Priming with gold nanoparticles leads to changes in the photosynthetic apparatus and improves the cold tolerance of wheat.

Nanotechnologies provide a great platform for researching nanoparticles effects on living organisms including plants. This work shows the stimulating effect of seed priming with gold nanoparticles (AuNPs) on photosynthetic apparatus of Triticum aestivum seedlings. It was found using inductively coupled plasma-atomic emission and mass spectrometry that AuNPs (the average diameter of 15.3 nm, concentration of 20 µg ml-1) penetrated into the seeds, but were not found in seedling leaves. Ultrastructural changes in chloroplasts were found using transmission electron microscopy in plants grown from treated seeds: increases in the size of plastids, starch grains, grana in chloroplasts, and the number of thylakoids in grana. The intensity of photosynthesis, the content of chlorophylls, and the portion of unsaturated fatty acids in the composition of total leaf lipids were increased in treated AuNPs plants. This study demonstrates that revealed changes determined the increased tolerance of wheat to low temperature. The adaptive significance of these changes, possible mechanisms of the AuNPs effects on plants and future perspectives of study are discussed. This is the first report showing nanopriming with AuNPs as a new method to study the mechanisms of stress tolerance.

Microtubule organization changes severely after mannitol and n-butanol treatments inducing microspore embryogenesis in bread wheat.

BACKGROUND: A mannitol stress treatment and a subsequent application of n-butanol, known as a microtubule-disrupting agent, enhance microspore embryogenesis (ME) induction and plant regeneration in bread wheat. To characterize changes in cortical (CMT) and endoplasmic (EMT) microtubules organization and dynamics, associated with ME induction treatments, immunocytochemistry studies complemented by confocal laser scanning microscopy (CLSM) were accomplished. This technique has allowed us to perform advanced 3- and 4D studies of MT architecture. The degree of MT fragmentation was examined by the relative fluorescence intensity quantification. RESULTS: In uni-nucleated mannitol-treated microspores, severe CMT and EMT fragmentation occurs, although a complex network of short EMT bundles protected the nucleus. Additional treatment with n-butanol resulted in further depolymerization of both CMT and EMT, simultaneously with the formation of MT aggregates in the perinuclear region. Some aggregates resembled a preprophase band. In addition, a portion of the microspores progressed to the first mitotic division during the treatments. Bi-nucleate pollen-like structures showed a high MT depolymerization after mannitol treatment and numerous EMT bundles around the vegetative and generative nuclei after n-butanol. Interestingly, bi-nucleate symmetric structures showed prominent stabilization of EMT. CONCLUSIONS: Fragmentation and stabilization of microtubules induced by mannitol- and n-butanol lead to new configurations essential for the induction of microspore embryogenesis in bread wheat. These results provide robust insight into MT dynamics during EM induction and open avenues to address newly targeted treatments to induce ME in recalcitrant species.

Relationship between polyamines conjugated to mitochondrion membrane and mitochondrion conformation from developing wheat embryos under drought stress.

The mitochondrion conformation and the contents of conjugated polyamines were investigated using the embryos of developing wheat (Triticum aestivum L.) grains of two cultivars differing in drought tolerance as experiment materials. After drought stress treatment for 7 days, the relative water content of embryo and relative increase rate of embryo dry weight of the drought-sensitive Yangmai No. 9 cv. decreased more significantly than those of the drought-tolerant Yumai No. 18 cv. Furthermore, the changes in mitochondrion conformation of Yangmai No. 9 were more marked. Meanwhile, the increases of the contents of conjugated non-covalently spermidine (CNC-Spd) and conjugated covalently putrescine (CC-Put) of Yumai No. 18 were more obvious than those of Yangmai No. 9. Treatment with exogenous Spd not only alleviated the injury of drought stress to Yangmai No. 9, but also enhanced the increase of CNC-Spd content and inhibited the change in the mitochondrion conformation of this cultivar. The treatments of Yumai No. 18 with two inhibitors, methylglyoxyl-bis (guanylhydrazone) and phenanthrolin, significantly inhibited the drought stress-induced increases of CNC-Spd and CC-Put contents of the cultivar, respectively. Meanwhile, the treatments with the two inhibitors aggravated the injury of drought stress to Yumai No. 18 and enhanced the change in the mitochondrion conformation of this cultivar. These results mentioned above suggested that the CNC-Spd and CC-Put in embryo mitochondrion membrane isolated from developing grains could enhance the wheat tolerance to drought stress by maintaining the mitochondrion conformation.

