This study aims to explore the effect of deformation parameters on microstructure evolution during the new two-stage annealing method composed of an aging treatment (AT) and a cooling recrystallization annealing treatment (CRT). Firstly, the hot compressive tests with diverse deformation parameters were finished for an initial aged deformed GH4169 superalloy. Then, the same two-stage annealing method was designed and carried out for the deformed samples. The results show that the deformation parameters mainly affect the grain microstructure during CRT by influencing the content, distribution and morphology of the δ phase after deformation. The reason for this is that there is an equilibrium of the content of the δ phase and Nb atom. When the deformation temperature is high, the complete dissolution behavior of the δ phase nuclei promotes the dispersion distribution of the δ phase with rodlike and needle-like shapes during AT. Thus, the fine and heterogeneous microstructure is obtained after annealing because the recrystallization nucleation is enhanced in those dispersed δ phases during CRT. However, when the retained content of δ phase nuclei is high after deformation, the clusters of intragranular δ phases will form during AT, resulting in the pinning of the motion for dislocation. The elimination of the mixed grain microstructure is slowed down due to the low static recrystallization (SRX) nucleation rate within the deformed grain. GH4169 superalloy is a crucial material for modern energy and aerospace industries owing to its fine corrosion resistance and prominent mechanical properties under high temperatures ,3,4. Thu3 Nb) is often used to pin grain boundary movement and act as nucleation sites for recrystallization at high temperatures [Owing to the existence of meta-dynamic recrystallization (MDRX) and SRX behaviors during annealing, the annealing treatment could serve as an efficient method to eliminate the mixed grain microstructure ,19,20. Aeratures ,22,23,24eratures , it couleratures ,27. In aeratures . Hence, In this study, for an initial aged GH4169 superalloy, a set of hot deformation experiments with varying deformation parameters were finished, and the two-stage annealing including AT and CRT was subsequently performed. Afterward, the role of different deformation parameters on the microstructure evolution during deformation was analyzed. Finally, the effects of deformation parameters on the evolution of the δ phase and grain microstructure during the two-stage annealing including AT and CRT were investigated.A commercial GH4169 superalloy was utilized, and its detailed chemical compositions (wt.%) are shown in The microstructure was characterized by optical microscope (OM), electron back-scattered diffraction (EBSD) and scanning electron microscope (SEM) technologies. The detailed preparation approaches for OM, EBSD and SEM observation can be found in Refs. ,31. AddiFrom Deformed microstructures with different deformation temperatures are shown in As demonstrated in −1 to 0.01 s−1 and 0.001 s−1. In addition, for the microstructure deformed at the strain rate of 0.001 s−1, the content of the unbroken long needle-like δ phases was lower than that at the high strain rate, and the content of the spherical and rod-like δ phases was higher. This phenomenon is mainly due to the high strain rate prolonging the dissolution time of the δ phases. Meanwhile, in −1 to 0.01 s−1 and 0.001 s−1. In addition, the size of the DRX grains increased significantly with the decrease in the strain rate due to the prolongation of the recrystallized grain growth time at a low strain rate. Meanwhile, the average KAM value decreased with the reduction in strain rate  [Additionally, it can be seen that deformation temperature imposes significant influences on the morphology of the δ phase. For the sample with a deformation temperature of 950 °C, the δ phases formed during AT principally nucleate inside deformed grains and appear mainly in the form of clusters with rodlike morphology g. The re3 Nb) . TherefoAs shown in −1 to 0.01 s−1 and 0.001 s−1. Apparently, the content of the δ phase of specimens deformed at the strain rates with 0.01 s−1 and 0.001 s−1 after AT is almost the same. However, the morphology of the δ phase was significantly diverse at different strain rates. For the specimens with the strain rates of 0.1 s−1 and 0.01 s−1, the rodlike δ phase precipitated during AT occurred mainly in the form of clusters inside the deformed grains. This is because the specimens deformed at strain rates of 0.1 s−1 and 0.01 s−1 have a higher residual content of δ phases after deformation  are used. As reported by Chen et al. [i is the size of the grain i, and h value represents the more homogeneous microstructure.To further compare the microstructure after CRT with diverse deformation parameters, the average grain size  The precipitation and dissolution of small size intragranular-δ phase  after deformation have a great influence on the distribution of the δ phase during AT. When the dissolution of the intragranular-δ phase nuclei is complete under a high deformation temperature, the dispersion distribution of δ phase forms during AT. However, when the dissolution of the intragranular-δ phase nuclei is relatively weak compared to the precipitation behavior during deformation, the clusters of the intragranular δ phases will generate during AT.(2) The morphology of the δ phases forming during AT is strongly related to the retained content of the δ phases after deformation. The size of the δ phases will be large and the length of the δ phases will be long during AT when the retained content of the δ phases is low after deformation. The reason is that there is an equilibrium of the content of the δ phase and Nb atom. The Nb atom content in the matrix becomes high as the content of the δ phase decreases during deformation, resulting in more Nb atoms retained for the precipitation of the the δ phase during AT.(3) The distribution and morphology of the δ phase after AT have a more positive impact on the elimination of the mixed grain microstructure during CRT compared to the deformation energy. The reason is that the distribution and morphology of the δ phase play a leading role in the thermal motion of dislocation and grain boundary. The clusters of intragranular δ phases with spherical and rodlike shapes after AT hinder the dislocation movement within deformed grains during CRT, which is bad for the accumulation of dislocation. Thus, the recrystallization nucleation rate within deformed grains is low, thereby slowing down the elimination of the deformed grains. However, the dispersion distribution of the δ phase with rodlike and needle-like morphology after AT is good for the formation of high-density dislocation cells during CRT. Consequently, the annealed microstructure is uniformly refined due to the strong PSN effect of those large δ phases.(4) The refining mechanism for the deformed samples of diverse deformation parameters during annealing is different and can be summarized as follows: High deformation temperature can promote the scattered distribution of the δ phase during AT, thereby evenly refining the mixed grain microstructure to an average grain size of 6.52 μm after CRT. A large true strain can contribute to the occurrence of a large quantity of DRX nuclei and the precipitation of many small the δ phases. Hence, the growth of those DRX grains is efficiently pinned during CRT. Eventually, an even microstructure with an average grain size of 5.93 μm is obtained. Additionally, a high recrystallization degree and proper deformation energy can be retained in the microstructure with a suitable strain rate, which helps to refine the mixed grain microstructure to an average grain size of 8.63 μm after CRT.


