fungi gall. Conclusions The unexpected labelling of AGPs, HGs, and arabinans in fungi cell walls is a novelty regarding the plant–fungi interaction. The primary metabolites detected in rust fungi support hyphae growth and spore maturation. Implications The immunolabelling of host plant cell wall components on fungi cell walls indicates the integrative role of some plant cell wall components in the biological process of pathogen colonisation in leaf tissues.

SSR genotypes of Puccinia triticina in 10 provinces of China reveal a possible source of the wheat leaf rust fungus

AbstractWheat leaf rust is a fungal disease caused by Puccinia triticina and spread by wind. In recent years, with changes in the climate and farming systems, damage caused by wheat leaf rust has become increasingly severe. To study the genetic structure of P. triticina populations in China and provide a basis for control of wheat leaf rust, collections of P. triticina were obtained from Xinjiang, Inner Mongolia, Hebei, Henan, Jiangsu, Anhui, Hubei, Sichuan, Guizhou and Yunnan provinces in 2019. A total of 177 single uredinial isolates were tested with 13 simple‐sequence repeat (SSR) primers. The results showed that there was frequent gene exchange within P. triticina populations in China, but a distinct population divergence was also revealed between those in Xinjiang and Yunnan provinces. Results also suggested that Inner Mongolia might be a source of P. triticina for other regions. In addition, the linkage disequilibrium test indicated that the P. triticina populations reproduced by clonal production of urediniospores.

A decade after the first Pucciniales genomes: A bibliometric snapshot of (post) genomics studies in three model rust fungi.

Pucciniales (rust fungi) are one of the largest fungal order of plant pathogens. They collectively infect key crops such as wheat and soybean, and threaten global food security. In the early 2010s, the genome sequences of three rust fungi were released: Melampsora larici-populina (the poplar leaf rust fungus), Puccinia graminis f. sp. tritici (the wheat stem rust fungus), and Puccinia striiformis f. sp. triciti (the wheat stripe rust or wheat yellow rust fungus). The availability of those genomes has forwarded rust biology into the post-genomic era, sparking a series of genomics, transcriptomics, in silico, and functional studies. Here, we snapshot the last 10 years of post-genomics studies addressing M. larici-populina, P. graminis f. sp. tritici, and/or P. striiformis f. sp. tritici. This mini-review notably reveals the model species-centered structure of the research community, and highlights the drastic increase of the number of functional studies focused on effectors since 2014, which notably revealed chloroplasts as a central host compartment targeted by rust fungi. This mini-review also discusses genomics-facilitated studies in other rust species, and emerging post-genomic research trends related to fully-phased rust genomes.

Virulence Phenotypes of the Wheat Leaf Rust Pathogen, Puccinia triticina, in the United States from 2018 to 2020.

Collections of wheat leaves infected with the leaf rust fungus, Puccinia triticina, were obtained from the southeastern states, the Ohio Valley, the Great Plains, and Washington in 2018, 2019, and 2020 to determine the prevalent virulence phenotypes in the wheat-growing regions of the United States. In the hard red winter wheat region of the southern and mid Great Plains, MNPSD and MPPSD were the two most common phenotypes in 2018 and 2019. In 2020, BBBQD with high virulence to durum wheat was the most common phenotype in the southern Great Plains. In the hard red spring wheat region of the northern Great Plains, MNPSD, MPPSD, MBDSD, and TBBGS were the predominant phenotypes. In the soft red winter wheat region of the southeastern states and Ohio Valley region, MBTNB, MCTNB, and MNPSD were the three most common phenotypes. Collections in Washington had phenotypes LBDSG, LCDSG, LCDJG, and MBDSB that were not found in any other region. Isolates with virulence to the leaf rust resistance (Lr) gene Lr11 were most frequent in the southeastern states and Ohio Valley regions. The frequency of isolates with virulence to the Lr39 gene was highest in the Great Plains region and frequency of isolates with virulence to the Lr21 gene was highest in the northern Great Plains region. Selection of virulence phenotypes by Lr genes in the different market classes of wheat, combined with the effects of clonal reproduction, overwintering in southern regions, and low migration between the Great Plains region and eastern wheat-producing regions, has maintained the different P. triticina populations in the United States.

