Grain filling and proper grain development are essential biological processes in the plant's life cycle, eventually contributing to the final seed yield and quality in all cereal crops. How the different wheat grain components contribute to the overall development of the seed is very scarcely found in the literature. We performed a proteomics and metabolomics analysis in four different developing components of the wheat grain (seed coat, embryo, endosperm and cavity fluid) to characterize molecular processes during early and late grain development. In-gel shotgun proteomics analysis in 12, 15, 20 and 25 days after anthesis (DAA) revealed 15,484 identified and quantified proteins out of which 410 differentially expressed proteins (DEPs) were identified in the seed coat, 815 in embryo, 372 in endosperm and 492 in cavity fluid. The abundance of selected protein candidates revealed spatially and temporally resolved protein functions associated with development and grain filling. Multiple wheat protein isoforms involved in starch synthesis such as sucrose synthases, starch phosphorylase, granule bound and soluble starch synthase, pyruvate phosphate dikinase (PPDK), 14-3-3 proteins as well as sugar precursors undergo a major tissue-dependent change in abundance during wheat grain development suggesting an intimate interplay of starch biosynthesis control. Different isoforms of the protein disulphide isomerase (PDI) family as well as glutamine levels both involved in glutenin macropolymer (GMP) pattern showed distinct spatially and temporally abundance revealing their specific role as indicators of wheat gluten quality. Proteins binned into the functional category of cell growth /division and protein synthesis/degradation were more abundant in the early stages (12 and 15 DAA). At the metabolome level all tissues and especially the cavity fluid showed highly distinct metabolite profiles. The tissue specific data are integrated with biochemical networks to explore a comprehensive map of molecular processes during grain filling and developmental processes.This article is protected by copyright. All rights reserved.

The critical node (CN, 85% germination) of seed viability is an important threshold for seed regeneration decisions after long-term conservation. Dependent on the germplasm, the storage period until CN is reached varies and information on the divergence of the proteomic profiles is limited. Therefore, the study aims to identify key proteins and mechanisms relevant for a long plateau phase and a late CN during artificial seed aging of wheat. Seeds of the storage-tolerant genotype (ST) TRI 23248, and the storage-sensitive genotype (SS) TRI 10230 were exposed to artificial ageing (AA) and extracted embryos of imbibed seeds were analyzed using an iTRAQ-based proteomic technique. ST and SS required AA for 24 and 18 days to reach the CN, respectively. Fifty-seven and 165 differentially abundant proteins (DAPs) were observed in the control and aged groups, respectively. Interestingly, a higher activity in metabolic processes, protein synthesis, transcription, cell growth/division, and signal transduction were already found in imbibed embryos of control ST seeds. After AA, 132 and 64 DAPs were accumulated in imbibed embryos of both aged ST and SS seeds, respectively, which were mainly associated with cell defense, rescue, and metabolism. Moreover, 78 DAPs of ST appeared before CN and were mainly enriched in biological pathways related to the maintenance of redox and carbon homeostasis and they presented a stronger protein translation ability. In contrast, in SS, only 3 DAPs appeared before CN and were enriched only in the structural constituents of the cytoskeleton. In conclusion, a longer span of plateau phase might be obtained in seeds when proteins indicate an intense stress response before CN and include the effective maintenance of cellular homeostasis, and avoidance of excess accumulation of cytotoxic compounds. Although key proteins, inherent factors and the precise regulatory mechanisms need to be further investigated, the found proteins may also have functional potential roles during long-term seed conservation.

Lysine acetylation (Kac) is a protein post-translational modification (PTM) widely found in plants that plays vital roles in metabolic pathways. Although seed germination and development are regulated by Kac, its potential function in seed ageing remains to be investigated. Our preliminary study demonstrated that Kac levels were altered during wheat seed artificial ageing. However, its specific role in this process still needs to be elucidated. Here, we performed quantitative acetylation proteomics analysis of soft wheat seeds with different germination rates during artificial ageing. A total of 175 acetylation proteins and 255 acetylation modification sites were remarkably changed. The differentially acetylated proteins were enriched in metabolism; response to harsh intracellular environment, such as ROS; protein storage and processing. Notably, expression, point mutation to mimic Kac by K to Q mutation at K80 and K138, protein purification and enzyme activity detection revealed that the Kac of ROS-scavenging glutathione transferase attenuated its activity, indicating that the defense ability of wheat seeds to stress gradually diminished, and the ageing process was inevitable. Collectively, our data provide a basis for further understanding the roles of Kac in seed ageing and might aid in the development of new techniques to prolong seed viability and food quality.