Multiseriate cortical sclerenchyma enhance root penetration in compacted soils.

Mechanical impedance limits soil exploration and resource capture by plant roots. We examine the role of root anatomy in regulating plant adaptation to mechanical impedance and identify a root anatomical phene in maize (Zea mays) and wheat (Triticum aestivum) associated with penetration of hard soil: Multiseriate cortical sclerenchyma (MCS). We characterize this trait and evaluate the utility of MCS for root penetration in compacted soils. Roots with MCS had a greater cell wall-to-lumen ratio and a distinct UV emission spectrum in outer cortical cells. Genome-wide association mapping revealed that MCS is heritable and genetically controlled. We identified a candidate gene associated with MCS. Across all root classes and nodal positions, maize genotypes with MCS had 13% greater root lignin concentration compared to genotypes without MCS. Genotypes without MCS formed MCS upon exogenous ethylene exposure. Genotypes with MCS had greater lignin concentration and bending strength at the root tip. In controlled environments, MCS in maize and wheat was associated improved root tensile strength and increased penetration ability in compacted soils. Maize genotypes with MCS had root systems with 22% greater depth and 49% greater shoot biomass in compacted soils in the field compared to lines without MCS. Of the lines we assessed, MCS was present in 30 to 50% of modern maize, wheat, and barley cultivars but was absent in teosinte and wild and landrace accessions of wheat and barley. MCS merits investigation as a trait for improving plant performance in maize, wheat, and other grasses under edaphic stress.

Comparison of Light Condition-Dependent Differences in the Accumulation and Subcellular Localization of Glutathione in Arabidopsis and Wheat.

This study aimed to clarify whether the light condition-dependent changes in the redox state and subcellular distribution of glutathione were similar in the dicotyledonous model plant Arabidopsis (wild-type, ascorbate- and glutathione-deficient mutants) and the monocotyledonous crop species wheat (Chinese Spring variety). With increasing light intensity, the amount of its reduced (GSH) and oxidized (GSSG) form and the GSSG/GSH ratio increased in the leaf extracts of both species including all genotypes, while far-red light increased these parameters only in wheat except for GSH in the GSH-deficient Arabidopsis mutant. Based on the expression changes of the glutathione metabolism-related genes, light intensity influences the size and redox state of the glutathione pool at the transcriptional level in wheat but not in Arabidopsis. In line with the results in leaf extracts, a similar inducing effect of both light intensity and far-red light was found on the total glutathione content at the subcellular level in wheat. In contrast to the leaf extracts, the inducing influence of light intensity on glutathione level was only found in the cell compartments of the GSH-deficient Arabidopsis mutant, and far-red light increased it in both mutants. The observed general and genotype-specific, light-dependent changes in the accumulation and subcellular distribution of glutathione participate in adjusting the redox-dependent metabolism to the actual environmental conditions.

Three-dimensional imaging reveals that positions of cyst nematode feeding sites relative to xylem vessels differ between susceptible and resistant wheat.