Incidence of Coffee Leaf Rust in Vietnam, Possible Original Sources and Subsequent Pathways of Migration.

This research focused on the incidence and population genetics of coffee leaf rust (CLR) fungus, Hemileia vastatrix, to estimate the possible original source(s) and subsequent migration pathways of wind-borne and human-aided spores in three main coffee production regions (Northwest, Central Highlands, and Southeast) in Vietnam. In southern Vietnam (Central Highlands and Southeast), Coffea canephora covers the majority area, while Catimor lines of C. arabica accounts for 95% of the coffee plantations in northwestern Vietnam. Field surveys conducted at eighty-five plantations, show coffee leaf samples infected by the rust fungus across forty-one plantations. Catimor varieties exhibited high levels of susceptibility with severe rust symptoms, while robusta varieties had varying degrees of susceptibility. We analyzed 863−869 base pairs of internal transcribed spacer (ITS) region from 83 samples (41 sequences from Vietnam, 2 from Thailand, and the remaining 40 from American countries); and fifty-two haplotypes consisting of 123 polymorphic sites were detected. Although the analysis of molecular variance (AMOVA) indicates significant genetic differentiation in the H. vastatrix populations in Vietnam, there was no clear genetic structure with respect to the three geographic areas surveyed. Based on the haplotype network, NeighborNet analysis, and geographical distribution patterns of the haplotypes, five haplotypes were identified as early established, from which most other haplotypes in Vietnam were derived. The early established haplotypes were found in the highest frequency in Northwest Vietnam. This finding corresponds to the earliest record of CLR in Vietnam. The phylogenetic network analysis also illustrated that H. vastatrix had expanded from the northwest to southern Vietnam. Pairwise genetic distance analysis and the geophylogenetic tree also suggests that CLR was first established in the Northwest. In addition, some scattered individuals on the principal coordinate analysis (PCoA) diagram and several separated haplotypes in the phylogenetic networks indicated that other branches of CLR in Vietnam were initiated in the Central Highlands. Hemileia vastatrix from these branches have been spreading in southern Vietnam.

Genotype Groups of the Wheat Leaf Rust Fungus Puccinia triticina in the United States as Determined by Genotyping by Sequencing.

Wheat leaf rust caused by Puccinia triticina is a widespread disease of wheat in the United States and worldwide. Populations of P. triticina are characterized by virulence phenotypes that change rapidly because of selection by wheat cultivars with leaf rust resistance genes. The objective of this study was to genotype collections of P. triticina from 2011 to 2018 in the United States using restriction site-associated genotyping by sequencing (GBS) to determine if recently identified new virulence phenotypes belong to established genotype groups or to groups previously not detected. A total of 158 isolates were phenotyped for virulence on 20 lines of Thatcher wheat that are isogenic for leaf rust resistance genes and also genotyped for single nucleotide polymorphism. Eight distinct groups of P. triticina genotypes from common wheat were described based on coancestry, nucleotide divergence, and principal coordinate plots. A separate genotype group had isolates with virulence to durum wheat. Isolates within groups had similar virulence phenotypes, and the overall population had high levels of heterozygosity and high levels of linkage disequilibria, which were all indicators of clonality. Two new genotype groups were described, thereby raising the possibility of new introductions of P. triticina; however, genotypes in these groups may have also originated from somatic nuclear exchange and recombination. A genome-wide association study detected 19 single nucleotide polymorphisms that were highly associated with virulence to 11 resistance genes in the Thatcher near-isogenic lines.

