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Summary: The biotrophic fungal pathogen Blumeria graminis f. sp. tritici (Bgt) is a crucial factor causing reduction in global wheat production. Wild wheat relatives, for example Thinopyrum intermedium, is one of the wild‐used parents in wheat disease‐resistant breeding. From T. intermedium line, we identified the aspartic protease gene, TiAP1, which is involved in resistance against Bgt. TiAP1 is a secreted protein that accumulates in large amounts at the infection sites of Bgt and extends to the intercellular space. Yeast two‐hybrid, luciferase complementation imaging and bimolecular florescent complimentary analysis showed that TiAP1 interacted with the chitin deacetylase (BgtCDA1) of Bgt. The yeast expression, purification and in vitro test confirmed the chitin deacetylase activity of BgtCDA1. The bombardment and VIGS‐mediated host‐induced gene silencing showed that BgtCDA1 promotes the invasion of Bgt. Transcriptome analysis showed the cell wall xylan metabolism, lignin biosynthesis‐related and defence genes involved in the signal transduction were up‐regulated in the transgenic TiAP1 wheat induced by Bgt. The TiAP1 in wheat may inactivate the deacetylation function of BgtCDA1, cause chitin oligomers expose to wheat chitin receptor, then trigger the wheat immune response to inhibit the growth and penetration of Bgt, and thereby enhance the resistance of wheat to pathogens. [ABSTRACT FROM AUTHOR]

Germin and germin-like proteins (GLPs) play multifaceted roles in plants and participate in signaling processes associated with host–pathogen interactions and response to abiotic stress signaling. Despite the important roles in plant survival and response to the environment, few reports exist about the activity and function of the GLP family in common wheat (Triticum aestivum L.). In this study, we performed a genome-wide identification of the GLP family in wheat and identified 258 GLP genes. Through phylogenetic analysis, the GLPs of Arabidopsis, rice, and wheat were divided into six groups, and each group had a similar genetic structure and a conserved motif. Chromosome 4B has the largest cluster region of TaGLPs among all chromosomes. The cis-regulatory elements in the promoter of TaGLPs, which are critical for the function of wheat GLPs and related to stress and development, are detected. The expression profiles of wheat GLPs under Blumeria graminis f. sp. tritici (Bgt) invasion were investigated using the RNA sequencing analysis and validated using quantitative real-time polymerase chain reaction (qRT-PCR) method. Most of the Bgt resistance-related TaGLP genes are located on the fourth homologous group chromosomes, indicating that pathogens resistance-related TaGLP genes repeat in large tandem in specific chromosomal regions during evolution. These results will help us understand the function of the GLP family in wheat and explore the mechanism of wheat GLPs genes in biotic resistance. [ABSTRACT FROM AUTHOR]

Key message: Present study revealed that specific expression of TaYUC10.3 in wheat young seeds could increase the content of auxin, and protein. Auxin is a vital endogenous hormone in plants, which is involved in the regulation of various physiological and biochemical processes in plants. The flavin-containing monooxygenase encoded by the YUCCA gene is a rate-limiting enzyme in the tryptophan-dependent pathway of auxin synthesis. TaYUC10.3 was identified, cloned and found that it was abundantly expressed in wheat young seeds. In this study, a seed-specific expression vector of TaYUC10.3 was constructed with the promoter of 1Bx17 glutenin subunit gene and transformed wheat using the particle bombardment method. The quantitative RT-PCR showed that TaYUC10.3 was expressed in a large amount in young seeds of the transgenic lines. Plant hormone-targeted metabolomics showed that the auxin content of the transgenic lines was significantly increased compared with controls. The GC / MS non-targeted metabolite multiple statistical analyses showed that the variable importance in projection (VIP) of tryptophan reduced in the transgenic lines. Simultaneously, the VIP of indole acetic acid increased. The precursor amino acids for synthesizing some proteins and carbohydrates were upregulated in the transgenic lines. Subsequently, it was found that the protein content of the seeds of the transgenic TaYUC10.3 wheat was significantly higher than that of the control. The wet gluten content and sedimentation value of the transgenic TaYUC10.3 wheat were also high. This result indicated that TaYUC10.3 might participate in auxin synthesis and affects the protein content of wheat seeds. [ABSTRACT FROM AUTHOR]