The bread samples with a 2.5% and 5% addition had a more dense structure of the crumb pores. Regarding sensory evaluation, the bread samples with LGF addition in the wheat flour were well appreciated by the consumers. The addition also was desirable due to the fact that it supplemented bread with a greater amount of protein and minerals due to the composition of lentil grains. Therefore, LGF could be successfully used as an ingredient for bread making in order to obtain bread with an improved quality.The present study analyzed the effects of germinated lentil flour (LGF) addition at different levels in wheat flour , on dough rheological behavior, dough microstructure, and bread quality. Creep-recovery tests showed that the dough samples with high levels of LGF addition presented a higher resistance to flow deformability of the dough. Dough microstructure as analyzed using EFLM showed an increase in the protein area (red color) and a decrease in the starch (green color) amount with the increased level of LGF addition in the wheat flour. It was found that the LGF addition led to the improvement of the porosity, specific volume, and elasticity of the bread samples. The breads with LGF addition were darker and had a slightly reddish and yellowish tint. The bread textural parameters highlighted significant ( Currently, consumers demand an increasing amount of healthy food products . The conWorldwide, the most consumed food product in various forms is bread, for which the base raw material is wheat flour . AlthougLens culinaris Merr.) to the germination process for four days led to an increase in the amount of proteins and ash and a decrease in the amount of lipids and carbohydrates. Moreover, it was observed that due to germination, the amount of sodium, magnesium, iron, and zinc in the lentil grains increased [Lentils have an important place among vegetables. They are also an important source of protein in vegetarian diets . Accordincreased .While different studies have reported on the possibility of using lentil flour as an ingredient in the bread-making process , very feThe present study analyzed the effect of LGF addition in wheat flour on dough rheological behavior, its microstructure, and bread quality in terms of the physicochemical, textural, structural, and sensory characteristics. Thus, by highlighting the optimal level of LGF addition, it was possible to quantify its positive effect on the quality of the bread and, implicitly, its technological role in bread making.Lens culinaris Merr. lentils. For the germination of lentils, the following parameters were followed: a temperature of 25 °C, a constant humidity of 80%, and a time of germination of four days. Germination was performed in exclusive dark conditions according to the procedure described in previous study [The main raw material used for the bread-making process was soft wheat flour (type 650). This was purchased from a bakery factory . Germinated lentil flour (LGF) was obtained from us study . After tThe wheat flour and LGF were analyzed in accordance with ICC standard methods  as folloThe dough rheological properties in the case of all samples were analyzed by performing creep and recovery tests at 25 °C using a HAAKE MARS 40 rheometer . For these tests, a nonserrated parallel plate geometry with a diameter of 40 mm and a gap width of 2 mm was used. The samples were mixed using an Alveo Consistograph while taking into account the optimum Consistograph water absorption. After preparing the samples, they were placed in the rheometer plate. The samples were rested for 5 min before analysis for relaxation and for temperature stabilization. The determinations were made at constant shear stress of 50 Pa following a frequency sweep from 1 to 20 Hz in the range of linear viscoelasticity. The times for creep and recovery stage were 60 s and 120 s, respectively. The equations that describe the specific parameters for these determinations were previously described in .The dough samples with or without LGF addition were analyzed from the microstructural point of view using a Motic AE 31  equipped with LWD PH 203 catadioptric objectives (N.A. 0.4). The preparation of the samples and the epifluorescence light microscopy (EFLM) analysis were performed according to the method described in our previous study . Thus, aSaccharomyces cerevisiae, and water according to the water absorption capacity value of wheat-germinated lentil flours. LGF replaced white wheat flour in various proportions that were previously specified. The amount of water used was determined according to the values of water absorption that were determined using the Alveo Consistograph. The water additions were as follows: 54.3% for the control sample and 53.7%, 53.1%, 52.6%, and 52.2% for the samples with 2.5%, 5%, 7.5%, and 10% LGF addition, respectively, in the wheat flour. To prepare the bread samples, the following steps were followed: dosing of the ingredients, mixing, dividing the dough obtained, leavening the dough, and baking the samples. A heavy-duty mixer  was used to mix the ingredients. The ingredients were mixed for 15 min. After kneading, the dough obtained was divided into three parts of 400 g each. The fermentation of the samples resulting from the modeling step was done in a fermentation chamber  at a temperature of 30 °C for 60 min at 85% relative humidity. The samples were baked in an electrical bakery convection oven with steam production, ventilation, and humidification  at a temperature of 220 °C for 30 min using the steam system of the oven in the first 2 min of baking.The ingredients used to prepare the bread samples were: white wheat flour (type 650), LGF , 1.5% sodium chloride, 3% compressed yeast of The physical characteristics of the bread samples without and with LGF addition were analyzed by determining the values for the specific volume, porosity, and elasticity of the samples. The rapeseed displacement method (AACC Method 10–05.01)  was usedL* (darkness/brightness), b* (shade of blue/yellow), and a* (shade of red/green). These determinations were based on the CIE Lab* color system and were made using UV–vis technology. For these analyses, before starting the determinations, the colorimeter was calibrated by scanning the standard white surface calibration plate , which used the standard illuminant D65 (working in daylight) and a 0° viewing angle.The color characteristics (for crumb and crust of the bread samples) were determined using a Konica Minolta CR-400 colorimeter . With the help of this colorimeter, the values for the following parameters were measured: A TVT-6700 texturometer device  was used to determine the textural characteristics of the bread samples. For the determination, the texturometer used was equipped with a 10 kg load cell. Slices of bread of that were 50 mm tall were used for the determination. These were subjected to two compression cycles to up to 20% of their initial height. The analysis was conducted using a 45 mm cylindrical probe. The protocol used was: a trigger force of 5 g, a speed of 1.0 mm/s, and a recovery period between compressions of 15 s. The textural parameters that were measured in this test were: firmness, gumminess, cohesiveness, and resilience.To analyze how the addition of LGF influenced the crumb structure of the bread samples, a Motic SMZ-140 stereo microscope  was used at a resolution of 2048 × 1536 pixels with a 20× objective.The characteristics that were taken into account in the sensory analysis were: appearance, color, aroma, taste, smell, texture, and global acceptability. The sensory analysis of the bread samples was conducted with the help of a panel of 20 semitrained judges using a 9-point hedonic scale from 1 to 9 in which 1 = dislike extremely, 5 = neither like nor dislike, and 9 = like extremely. At the end, the arithmetic average was calculated for the points awarded and the average score for each judge was established.Statistical processing of data obtained from specific determinations for all bread samples was done using the Statistical Package for Social Science . Data were expressed as the mean ± standard deviation and a one-way analysis of variance (ANOVA) was used with a Tukey’s test at a level of 5% of significant differences.The wheat flour presented the following characteristics: 0.66% ash content, 14.6% moisture, 1.12% fat, 12.3% protein, 30.4% wet gluten, and a 3 mm gluten deformation index. The value of the falling number index was 356 s. According to these data, the white wheat flour had a low α-amylase activity and was one of strong quality for bread making . LGF pre2 > 0.97) to the Burger’s model, the parameters for which are presented in Co parameter. At an LGF addition of 10%, the value of this parameter significantly increased (p < 0.05) but did not exceed that for the control sample. In general, the creep compliance  values decreased with an increased addition of LGF. Only at an addition of 10% LGF in the wheat flour did the value of these parameters significantly increase (p < 0.05), but without exceeding the values for the control sample. In the case of the λC parameter (creep retardation time), we observed that its value was significantly increased (p < 0.05) by an addition of 5% LGF, then its value significantly decreased (p < 0.05) but was not lower than in the case of the control sample. Regarding the μCo parameter, we observed that it generally registered a higher value for the samples with LGF addition.The experimental data for creep compliance were well adjusted (Rp < 0.05) depending on the LGF level addition in the dough sample. Considering the sample without LGF addition, the value of the λR parameter was lower in the case of the addition of 5% and 10% LGF and higher in the cases of an addition level of 2.5% and 7.5% LGF. The Jr/Jmax ratio had higher values for the bread containing LGF.In the case of the recovery test we noticed that the compliance parameter was significantly influenced (p < 0.05) with an increasing level of LGF and reached a maximum value at 7.5% LGF incorporated in the bread recipe. At an addition level of 10% LGF, the specific volume of the bread samples decreased but was not lower than in the case of the sample without LGF addition in the bread recipe. Regarding the porosity of the samples, it can be seen in our data that the addition of LGF significantly improved (p < 0.05) this parameter’s value. At an addition level of 10% LGF in wheat flour, the value of the porosity began to decrease significantly (p < 0.05). The bread elasticity was improved due to the LGF addition even at an addition level of 7.5% LGF in the wheat flour.The physical characteristics of the bread samples used in L* parameter (darkness/brightness) significantly decreased (p < 0.05) due to the addition of LGF in the bread recipe, which meant that the samples with LGF addition had a darker crust. The same trend could seen for the bread crumb. Regarding to the a* parameter (shade of green/red), we observed that its value was significantly increased (p < 0.05) due to the LGF addition, both in the case of the crust and the crumb. This meant that the LGF addition gave the bread samples a slightly reddish color. The b* parameter (shade of blue/yellow) showed a significantly increasing value (p < 0.05) as the LGF addition value increased, which meant that the yellow hue of the bread samples was intensified due to the addition of LGF in the bread recipe.p < 0.05) with an increase in the level of LGF in the wheat flour. The gumminess parameter significantly increased (p < 0.05) at an addition level of 2.5% and 5% LGF. Then, its value decreased but did not go lower than in the case of the control sample without LGF addition. The cohesiveness of the bread samples showed a decreasing trend as the level of LGF addition increased. The resilience also decreased with an increasing level of LGF in the wheat flour.In general, the addition of LGF in the bread recipe was well appreciated by panelists. In Co parameter could be correlated with a significant (p < 0.05) decrease in the instantaneous elasticity of the dough. The lowest value for the JCo parameter was obtained in the case of the incorporation of 7.5% LGF in the wheat flour. An increase in the value of the λC parameter indicated the fact that the retarded elastic creep took place more slowly. A lower value of the λR parameter at an addition level of 5% and 10% LGF compared to the control sample highlighted the fact that the retarded elastic recovery took place more rapidly for these samples. A higher value for the μCo parameter for the samples with high levels of LGF addition indicated a higher resistance to flow deformability by the dough compared to the control sample. However, the sample with a 2.5% LGF addition in the wheat flour had a lower value for the μCo parameter, which meant that it presented a lower opposition to deformation. In general, the effect of the LGF addition in the wheat flour on the dough behavior could be explained by the dilution of gluten, which changed its elasticity and viscosity. The Jmax parameter presented a decreasing trend due to the LGF addition in the dough recipes. This trend was similar to those reported in other studies that concluded that the addition of 15% and 20% roasted chickpea flour in the dough recipe had the effect of decreasing the Jmax value [In the case of the creep test, the increase in the Jax value .Ro parameter. The fact that the value of the compliance elasticity during the recovery parameter was higher indicated a higher recoverable energy. This recoverable energy was stored by more cross-linked gluten than in the case of the sample without LGF addition in the dough recipe. This meant that with the LGF addition, the aggregation between gluten proteins increased. The observed increase in the values for the λR parameter for the samples with LGF addition meant that the retarded elastic recovery of dough was slower due to the LGF addition in the wheat flour. The Jr/Jmax ratio provided information on the elasticity of the dough samples. According to Moreira et al. [The results of the recovery test highlighted the effect of LGF addition on the dough elasticity. According to our data, the control sample had the lowest elasticity while the sample with 10% LGF addition had the highest elasticity and presented the highest value for the Ja et al. , the incAs the EFLM images show, the control sample without LGF addition had the highest amount of green area, which indicated a high starch amount. The samples with the LGF addition in the dough system were highlighted by slightly different EFLM images due to a more nuanced red coloration specific to rhodamine B that was correlated with the amount of protein in the system. Thus, the EFLM images showed a higher protein content and a lower amount of starch once the LGF addition level was increased. This was expected due to the fact that white wheat flour contains a much lower amount of protein than lentils . MoreoveThe incorporation of LGF in the technological process of bread making at a level of 2.5–10% in the wheat flour influenced each physical parameter of the bread samples. The improvement in specific volume was similar to the porosity and elasticity increases. This was desirable because these parameters are indicators of bread quality. According to other studies, the use of different leguminous flours as ingredients in bread making decreased these values . However2 [Saccharomyces cerevisiae baking yeast. Moreover, amylases also influenced the process of slowing the staling of bread [p < 0.05) than that of the control sample. The significant decrease (p < 0.05) in the bread sample with 10% LGF addition may have been due to a slight dough weakening as a consequence of the gluten amount decreasing in the dough system while taking into account that the LGF did not contain gluten.Regarding the improvement in the bread volume due to the addition of LGF, we can state that this was due to the changes that occurred during germination in the gelatinization of starch and in the aggregation of proteins. Thus, in the germination process there was an increase in protein solubility that coincided with the improvement in the ability of the lentil flour to foam and emulsify. The germination process led to an increase in the proteolytic activity, which led to the degradation of the lentils’ storage proteins. This led to shorter peptides and free amino acids. As a result, the solubility of the proteins increased . The cha2 . The fin2 . In addi2 , becauseof bread . Similarof bread . Accordip < 0.05) in the bread porosity due to the LGF addition may be explained by the fact that the capacity to produce and retain gases in the dough proofing stage increased, leading to a denser porous structure. Similar results in the sense that the germination process could be successfully used to improve the porosity of the bread were reported by Cardone et al., who concluded that the germination process under controlled conditions for durum wheat grains had the effect of improving the quality characteristics of bread of volume and porosity [p < 0.05) in porosity values at high levels of LGF addition in the wheat flour may be attributed to the decrease in the amount of gluten from the dough matrix. Due to its substitution with LGF flour, there was a decrease in the amount of gluten in the dough system. This also led to the weakening of the dough network, which may have presented a lower capacity to form pores in the proofing stage and a lower capacity to retain carbon dioxide in the dough system.The addition of LGF in the bread-making recipe had the effect of improving the porosity of the bread. However, after exceeding a 7.5% level of LGF addition, the porosity began to decrease slightly but did not fall below the value for the control sample. The significant increase (porosity . The sigp < 0.05) in the elasticity value of the bread samples can be explained by the presence of the enzyme α-amylase in a larger amount when the addition level of LGF in the bread recipe was increased. These data were similar to those from another study that concluded that the amylolytic enzymes improved the physical characteristics of bread and the viscoelastic properties of wheat dough [Regarding the elasticity value, in this study we observed that the LGF addition led to an improvement in this parameter even at an addition level of 7.5% in the wheat flour. The significant increase (at dough .p < 0.05) in the L* parameter value with an increased level of LGF in the wheat flour indicated a darkening of the color of the bread. This can be attributed to the fact that the LGF contained a higher amount of protein than the white wheat flour. Similar data were also reported by Xing et al., 2021, who concluded that protein addition in wheat bread had the effect of darkening the bread [a* (shade of green/red) increased in the cases of the crust and of the bread crumb, which indicated that the samples acquired a slightly reddish hue. This was particularly due to the specific pigments in the red lentils used as an addition in this study. Studies have shown that red lentils contain a significant amount of carotenoids [b* (shade of blue/yellow) due to the LGF addition indicated that it had given a yellow hue to the bread samples. This was due to the increase in the amount of protein in the nutritional composition of the bread once the addition level of LGF had increased. Along with sugars, amino acids are precursors for Maillard reactions [The incorporation of LGF in the bread making changed all three color parameters (for the crust and for the bread crumb). The significant decrease  for the gumminess parameter compared to the control sample may be explained by the fact that the LGF addition introduced an additional amount of fiber. A higher amount of fiber led to an increase in the gumminess of the bread, as was reported in a previous study [The firmness parameter of the bread samples was influenced by the LGF addition in the bread recipe. Its value decreased with an increase in the level of LGF addition in the wheat flour. This can be explained by the fact that once the quantity of proteins in the bread matrix had increased, the interactions between them and the starch changed . The incus study . Higher us study . The decus study .2 and a denser distribution of pores. An uneven pore distribution can be attributed to the substitution of gluten in the dough matrix due to LGF incorporation, which led to a lower capacity of the dough to retain the released gas [The crumb structure of bread samples is an important sensory feature for consumers. The addition of LGF in the wheat flour modified both the structure of the pores and their density. The change in the structure of the pores could be explained by the fact that the LGF addition in the wheat flour modified the gluten matrix: the dough had a lower capacity for gas expansion during the proofing and prebaking stages. Another explanation could be the fact that a larger amount of fiber due to the LGF addition led to a decrease in the bread’s ability to retain gases in the dough matrix . A denseased gas .The sensory characteristics of the bread samples with the LGF incorporation in the wheat flour were more appreciated by the panelists. The appearance of the bread was better evaluated by the panelists, probably due to the fact that LGF addition in the bread recipe improved the loaf volume and porosity. The color of the bread became darker with the increase in the level of LGF addition in the wheat flour, which led to the idea of wholemeal wheat bread correlated by the panelists with a superior nutritional value compared to the white wheat flour. The LGF addition in the wheat flour also intensified the Maillard reaction, which provided more copper-colored samples that were more pleasing visually. Regarding the taste of the samples, we can state that it was improved because, as already highlighted in other previous studies, germination had the effect of improving the taste and aroma profile of the grains subjected to this process. During germination, there was a release of the reducing sugars and amino acids from the grains. In the bread-baking stage, they reacted and formed melanoidins . In addiC) and a lower elastic deformation at constant stress (J) for dough samples with LGF addition. The EFLM images showed a homogeneous, compact dough matrix for all the