Fusarium crown rot is major disease in wheat. However, the wheat defense mechanisms against this disease remain poorly understood.

Wheat stripe rust and powdery mildew are important worldwide diseases of wheat (Triticum aestivum). The wheat cultivar Xingmin318 (XM318) is resistant to both wheat stripe rust and powdery mildew, which are caused by Puccinia striiformis f. sp. tritici (Pst) and Blumeria graminis f. sp. tritici (Bgt), respectively. To explore the difference between wheat defense response against Pst and Bgt, quantitative proteomic analyses of XM318 inoculated with either Pst or Bgt were performed using tandem mass tags (TMT) technology. A total of 741 proteins were identified as differentially accumulated proteins (DAPs). Bioinformatics analyses indicated that some functional categories, including antioxidant activity and immune system process, exhibited obvious differences between Pst and Bgt infections. Intriguingly, only 42 DAPs responded to both Pst and Bgt infections. Twelve DAPs were randomly selected for RT-qPCR analysis, and the mRNA expression levels of 11 were consistent with their protein expression. Furthermore, gene silencing using the virus-induced gene silencing system indicated that glutathione S-transferase (TaGSTU6) has an important role in resistance to Bgt but not to Pst. TaGSTU6 interacted with the cystathionine beta-synthase (CBS) domain-containing protein (TaCBSX3) in both Pst and Bgt infections. Knockdown of TaCBSX3 expression only reduced wheat resistance to Bgt infection. Overexpression of TaGSTU6 and TaCBSX3 in Arabidopsis (Arabidopsis thaliana) promoted plant resistance to Pseudomonas syringae pv. Tomato DC3000. Our results indicate that TaGSTU6 interaction with TaCBSX3 only confers wheat resistance to Bgt, suggesting that wheat has different response mechanisms to Pst and Bgt stress.© The Author(s) 2022. Published by Oxford University Press on behalf of American Society of Plant Biologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

Fusarium head blight (FHB) is a devastating disease of wheat, causing yield losses and quality reduction as a result of mycotoxin production. In this study, iTRAQ (isobaric tags for relative and absolute quantification)-labeling-based mass spectrometry was employed to characterize the proteome in wheat cultivars Xinong 538 and Zhoumai 18 with contrasting levels of FHB resistance as a means to elucidate the molecular mechanisms contributing to FHB resistance. A total of 13,669 proteins were identified in the two cultivars 48 h after Fusarium graminearum inoculation. Among these, 2,505 unique proteins exclusively accumulated in Xinong 538 (resistant) and 887 proteins in Zhoumai 18 (susceptible). Gene Ontology enrichment analysis showed that most differentially accumulated proteins (DAPs) from both cultivars were assigned to the following categories: metabolic process, single-organism process, cellular process, and response to stimulus. Kyoto Encyclopedia of Genes and Genomes analysis showed that a greater number of proteins belonging to different metabolic pathways were identified in Xinong 538 compared with Zhoumai 18. Specifically, DAPs from the FHB-resistant cultivar Xinong 538 populated categories of metabolic pathways related to plant–pathogen interaction. These DAPs might play a critical role in defense responses exhibited by Xinong 538. DAPs from both genotypes were assigned to all wheat chromosomes except chromosome 6B, with approximately 30% mapping to wheat chromosomes 2B, 3B, 5B, and 5D. Twenty single nucleotide polymorphism markers, flanking DAPs on chromosomes 1B, 3B, 5B, and 6A, overlapped with the location of earlier mapped FHB-resistance quantitative trait loci. The data provide evidence for the involvement of several DAPs in the early stages of the FHB-resistance response in wheat; however, further functional characterization of candidate proteins is warranted.

Dwarf bunt and common bunt diseases of wheat are caused by Tilletia controversa Kühn and Tilletia foetida Kühn, respectively, and losses caused by these diseases can reach 70–80% in favourable conditions. T. controversa and T. foetida are fungal pathogens belonging to the Exobasidiomycetes within the basidiomycetous smut fungi (Ustilaginomycotina). In order to illuminate the proteomics differences of wheat spikes after the infection of T. controversa and T. foetida, the isobaric tags for relative and absolute quantification (iTRAQ) technique was used for better clarification. A total of 4553 proteins were differentially detected after T. controversa infection; 4100 were upregulated, and 453 were downregulated. After T. foetida infection, 804 differentially expressed proteins were detected; 447 were upregulated and 357 were downregulated. In-depth data analysis revealed that 44, 50 and 82 proteins after T. controversa and 9, 6 and 16 proteins after T. foetida were differentially expressed, which are antioxidant, plant-pathogen interaction and glutathione proteins, respectively, and 9 proteins showed results consistent with PRM. The top 20 KEGG enrichment pathways were identified after pathogen infection. On the basis of gene ontology, the upregulated proteins were linked with metabolic process, catalytic activity, transferase activity, photosynthetic membrane, extracellular region and oxidoreductase activity. The results expanded our understanding of the proteome in wheat spikes in response to T. controversa and T. foetida infection and provide a basis for further investigation for improving the defense mechanism of the wheat crops.