Extracts of the Native Brazilian Tree Garcinia gardneriana Inhibit Urediniospore Germination of Coffee Leaf Rust Fungus

The fungal Hemileia vastatrix is the causal agent of coffee leaf rust, one of the worst and devastating disease in coffee cultures worldwide. As a result of our research on natural products for the development of novel agrochemicals, we found that the hexane extract from leaves of the Brazilian medicinal plant Garcinia gardneriana, at 500 μg mL–1, inhibited in 98% the germination of H. vastatrix urediniospores. This extract showed no phytotoxicity when tested for seed germination and seedling growth inhibitory activity using sensible plant species. Also, the hexane extract from leaves was tested for anti-acetylcholinesterase activity, which constitutes a mechanism of action of major commercial insecticides used in agriculture, and showed low activity even at concentrations about two times higher than the half maximal inhibitory concentration (IC50) found in the antifungal assays. Gas chromatography-mass spectrometry (GC-MS) analysis showed that the hexane extract is constituted mainly by the pentacyclic triterpene lupeol, together with a series of sesquiterpenes as minor components. This is the first report on the investigation of antifungal, phytotoxic and acetylcholinesterase activities of extracts from leaves of G. gardneriana. These findings indicate that G. gardneriana may constitute a promising source of natural products for controlling the coffee leaf rust fungus.

Whole-genome sequencing of multiple isolates of Puccinia triticina reveals asexual lineages evolving by recurrent mutations.

The wheat leaf rust fungus, Puccinia triticina Erikss., is a worldwide pathogen of tetraploid durum and hexaploid wheat. Many races of P. triticina differ for virulence to specific leaf rust resistance genes and are found in most wheat-growing regions of the world. Wheat cultivars with effective leaf rust resistance exert selection pressure on P. triticina populations for virulent race types. The objectives of this study were to examine whole-genome sequence data of 121 P. triticina isolates and to gain insight into race evolution. The collection included isolates comprising of many different race phenotypes collected worldwide from common and durum wheat. One isolate from wild wheat relative Aegilops speltoides and two from Ae. cylindrica were also included for comparison. Based on 121,907 informative variants identified relative to the reference Race 1-1 genome, isolates were clustered into 11 major lineages with 100% bootstrap support. The isolates were also grouped based on variation in 1311 predicted secreted protein genes. In gene-coding regions, all groups had high ratios of nonsynonymous to synonymous mutations and nonsense to readthrough mutations. Grouping of isolates based on two main variation principle components for either genome-wide variation or variation just within the secreted protein genes, indicated similar groupings. Variants were distributed across the entire genome, not just within the secreted protein genes. Our results suggest that recurrent mutation and selection play a major role in differentiation within the clonal lineages.

Functional Analysis of Wheat NAC Transcription Factor, TaNAC069, in Regulating Resistance of Wheat to Leaf Rust Fungus.

NAC transcription factors are one of the largest transcription factor families having functions in a variety of stress responses. Few NACs have been reported for interactions between wheat and the wheat rust fungus Puccinia triticina (Pt). In this study, based on analysis of RNA-seq data from wheat line TcLr19 inoculated by Pt, the NAC transcription factor TaNAC069 was cloned from wheat, and its transcriptional activity and homologous dimer formation were verified. Quantitative real-time PCR analysis showed that the expression of TaNAC069 was induced by Pt and associated signaling molecules. To further characterize the function of the TaNAC069 gene in wheat resistance to Pt, virus-induced gene silencing (VIGS) was utilized, and it revealed that Pt resistance in TaNAC069-silenced plants was significantly reduced. Potential interaction targets of TaNAC069 from wheat and Pt were screened and identified by yeast two-hybrid technology. Eukaryotic elongation factor eEF1A, CBSX3 protein, and cold acclimation protein WCOR410c were screened by yeast one-hybrid technology. The results indicate that the TaNAC069 gene plays a positive regulatory role in wheat resistance to Pt, laying a good foundation to analyze the molecular mechanisms of TaNAC069 and its functional role in wheat resistance to Pt.