analyzed dough samples even when high levels of LGF were incorporated in the dough recipe. The bread quality was improved by LGF addition in the bread recipe. The specific volume, porosity, and elasticity all presented higher values for bread samples with LGF addition compared to the control sample. The color parameters indicated a darkening of the bread crumb and crust via a decrease in the L* value and an increase in the b* and a* values. For all the bread samples with LGF incorporated in bread recipe, the textural parameters of firmness and gumminess presented higher values compared to the control sample, whereas the cohesiveness and resilience presented lower values. The bread crumb structure with LGF incorporated in the bread recipe presented changes, especially for the bread samples in which high levels of LGF were incorporated in the wheat flour. When considering the results of the sensory analysis, we concluded that the bread samples with LGF addition in the bread recipe were better appreciated than the control sample. When taking into account the results obtained for the dough samples with different levels of LGF addition and also those for the bread samples, a value of 7.5% is recommended as the optimal level for LGF addition in bread recipe. This LGF addition level had the best overall acceptability. According to our study, LGF proved to be a valuable functional ingredient for food producers that provide natural bakery products without other additives or enzymatic addition and of a very good quality. In addition, it may improve the nutritional quality of bakery products due to the nutrients provided by LGF.The germinated lentil flour addition in wheat flour led to significant changes in the dough microstructure, rheological behavior, and bread quality. Depending on the percentage of LGF addition in the wheat flour, its influences on dough rheology, bread quality, and consumer acceptability were different. Creep and recovery tests showed a higher viscosity at steady state (μ


The p53 protein family is the most studied protein family of all. Sequence analysis and structure determination have revealed a high similarity of crucial domains between p53, p63 and p73. Functional studies, however, have shown a wide variety of different tasks in tumor suppression, quality control and development. Here we review the structure and organization of the individual domains of p63 and p73, the interaction of these domains in the context of full-length proteins and discuss the evolutionary origin of this protein family.Distinct physiological roles/functions are performed by specific isoforms.The non-divided transactivation domain of p63 has a constitutively high activity while the transactivation domains of p53/p73 are divided into two subdomains that are regulated by phosphorylation.Mdm2 binds to all three family members but ubiquitinates only p53.TAp63α forms an autoinhibited dimeric state while all other vertebrate p53 family isoforms are constitutively tetrameric.The oligomerization domain of p63 and p73 contain an additional helix that is necessary for stabilizing the tetrameric states. During evolution this helix got lost independently in different phylogenetic branches, while the DNA binding domain became destabilized and the transactivation domain split into two subdomains.Is the autoinhibitory mechanism of mammalian TAp63α conserved in p53 proteins of invertebrates that have the same function of genomic quality control in germ cells?What is the physiological function of the p63/p73 SAM domains?Do the short isoforms of p63 and p73 have physiological functions?What are the roles of the N-terminal elongated TAp63 isoforms, TA* and GTA? The world of the tumor suppressor p53 was firmly anchored within the field of cancer biology but in the late 1990s this exclusive connection was questioned by the discovery of two proteins, p73 –5, with For p73 the assignment of physiological functions to well defined isoforms is less clear. In the brain both TAp73 and ΔNp73 isoforms are often expressed in the same neuronal cell types . In genRecent progress in our understanding of the specific conformations adopted by different isoforms have advanced our insight into the regulation of their distinct functions. Here we review the structures of the individual domains, their structural and functional interplay with the sequences surrounding these domains and discuss a potential evolutionary path. As the conformation—function relationship is better understood for p63 than for p73, a focus of this review will be on p63. As the structure and function of p53 domains have been reviewed many times –50, we iOf all domains of p63 the N-terminal TA domain shows the lowest sequence identity to p53 but also to p73. Structurally the TA domains of all three family members are intrinsically unfolded when isolated , 52, buto relative to each other. Since the TA domain of p63 is longer by one helical turn it forms additional hydrophobic contacts with the Taz2 domain.In p63 residues 8–25 form a single long helix that comprises the entire TA domain and that combines elements of both p53 subdomains . No furtOf all three family members the affinity of the TA domain of p73 to the Taz2 domain is the lowest, but can be enhanced by phosphorylation of T14 . SimilarIn addition to interaction with domains of p300 and CBP the TA domains of all three family members also bind to the N-terminal domain of the E3 ligase Mdm2 (Mouse double minute 2 homolog). This interaction is very well characterized for p53 where it leads to ubiquitination via Mdm2’s RING finger domain and proteasomal degradation . For thip53, p73 and p63 share the highest sequence identity in their DBD reaching 65%. Despite this high sequence conservation transcriptomic analysis has revealed differences in the genes regulated by the three proteins –92. FurtThe structure of the p63 DBD has been solved in complex with DNA with both a 10-bp DNA half-site response element (5′AAACATGTTT3′) as well as with a 22-bp DNA full response element containing a 2-bp spacer between both half-sites (5′AAACATGTTTTAAAACATGTTT3′) . Both DNIn principle, the OD ensures that all family members interact with the DNA as tetramers, thus increasing binding affinity . In addition, the individual DBDs can interact with each other using different interfaces. Cooperative binding of two DBDs to one half site is known from p53 where R181 and E180 in the H1 α-helix form two salt bridges between two monomers  to whichCrystal structures of the p53 DBD bound to various DNA oligomers have shown that protein-protein contacts also exist between two dimers, thus forming an additional tetramerization interface , 107, thThe structure of the isolated DBD has been solved by liquid state NMR spectroscopy  as well Crystal structures were also published for the isolated p73 DBD , its com2:p732 tetramer being the thermodynamically most stable state (even more stable than both homo-tetramers) [The ODs of all three members of the p53 family are dimers of dimers , 120. Thtramers) , 122. Thtramers) . Since ptramers) , 46. Thetramers) , 122.Fig10-helix; Fig. The SAM domain which forms a compact structure consisting of five helices  syndrome , 141. PaThe α-C-terminus of p63 contains a functional domain (amino acids 597–614) that is important for regulating the transcriptional activity of TAp63α . The pepp73 contains a very similar sequence with a classical sumoylation site at its C-terminus (IKEEFT) as well. This site was shown to be the main modification site with SUMO in p73α by PIAS-1  and sumoThe TI domain of p63 also plays a role in patients with the AEC syndrome . The TI domains of both p63 and p73 have a low intrinsic aggregation propensity . Some ofCcgn1 (p21) or Bax [K14 [ΔNp63α is an isoform that is expressed from a promoter situated upstream of exon 3′. Due to the shifted start site the N-terminal 69 residues of TAp63α are replaced with 14 unique amino acids . The lac) or Bax . TranscrBax [K14 , 160. ThBax [K14 –166. ChIBax [K14 , 167–174Bax [K14 . To fulfBax [K14 . This taBax [K14 . SimilarBax [K14 .TAp63α is highly expressed in mammalian oocytes that are arrested in prophase of meiosis I , 32. In Activation of this closed dimeric state to the open tetrameric state that can bind to DNA and initiate a pro-apoptotic program  requiresThis irreversibility of the activation paired with the importance of the process  requires tight regulation. Surprisingly, only Chk2 is activated (by ATM) while CK1 is thought to be a constitutively active kinase . This woMost likely, the original role of TAp63α was to eliminate all oocytes that had not repaired the Spo11 induced DNA DSBs at the beginning of the dictyate arrest phase. However, TAp63α remains expressed in oocytes throughout the entire dictyate arrest phase and DSBs caused by other processes during that period result in activation of TAp63α. This is in particular a problem for patients treated for cancer or autoimmune diseases with chemotherapy or irradiation , 188–193The high sequence identity between the p63 and p73 TI domains suggested that TAp73α might also adopt a closed and only dimeric conformation. Analysis of the oligomeric state, however, revealed that all p73 isoforms are constitutively tetrameric . Domain TAp63α is not the longest p63 isoform. Using a start site in exon 1, TA*p63α has an N-terminal extension of 39 amino acids . BiophyA similar isoform is GTAp63α. It differs from TA*p63α by replacing the first 21 amino acids with 19 unique residues . Both isFor p73 similar N-terminal extensions have so far not been reported.Trp73Δ13/Δ13) suffers from severe hippocampal dysgenesis, reduced synaptic functionality and impaired learning and memory capabilities caused by the depletion of CR cells [The β-isoforms of p63 and p73 are created by alternative splicing at the 3′ end of the p63 mRNA which removes at the protein level the SAM domain except for the three N-terminal amino acids of the first α-helix. This splicing also removes the inhibitory TI domain and thus makes all β-isoforms constitutively tetrameric. While no physiological functions for the p63β proteins are known , the TACR cells . In contCR cells .The γ-C-terminus is created by alternatively splicing of exon 10 to the γ-specific exon 10′, thereby adding 38 γ-specific residues directly C-terminal to the OD . For p63C. elegans, Cep-1 [The identification of p53 homologs in short lived invertebrates has sparked the interest in the evolution of the p53 family , 207–212s, Cep-1 , 214, foC. elegans p53 homolog Cep-1 support the hypothesis of the germ cell quality control by p63 being the evolutionary origin. These investigations have shown that the DBD is again very similar to the DBDs of p53 and p63 with a similar DNA sequence preference [This hypothesis also means that the function of TAp63α in oocytes evolutionary precedes the function of ΔNp63α in epithelial stem cells. Structure determination of the folded domains of the eference . The OD,eference . Insteadeference .Fig. 8StDrosophila melanogaster, each secondary structure element of the p53 OD is doubled: each monomer not only comprises an additional C-terminal helix as is the case in the ODs of p63 and p73, it contains also an additional N-terminal β-strand  [While the existence of the N-terminal β-strand in the Dmp53 OD and the SAM domain at the C-terminus of the Cep-1 OD are so far unique features of these proteins, a second helix within the OD has been found in many other members of the p53 family . Vertebr fishes) . As the  fishes)  and repl fishes) –226. Sta fishes) .Ciona intestinalis [Gene duplication and evolutionary loss of the second helix occurred also in invertebrates. Structural information on the OD is available for the two p53 family members found in stinalis , 227, anstinalis . The C.istinalis . This sestinalis : the N-tstinalis . Interesstinalis . As heli


