In Arabidopsis, both pathogen invasion and benzothiadiazole (BTH) treatment activate the nonexpresser of pathogenesis-related genes 1 (NPR1)-mediated systemic acquired resistance, which provides broad-spectrum disease resistance to secondary pathogen infection. However, the BTH-induced resistance in Triticeae crops of wheat and barley seems to be accomplished through an NPR1-independent pathway. In the current investigation, we applied transcriptome analysis on barley transgenic lines overexpressing wheat wNPR1 (wNPR1-OE) and knocking down barley HvNPR1 (HvNPR1-Kd) to reveal the role of NPR1 during the BTH-induced resistance. Most of the previously designated barley chemical-induced (BCI) genes were upregulated in an NPR1-independent manner, whereas the expression levels of several pathogenesis-related (PR) genes were elevated upon BTH treatment only in wNPR1-OE. Two barley WRKY transcription factors, HvWRKY6 and HvWRKY70, were predicted and further validated as key regulators shared by the BTH-induced resistance and the NPR1-mediated acquired resistance. Wheat transgenic lines overexpressing HvWRKY6 and HvWRKY70 showed different degrees of enhanced resistance to Puccinia striiformis f. sp. tritici pathotype CYR32 and Blumeria graminis f. sp. tritici pathotype E20. In conclusion, the transcriptional changes of BTH-induced resistance in barley were initially profiled, and the identified key regulators would be valuable resources for the genetic improvement of broad-spectrum disease resistance in wheat. [ ] Copyright © 2020 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Systemic acquired resistance (SAR) in Arabidopsis is established beyond the initial pathogenic infection or is directly induced by treatment with salicylic acid or its functional analogs (SA/INA/BTH). NPR1 protein and WRKY transcription factors are considered the master regulators of SAR. Our previous study showed that NPR1 homologs in wheat (Triticum aestivum L.) and barley (Hordeum vulgare L.) regulated the expression of genes encoding pathogenesis-related (PR) proteins during acquired resistance (AR) triggered by Pseudomonas syringae pv. tomato DC3000. In the present examination, AR induced by P. syringae DC3000 was also found to effectively improve wheat resistance to Puccinia triticina (Pt). However, with more complex genomes, genes associated with this SAR-like response in wheat and barley are largely unknown and no specific WRKYs has been reported to be involved in this biological process. In our subsequent analysis, barley transgenic line overexpressing wheat wNPR1 (wNPR1-OE) showed enhanced resistance to Magnaporthe oryzae isolate Guy11, whereas AR to Guy11 was suppressed in a barley transgenic line with knocked-down barley HvNPR1 (HvNPR1-Kd). We performed RNA-seq to reveal the genes that were differentially expressed among these transgenic lines and the wild-type barley plants during the AR. Several PR and BTH-induced (BCI) genes were designated as downstream genes of NPR1. The expression of few WRKYs was significantly associated with NPR1 expression during the AR events. The transient expression of three WRKY genes, including HvWRKY6, HvWRKY40, and HvWRKY70, in wheat leaves by Agrobacterium-mediated infiltration enhanced the resistance to Pt. In conclusion, a profile of genes associated with NPR1-mediated AR in barley was drafted and WRKYs discovered in the current study showed a substantial potential for improving wheat resistance to Pt.

Wheat leaf rust, caused by Puccinia triticina, is an important fungal disease of wheat in China. To study races of the pathogen in China, leaf rust samples were collected from 14 provinces in 2014 and 15 provinces in 2015. From the samples, 494 single-uredinial isolates were derived from the 2014 collection and 649 from the 2015 collection. These isolates were tested on 40 near-isogenic lines of Thatcher carrying single leaf rust resistance genes. From the isolates, 84 races were identified in 2014 and 65 races in 2015. Races THTT (22.1%), THTS (19.6%), THJT (8.7%), PHTT (4.9%), and PHJT (3.6%) were the most common races in 2014, and THTT (28.4%), THTS (12.8%), THJT (11.6%), THJS (9.9%), and PHTT (9.7%) were the most frequent in 2015. All of these races were avirulent to resistance genes Lr9 and Lr24. THTT and THTS, the most frequent races in both years, were widely distributed throughout the country. The frequencies of isolates with virulence to Lr1, Lr2a, Lr2c, Lr3, Lr16, Lr26, Lr11, Lr17, LrB, Lr10, Lr14a, Lr2b, Lr3bg, Lr14b, Lr32, Lr33, and Lr50 were over 80%, whereas the frequencies of virulence to Lr9, Lr19, Lr25, Lr28, Lr29, and Lr47 were less than 3.5%. In the present study, all isolates were avirulent to Lr24 and Lr38. The race analysis and individual virulence frequencies provide guidance to breeders in choosing leaf rust resistance genes for use in breeding programs.