The fungal pathogen Magnaporthe oryzae causes devastating blast disease in various cereals, including rice (Oryza sativa), wheat (Triticum aestivum), maize (Zea mays), and barley (Hordeum vulgare). Despite previous reports on fungal host specificity, the mechanisms underlying differential host infection strategies remain unclear. This study aimed to identify differentially abundant proteins (DAPs) in the apoplast of rice, barley, and wheat following infection with two M. oryzae pathovars using liquid chromatography-tandem mass spectrometry (LC–MS/MS). LC–MS/MS analysis revealed an enrichment of both M. oryzae and host proteins in the apoplast during the compatible reaction compared to the incompatible reaction. DAPs from M. oryzae involved in the host interaction included secreted extracellular enzymes (e.g., hydrolases), which were significantly increased in the M. oryzae Oryzae (MoO)-infected rice apoplast. Among host proteins, the proportion of protein-modifying enzymes increased in the M. oryzae Triticum (MoT)-infected rice and MoO-infected wheat apoplastic fluids, particularly rice glycosidases, peroxidases, and serine proteases, as well as wheat serine proteases. Furthermore, DAPs from MoL-infected rice were enriched in carbohydrate metabolism, suggesting that carbohydrate metabolism-related proteins may play a vital role in rice resistance to MoL. Additionally, protein-modifying and cytoskeletal proteins, as well as stress-responsive proteins, were enriched in the MoO-infected wheat apoplastic fluid. Finally, DAPs from both MoO- and MoL-infected barley were enriched in hydrogen peroxide catabolism, suggesting that peroxidases may be vital for barley resistance to M. oryzae. The identification of DAPs from both M. oryzae strains and the three host plants offers valuable insights into the host specificity mechanisms of M. oryzae in cereal crops.

Chinese wheat mosaic virus (CWMV) is a severe threat to winter wheat and is transmitted by Polymyxa graminis. The mechanisms of interactions between CWMV and plants are poorly understood. In this study, a comparative proteomics analysis based on nanoliquid chromatography mass spectrometry (MS)/MS was conducted to characterize proteomic changes in plants responding to CWMV infection.

\n Hydrogen sulfide (H\n 2\n S) is a key gaseous signaling molecule involved in plant growth and stress responses, yet its role in wheat resistance to stripe rust remains poorly understood. Here, we show that exogenous H\n 2\n S enhances resistance of wheat (\n Triticum aestivum\n L.) to\n Puccinia striiformis\n f. sp\n. tritici\n (\n Pst\n ), the causative agent of stripe rust. Comparative persulfidation proteomics identified the autophagy-related protein TaATG6c as a\n Pst\n -responsive H₂S target. Site-specific mass spectrometry and a modified biotin-switch assay demonstrated that Cys177 and Cys180 of TaATG6c undergo H₂S-induced persulfidation. Structural modeling based on AlphaFold predicted that these two site mutations reduced the binding activity of ATG6c to ATG14. Functional characterization using virus-induced gene silencing (VIGS) revealed that TaATG6 positively regulates wheat immunity against\n Pst\n, as silencing\n TaATG6\n promoted fungal growth. Moreover,\n TaATG6\n expression was markedly induced during\n Pst\n infection. Notably, the resistance-promoting effect of NaHS was compromised in\n TaATG6\n -silenced plants. Conversely, transient overexpression of\n TaATG6\n enhanced wheat resistance to stripe rust, whereas mutation of Cys177 and Cys180 attenuated this effect. Endogenous biotin-switch assays further showed that TaATG6c persulfidation exhibits pathogen-responsive and dynamic characteristics, which were abolished in the\n TaATG6\n C177A/C180A\n mutant. Consistently, H₂S treatment and\n Pst\n infection stimulated the accumulation of lipidated ATG8 (ATG8–PE), indicating activation of autophagy, while this response was largely abolished in\n TaATG6\n -silenced plants. Together, these results suggest that H₂S promotes autophagy initiation through persulfidation of TaATG6c, thereby enhancing wheat resistance to stripe rust and highlighting a redox-regulated mechanism underlying plant stress adaptation.\n