Auxin is an important endogenous hormone in plants. The YUCCA gene encodes a flavin monooxygenase, which is an important rate-limiting enzyme in the auxin synthesis pathway and involved in the regulation of plant growth and development. In the study, we identified 63 wheat TaYUCCA genes; among them, some genes appeared in clusters. By constructing phylogenetic trees, we found that the TaYUCCA genes could be divided into six groups. In the WheatExp database, there were 22 differential expressed TaYUCCA genes, among which the TaYUCCA10 gene was abundantly expressed in the endosperm and medium milk stage, the TaYUCCA2 gene was abundantly expressed in the roots of three leaves and meiosis and transfer cells at 20 days post anthesis and the others 16 TaYUCCA genes had different expression level at different developmental stages in wheat, and there were 15 TaYUCCA genes induced by drought and heat stress, among which the TaYUCCA2-D, TaYUCCA3-B, and TaYUCCA9-D might be upregulated induced by drought stress, TaYUCCA10.1 might be upregulated induced drought and heat stress, TaYUCCA6-A was upregulated induced both drought and heat stress and the others 9 TaYUCCA genes were downregulated induced by drought and heat stress. Transcriptome and qRT-PCR analysis showed that TaYUCCA7-A was upregulated significantly after induced by powdery mildew. The comprehensive annotation and expression profiling of the TaYUCCA genes in this study enhanced our understanding of TaYUCCA family gene expression in wheat growth and development and laid the foundation for the further study of TaYUCCA gene mechanism. [ABSTRACT FROM AUTHOR]

Aspartic proteases (APs) are one of the four main protease super families. In plants, they are involved in many biological processes, such as biotic and abiotic stress resistance, protein processing and degradation, senescence, and programmed cell death. By performing a database (TGACv1) search and domain prediction, we identified 263 wheat AP (TaAP) proteins and observed 38 TaAP genes exhibiting alternative splicing. Moreover, by constructing a phylogenetic tree, we found that the TaAP proteins can be divided into three families and have a certain close evolutionary relationship to Arabidopsis thaliana and rice AP proteins. Transcriptome analysis showed that 29 genes in the TaAP family were up-regulated after being induced by powdery mildew. The expression of TaAP224 showed the most significant difference in transcriptome and qRT-PCR analyses. Subsequently, the promoters of these 29 genes were analysed, and we found that they contained multiple disease resistance and hormone elements, such as WRKY71OS, a common disease resistance element that is also involved in the GA signalling pathway and inhibits starch hydrolysis. The comprehensive annotation and expression profiling performed in this study increased our understanding of the TaAP family genes in wheat growth and development, and the results can be used as a basis for further study of candidate TaAP genes involved in powdery mildew resistance mechanisms. [ABSTRACT FROM AUTHOR]

Key message: Expression of TaRUB1 gene in Arabidopsis thaliana elevates the level of disease-related genes in response to pathogen invasion through the accumulation of callose, necrotic cells, and the outbreak of ROS. Abstract: Ubiquitin (Ub) and ubiquitin-like proteins are highly conserved in sequence and can covalently bind and modify many intracellular proteins which can be recognized and degraded by 26S proteasome. Post-translational modification of proteins has become a hot research spot today. In the previous study, a cDNA of related-to-ubiquitin protein belonged to ubiquitin-like proteins, whose spatial structure comprised Ub and NEDD8, was obtained from wheat SN6306 by suppression-subtractive hybridization and was named TaRUB1. TaRUB1 is induced by wheat powdery mildew and significantly upregulated in resistant wheat SN6306. In this study, heterologous expression of TaRUB1 in A. thaliana was used to study the function of this gene in response to pathogen Pseudomonas syringae pv. Tomato DC3000 ( Pst DC3000). Transgenic A. thaliana showed relatively fewer disease symptoms, accompanied by common inhibition of living body parasitic defense responses, accumulation of more callose and reactive oxygen species (ROS), and concentrated cell death, simultaneously antioxidant enzyme activities of superoxide dismutase, peroxidase, catalase, and ascorbate peroxidase were higher than those in wild-type (WT) plant after infection with Pst DC3000. Meanwhile, hypersensitive cell death, which was possibly ROS burst, was also observed in transgenic A. thaliana. By quantitative reverse transcription-polymerase chain reaction analysis, some marker genes for hypersensitive response showed significantly higher transcriptional expression level in transgenic A. thaliana, which activates system-acquired resistance, than that of WT plants. Heterologous expression of TaRUB1 can significantly enhance resistance to Pst DC3000 in A. thaliana, suggesting that TaRUB1 is related to plant disease resistance. [ABSTRACT FROM AUTHOR]