Taxonomy of Southeast Asian-Australasian grapevine leaf rust fungus and its close relatives

Rust fungi on commercial Vitis cultivars, wild Vitaceae, and Meliosma (Sabiaceae), potential alternate hosts of a grapevine leaf rust (GLR) fungus, were surveyed in Thailand and Vietnam. Molecular phylogenetic analyses using the internal transcribed spacer 2 and the large-subunit rRNA gene (D1/D2 region) and morphological examinations confirmed that the GLR fungus distributed in Southeast Asia and Australasia is distinct from Neophysopella species involved in GLRs in East Asia and the Americas. Therefore, the Southeast Asian-Australasian GLR fungus is recognized as a new Neophysopella species and named as N. tropicalis. Two fungi on Meliosma species in Thailand were aecial anamorph of Neophysopella species. One parasitic on M. simplicifolia and M. simplicifolia subsp. fordii was previously named as Aecidium wareoense, and a new name, N. wareoensis, is proposed for it. Another on M. arnottiana in Thailand was phylogenetically and morphologically distinct from all other Meliosma rust fungi, and, therefore, it is recognized as a new species, N. sriphumensis. A uredinial-telial fungus on Ampelocissus araneosa (Vitaceae) in Thailand was morphologically similar to Phakopsora cronartiiformis on Parthencissus semicordata (Vitaceae) in the Himalayas. However, the former fungus was distinct from the latter in producing characteristic urediniospores with labyrinthiform surface structure and cone-shaped projections. It is, therefore, recognized as a new species, N. doipuiensis.

Genome-Wide Analysis of the PIN Auxin Efflux Carrier Gene Family in Coffee.

Coffee is one of the most popular beverages around the world, which is mainly produced from the allopolyploid Coffea arabica. The genomes of C. arabica and its two ancestors C. canephora and C. eugenioides have been released due to the development of next generation sequencing. However, few studies on C. arabica are related to the PIN-FORMED (PIN) auxin efflux transporter despite its importance in auxin-mediated plant growth and development. In the present study, we conducted a genome-wide analysis of the PIN gene family in the three coffee species. Totals of 17, 9 and 10 of the PIN members were characterized in C. Arabica, C. canephora and C. eugenioides, respectively. Phylogenetic analysis revealed gene loss of PIN1 and PIN2 homologs in C. arabica, as well as gene duplication of PIN5 homologs during the fractionation process after tetraploidy. Furthermore, we conducted expression analysis of PIN genes in C. arabica by in silico and qRT-PCR. The results revealed the existence of gene expression dominance in allopolyploid coffee and illustrated several PIN candidates in regulating auxin transport and homeostasis under leaf rust fungus inoculation and the tissue-specific expression pattern of C. arabica. Together, this study provides the basis and guideline for future functional characterization of the PIN gene family.

TaTLP1 interacts with TaPR1 to contribute to wheat defense responses to leaf rust fungus.

Thaumatin-like proteins (TLPs), which are defined as pathogenesis-related protein family 5 (PR5) members, are common plant proteins involved in defense responses and confer antifungal activity against many plant pathogens. Our earlier studies have reported that the TaTLP1 gene was isolated from wheat and proved to be involved in wheat defense in response to leaf rust attack. The present study aims to identify the interacting proteins of TaTLP1 and characterize the role of the interaction between wheat and Puccinia triticina (Pt). Pull-down experiments designed to isolate the molecular target of TaTLP1 in tobacco resulted in the identification of TaPR1, a pathogenesis-related protein of family 1, and the interaction between TaTLP1 and TaPR1 was confirmed by yeast two-hybrid experiments (Y2H), bimolecular fluorescence complementation (BiFC), and co-immunoprecipitation (Co-IP). In vitro, TaTLP1 and TaPR1 together increased antifungal activity against Pt. In vivo, the disease resistance phenotype, histological observations of fungal growth and host responses, and accumulation of H2O2 in TaTLP1-TaPR1 in co-silenced plants indicated that co-silencing significantly enhanced wheat susceptibility compared to single knockdown TaTLP1 or TaPR1 plants. The accumulation of reactive oxygen species (ROS) was significantly reduced in co-silenced plants compared to controls during Pt infection, which suggested that the TaTLP1-TaPR1 interaction positively modulates wheat resistance to Pt in an ROS-dependent manner. Our findings provide new insights for understanding the roles of two different PRs, TaTLP1 and TaPR1, in wheat resistance to leaf rust.