Drought stress severely affects crop productivity and threatens food security. As current trends of global warming are predicted to exacerbate droughts, developing drought-resilient crops becomes urgent. Here, we used the drought-tolerant (BW35695) and drought-sensitive (BW4074) wheat varieties to investigate the physiological, biochemical, and leaf proteome responses underpinning drought tolerance. In response to drought, the tolerant variety had higher osmolyte accumulation and maintained higher leaf water content than the sensitive variety. BW35695 also had an enhanced antioxidant enzyme capacity and reduced reactive oxygen species (ROS), resulting in diminished membrane lipid damage, as reflected by malondialdehyde content. Proteomic analysis revealed that drought-induced differential expression of proteins involved in diverse biological processes in both wheat varieties, including primary and secondary metabolism, protein synthesis/folding/degradation, defense/ROS detoxification, energy, transcription, and cell structure. Notably, photosynthesis emerged as the most enriched biochemical process targeted for suppression in the drought-tolerant BW35695 wheat, but not in drought-sensitive BW4074, possibly as a survival strategy for averting cell damage inflicted by photosynthesis-derived ROS. Additionally, protein synthesis-related proteins were highly upregulated in BW35695, presumably to drive cell-wide stress-adaptive responses. The protein network identified here will be useful in further studies to understand the molecular basis for divergent drought response phenotypes in crops.

Climate warming has become an indisputable fact, and wheat is among the most heat-sensitive cereal crops. Heat stress during grain filling threatens global wheat production and food security. Here, we analyzed the physiological and proteomic changes by delayed sowing on the photosynthetic capacity of winter wheat leaves under heat stress. Our aim is to provide a new cultivation way for the heat stress resistance in wheat.

High-temperature (HT) stress can induce male sterility in wheat; however, the underlying mechanisms remain poorly understood. This study examined proteomic alterations across three developmental stages between normal and HT-induced male-sterile (HT-ms) anthers in wheat. Utilizing tandem mass tags-based proteomics, we identified 2532 differentially abundant proteins (DAPs): 27 in the tetrad stage, 157 in the binuclear stage, and 2348 in the trinuclear stage. Analyses through Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathways indicated significant enrichment of these DAPs in seven pathways, namely phenylpropanoid biosynthesis, flavonoid biosynthesis, sphingolipid metabolism, MAPK signaling pathway, starch and sucrose metabolism, response to heat, and response to reactive oxygen species (ROS). Our results indicated the downregulation of DAPs associated with phenylpropanoid biosynthesis and starch and sucrose metabolism, which aligns with anther indehiscence and the lack of starch in HT-ms anthers. By contrast, DAPs in the ROS pathway were upregulated, which aligns with excessive ROS accumulation in HT-ms anthers. Additionally, we conducted protein–protein interaction analysis for the DAPs of these pathways, identifying 15 hub DAPs. The abundance of these hub proteins was confirmed through qRT-PCR, assessing mRNA expression levels of the corresponding transcripts. Collectively, these results offer insights into the molecular mechanisms underlying HT-induced male sterility in wheat at the proteomic level, providing a valuable resource for further research in plant sexual reproduction.

<p class="MsoNormal"> <div style="text-align:justify;"> Low temperature (LT) in spring has become one of the principal abiotic stresses that restrict the growth and development of wheat.&nbsp; Diverse analyses were performed to investigate the mechanism underlying the response of wheat grain development to LT stress during booting.&nbsp; These included morphological observation, measurements of starch synthase activity, and determination of amylose and amylopectin content of wheat grain after exposure to treatment with LT during booting.&nbsp; Additionally, proteomic analysis was performed using tandem mass tags (TMT).&nbsp; Results showed that the plumpness of wheat grains decreased after LT stress.&nbsp; Moreover, the activities of sucrose synthase (SuS, EC 2.4.1.13) and ADP-glucose pyrophosphorylase (AGPase, EC 2.7.7.27) exhibited a significant reduction, leading to a significant reduction in the contents of amylose and amylopectin.&nbsp; A total of 509 differentially expressed proteins (DEPs) were identified by proteomics analysis.&nbsp; The Gene Ontology (GO) enrichment analysis showed that the protein difference multiple in the nutritional repository activity was the largest among the molecular functions, and the up-regulated seed storage protein (SSP) played an active role in the response of grains to LT stress and subsequent damage.&nbsp; The Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed that LT stress reduced the expression of DEPs such as sucrose phosphate synthase (SPS), glucose-1-phosphate adenylyltransferase (glgC), and β-fructofuranosidase (FFase) in sucrose and starch metabolic pathways, thus affecting the synthesis of grain starch.&nbsp; In addition, many heat shock proteins (HSPs) were found in the protein processing in endoplasmic reticulum pathways, which can resist some damage caused by LT stress.&nbsp; These findings provide a new theoretical foundation for elucidating the underlying mechanism governing wheat yield development after exposure to LT stress in spring. </div> <div> <br> </div></p>