Wheat powdery mildew, caused by the fungal pathogen Blumeria graminis f. sp. tritici ( Bgt), is a serious wheat disease both in China and worldwide. Finding and cloning the key resistance genes to Bgt are important for wheat breeding. In this study, 23 differentially expressed genes were isolated by GeneFishing from the hybrid wheat alien disomic addition line germplasm SN6306, which is highly resistant to powdery mildew. SN6306 originated from the intergeneric hybridization between wheat Yannong 15 and Elytrigia intermedium (Host) Nevski. Among the 23 genes, a gene encoding for auxin-repressed protein ( TaARP1) was selected for further study, and its full-length complementary DNA (cDNA) and DNA sequences were obtained. The function of TaARP1 was investigated by real-time quantitative reverse transcription PCR and Barley Stripe Mosaic Virus-mediated gene silencing. Results showed that, after inoculation with Bgt, the expression of TaARP1 was upregulated more than ten times compared with non-inoculated controls. Ten days after inoculation with powdery mildew, BSMV: TaARP1 leaves clearly showed higher susceptibility to mildew infections than BSMV:00 leaves. Powdery mildew colonies and mycelia development were analyzed by microscopic observation. These data suggested that TaARP1 may be involved in the response to Bgt and may be a potential resistance gene to wheat powdery mildew. [ABSTRACT FROM AUTHOR]

Protein modification by ubiquitin (Ub) and ubiquitin-like proteins (Ubls) is among the most complex and heavily investigated mechanisms of posttranslational protein regulation in eukaryotes. Moreover, Ub plays diverse roles in the regulation of numerous biological pathways. In this study, a suppression-subtractive hybridization cDNA library has been constructed using RNA isolated from leaves of the wheat line SN6306 inoculated with Blumeria graminis as the tester and RNA from non-inoculated leaves as the driver. Eighteen expressed sequence tags (ESTs) highly homologous with known proteins involved in disease resistance and defense reactions have been identified. Among these, a 648 bp EST with a high degree of sequence similarity in both nucleic and amino acid levels to other members of the gene family of plant related-to-ubiquitin (RUB) proteins has been selected, and its genomic sequence obtained. The cDNA sequence of this EST is interrupted by two introns at 111 and 1450 bp, with an open reading frame of 462 bp that encodes 153 amino acids. The predicated protein is a fusion protein of an ubiquitin-coding region and an RUB region. The newly screened gene, designated as TaRUB1 (GenBank Accession No. JF927719) belongs to the family of wheat RUB genes. Quantitative real-time polymerase chain reaction revealed that TaRUB1 is upregulated in leaves infected with powdery mildew during early stages of infection. Furthermore, the overexpression of TaRUB1 in transgenic Arabidopsis showed that TaRUB1 enhances plant tolerance to both osmotic and salt stresses. This finding suggests that TaRUB1 has multiple functions in plant abiotic and biotic stress tolerance. [ABSTRACT FROM AUTHOR]

Grain protein content in wheat ( Triticum aestivum L.) is generally considered a highly heritable character that is negatively correlated with grain yield and yield-related traits. Quantitative trait loci (QTL) for protein content was mapped using data on protein content and protein content conditioned on the putatively interrelated traits to evaluate possible genetic interrelationships between protein content and yield, as well as yield-related traits. Phenotypic data were evaluated in a recombinant inbred line population with 302 lines derived from a cross between the Chinese cultivar Weimai 8 and Luohan 2. Inclusive composite interval mapping using IciMapping 3.0 was employed for mapping unconditional and conditional QTL with additives. A strong genetic relationship was found between protein content and grain yield, and yield-related traits. Unconditional QTL mapping analysis detected seven additive QTL for protein content, with additive effects ranging in absolute size from 0.1898% to 0.3407% protein content, jointly accounting for 43.45% of the trait variance. Conditional QTL mapping analysis indicated two QTL independent from yield, which can be used in marker-assisted selection for increasing yield without affecting grain protein content. Three additional QTL with minor effects were identified in the conditional mapping. Of the three QTLs, two were identified when protein content was conditioned on yield, which had pleiotropic effects on those two traits. Conditional QTL mapping can be used to dissect the genetic interrelationship between two traits at the individual QTL level for closely correlated traits. Further, conditional QTL mapping can reveal additional QTL with minor effects that are undetectable in unconditional mapping. [ABSTRACT FROM AUTHOR]