Nuclear m6A reader protein YTHDC1 plays a significant role in RNA metabolism including some non-coding RNA such as LINE or circRNA. It is also known to regulate mRNA splicing through recruiting SRSF3 to the targeted mRNAs, which then mediates export of YTHDC1-bound RNA to the cytoplasm. Additionally, it has been indicated that SRSF3 binding to YHTDC1 may be mediated by its dephosphorylated status. However, their binding mechanism, including the positions of dephosphorylated residues of SRSF3, has not been sufficiently investigated. Thus, we explored the mechanism of interaction between SRSF3 and YTHDC1 in human cells. We used co-immunoprecipitation to examine the binding of YTHDC1/SRSF3 through their N- and C-terminal amino-acid residues. Furthermore, dephosphorylation-mimic serine to alanine mutants of SRSF3 indicated the position of phosphorylated residues. Cumulatively, our results demonstrate that YTHDC1 binding to SRSF3 is regulated by not only hypo-phosphorylated residues of arginine/serine-rich (RS) domain of SRSF3 but also other parts of SRSF3 via YTHDC1 N- or C-terminal residues. Our results contribute to the understanding of the complex mechanism of binding between SR protein SRSF3 and the m6A reader YTHDC1 to regulate the expression of mRNA and non-coding RNAs.N6-methyladenosine (m Recent studies have identified multiple components belonging to these categories. METTL3 and METTL14 are core components of “writers” and add methyl residues at position N6 of adenosine, together with the accessory components WTAP, VIRMA, RBM15, and ZC3H13 [6A by the YT521-B homology (YTH) domain, and subsequently direct RNA metabolism. In addition to YTH domain family proteins, other proteins, including several heterogeneous nuclear ribonucleoproteins, have been known to work as “readers” [In md ZC3H13 ,17. The d ZC3H13 ,18. “Reareaders” ,19. The readers” . For insAmong YTH domain-containing family proteins, only YTHDC1 localizes in the nuclear compartment and has significant roles in splicing, export of RNA, reshaping nuclear speckles with MALAT1, or regulation of scaffold function of LINE1 ,23,24,25As for SRSF3, it has been reported that SRSF3 has great influence on various type of cancers via splicing regulation ,27,28,292.The HeLa cell line was acquired from RIKEN Bioresource Center and was cultured in Dulbecco’s modified Eagle medium , with 4.5 g/L glucose supplemented with 10% fetal bovine serum , 100 mg/mL penicillin, and 100 U/mL streptomycin (Wako) at 37 °C in 5% COSRSF3 or YTHDC1 expression vectors were constructed from the pReceiver vector (Genecopoeia) and PCR amplified SRSF3 and YTHDC1 sequences from extracted Hela mRNA. The SRSF3 serine-to-alanine mutant vector was constructed as previously described ,31. Eachg for 15 min. The supernatant was incubated with RNase A (1 mg/mL)  for 40 min at 20 °C and subsequently cleared by centrifugation. The lysate was then incubated with rabbit anti-Myc or anti-GFP antibodies  with gentle rotation overnight at 4 °C. Agarose G beads  were added and rotated for 2 h at 4 °C. Beads were washed with a lysis buffer six times to remove non-specific proteins. After washing, beads were boiled in 2 × SDS loading buffers. The immunoprecipitants were analyzed by SDS-PAGE/western blot analysis.Co-immunoprecipitation was performed as previously described  with minTo elucidate the phosphorylation status, we used phos-tag SDS-PAGE, which separates phosphorylated and non-phosphorylated proteins, based on their levels by capturing phosphate residues. Phos-tag gels were self-made using Phos-tag acrylamide reagent . Cell lysates or precipitates from co-immunoprecipitation assays were used for western blotting. Each sample was mixed with a Laemmli sample buffer , resolved on SDS-PAGE or phos-tag SDS-PAGE, and transferred to PVDF membranes. Protein-bound membranes were blocked with a 5% skim milk buffer and incubated with a primary antibody diluted in 0.5% skim milk buffer. Mouse anti-Myc antibody  and mouse anti-GFP antibody  were used as primary antibodies. Horseradish peroxidase-conjugated anti-mouse IgG antibody was used as the secondary antibody . The membrane was subsequently washed and developed with ImmunoStar Zeta chemiluminescent reagents  and chemiluminescence was visualized using the Fusion .We first focused on the arginine/serine-rich (RS) domain of SRSF3 as this contains two serial RS repeats similar to those of SRSF1 a. As witNext, their phosphorylation states were elucidated by western blotting using phos-tag gel SDS-PAGE c, right.We next explored the position of serine phosphorylation on the RS domain of SRSF3 as previously described in . SRSF3 pFrom the phos-tag analysis a, resultTo reveal the multiple phosphorylation status of SRSF3, we then examined the phosphorylation status with multiple replacement mutants a. As a rBased on phos-tag gel electrophoresis, it was suggested that phosphorylation occurred on SRSF3, at positions 108, 115, and near 124–130. Furthermore, hyperphosphorylation from replacement at positions 154–160 disappeared when N-terminal phosphorylated serine residues were replaced with alanine. Thus, the replacement of the C-terminal serine residues implied that the hyperphosphorylated site existed at positions 108, 115, and/or 124–130. Therefore, we concluded that endogenous SRSF3 was phosphorylated at positions 108, 115, and near 124–130.Next, to verify the SRSF3 binding sites of YTHDC1 a, we perTherefore, we examined SRSF3 binding with the C-terminal fragments of YTHDC1 . We predThese results posed questions concerning how the described N-terminal YTHDC1 binding to RS domain of SRSF3 had an effect on the interaction between YTHDC1 and SRSF3, or which sites of RS domain were used for binding. Thus, to clarify these questions, we generated constructs comprising the SRSF3 RRM with the N-terminal or C-terminal half of RS domain, SRSF3(1–132) or SRSF3(Δ84–132), respectively. In addition, we also constructed an SA mutant, as for SRSF3(1–132), where mutations were inserted at positions 108, 115, and 124–130.From the results of co-immunoprecipitation and western blotting, we confirmed YTHDC1 binding to SRSF3(1–132) and the SA mutant a. The 1-Investigations also confirmed the binding of YTHDC1(1–353) to the SRSF3(1–132) SA mutant . However6A reader proteins, various associators of YTH proteins have been found to be related to processes in RNA metabolism, such as mRNA translation or RNA stability. However, as initial investigations focused on YTHDF2 in the cytoplasm, the described physiological functions of the YTH proteins have mainly been cytoplasmic [6A binding protein YTHDC1 remains largely unknown. However, YTHDC1 is known to affect alternative splicing [Following the identification of moplasmic . Thus, tsplicing ,36 and hsplicing , and sevsplicing ,23, wherIn this study, we focused on the binding between SRSF3 and YTHDC1. Previously, the RS domain of SRSF3 was reported to be phosphorylated ; the phoNext, we investigated the phosphorylated serines in succession using serine to alanine replaced mutants. Based on phos-tag gel electrophoresis, it was supposed that phosphorylation was induced at positions 108, 115, and near 124–128. Unexpectedly, the SA mutants in the C-terminal half of the RS domain displayed hyperphosphorylation. Therefore, at least, the C-terminal half of the RS domain was not supposed to be phosphorylated, but different from result of C-terminal RS domain fragment.Subsequent phos-tag analysis with multiple replacement mutants specifically identified the phosphorylated serine residues and demonstrated that phosphorylation of SRSF3 occurred only within the N-terminal RS domain, and hyperphosphorylated residues came from the C-terminal SA replacement and existed at the N-terminal half of the RS domain. Therefore, it was suggested that the C-terminal RS domain had potential to be phosphorylated but phosphorylation was restricted by the N-terminal half of the RS domain, and might have another role, such as a scaffold for CLK1, as in the case of SRSF1, in regulating phosphorylation in the N-terminal half of the RS domain . In relaThe RS domain was successfully co-immunoprecipitated with Myc-tagged YTHDC1, consistent with previously published data . UnexpecSurprisingly, the results of co-immunoprecipitation with YTHDC1(493–727)-truncated protein resembled that of the full-length YTHDC1 protein, indicating that the C-terminal region of YTHDC1, which had an arginine-rich sequence, may bind to SRSF3, even though this lacks a methylated RNA-binding YTH domain. Although the results from the immunoprecipitation of YTHDC1(493–727) were unexpected, it might be possible since the arginine-rich sequence of YTHDC1 contains a phosphorylation that mimics RS repeat-like sequence, SRPRERDRERERD, and it has been reported that some of the RS domains can interact with each other .In addition, the interaction between RRM and RS domain or RS repeats like sequence is plausible since SRSF1 shows an intra-molecular interaction between RRM and RS domain . The ful6A with an adjacent SRSF3 binding site and subsequent RNA splicing. As the binding conformation with SRSF3 was shown to be different between N-terminal YTHDC1 and C-terminal YTHDC1 regions, m6A bound YTHDC1 may direct mRNA-binding hypo- or de-phosphorylated SRSF3, which has already completed the splicing reaction and reside at proper position for NFX1 binding, resulting in mRNA export.Additionally, phosphorylated SRSF3 was precipitated to a minor extent by co-immunoprecipitation by YTHDC1 . These r6A modification machinery and the effect on oncogenic factors, such as BRCA2 or PGR [6A-deposited RNA by SR protein, and not all SR protein expression is reported to be controlled by m6A modification. With the exception of SRSF1, the phosphorylated sites or mechanisms for SR proteins are mostly unreported. Therefore, it is possible that part of the proteomic diversity in cancer cells may be regulated by unknown mechanisms that derive from m6A- deposited RNA splicing or export with YTHDC1 or SR proteins, including their various phosphorylation statuses.Several studies have reported the difference in YTHDC1 expression in a variety of cancers ,43,44. A2 or PGR , it has 2 or PGR . Thus, Y2 or PGR ,28,29,466A by SR proteins or proteins that possess RS repeats.In conclusion, the RS domain of SRSF3 plays a role in both splicing and nuclear export via interactions with the N- or C-terminal sequence of YTHDC1. In addition, the SRSF3 RS domain and RRM domain were revealed to interact with YTHDC1 through its C-terminal region. We have clarified the phosphorylation sites of SRSF3 and provided a mechanistic basis for the binding between YTHDC1 and SRSF3. Previous reports did not indicate a preference in splicing regulation with YTHDC1-interacting SR proteins, such as SRSF1 or SRSF7, since the phosphorylation status of the RS domain or similar sequences was not completely elucidated; therefore, further investigation should be performed to reveal the regulation of m