The development of a salt-tolerant hexaploid triticale cultivar offers an economical and efficient solution for utilizing marginal land. Understanding how hexaploid triticales respond to salt stress is essential if this goal is to be achieved. A genome-wide association study (GWAS), along with transcriptome and proteome analyses, were used in the present study to determine the molecular responses to salt stress in hexaploid triticale. In total, 81 marker-trait associations for 10 salt-tolerance traits were identified in 153 hexaploid triticale accessions, explaining 0.71% to 56.98% of the phenotypic variation, and 54 GWAS-associated genes were uncovered. A total of 67, 88, and 688 differential expression genes were co-expressed at both the transcriptomic and proteomic levels after 4, 12, and 18 h of salt stress, respectively. Among these differentially expressed genes, six appeared in the coincident expression trends for both the transcriptomic and proteomic levels at the seed germination stage. A total of nine common KEGG pathways were enriched at both the transcriptomic and proteomic levels at 4, 12, and 18 h. After integrating GWAS-target genes with transcriptomics and proteomics approaches that the candidate gene late embryogenesis abundant protein 14 (LEA14) was up-regulated at the transcriptomic and proteomic levels. LEA14 contained important stress-responsive cis-acting regulatory elements that could be dynamically regulated by the binding of transcription factors (TFs). This suggested that LEA14 was a key gene associated with salt tolerance in hexaploid triticale and could respond quickly to salt stress. This study improved understanding about the potential molecular mechanisms associated with hexaploid triticale salt tolerance and contributed to the breeding of salt-tolerant germplasms and the utilization of saline soils.

Here we document physiological and molecular attributes of three wheat cultivars (ZM9023, YM158 and FM1228) under low light intensity with advanced technologies, including non-standard quantitative technology and quantitative proteomics technology. We found lower dry matter accumulation of YM158 compared with ZM 9023 and FM1228 under low light intensities due to up-regulation of photosynthetic parameters electron transport rate (ETR), Y(II), Fv/Fm, Chl (a + b) of YM158 and down-regulation of Chl a/b. ETR, Y(II) and Fv/Fm significantly decreased between ZM9023 and FM1228. The ETR between PSII and PSI of YM158 increased, while light use efficiency (LUE) of ZM9023 and FM1228 decreased. We found that YM158 had greater propensity to adapt to low light compared with ZM9023, as the former was able to increase photochemical electron transfer rate, enhance photosystem activity, and increase the light energy under low light. This meant that the YM158 flag leaf has stronger regulatory mechanism under low light environment. Through proteomic analysis, we found LHC protein (LHCB1, LHCB4, LHCA2, LHCA3) for YH158 was significantly up-regulated, while the PSII subunit protein of FM1228 and ZM9023 b559 subunit protein were down-regulated. We also documented enhanced light use efficiency (LUE) due to higher light capture pigment protein complex (LHC), photosystem II (PSII), PSI and cytochrome B6F-related proteins, with dry matter accumulation being positively correlated with Fv/Fm, ETR, and ΦPS(II), and negatively correlated with initial fluorescence F0. We suggest that Fv/Fm, ETR, and ΦPS(II) could be considered in shade tolerance screening to facilitate wheat breeding.