KEY MESSAGE: Resistance conferred by the Cre8 locus of wheat prevents cereal cyst nematode feeding sites from reaching and invading root metaxylem vessels. Cyst nematodes develop syncytial feeding sites within plant roots. The success of these sites is affected by host plant resistance. In wheat (Triticum aestivum L.), 'Cre' loci affect resistance against the cereal cyst nematode (CCN) Heterodera avenae. To investigate how one of these loci (Cre8, on chromosome 6B) confers resistance, CCN-infected root tissue from susceptible (-Cre8) and resistant (+Cre8) wheat plants was examined using confocal microscopy and laser ablation tomography. Confocal analysis of transverse sections showed that feeding sites in the roots of -Cre8 plants were always adjacent to metaxylem vessels, contained many intricate 'web-like' cell walls, and sometimes 'invaded' metaxylem vessels. In contrast, feeding sites in the roots of +Cre8 plants were usually not directly adjacent to metaxylem vessels, had few inner cell walls and did not 'invade' metaxylem vessels. Models based on data from laser ablation tomography confirmed these observations. Confocal analysis of longitudinal sections revealed that CCN-induced xylem modification that had previously been reported for susceptible (-Cre8) wheat plants is less extreme in resistant (+Cre8) plants. Application of a lignin-specific stain revealed that secondary thickening around xylem vessels in CCN-infected roots was greater in +Cre8 plants than in -Cre8 plants. Collectively, these results indicate that Cre8 resistance in wheat acts by preventing cyst nematode feeding sites from reaching and invading root metaxylem vessels.

Plant Cell Wall Changes in Common Wheat Roots as a Result of Their Interaction with Beneficial Fungi of Trichoderma.

Plant cell walls play an important role in shaping the defense strategies of plants. This research demonstrates the influence of two differentiators: the lifestyle and properties of the Trichoderma species on cell wall changes in common wheat seedlings. The methodologies used in this investigation include microscopy observations and immunodetection. In this study was shown that the plant cell wall was altered due to its interaction with Trichoderma. The accumulation of lignins and reorganization of pectin were observed. The immunocytochemistry indicated that low methyl-esterified pectins appeared in intercellular spaces. Moreover, it was found that the arabinogalactan protein epitope JIM14 can play a role in the interaction of wheat roots with both the tested Trichoderma strains. Nevertheless, we postulate that modifications, such as the appearance of lignins, rearrangement of low methyl-esterified pectins, and arabinogalactan proteins due to the interaction with Trichoderma show that tested strains can be potentially used in wheat seedlings protection to pathogens.

Analysis of development, accumulation and structural characteristics of starch granule in wheat grain under nitrogen application.

Nitrogen is one of the most important nutrients for wheat growth and has a critical influence on yield and quality. This study aims to examine how medium nitrogen level (240 kg/hm2) affects the starch granule development, starch accumulation, and structural characteristics of wheat starch. The results showed that nitrogen treatment could reduce the biosynthesis of starch and amylose, delay the degradation of starch in pericarp, and promote the proportion of B-type small starch granule in endosperm compared with those in the N0. In addition, the composition and distribution of starch granules were changed, the crystal structure in the inner lamellae and ordered structure in the external region of starch granules were affected, and the swelling power and solubility of starch during wheat development were increased. The effect of nitrogen treatment on the mRNA expression of enzymes related to starch biosynthesis or degradation varied in different developmental stages. During middle and later grain filling stages, AGPase, GBSSI, and GBSSII were lower, and SSS, SBE, and DBE were higher in N240 than in N0. This study indicated that nitrogen application at booting stage significantly affected the structural characteristics of starch, and ultimately determines its quality.

Source-sink modifications affect leaf senescence and grain mass in wheat as revealed by proteomic analysis.