RING  zinc finger (RING-zf) proteins belong to an important subclass of zinc fingers superfamily, which play versatile roles during various developmental stages and in abiotic stress responses. Based on the conserved cysteine and histidine residues, the RING-zf domains are classified into RING-HC (C3HC4), RING-H2 (C3H2C3), RING-v, RING-D, RING-S/T, RING-G, and RING-C2. However, little is known about the function of the RING-zfs of wheat.T. aestivum, and might represent the intermediate other states between RING-zf domain and other modified domains. The consensus sequence of the RING-M domain can be described as M-X2-R-X14-Cys-X1-H-X2-Cys-X2-Cys-X10-Cys-X2-Cys. Further interspecies collinearity analyses showed that TaRING-zfs were more closely related to the genes in Poaceae. According to the public transcriptome data, most of the TaRING-zfs were expressed at different 15 stages of plant growth, development, and some of them exhibited specific responses to drought/heat stress. Moreover, 4 RING-HC  and 2 RING-H2 (TraesCS3A02G288900.1 and TraesCS4A02G174600.1) were significantly expressed at different development stages and under drought stress. These findings provide valuable reference data for further study of their physiological functions in wheat varieties.In this study, 129 (93.5%) of 138 members were found in nucleus, indicating TaRING-zf were primarily engaged in the degradation of transcription factors and other nuclear-localized proteins. 138 TaRING-zf domains can be divided into four canonical or modified types . The RING-M was newly identified in Taken together, the characterization and classifications of the TaRING-zf family were extensively studied and some new features about it were revealed. This study could provide some valuable targets for further studies on their functions in growth and development, and abiotic stress responses in wheat.The online version contains supplementary material available at 10.1186/s12864-022-08905-x. The RING-zf domain is a Cys-rich domain with 8 metal ligand positions and four pairs of Cys/His residues that coordinate binding of two zinc ions in an across-brace structure  approach  was used to calculate the expression levels [Based on the manufacturer’s instructions. The total RNAs from different tissues and differently treated materials under heat, drought, and heat and drought stress were extracted using Trizol reagent (Invitrogen). RNA integrity and quality were detected by NanoDrop 1000 and electrophoresis. First-strand cDNA was synthesized by the SuperScript II Reverse Transcriptase (Invitrogen). qRT-PCR research revealed 18 tissue-specific TaRING-zf gene expression profiles and 12 TaRING-zf genes that respond strongly to heat and drought conditions. Table Sn levels .Additional file 1: Figure S1. The alignment of 138 TaRING-zf protein sequences.Additional file 2: Figure S2. Phylogenetic relationships, gene structure and architecture of conserved protein motifs in 138 TaRING-zfs from T. aestivum.a The name of 138 TaRING-zf genes. b Exon, intron, and UTR structure of 138 TaRING-zf genes. Yellow boxes indicate untranslated 5’ and 3’ regions, green boxes indicate exon, and black lines indicate introns. The gene full length and protein length can be estimated by using the scale at the bottom. c The motif composition of TaRING-zf proteins. The motifs, numbers 1-15, are displayed in different colored boxes.Additional file 3: Figure S3. Domain-based classification of T. aestivum RING-zf proteins. a Additional TM domain contained in the RING-zf protein. b Additional IBR domain contained in the RING-zf protein. c Additional coiled coil domain contained in the RING-zf protein. d Additional RPT1 domain contained in the RING-zf protein. e Additional RING-ZnF-CHY, RING-Zinc_ribbon_6 and pfam Lon-substr-bdg domains contained in the RING-zf protein. f Additional DEXDc, HIRAN and HELICc domains contained in the RING-zf protein. g Additional WD40 domain contained in the RING-zf protein. h 2 TaRING-zf proteins were only contained 2 or 3 RING-zf domains. The additional domains architecture was predicted by on-line SMART program (http://smart.embl-heidelberg.de/).Additional file 4: Table S1. Detailed information of all 138 TaRING zinc finger genes identified in tomato genome.Additional file 5: Table S2. One-to-one orthologous relationships among T.aestivum and T.aestivum, A.thaliana, A. tauschii, B. distachyon, H. vulgare, rice, S.italica and Zea mays, respectively.Additional file 6: Table S3. The information of cis-acting elements on the promoters of T. aestivum RING-zf genes.Additional file 7: Table S4. The information of transcriptome data of T. aestivum RING-zf genes.Additional file 8: Table S5. The list of primers.


Bread is one of the most widely consumed foods in all regions of the world. Wheat flour being its principal ingredient is a cereal crop low in protein. The protein content of a whole grain of wheat is about 12–15% and is deficit in some essential amino acids, for example, lysine. Conversely, the protein and fibre contents of legume crops are between 20 and 35% and 15 and 35%, respectively, depending on the type and cultivar of the legume. The importance of protein-rich diets for the growth and development of body organs and tissues as well as the overall functionality of the body is significant. Thus, in the last two decades, there has been a greater interest in the studies on the utilization of legumes in bread production and how the incorporation impacts the quality characteristics of the bread and the breadmaking process. The addition of plant-based protein flours has been shown to produce an improved quality characteristic, especially the nutritional quality aspect of bread. The objective of this review is to synthesize and critically investigate the body of research on the impact of adding legume flours on the rheological attributes of dough and the quality and baking characteristics of bread. Triticum aestivum) is a cereal crop which constitutes about 20% of the calories consumed by humans all over the world [Breadmaking has been described as one of the oldest practices known to mankind. It is believed to be a major part of the diet of the people of Babylon, Egypt, Greece and Rome for many decades before the present era. The first bread was produced around 10,000 years BC or over 12,000 years ago and was a result of the deliberate test with water and grain (wild wheat and wild barley) and plant roots flours . Bread ihe world . The twohe world ,5,6,7. Whe world .People in different parts of the world consume bread in different shapes and forms with an average consumption of 70 kg per year per capita. There are thousands of recipes for bread making and this accounts for the wide array of bread types available . The typIn the last few decades, there is a progressive increase in the consumption of bread in many developing countries. This trend can be attributed to changing eating habits and a constant increase in the population . In manyThis trend has birthed a greater interest among researchers, policymakers and food product developers in the studies on the utilization  of legumes or pulses flour in bread making and how this impacts bread quality characteristics ,16. TherThe English word “flour” was derived from the Old French word fleur or flour, which means “the finest”. Flours are obtained upon the elimination of coarse and unwanted substances from grain during the milling process. They are fine powdery materials produced from raw grains, roots, beans, nuts or seeds after the grinding and sifting operations. Traditionally, the milling process is accomplished with a grinding stone or steel wheel. However, in the modern era, this has been replaced with roller mills. Flours contain a high amount of carbohydrates; also called polysaccharides, with starches dominating ,18. Wheat is a cereal crop which accounts for one-fifth of the calories consumed by humans all over the world ,19. The The milling process is designed to produce mainly white flour. White or refined flours are generally obtained from wheat grains containing only the endosperm . Whole wheat flour on the other hand is made from whole wheat grains . Germ flours are made from both the endosperm and germ with the exclusion of the bran . HardnesThe endosperm, which is the main component of white flour consists of two types of proteins: gluten proteins and water-soluble proteins. The gluten proteins account for about 85% of wheat protein and the remainder is the water-soluble proteins. This combination is responsible for the unique bread-making properties of wheat flour . The twoWheat flour has been used for different purposes. The presence of gluten, the dough-forming constituent of wheat flour, accounts for its applications in the production of leavened products . With thDespite the great properties of wheat flour and its suitability for bread making, research has shown that it contains a lower amount of health-promoting bioactive molecules like vitamins, β-carotene, polyphenols, dietary fibre and flavonoids . AdditioComposite flour is described as a mix of flours, starches and other ingredients in which wheat flour is completely or partly replaced with flours from other sources in bakery and pastry products. This can be a mixture of two or three kinds of flour from either plant or animal sources with or without the addition of wheat flour . These fpuls, meaning ‘a pottage made of meal’. One of the most important characteristics of legumes is the presence of high amounts of proteins. These proteins have high lysine content, an essential amino acid needed by the body. Proteins in legumes are deficient in Sulphur-containing amino acids, making them an excellent addition to other commonly used cereal proteins  which are low in lysine, but rich in Sulphur amino acid content [Legumes are defined as the edible seeds of the family Leguminosae or Fabaceae, the second-largest family of seed plants containing 600 genera and about 13,000 species. The word ‘pulses’  is derived from the Latin word,  content ,30,42. T content . Nutritionally, pulses contain a relatively high amount of protein (20–35%) and fibre (15–20%) as compared with cereal grains and lower amounts of fats (below 10%) . The nutDifferent legumes have been blended with wheat and other cereal flour to produce different kinds of bakery and pastry products. Notably include: Vicia faba)  which has been used to produce bread [Faba beans   is used in the production of bread [Cowpea (of bread  and macaof bread .Glycine max) has high lysine content and this makes it an ideal crop to improve the essential amino acids profile when blended with cereal crops. It has been used to produce bread [Soybean  has been used in the production of bread [Bambara