To identify differential proteins induced by N-(2-chloro-4-pyridy)-N′-phenylurea or forchlorfenuron (CPPU), two-dimensional electrophoresis was used to study the variations in protein expression of wheat seedlings induced with CPPU. The expressed protein spots were identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry and protein data bank query. About 800 spots were reproducibly detected in three replicates of each sample. A total of 10 differential proteins were identified, including tRNA delta (2)-isopentenylpyrophosphate transferase (S1), probable peptide ABC transporter ATP-binding protein y4tS (S2), ribosomal RNA large subunit methyltransferase N (S3), microtubule-associated protein 6 (S4), actin-66 (Fragment) (S5), mitochondrial ATP synthase subunit alpha (S6), S-adenosylmethionine synthase 1 (S7), flavin-containing monooxygenase YUCCA4 (S8), GTPase obg (S9), and ribosomal RNA small subunit methyltransferase G (S10). These proteins were closely related to phytohormone synthesis, plant stress response, energy metabolism, protein synthesis, cytoskeleton, and transport. All of these processes may play important roles in wheat seedling development. These findings could provide valuable information for the further study of the functions and underlying mechanism of the action of CPPU. Thus, proteomic analysis revealed that CPPU treatment induces variations in many proteins involved in plant growth and development. [ABSTRACT FROM AUTHOR]

Kernel dimensions (KD) contribute greatly to thousand-kernel weight (TKW) in wheat. In the present study, quantitative trait loci (QTL) for TKW, kernel length (KL), kernel width (KW) and kernel diameter ratio (KDR) were detected by both conditional and unconditional QTL mapping methods. Two related F8:9 recombinant inbred line (RIL) populations, comprising 485 and 229 lines, respectively, were used in this study, and the trait phenotypes were evaluated in four environments. Unconditional QTL mapping analysis detected 77 additive QTL for four traits in two populations. Of these, 24 QTL were verified in at least three trials, and five of them were major QTL, thus being of great value for marker assisted selection in breeding programmes. Conditional QTL mapping analysis, compared with unconditional QTL mapping analysis, resulted in reduction in the number of QTL for TKW due to the elimination of TKW variations caused by its conditional traits; based on which we first dissected genetic control system involved in the synthetic process between TKW and KD at an individual QTL level. Results indicated that, at the QTL level, KW had the strongest influence on TKW, followed by KL, and KDR had the lowest level contribution to TKW. In addition, the present study proved that it is not all-inclusive to determine genetic relationships of a pairwise QTL for two related/causal traits based on whether they were co-located. Thus, conditional QTL mapping method should be used to evaluate possible genetic relationships of two related/causal traits. [ABSTRACT FROM AUTHOR]

Plant height (PH) in wheat is a complex trait; its components include spike length (SL) and internode lengths. To precisely analyze the factors affecting PH, two F recombinant inbred line (RIL) populations comprising 485 and 229 lines were generated. Crosses were performed between Weimai 8 and Jimai 20 (WJ) and between Weimai 8 and Yannong 19 (WY). Possible genetic relationships between PH and PH components (PHC) were evaluated at the quantitative trait locus (QTL) level. PH and PHC (including SL and internode lengths from the first to the fourth counted from the top, abbreviated as FIITL, SITL, TITL, and FOITL, respectively) were measured in four environments. Individual and the pooled values from four trials were used in the present analysis. A QTL for PH was mapped using data on PH and on PH conditioned by PHC using IciMapping V2.2. All 21 chromosomes in wheat were shown to harbor factors affecting PH in two populations, by both conditional and unconditional QTL mapping methods. At least 11 pairwise congruent QTL were identified in the two populations. In total, ten unconditional QTL and five conditional QTL that could be detected in the conditional analysis only have been verified in no less than three trials in WJ and WY. In addition, three QTL on the short arms of chromosomes 4B, 4D, and 7B were mapped to positions similar to those of the semi-dwarfing genes Rht- B1, Rht- D1 and Rht13, respectively. Conditional QTL mapping analysis in WJ and WY proved that, at the QTL level, SL contributed the least to PH, followed by FIITL; TITL had the strongest influence on PH, followed by SITL and FOITL. The results above indicated that the conditional QTL mapping method can be used to evaluate possible genetic relationships between PH and PHC, and it can efficiently and precisely reveal counteracting QTL, which will enhance the understanding of the genetic basis of PH in wheat. The combination of two related populations with a large/moderate population size made the results authentic and accurate. [ABSTRACT FROM AUTHOR]