BACKGROUND: The grain yield of cereals is determined by the synergistic interaction between source activity and sink capacity. However, source-sink interactions are far from being fully understood. Therefore, a field experiment was performed in wheat to investigate the responses of flag leaves and grains to sink/source manipulations. RESULTS: Half-degraining delayed but partial defoliation enhanced leaf senescence. Sink/source manipulations influenced the content of reactive oxygen species in the flag leaf and the concentration of phytohormones, including cytokinins, indoleacetic 3-acid and jasmonic acid, in the flag leaves (LDef) and grains (GDef) in defoliated plants and flag leaves (LDG) and grain (GDG) in de-grained plants. Isobaric tag for relative and absolute quantitation (iTRAQ)-based quantitative proteomic analysis indicated that at 16 days after manipulation, a total of 97 and 59 differentially expressed proteins (DEPs) from various functional categories were observed in the LDG and LDef groups, respectively, compared with the control, and 115 and 121 DEPs were observed in the GDG and GDef groups, respectively. The gene ontology annotation terms of the DEPs mainly included carbon fixation, hydrogen peroxide catabolic process, chloroplast and cytoplasm, oxidoreductase activity and glutamate synthase activity in the flag leaves of manipulated plants and organonitrogen compound metabolic process, cytoplasm, vacuolar membrane, CoA carboxylase activity, starch synthase activity and nutrient reservoir activity in the grains of manipulated plants. KEGG pathway enrichment analysis revealed that photosynthesis, carbon, nitrogen and pyruvate metabolism and glycolysis/gluconeogenesis were the processes most affected by sink/source manipulations. Sink/source manipulations affected the activities of amylase and proteinases and, ultimately, changed the mass per grain. CONCLUSIONS: Manipulations to change the sink/source ratio affect hormone levels; hydrolytic enzyme activities; metabolism of carbon, nitrogen and other main compounds; stress resistance; and leaf senescence and thus influence grain mass.

Diter von Wettstein and The Meiotic Program of Pairing and Recombination.

Recombination and pairing are prominent features of meiosis where they play an important role in increasing genetic diversity. In most organisms recombination also plays mechanical roles in mediating pairing of homologous chromosomes during prophase and in ensuring regular segregation of homologous pairs at the first meiotic division. The laboratory directed by D. von Wettstein identified six key steps in the meiotic process: (1) Recombination mediated processes occur in physical and functional linkage with the synaptonemal complex (SC), a highly conserved, meiosis-specific structure that links homologous axes along their lengths. (2) The pairing process involves formation and resolution of chromosomal entanglements/interlockings. (3) The SC normally forms specifically between homologous chromosomes, but in unusual situations can form between nonhomologous chromosomes or regions resulting in two-phase SC formation. (4) In hexaploid common wheat, extensive multivalents form with multiple, pairing partner shifts, indicating homology recognition and SC formation among homoeologs as well as homologs. (5) Linkage between recombination and the SC is revealed by crossover-correlated nodules localized in the SC central region. (6) Modified SCs sometimes play a direct role in homolog segregation, providing the required connection between homologs in absence of crossovers/chiasmata.

The Major Factors Causing the Microspore Abortion of Genic Male Sterile Mutant NWMS1 in Wheat (Triticum aestivum L.).

Male sterility is a valuable trait for genetic research and production application of wheat (Triticum aestivum L.). NWMS1, a novel typical genic male sterility mutant, was obtained from Shengnong 1, mutagenized with ethyl methane sulfonate (EMS). Microstructure and ultrastructure observations of the anthers and microspores indicated that the pollen abortion of NWMS1 started at the early uninucleate microspore stage. Pollen grain collapse, plasmolysis, and absent starch grains were the three typical characteristics of the abnormal microspores. The anther transcriptomes of NWMS1 and its wild type Shengnong 1 were compared at the early anther development stage, pollen mother cell meiotic stage, and binucleate microspore stage. Several biological pathways clearly involved in abnormal anther development were identified, including protein processing in endoplasmic reticulum, starch and sucrose metabolism, lipid metabolism, and plant hormone signal transduction. There were 20 key genes involved in the abnormal anther development, screened out by weighted gene co-expression network analysis (WGCNA), including SKP1B, BIP5, KCS11, ADH3, BGLU6, and TIFY10B. The results indicated that the defect in starch and sucrose metabolism was the most important factor causing male sterility in NWMS1. Based on the experimental data, a primary molecular regulation model of abnormal anther and pollen developments in mutant NWMS1 was established. These results laid a solid foundation for further research on the molecular mechanism of wheat male sterility.

Association of wheat chaff derived silica fiber and esophageal cancer in north China.

