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Worldwide food security is severely threatened by the devastating wheat stripe rust disease. The utilization of resistant wheat cultivars represents the most cost‐effective and efficient strategy for combating this disease. However, the lack of resistant resources has been a major bottleneck in breeding for wheat disease resistance. Therefore, revealing novel gene resources for combating stripe rust and elucidating the underlying resistance mechanism is of utmost urgency. In this study, we identified that the soybean NF‐YB transcription factor GmNF‐YB20 in wheat provides resistance to the stripe rust fungus (Puccinia striiformis f. sp. tritici, Pst). Wheat lines with stable overexpression of the GmNF‐YB20 enhanced resistance against multiple Pst races. Transcriptome profiling of GmNF‐YB20 transgenic wheat under Pst infection unveiled its involvement in the lipid signaling pathway. RT‐qPCR assays suggested that GmNF‐YB20 increased transcript levels of multiple nonspecific lipid transfer protein (LTP) genes during wheat‐Pst interaction, luciferase reporter analysis illustrates that it activates the transcription of TaLTP1.50 in wheat protoplast, and GmNF‐YB20 overexpressed wheat plants had higher total LTP content in vivo during Pst infection. Overexpression of TaLTP1.50 in wheat significantly increased resistance to Pst, whereas knockdown of TaLTP1.50 exhibited the opposite trends, indicating that TaLTP1.50 plays a positive role in wheat resistance. Taken together, our findings provide perspective regarding the molecular mechanism of GmNF‐YB20 in wheat and highlight the potential use for wheat breeding. Summary statement: The soybean transcription factor GmNF‐YB20 confers resistance to stripe rust in transgenic wheat by regulating nonspecific lipid transfer protein genes. Our findings provide perspective regarding the molecular mechanism of GmNF‐YB20 in wheat and highlight the potential use for wheat breeding. [ABSTRACT FROM AUTHOR]

Ca2+ plays a crucial role as a secondary messenger in plant development and response to abiotic/biotic stressors. Calcium-dependent protein kinases (CDPKs/CPKs) are essential Ca2+ sensors that can convert Ca2+ signals into downstream phosphorylation signals. However, there is limited research on the function of CDPKs in the context of wheat–Puccinia striiformis f. sp. tritici (Pst) interaction. In this study, we aimed to address this gap by identifying putative CDPK genes from the wheat reference genome and organizing them into four phylogenetic clusters (I-IV). To investigate the expression patterns of the TaCDPK family during the wheat–Pst interaction, we analyzed time series RNA-seq data and further validated the results through qRT-PCR assays. Among the TaCDPK genes, TaCDPK7 exhibited a significant induction during the wheat–Pst interaction, suggesting that it has a potential role in wheat resistance to Pst. To gain further insights into the function of TaCDPK7, we employed virus-induced gene silencing (VIGS) to knock down its expression which resulted in impaired wheat resistance to Pst, accompanied by decreased accumulation of hydrogen peroxide (H2O2), increased fungal biomass ratio, reduced expression of defense-related genes, and enhanced pathogen hyphal growth. These findings collectively suggest that TaCDPK7 plays an important role in wheat resistance to Pst. In summary, this study expands our understanding of wheat CDPKs and provides novel insights into their involvement in the wheat–Pst interaction. [ABSTRACT FROM AUTHOR]

Plant 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. [ABSTRACT FROM AUTHOR]

DREB transcription factor can significantly improve the plant resistance to drought, high salt, low temperature, high temperature, and other abiotic stress. A previous study showed that DREB transcription factor gene GmDREB3 from soybean can improve drought resistance in tobacco, however, whether it can improve drought resistance in wheat needs to be further investigated. In the present study, GmDREB3 was transferred to a wheat variety, Shi366 and two positive lines of the T5 generation were obtained by additive culture and molecular identification. GmDREB3‐OE (overexpression) lines conferred drought tolerance characteristics, including higher survival rates, lower malondialdehyde (MDA) content, lower relative conductivity, higher chlorophyll content, and higher proline content than Shi366 wheat plants under drought stress. The SPAD, Fv/Fm, panicle number, and grain yield of GmDREB3‐OE lines were increased but the transpiration rate was decreased, compared to the Shi366 plants in the field condition under drought stress. Significant differences in the stress indices were observed between the GmDREB3–OE lines and Shi366 wheat plants, which suggested that the GmDREB3 significantly improved wheat tolerance to drought stress. In addition, RT‐qPCR analysis indicated that GmDREB3 could enhance the expression of drought‐related genes in wheat. Therefore, GmDREB3 significantly increased drought tolerance in wheat plants along with improved physiological and agronomic traits and grain yield. [ABSTRACT FROM AUTHOR]