Transient and chronic waterlogging constrains crop production in many regions of the world. Here, we invoke a novel iTRAQ-based proteomic strategy to elicit protein synthesis and regulation responses to waterlogging in tolerant (XM 55) and sensitive genotypes (YM 158). Of the 7,710 proteins identified, 16 were distinct between the two genotypes under waterlogging, partially defining a proteomic basis for waterlogging tolerance (and sensitivity). We found that 11 proteins were up-regulated and 5 proteins were down-regulated; the former included an Fe-S cluster assembly factor, heat shock cognate 70, GTP-binding protein SAR1A-like and CBS domain-containing protein. Down-regulated proteins contained photosystem II reaction center protein H, carotenoid 9, 10 (9′, 10′)-cleavage dioxygenase-like, psbP-like protein 1 and mitochondrial ATPase inhibitor. We showed that nine proteins responded to waterlogging with non-cultivar specificity: these included 3-isopropylmalate dehydratase large subunit, solanesyl-diphosphate synthase 2, DEAD-box ATP-dependent RNA helicase 3, and 3 predicted or uncharacterized proteins. Sixteen of the 28 selected proteins showed consistent expression patterns between mRNA and protein levels. We conclude that waterlogging stress may redirect protein synthesis, reduce chlorophyll synthesis and enzyme abundance involved in photorespiration, thus influencing synthesis of other metabolic enzymes. Collectively, these factors accelerate the accumulation of harmful metabolites in leaves in waterlogging-susceptible genotypes. The differentially expressed proteins enumerated here could be used as biological markers for enhancing waterlogging tolerance as part of future crop breeding programs.

Wheat growth and nitrogen (N) uptake gradually decrease in response to high NH4+/NO3–ratio. However, the mechanisms underlying the response of wheat seedling roots to changes in NH4+/NO3–ratio remain unclear. In this study, we investigated wheat growth, transcriptome, and proteome profiles of roots in response to increasing NH4+/NO3–ratios (Na: 100/0; Nr1: 75/25, Nr2: 50/50, Nr3: 25/75, and Nn: 0/100). High NH4+/NO3–ratio significantly reduced leaf relative chlorophyll content, Fv/Fm, and ΦII values. Both total root length and specific root length decreased with increasing NH4+/NO3–ratios. Moreover, the rise in NH4+/NO3–ratio significantly promoted O2–production. Furthermore, transcriptome sequencing and tandem mass tag-based quantitative proteome analyses identified 14,376 differentially expressed genes (DEGs) and 1,819 differentially expressed proteins (DEPs). The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis indicated that glutathione metabolism and phenylpropanoid biosynthesis were the main two shared enriched pathways across ratio comparisons. Upregulated DEGs and DEPs involving glutathione S-transferases may contribute to the prevention of oxidative stress. An increment in the NH4+/NO3–ratio induced the expression of genes and proteins involved in lignin biosynthesis, which increased root lignin content. Additionally, phylogenetic tree analysis showed that both A0A3B6NPP6 and A0A3B6LM09 belong to the cinnamyl-alcohol dehydrogenase subfamily. Fifteen downregulated DEGs were identified as high-affinity nitrate transporters or nitrate transporters. UpregulatedTraesCS3D02G344800andTraesCS3A02G350800were involved in ammonium transport. Downregulated A0A3B6Q9B3 is involved in nitrate transport, whereas A0A3B6PQS3 is a ferredoxin-nitrite reductase. This may explain why an increase in the NH4+/NO3–ratio significantly reduced root NO3–-N content but increased NH4+-N content. Overall, these results demonstrated that increasing the NH4+/NO3–ratio at the seedling stage induced the accumulation of reactive oxygen species, which in turn enhanced root glutathione metabolism and lignification, thereby resulting in increased root oxidative tolerance at the cost of reducing nitrate transport and utilization, which reduced leaf photosynthetic capacity and, ultimately, plant biomass accumulation.

Boron (B) is a trace element that is essential for normal wheat development, such as root growth. In wheat, roots are important organs that absorb nutrients and water. However, at present, there is insufficient research on the molecular mechanism underlying how short-term B stress affects wheat root growth.Here, the optimal concentration of B for wheat root growth was determined, and the proteomic profiles of roots under short-term B deficiency and toxicity were analyzed and compared by the isobaric tag for relative and absolute quantitation (iTRAQ) technique. A total of 270 differentially abundant proteins (DAPs) that accumulated in response to B deficiency and 263 DAPs that accumulated in response to B toxicity were identified. Global expression analysis revealed that ethylene, auxin, abscisic acid (ABA), and Ca signals were involved in the responses to these two stresses. Under B deficiency, DAPs related to auxin synthesis or signaling and DAPs involved in calcium signaling increased in abundance. In striking contrast, auxin and calcium signals were repressed under B toxicity. Twenty-one DAPs were detected under both conditions, including RAN1 that played a core role in the auxin and calcium signals. Overexpression of RAN1 was shown to confer plant resistance to B toxicity by activating auxin response genes, including TIR and those identified by iTRAQ in this research. Moreover, growth of the primary roots of tir mutant was significantly inhibited under B toxicity.Taken together, these results indicate that some connections were present between RAN1 and the auxin signaling pathway under B toxicity. Therefore, this research provides data for improving the understanding of the molecular mechanism underlying the response to B stress.© 2023. The Author(s), under exclusive licence to Springer Nature B.V.