BACKGROUND: Despite decades of research and intervention programs, the epidemic of esophageal squamous cell carcinoma (ESCC) in the Taihang Mountain area of north China has not seen convincing explanation by any risk factor yet and the incidence has not seen a substantial decrease. Based on recently disclosed association of aridity and wheat consumption with esophageal cancer, we revisited the hypothesis of biogenic silica in esophageal cancer development. METHODS: From the archives of the Pathology Department of Heping Hospital, Changzhi Medical College, we selected three pairs of formalin-fixed samples, tumor tissues and distant normal tissues, of three patients operated for ESCC who had no history of workplace exposure to silica dust. Two pairs of dried tissue samples were used for phytolith (silica body) analysis and another pair for microanalysis with Transmission Electron Microscope (TEM). RESULTS: One of the phytoliths in ESCC tumor tissue was similar to the prickle hair on the surface of wheat bract. In the mineral particles detected in the tumor tissue the predominant elements were Si, Ca, and P, whereas Si signals were not obvious in the distant normal tissue. CONCLUSIONS: The preliminary findings on the detection of phytoliths and the higher than normal Si concentration in ESCC tumor tissue warrants further testing the role of biogenic silica in esophageal cancer.

Btr1-A Induces Grain Shattering and Affects Spike Morphology and Yield-Related Traits in Wheat.

Spike brittleness represents an important domestication trait in crops. Although the brittle rachis of wild wheat was cloned, however, the molecular mechanism underlying spike brittleness is yet to be elucidated. Here, we identified a single dominant brittle rachis gene Br-Ab on chromosome arm 3AbS using an F2 population of diploid wheat and designated Btr1-Ab. Sequence analysis of the Btr1-A gene in 40 diploid wheat accessions, 80 tetraploid wheat accessions and 38 hexaploid wheat accessions showed that two independent mutations (Ala119Thr for diploid and Gly97* for polyploids) in the Btr1-A coding region resulting in the nonbrittle rachis allele. Overexpression of Btr1-Ab in nonbrittle hexaploid wheat led to brittle rachis in transgenic plants. RNA-Seq analysis revealed that Btr1-A represses the expression of cell wall biosynthesis genes during wheat rachis development. In addition, we found that Btr1-A can modify spike morphology and reduce threshability, grain size and thousand grain weight in transgenic wheat. These results demonstrated that Btr1-A reduces cell wall synthesis in rachis nodes, resulting in natural spikelet shattering, and that the transition from Btr1-A to btr1-A during wheat domestication had profound effects on evolution of spike morphology and yield-related traits.

Vortex-induced dispersal of a plant pathogen by raindrop impact.

Raindrop impact on infected plants can disperse micron-sized propagules of plant pathogens (e.g., spores of fungi). Little is known about the mechanism of how plant pathogens are liberated and transported due to raindrop impact. We used high-speed photography to observe thousands of dry-dispersed spores of the rust fungus Puccinia triticina being liberated from infected wheat plants following the impact of a single raindrop. We revealed that an air vortex ring was formed during the raindrop impact and carried the dry-dispersed spores away from the surface of the host plant. The maximum height and travel distance of the airborne spores increased with the aid of the air vortex. This unique mechanism of vortex-induced dispersal dynamics was characterized to predict trajectories of spores. Finally, we found that the spores transported by the air vortex can reach beyond the laminar boundary layer of leaves, which would enable the long-distance transport of plant pathogens through the atmosphere.

Effects of chitosan nanoparticles on seed germination and seedling growth of wheat (Triticum aestivum L.).

To investigate the effect and mechanism of chitosan nanoparticles (CSNPs) on the germination and seedling growth of wheat (Triticum aestivum L.), we conducted systematic research on the impact of different concentrations (1-100 µg/mL) of CSNPs and chitosan (CS). The result of energy-dispersive spectroscopy (EDS) and confocal laser scanning microscopy (CLSM) showed that adsorption of CSNPs on the surface of wheat seeds was higher than that of CS. CSNPs had growth promoting effect at a lower concentration (5 µg/mL) compared with CS (50 µg/mL). In addition, the application of 5 µg/mL CSNPs induced the auxin-related gene expression, accelerated indole-3-acetic acid (IAA) biosynthesis and transport, and reduced IAA oxidase activity resulting in the increase of IAA concentration in wheat shoots and roots. The results suggest that CSNPs have positive effect on seed germination and seedling growth of wheat at a lower concentration than CS due to higher adsorption on the surface of wheat seeds.