Wheat leaf rust caused by Puccinia triticina is one of the most common and serious diseases in wheat production. The constantly changing pathogens overcome the plant resistance to P. triticina. Plant pathogens secrete effector proteins that alter the structure of the host cell, interfere plant defenses, or modify the physiology of plant cells. Therefore, the identification of effector proteins is critical to reveal the pathogenic mechanism. We used SignalP v4.1, TargetP v1.1, TMHMM v2.0, and EffectorP v2.0 to screen the candidate effector proteins in P. triticina isolates – KHTT, JHKT, and THSN. As a result, a total of 635 candidate effector proteins were obtained. Structural analysis showed that effector proteins were small in size (50AA to 422AA) and of diverse sequences, and the conserved sequential elements or clear common elements were not involved, regardless of their secretion from the pathogen to the host. There were 427 candidate effector proteins that contain more than or equal to 4 cysteine residues, and 339 candidate effector proteins contained the known motifs. Sixteen families, 9 domains, and 53 other known functional types were found in 186 candidate effector proteins using the Pfam search. Three novel motifs were found by MEME. Heterogeneous expression system was performed to verify the functions of 30 candidate effectors by inhibiting the programmed cell death (PCD) induced by BAX (the mouse-apoptotic gene elicitor) on Nicotiana benthamiana. Hypersensitive response (HR) can be induced by the six effectors in the wheat leaf rust resistance near isogenic lines, and this would be shown by the method of transient expression through Agrobacterium tumefaciens infiltration. The quantitative reverse transcription PCR (qRT-PCR) analysis of 14 candidate effector proteins secreted after P. triticina inoculation showed that the tested effectors displayed different expression patterns in different stages, suggesting that they may be involved in the wheat–P. triticina interaction. The results showed that the prediction of P. triticina effector proteins based on transcriptomic analysis and multiple bioinformatics software is effective and more accurate, laying the foundation of revealing the pathogenic mechanism of Pt and controlling disease.

Puccinia triticina (Pt), the causal agent of wheat leaf rust, is one of the most destructive fungal pathogens threatening global wheat cultivations. The rational utilization of leaf rust resistance (Lr) genes is still the most efficient method for the control of such diseases. The Lr47 gene introgressed from chromosome 7S of Aegilops speltoides still showed high resistance to the majority of Pt races collected in China. However, the Lr47 gene has not been cloned yet, and the regulatory network of the Lr47-mediated resistance has not been explored. In the present investigation, transcriptome analysis was applied on RNA samples from three different wheat lines (“Yecora Rojo”, “UC1037”, and “White Yecora”) carrying the Lr47 gene three days post-inoculation with the epidemic Pt race THTT. A comparison between Pt-inoculated and water-inoculated “Lr47-Yecora Rojo” lines revealed a total number of 863 upregulated (q-value < 0.05 and log2foldchange > 1) and 418 downregulated (q-value < 0.05 and log2foldchange < −1) genes. Specifically, differentially expressed genes (DEGs) located on chromosomes 7AS, 7BS, and 7DS were identified, ten of which encoded receptor-like kinases (RLKs). The expression patterns of these RLK genes were further determined by a time-scale qRT-PCR assay. Moreover, heatmaps for the expression profiles of pathogenesis-related (PR) genes and several transcription factor gene families were generated. Using a transcriptomic approach, we initially profiled the transcriptional changes associated with the Lr47-mediated resistance. The identified DEGs, particularly those genes encoding RLKs, might serve as valuable genetic resources for the improvement of wheat resistance to Pt.

Wheat leaf rust, caused by Puccinia triticina, is a common fungal disease of wheat in China. In order to identify races and determine the individual virulence of isolates in different wheat-growing regions in China, leaf rust samples collected from 18 provinces in 2011 to 2013 were tested on 37 Thatcher near-isogenic lines each carrying a different single leaf rust resistance gene. A total of 158 races were identified. Races THTT (19.5%), THTS (16.9%), PHTT (7.7%), THJS (5.0%), THJT (4.2%), and PHTS (4.0%) were the most predominant races in 2011 to 2013. All of these races were avirulent to resistance genes Lr9 and Lr24. The two most frequent races, THTT and THTS, were widely distributed. The frequencies of the isolates with virulence to Lr1, Lr2c, Lr3, Lr16, Lr26, Lr17, LrB, Lr10, Lr14a, Lr3bg, Lr14b, Lr33, Lr37, and Lr50 exceeded 90%. Frequencies of virulence to Lr2a, Lr3ka, Lr11, Lr30, Lr2b, and Lr32 exceeded 70% but were less than 90%. Frequencies of virulence to Lr18, Lr21, Lr15, Lr23, Lr33+34, Lr36, Lr39, and Lr44 were below 70%, whereas the frequency of virulence to Lr25 was less than 1%. All isolates were avirulent to Lr9, Lr19, Lr24, Lr28, Lr42, Lr29, Lr38, and Lr47. The identified races and individual virulence frequencies provide a basis for selection of effective leaf rust resistance genes for use in breeding programs and can also provide information for the study of race evolution of P. triticina.