Optimizing canopy light distribution (CLD) has manifested improved light utility and yield without modifying other inputs. Nonetheless, molecular mechanisms working at cellular and organelle level remain to be elucidated. The current study aimed to assess the effect of CLD on photosynthetic performance and yield of wheat, and to investigate into the molecular mechanism underlying the photosynthetically active radiation (PAR)–use efficiency (PUE) at optimized CLD. Wheat was planted in two rows having different spacing [R1 (15 cm) and R2 (25 cm)] to simulate different CLD. Flag and penultimate leaves were subjected to chloroplast proteomics analysis. An increase in row spacing positively affects CLD. A decrease (16.64%) of PAR interception in the upper layer, an increase (19.76%) in the middle layer, improved PUE (12.08%), and increased yield (9.38%) were recorded. The abundance of proteins associated with photosynthetic electron transport, redox state, and carbon-nitrogen assimilation was differentially altered by CLD optimization. In the penultimate leaves, R2 reduced the abundance of photosystem II (PSII) light-harvesting proteins, PSII-subunits, and increased the photosystem I (PSI) light-harvesting proteins, NAD(P)H quinone oxidoreductase (NQO) and enzymes involved in carbon assimilation compared to R1. Additionally, leaf stomatal conductance increased. Altogether, these findings demonstrated that the regulation of chloroplast proteome is intimately linked to light utilization, which provide basis for genetic manipulation of crop species for better adaptation and improvement of cultivation strategies.

Novel genetic variations can be obtained by inducing mutations in the plant which help to achieve novel traits. The useful mutant can be obtained through radiation mutation in a short period which can be used as a new material to produce new varieties with high yield and good quality wheat. In this paper, the proteomic analysis of wheat treated with different doses of 12C and 7Li ion beam radiation at the seedling stage was carried out through a Tandem Mass Tag (TMT) tagging quantitative proteomic analysis platform based on high-resolution liquid chromatography-mass spectrometry, and the traditional 60Co-γ-ray radiation treatment for reference. A total of 4,764 up-regulated and 5,542 down-regulated differentially expressed proteins were identified. These proteins were mainly enriched in the KEGG pathway associated with amino acid metabolism, fatty acid metabolism, carbon metabolism, photosynthesis, signal transduction, protein synthesis, and DNA replication. Functional analysis of the differentially expressed proteins showed that the oxidative defense system in the plant defense system was fully involved in the defense response after 12C ion beam and 7Li ion beam radiation treatments. Photosynthesis and photorespiration were inhibited after 12C ion beam and 60Co-γ-ray irradiation treatments, while there was no effect on the plant with 7Li ion beam treatment. In addition, the synthesis of biomolecules such as proteins, as well as multiple signal transduction pathways also respond to radiations. Some selected differentially expressed proteins were verified by Parallel Reaction Monitoring (PRM) and qPCR, and the experimental results were consistent with the quantitative results of TMT. The present study shows that the physiological effect of 12C ion beam radiation treatment is different as compared to the 7Li ion beam, but its similar to the 60Co-γ ray depicting a significant effect on the plant by using the same dose. The results of this study will provide a theoretical basis for the application of 12C and 7Li ion beam radiation in the mutation breeding of wheat and other major crops and promote the development of heavy ion beam radiation mutation breeding technology.

Lysine 2-hydroxyisobutyrylation (Khib) is a novel naturally occurring post-translational modification. The system Khib identification at proteomics level has been performed in various species and tissues to characterize the role of Khib in biological activities. However, the study of Khib in plant species is relatively less. In the present study, the first plant root tissues lysine 2-hydroxyisobutyrylome analysis was performed in wheat with antibody immunoprecipitation affinity, high resolution mass spectrometry-based proteomics and bioinformatics analysis. In total, 6328 Khib sites in 2186 proteins were repeatedly identified in three replicates. These Khib proteins showed a wide subcellular location distribution. Function and pathways characterization of these Khib proteins indicated that many cellular functions and metabolism pathways were potentially affected by this modification. Protein and amino acid metabolism related process may be regulated by Khib, especially ribosome activities and proteins biosynthesis process. Carbohydrate metabolism and energy production related processes including glycolysis/gluconeogenesis, TCA cycle and oxidative phosphorylation pathways were also affected by Khib modification. Besides, root sulfur assimilation and transformation related enzymes exhibited Khib modification. Our work illustrated the potential regulation role of Khib in wheat root physiology and biology, which could be used as a useful reference for Khib study in plant root.