The constitutive photomorphogenesis 9 (COP9) signalosome (CSN) is a versatile regulator of plant growth, development, and response to diverse pathogens. However, little research has been done to understand the function of those CSN genes in broad‐spectrum resistance to pathogens. In this study, we found that the transcript levels of wheat TaCSN5 were induced in response to inoculation with Puccinia striiformis f. sp. tritici (Pst) and treatment with salicylic acid (SA). Overexpression of TaCSN5 in Arabidopsis resulted in increased susceptibility to Pseudomonas syringae pv. tomato DC3000 infection accompanied by down‐regulation of AtPR1 expression. Overexpression of TaCSN5 in wheat lines significantly increased susceptibility to Pst accompanied by decreased SA accumulation, whereas TaCSN5‐RNAi wheat lines exhibited opposite trends. Moreover, we found that TaCSN5 negatively regulated TaG3NPR1 genes involved in the SA signalling pathway. In addition, TaCSN5‐RNAi lines showed increased resistance to multiple races of Pst. Taken together, we demonstrate that TaCSN5 contributes to negative regulation of wheat resistance to Pst in an SA‐dependent manner. [ABSTRACT FROM AUTHOR]

A biotrophic fungus, Puccinia striiformis f.sp. tritici (Pst), which causes stripe rust disease in wheat is the most yield-limiting factor in wheat production. Plants have complex defense mechanisms against invading pathogens. Hypersensitive response (HR), a kind of programmed cell death (PCD) at the infection site, is among these defense mechanisms. Transcription factors (TFs) play a crucial role in plant defense response against invading pathogens. Myeloblastosis (MYB) TFs are among the largest TFs families that are involved in response to both biotic and abiotic stresses. However, little is known about the mechanisms of MYB TFs during the interaction between wheat and the stripe rust fungus. Here, we identified an R2R3 MYB TF from wheat, designated as TaMYB391, and characterized its functional role during wheat–Pst interaction. Our data indicated that TaMYB391 is induced by Pst infection and exogenous application of salicylic acid (SA) and abscisic acid (ABA). TaMYB391 is localized in the nucleus of both wheat and Nicotiana benthamiana. Transient overexpression of TaMYB391 in N. benthamiana triggered HR-related PCD accompanied by increased electrolyte leakage, high accumulation of reactive oxygen species (ROS), and transcriptional accumulation of SA defense-related genes and HR-specific marker genes. Overexpression of TaMYB391 in wheat significantly enhanced wheat resistance to stripe rust fungus through the induction of pathogenesis-related (PR) genes, ROS accumulation and hypersensitive cell death. On the other hand, RNAi-mediated silencing of TaMYB391 decreased the resistance of wheat to Pst accompanied by enhanced growth of the pathogen. Together our findings demonstrate that TaMYB391 acts as a positive regulator of HR-associated cell death and positively contributes to the resistance of wheat to the stripe rust fungus by regulating certain PR genes, possibly through SA signaling pathways. [ABSTRACT FROM AUTHOR]

SUMMARY: Puccinia striiformis f. sp. tritici (Pst), the causal agent of stripe rust, is a destructive pathogen of Triticum aestivum (wheat), threatening wheat production worldwide. Pst delivers hundreds of effectors to manipulate processes in its hosts during infection. The SGT1 (suppressor of the G2 allele of skp1), RAR1 (required for Mla12 resistance) and HSP90 (heat‐shock protein 90) proteins form a chaperone complex that acts as a core modulator in plant immunity. However, little is known about how Pst effectors target this immune component to suppress plant immunity. Here, we identified a Pst effector PstSIE1 that interacts with TaSGT1 in wheat and is upregulated during the early infection stage. Transient expression of PstSIE1 suppressed cell death in Nicotiana benthamiana induced by VmE02 and PcNLP2. Transgenic expression of PstSIE1‐RNAi constructs in wheat significantly reduced the virulence of Pst. Overexpression of PstSIE1 in wheat increased the number of rust pustules and reduced the accumulation of reactive oxygen species (ROS), indicating that PstSIE1 functions as an important pathogenicity factor in Pst. PstSIE1 was found to compete with TaRAR1 to bind TaSGT1, thus disrupting the formation of the TaRAR1–TaSGT1 subcomplex. Taken together, PstSIE1 is an important Pst effector targeting the immune component TaSGT1 and involved in suppressing wheat defense. Significance Statement: PstSIE1 is an important Pst effector targeting the immune component TaSGT1 and involved in suppressing wheat defense. This work broadens our understanding of how the molecular mechanisms of fungal effectors suppress plant immunity. [ABSTRACT FROM AUTHOR]