Proteomic analysis for phenanthrene-elicited wheat chloroplast deformation.

The exposure of polycyclic aromatic hydrocarbons (PAHs) can cause wheat leaf chlorosis. Thus, we hypothesize that chloroplast inner structure damage is the reason for leaf chlorosis. This study was conducted with the wheat seedlings exposed to Hoagland nutrient solution containing 1.0 mg L-1 phenanthrene for 9 days. Subcellular observation showed that chloroplast turns round and loses its structural integrity. Herein, iTRAQ (isobaric tag for relative and absolute quantification) was applied to analyze the changes of protein profile in chloroplast exposed to phenanthrene. A total of 517 proteins are identified, 261 of which are up-regulated. Eight proteins related with thylakoid (the structural component of chloroplast) are down-regulated and the expression of related genes further confirms the proteomic results through real-time PCR under phenanthrene treatment, suggesting that the thylakoid destruction is the reason for chloroplast deformation. Four proteins related with envelope and stroma are up-regulated, and this is the reason why chloroplast remains round. This study is useful in discussing the carcinogenic and teratogenic effects of PAHs in plant cells in the environment, and provides necessary knowledge for improving crop resistance to PAH pollution.

In Vitro Wound Healing Activity of Wheat-Derived Nanovesicles.

Triticum aestivum plant extracts are often used as a natural healer in traditional medicine but which particles mainly have role in these processes are not scientifically proven. In other words, no attempts have been made to investigate the effects of wheat exosomes in regenerative medicine applications or drug development up to now. The current study was first time performed to demonstrate the activity of wheat exosomes in wound healing process using in vitro approaches. Although its fundamental wound healing process remains a mystery, in the current study, the efficiency of wheat grass juice-derived exosomes on cell viability and migration was examined. Increasing concentrations up to 200 µg/mL of the wheat exosome have yielded astonishing proliferative and migratory effects on endothelial, epithelial, and dermal fibroblast cells. RT-PCR analysis also showed collagen type I; mRNA levels were approximately twofold higher in expression after treating with 200 µg/mL wheat exosome. Additionally, Annexin V staining of apoptotic cells accompanied with the cell cycle analysis resulted with the reduction of the apoptotic cell number with no dispersion to the cell cycle analysis while plant exosomes have also increased tube-like structure formation of the endothelial cells. All in all, this research suggests a brand-new opening for skin wound healing therapy strategy by using wheat-derived exosomes due to its proliferative and migratory characteristics. Plant exosomes require a further research both clinically and in in vivo for wound healing drug development. Moreover, plant exosome therapy strategies would be safer and economical alternative for clinical wound healing.

Ice segregation in the crown of winter cereals: Evidence for extraorgan and extratissue freezing.

Meaningful improvements in winter cereal cold hardiness requires a complete model of freezing behaviour in the critical crown organ. Magnetic resonance microimaging diffusion-weighted experiments provided evidence that cold acclimation decreased water content and mobility in the vascular transition zone (VTZ) and the intermediate zone in rye (Secale cereale L. Hazlet) compared with wheat (Triticum aestivum L. Norstar). Differential thermal analysis, ice nucleation, and localization studies identified three distinct exothermic events. A high-temperature exotherm (-3°C to -5°C) corresponded with ice formation and high ice-nucleating activity in the leaf sheath encapsulating the crown. A midtemperature exotherm (-6°C and -8°C) corresponded with cavity ice formation in the VTZ but an absence of ice in the shoot apical meristem (SAM). A low-temperature exotherm corresponded with SAM injury and the killing temperature in wheat (-21°C) and rye (-27°C). The SAM had lower ice-nucleating activity and freezing survival compared with the VTZ when frozen in vitro. The intermediate zone was hypothesized to act as a barrier to ice growth into the SAM. Higher cold hardiness of rye compared with wheat was associated with higher VTZ and intermediate zone desiccation resulting in the formation of ice barriers surrounding the SAM.