Cytoplasmic male sterility (CMS) plays an important role in the application of heterosis in wheat (Triticum aestivum L.). However, the molecular mechanism underlying CMS remains unknown. This study provides a comprehensive morphological and proteomic analysis of the anthers of a P-type CMS wheat line (P) and its maintainer line, Yanshi 9 hao (Y). Cytological observations indicated that the P-type CMS line shows binucleate microspore abortion. In this line, the tapetum degraded early, leading to anther cuticle defects, which could not provide the nutrition needed for microspore development in a timely manner, thus preventing the development of the microspore to the normal binucleate stage. Proteomic analysis revealed novel proteins involved in P-type CMS. Up to 2576 differentially expressed proteins (DEPs) were quantified in all anthers, and these proteins were significantly enriched in oxidative phosphorylation, glycolysis/gluconeogenesis, citrate cycle (TCA cycle), starch and sucrose metabolism, phenylpropanoid biosynthesis, and pyruvate metabolism pathways. These proteins may comprise a network that regulates male sterility in wheat. Based on the function analysis of DEPs involved in the complex network, we concluded that the P-type CMS line may be due to cellular dysfunction caused by disturbed carbohydrate metabolism, inadequate energy supply, and disturbed protein synthesis. These results provide insights into the molecular mechanism underlying male sterility and serve as a valuable resource for researchers in plant biology, in general, and plant sexual reproduction, in particular.

Thermosensitive sterile lines are natural materials for exploring the effects of anther development on male fertility. To study the possible molecular mechanisms regulating protein activity during the induction of male sterility, proteomic and phosphoproteomic analyses with tandem mass tags (TMTs) were used to study the binucleate anther of the thermosensitive sterile wheat line YS3038. A total of 9072 proteins, including 5019 phosphoproteins, were identified. Enrichment analyses of differentially abundant proteins (DAPs) and phosphoproteins (DAPPs) in metabolic pathways showed that both were mainly related to energy metabolism. Soluble sugar and ATP content were significantly decreased, free fatty acid content was significantly increased, and ROS was abnormally accumulated in male sterile YS3038-A. In addition, 233 kinase–substrate pairs involved in potential phosphorylation control networks were predicted to regulate fertility. Candidate proteins were identified, and a quantitative real-time polymerase chain reaction (qRT-PCR) analysis was used to validate the TMT results. TaPDCD5 is likely to be involved in fertility conversion of YS3038 by barley stripe mosaic virus-induced gene silencing (BSMV-VIGS). Our data provide new insights into the mechanism of TCMS, which has value for identifying potential candidate proteins associated with the formation or abortion of pollen and promotion of wheat heterosis utilization.

Triticum aestivum is an important crop whose reference genome (International Wheat Genome Sequencing Consortium (IWGSC) RefSeq v2.1) offers a valuable resource for understanding wheat genetic structure, improving agronomic traits, and developing new cultivars. A key aspect of gene model annotation is protein-level evidence of gene expression obtained from proteomics studies, followed up by proteogenomics to physically map proteins to the genome. In this research, we have retrieved the largest recent wheat proteomics datasets publicly available and applied the Basic Local Alignment Search Tool (tBLASTn) algorithm to map the 861,759 identified unique peptides against IWGSC RefSeq v2.1. Of the 92,719 hits, 83,015 unique peptides aligned along 33,612 High Confidence (HC) genes, thus validating 31.4% of all wheat HC gene models. Furthermore, 6685 unique peptides were mapped against 3702 Low Confidence (LC) gene models, and we argue that these gene models should be considered for HC status. The remaining 2934 orphan peptides can be used for novel gene discovery, as exemplified here on chromosome 4D. We demonstrated that tBLASTn could not map peptides exhibiting mid-sequence frame shift. We supply all our proteogenomics results, Galaxy workflow and Python code, as well as Browser Extensible Data (BED) files as a resource for the wheat community via the Apollo Jbrowse, and GitHub repositories. Our workflow could be applied to other proteomics datasets to expand this resource with proteins and peptides from biotically and abiotically stressed samples. This would help tease out wheat gene expression under various environmental conditions, both spatially and temporally.