Puccinia striiformis f. sp. tritici (Pst) secretes an array of specific effector proteins to manipulate host immunity and promote pathogen colonization. In a previous study, we functionally characterized a glycine-serine-rich effector PstGSRE1 with a glycine-serine-rich motif (m9). However, the mechanisms of glycine-serine-rich effectors (GSREs) remain obscure. Here we report a new glycine-serine-rich effector, PstGSRE4, which has no m9-like motif but inhibits the enzyme activity of wheat copper zinc superoxide dismutase TaCZSOD2, which acts as a positive regulator of wheat resistance to Pst. By inhibiting the enzyme activity of TaCZSOD2, PstGSRE4 reduces H2O2 accumulation and HR areas to facilitate Pst infection. These findings provide new insights into the molecular mechanisms of GSREs of rust fungi in regulating plant immunity. Author summary: Pst secretes numerous effectors to modulate host defense systems. However, the mechanisms of these effectors, especially for glycine-rich or serine-rich effectors, remain obscure. In this study, we identified a new glycine-serine-rich effector, PstGSRE4, which exhibits unusual biochemical properties and is highly induced during early stages of infection. Transgenic expression of PstGSRE4-RNAi constructs in wheat significantly reduced virulence of Pst and increased H2O2 accumulation in wheat. Overexpression of PstGSRE4 in wheat significantly increased virulence of Pst and reduced H2O2 accumulation in wheat. PstGSRE4 was shown to target the ROS-associated regulatory factor TaCZSOD2, which was proved as a positive regulator of wheat immunity in this study. Further study revealed that PstGSRE4 inhibited the enzyme activity of TaCZSOD2 and thus compromises the host immune systems. This work reveals a novel strategy that rust fungi exploit to modulate host defense and facilitate pathogen infection. [ABSTRACT FROM AUTHOR]

AP2 transcription factors play a crucial role in plant development and reproductive growth, as well as response to biotic and abiotic stress. However, the role of TaAP2-15, in the interaction between wheat and the stripe fungus, Puccinia striiformis f. sp. tritici (Pst), remains elusive. In this study, we isolated TaAP2-15 and characterized its function during the interaction. TaAP2-15 was localized in the nucleus of wheat and N. benthamiana. Silencing of TaAP2-15 by barley stripe mosaic virus (BSMV)-mediated VIGS (virus-induced gene silencing) increased the susceptibility of wheat to Pst accompanied by enhanced growth of the pathogen (number of haustoria, haustorial mother cells and hyphal length). We confirmed by quantitative real-time PCR that the transcript levels of pathogenesis-related genes (TaPR1 and TaPR2) were down-regulated, while reactive oxygen species (ROS)-scavenging genes (TaCAT3 and TaFSOD3D) were induced accompanied by reduced accumulation of H2O2. Furthermore, we found that TaAP2-15 interacted with a zinc finger protein (TaRZFP34) that is a homolog of OsRZFP34 in rice. Together our findings demonstrate that TaAP2-15 is positively involved in resistance of wheat to the stripe rust fungus and provides new insights into the roles of AP2 in the host-pathogen interaction. [ABSTRACT FROM AUTHOR]

Yellow stripe-like (YSL) transporters are required for the transportation of metal-phytosiderophores and are structurally related to metal-nicotianamine complexes. Some studies also reported the involvement of YSL transporters in pathogen-induced defense. However, the molecular mechanisms of YSL genes involved in biotic stress responses are still not clear, especially in cereal crops. This study aimed to functionally characterize TaYS1A during the interaction of wheat and Puccinia striiformis f. sp. tritici (Pst), the causal agent of stripe rust disease. TaYS1A was localized in the cell membrane of wheat protoplasts and Nicotiana benthamiana cells. TaYS1A was significantly up-regulated in wheat leaves after being infected with the avirulent Pst isolate CYR23 and after treatment with salicylic acid (SA). Silencing of TaYS1A by the virus-induced gene silencing method enhanced the susceptibility of wheat to Pst accompanied by reducing the accumulation of SA and H2O2 and down-regulating the transcriptions of TaPR1 and TaPR2. In addition, TaYS1A was found to interact with TaNH2, a homolog of OsNH2, by yeast-two-hybrid assay, and silencing of TaYS1A diminished the expression of TaNH2. Our findings suggested the existence of positive regulation of TaYS1A in providing resistance against Pst by modulating SA-induced signaling and offered new insight into the biological role of YSL in wheat against pathogens. [ABSTRACT FROM AUTHOR]