groundnut (of bread  and pastof bread .Prosopis spp.). Bigne et al. (2018) used mesquite flour at 150–350 g/kg mixed with wheat flour at 850–650 g/kg to produce composite sweet bread [Mesquite flour is defined as a sweet and aromatic product produced from milling the whole ripe fruit of the mesquite tree (et bread .Lens culinaris) was used by Perri et al. (2021) to produce bread. They used a blend of wheat flour and sourdough made from whole and sprouted lentil flour. They reported that processes like fermentation and germination are potential approaches to enhance the use of legumes in novel foods [Lentil (el foods . Other pel foods  and cookel foods .Pisum sativum) has been used to produce biscuits [Yellow pea flour (biscuits .Cicer arietinum) has been used to produce bread [Chickpea  was used to produce spaghetti pasta with a blend of Mexican common bean flour and semolina [Common bean (semolina . Cajanus cajan) is also one of the nutritionally important legumes of the tropical and subtropical regions of the world. Its application for pastry products was explored by Rafiq et al. (2017) who produced pasta from semolina flour substituted with legume flour and brown rice flour using a hot extrusion process (twin screw extruder) [Pigeon pea (xtruder) . Telfairia occidentalis) seed flour was used to produce bread with a blend of wheat flour and fluted pumpkin seed flour [Fluted pumpkin  among the protein molecules significantly contribute to the rheological characteristics of dough . The rheThe dough rheology and quality of bread depend largely on the starch-protein complex, and most importantly, the presence of gluten. The addition of non-wheat flour for baked goods can have negative effects on the gluten network, leading to weakened bread dough and degradation in bread quality characteristics . The majWater absorption capacity is an essential property that indicates a flour’s ability to absorb water and produce dough of excellent consistency. The impact of legumes/pulses addition on the water absorption capacity of dough has been extensively studied. The addition of chickpea, soybean, common bean, fava bean and lentil flours to wheat flour has been reported to increase the water absorption capacity of dough compared to wheat dough ,73,74,75The addition of the enzyme transglutaminase has been reported to reinforce the protein network and induced a significant increase in the water absorption capacity of rice flour, soy flour and pea protein isolate blends, producing a synergetic effect and a reduction in the storage (G′) and viscous (G′′) moduli. The main function of the enzyme transglutaminase is to covalently crosslink proteins through the association between an ε-amino group on protein-bound lysine residues and a γ-carboxamide group on protein-bound glutamine residues. . Differep < 0.05) in the functional characteristics, excluding the bulk density and swelling capacity of composite flour produced from wheat flour partially substituted with 10% acha flour and 0–15% cowpea flour compared with that produced from 100% wheat flour [The inclusion of wheat-lupin protein isolates has been reported to enhance the development time of dough, its strength and resistance to deformation and extensibility. This was a result of the lupin particle entrapped inside the gluten network structure, and a likely correlation between the gluten and some of the lupin proteins present in the outer part of the moistened particles . The incat flour . According to Kahraman et al. (2018), raw, roasted and dehulled chickpea flours increased the viscous and elastic moduli of rice-based dough, leading to a good structuring of the dough. A reduced retrogradation tendency of the slurry comprising chickpea flours was also confirmed by the viscoamylographic test. This is a promising outcome for baking food applications . Baiano Artemisia sphaerocephala Krasch gum (ASKG) (0.03–0.5%) caused a significant improvement in the viscoelastic characteristics of the composite dough system, followed by a reduced trend at a higher level of gum inclusion (0.8%). Addition of the gum at 0.03–0.5% increased dough G′/G″ values. The addition of 0.3% of the gum resulted in a relatively denser and more arranged network structure of the dough while 0.5% and 0.8% of the gum resulted in the disruption of the strong network with visible signs of starch deformation [Other processing techniques such as the usage of food hydrocolloids, enzymes, and sourdough fermentation have been demonstrated to improve the functionality of the dough and bread textural quality of pan-type bread produced from non–wheat flours . The incormation . TherefoW. confusa) to faba beans sourdough containing dextran improved the viscoelastic properties of the dough, improved the specific volume, and decreased crumb hardness of the bread produced as compared with the unblended sample [In addition, Wang et al. (2018) reported that the addition of microbial dextran , emulsifiers  and hydrocolloids   For examBaking creates sequences of chemical, physical, and biochemical reactions, producing different changes in the end product characteristics such as volume enlargement, moisture evaporation, formation of porous structure, protein denaturation, starch gelatinization, formation of crust and browning reaction, protein cross-linking, melting of fat and crystals and their incorporation into the outer layers of air cells, gas cells rupture and fragmentation of cell walls ,8,87. ThIn addition, functional characteristics of food proteins such as the ability to foam and water retention can be improved with heat treatment in the presence of sugars; complex carbohydrates by the process known as the Maillard reaction ,89. The Quality is defined as the combination of distinguishing characteristics and properties of a food commodity that can determine its degree of acceptance by a user or consumer . Bread qExamples of different attributes of bread that can be objectively measured and have been quantified to determine the quality of bread include volume (using rapeseed dis-placement), specific weight, specific volume, moisture content, water activity, crust and crumb colour, crust crispiness, crumb hardness, cell distribution within the loaf slice using image analysis, and volatile composition. The attributes related to sensory sensations of bread include visual appearance, taste, odour and tactile and oral texture . The impCereals are grass crops grown for the edible components of their grains. They are regarded as staple foods and an essential source of micronutrients such as vitamins, minerals and macronutrients such as proteins, carbohydrates, fibre, crude fats, and essential fatty acids, which perform important functions in the health of humans. The extensively consumed grains are wheat, maize, rice, and barley while minimally consumed grains are rye, sorghum, quinoa, oat, and millet ,103. CerAmong the commonly studied cereal crops used to produce bread and other pastry products is sorghum. The addition of sorghum flour has been reported to result in a gradual decrease in composite bread ,106. Thia*) values and reduced the cohesiveness and luminosity (L*) values of instant noodles produced [Quinoa is also a widely studied cereal crop in the production of pastry products. Bilgicli (2013) reported that the addition of pseudo-cereals, e.g., quinoa at 25% of a recipe in gluten-free noodles can enhance nutrient content, for instance, proteins and minerals  . Giménezproduced . Alvarezproduced . Cárdenaproduced . Another cereal crop which has been used in the production of pastry products is millet. Chaitra et al. (2020) produced Belgian waffles with wheat flour substituted with finger millet and pearl millet flours and reported that the control sample (100% wheat flour) had a harder texture with a shear force of 35.86 N compared with the blended samples with a shear force ranging from 19.68 N to 27.02 N. The result of the sensory analysis proved that the samples containing millet flour were more accepted, up to 50% . IbidapoLin et al. (2012) produced bread from wheat flour partially substituted with micro-fluidized corn bran at 18, 20, and 22% of flour. They reported that for the three kinds of bran substitution when the moisture content was improved from its standard values of 38.3, 38.6, 38.8% to 40.8, 41.9, and 44.0%, correspondingly, the loaves obtained showed comparable microstructure, specific loaf volume, and textural characteristics as the unblended bread . Jeong eRenoldi et al. (2021) reported that pasta produced from psyllium seed husk was firmer and sticker than 100% durum wheat semolina. The cooking loss was reported to have increased with increasing levels of psyllium seed husk substitution above 25 g/kg with values below the technologically acceptable limit of 80 g/kg. Replacement of semolina with 50–100 g/kg psyllium seed husk was potent in reducing the predictive glycemic response of supplemented pasta in comparison with the unfortified sample and this was attributed to the formation of fibre aggregates limiting starch swelling after the scanning electron microscopy and dough rheology . Roots and tuber crops are an essential component of the human diet as they are the main source of energy in the form of carbohydrates for the body. There are enormous kinds of roots and tuber crops produced globally. Nevertheless, their extensive utilization in the food industry is limited to only a small number of common types such as potato, cassava, sweet potato, yams and taro ,122. OraIdowu et al. (1996) reported a reduction in oven springs and specific volumes when cocoyam flour was blended with wheat flour to produce bread . ChisengNyembwe et al. (2018) also reported that the defatted marama flour mixed with cassava flour at a ratio of 33:67 can yield a dough of similar strength, but with reduced stability compared with wheat flour dough . NindjinPulses are eco-friendly, nutrient-dense and widely cultivated crops all over the world. Due to their high nutrient profiles, they are useful tools to compact undernutrition in low-income countries and malnutrition in developed countries of the world. Wheat flour is the traditional flour used in bread making and other pastry products. This review has summarized reports on the recent applications of legumes/pulses in the production of baked and pastry products, impacts on the rheological qualities of dough and effects on the baking and quality attributes of bread. Wheat flour is a cereal crop and is a deficit in some nutrients highly required for body growth and functionality. Therefore, from this review, it has been shown that the addition of pulse flour to wheat flour in bread making is a viable means to enhance the nutritional profile of the products. However, from the reviewed literature, most of the studies reported focused on just the particle size of the legume flours and a limited study was reported on the impact of particle size on the quality and baking characteristics of the bread produced from the composite flours. Therefore, more research should be conducted on the potential effects of different flour particle sizes on the quality attributes of bread and the overall acceptability by consumers.