Out-of-position telomeres in meiotic leptotene appear responsible for chiasmate pairing in an inversion heterozygote in wheat (Triticum aestivum L.).

Chromosome pairing in meiosis usually starts in the vicinity of the telomere attachment to the nuclear membrane and congregation of telomeres in the leptotene bouquet is believed responsible for bringing homologue pairs together. In a heterozygote for an inversion of a rye (Secale cereale L.) chromosome arm in wheat, a distal segment of the normal homologue is capable of chiasmate pairing with its counterpart in the inverted arm, located near the centromere. Using 3D imaging confocal microscopy, we observed that some telomeres failed to be incorporated into the bouquet and occupied various positions throughout the entire volume of the nucleus, including the centromere pole. Rye telomeres appeared ca. 21 times more likely to fail to be included in the telomere bouquet than wheat telomeres. The frequency of the out-of-bouquet rye telomere position in leptotene was virtually identical to the frequency of telomeres deviating from Rabl's orientation in the nuclei of somatic cells, and was similar to the frequency of synapsis of the normal and inverted chromosome arms, but lower than the MI pairing frequency of segments of these two arms normally positioned across the volume of the nucleus. Out-of-position placement of the rye telomeres may be responsible for reduced MI pairing of rye chromosomes in hybrids with wheat and their disproportionate contribution to aneuploidy, but appears responsible for initiating chiasmate pairing of distantly positioned segments of homology in an inversion heterozygote.

Physiological and metabolome changes during anther development in wheat (Triticum aestivum L.).

This study used cytology, cytochemistry, and non-targeted metabolomics to investigate the distribution characteristic of polysaccharides, lipids, and all the metabolites present during five wheat (Triticum aestivum L.) anther developmental stages to provide insights into wheat anther development. Anthers were collected from the tetrad through trinucleate stages, and 1.5% (w/v) acetocarmine and 4',6-diamidino-2-phenylindole staining were used to confirm the developmental stage and visualize the nuclei, respectively. Polysaccharides and lipids were detected by staining with periodic acid-Schiff and Sudan Black B, respectively. The integrated optical density of the tapetum and microspores were calculated using IPP6.0 software. Furthermore, the metabolites were identified by gas chromatograph system coupled with a Pegasus HT time-of-flight mass spectrometer (GC-TOF-MS). The results indicated that the interior and exterior surface cells of anthers are orderly. Pollen was rich in numerous nutrient substances (e.g., lipids, insoluble carbohydrates, and others), and formed a normal sperm cell that contained three nuclei, i.e., one vegetative nuclei and two reproductive nuclei in the mature pollen. Semi-thin sectioning indicated that the tapetum cells degraded progressively from the tetrad to late uninucleate stage and disappeared from the bi-to trinucleate stages. Moreover, nutrient substances (lipids and insoluble carbohydrates) accumulated, were synthesized in the pollen, and gradually increased from the tetrad to trinucleate stages. Finally, the metabolomics results identified that 146 metabolites were present throughout the wheat anther developmental stages. Principal component analysis, hierarchical cluster analysis, and metabolite-metabolite correlation revealed distinct dynamic changes in metabolites. The metabolism of organic acids, amino acids, sugars, fatty acids, amines, polyols, and nucleotides were interrelated and involved in the tricarboxylic acid (TCA) cycle and glycolysis. Furthermore, their interactions were revealed using an integrated metabolic map, which indicated that the TCA cycle and glycolysis were very active during anther development to provide the required energy for anther and pollen development. Our study provides valuable insights into the mechanisms of substance metabolism in wheat anthers and can be used for possible application by metabolic engineers for the improvement of cell characteristics or creating new compounds and molecular breeders in improving pollen fertility or creating the ideal male sterile line, to improve wheat yield per unit area to address global food security.