The effective capture and storage of volatile molecular iodine from nuclear waste is of great significance. Covalent organic frameworks (COFs) are a class of extended crystalline porous polymers that possess unique architectures with high surface areas, long-range order, and permanent porosity. Substantial efforts have been devoted to the design and synthesis of COF materials for the capture of radioactive iodine. In this review, we first introduce research techniques for determining the mechanism of iodine capture by COF materials. Then, the influencing factors of iodine capture performance are classified, and the design principles and strategies for constructing COFs with potential for iodine capture are summarized on this basis. Finally, our personal insights on remaining challenges and future trends are outlined, in order to bring more inspiration to this hot topic of research. Additionally, although 131I has a short half-life (about 8 days), it is often combined with other hydrocarbons to yield organic compounds, such as methane iodide, which also make it extremely harmful to its ecological surroundings and human health −/[2I2Br]− are approximately twice that between TAPT-OH and I3−/I5−, suggesting a stronger affinity of TAPT-AB toward the iodine species. This is consistent with expectations and explains the experimental observations.Density functional theory (DFT) calculations can help researchers gain insight into how the COF frameworks bind iodine. It is well known that the binding energy can remarkably affect iodine capture. Normally, the stronger the interaction between iodine and the adsorption sites, the higher the iodine uptake. The binding energy between the model compounds was calculated via DFT and the corresponding conclusions were obtained by comparing the values. Han’s group  calculatPXRD and transmission electron microscopy (TEM) analysis can show whether the crystallinity and morphology of COFs can be maintained throughout the iodine capture process.Physical adsorption mainly depends on the surface areas, pore sizes and pore volumes of COFs. Thus, the functionalized architectures are designed to have high specific surface areas, and large pore sizes and pore volumes for efficient physisorption of iodine. For example, Liu et al. designed and synthesized four 2D COFs with different pore sizes for volatile iodine adsorption . Finally3·g−1 achieved a remarkable iodine adsorption capacity (6.26 g·g−1) only in physical adsorption, which was driven by van der Waals forces. They proved that I2 capture does not require specific functionalization of the porous skeleton, and that pores of any shape or size can be 100% occupied by physical adsorption. However, its adsorption kinetics were quite slow (0.13 g·g−1·h−1).Jiang’ group reported a series of 2D COFs with 1D open channels, which possessed various topologies from hexagonal to tetragonal and trigonal, and were free of specific binding sites and interchannel interpenetration . So, the2 with electron-rich groups  and pore channels results in a combination of physical and chemical adsorption for the I2 capture process. The iodine vapor adsorbed into the pores through physical adsorption can readily generate the polyiodide species I3− and I5− due to the strong interactions between the exposed electron-rich groups and I2, implying that a chemisorption process occurs. In the meantime, iodine molecules may fill the remaining pore channels through physical adsorption.As aforementioned, porosity plays an important role in the adsorption of iodine vapor. However, in addition to the amount of adsorption, the adsorption rate is also a significant factor that must be considered. As for the TPB-DMTP-COF, it has an ultrahigh uptake capacity but takes 100 h to reach saturation, which is particularly unfavorable in the case of emergency disasters. Studying only physical adsorption—whereby the iodine uptake capacity of COF materials is determined by pore volume, while the iodine uptake kinetics are determined by pore connectivity and size—limits the application of COFs for iodine capture. The combination of physical and chemical adsorption is of paramount importance in the designed synthesis of COF materials for the capture of radioiodine. It is commonly believed that the interaction of I2 via an electron-rich quinoline unit, but can also adsorb I2 via a chemical reaction with phenylacetylene moieties. Although the surface area and pore size of COF-PA were reduced compared with before functionalization, the adsorption rate was accelerated and the adsorption capacity was still high at low iodine concentrations. Very recently, Zhai et al. discovered two rare cationic COFs, C-TP-PDA-COFs and C-TP-BPDA-COFs , via a post-function process. The cationic C-TP-BPDA-COF exhibits a higher iodine capture value (6.11 g·g−1) than that of neutral COF . XP. XP54]. tral COF .2 and anion I3− signals were observed. This indicates the presence of both physical and chemical adsorption. Chang et.al reported that two tetrathiafulvalene (TTF)-based COFs, JUC-560 (A16-M4) and JUC-561(A16-M8), achieve excellent iodine adsorption capacity (8.19 g·g−1) and ultrafast adsorption kinetics (0.70 g·g−1·h−1) thiophene-2,5-dicarbaldehyde (A6) as the aldehyde monomer and tri(4-aminophenyl)benzene (M11) or tris(4-aminophenyl)amine (M8) as the amino monomer trithiophene-2,5,8-tricarbaldehyde (A11) and N-rich monomer 1,3,5-tris-(4-aminophenyl)triazine (M9) . BTT-TAP−1 for USTB-1c. The mechanism investigation unveiled the superiority of N atoms over S atoms for POCs in iodine vapor capture with the assistance of definite crystal structures. The results of the DFT calculations suggested a stronger binding interaction with the I3− and I5− of the former 2,2′-bipyridine moiety due to its N-rich moiety. At the same time, the work of Song’s group also proved that the binding energy would remarkably affect iodine capture [3− ions at the potential sites of -S-, -N-, and the conjugate sites were −31.23, −11.35, and −10.38 kcal·mol−1, respectively, which indicated that it was hard for I3− to bind at the -S- site. Therefore, the -S- site was not a good iodine adsorption site, which is consistent with Jiang’s conclusion.Moreover, Jiang’s group identified two N-rich 2D COFs, USTB-1 (A23-M7) and USTB-2 (A23-M14), which were transformed by BPPOC with a single-crystal X-ray diffraction structure . The cag capture . They em2·g−1), and a large π-conjugated skeleton [−1 and 1.30 g·g−1, respectively, suggesting that P-COF may be a potential candidate for radioiodine capture. FT-IR and XPS spectra indicate that iodine adsorption could occur simultaneously at the aromatic ring and C=N linkage. However, the mechanism of sorption sites in the adsorption process has not been revealed, and no PXRD diffraction peaks can be observed after the first recovery of P-COF.Additionally, Ke’s group reported a phosphine-based COF  with abundant heteroatom  adsorption sites, a large surface area trimethylammonium bromide, which can bind polyiodide anions via Coulomb interactions. Recently, Han’s group identified a series of ionic COFs (iCOFs) via the “multivariate” synthetic strategy combined with post-synthesis modification [2 adsorption capacities of COF-OH-50 and iCOF-AB-50 under static conditions were 6.49 and 10.21 g·g−1, respectively. Additionally, the I2 adsorption capacities of COF-OH-50 and iCOF-AB-50 under dynamic conditions and at room temperature were 1.70 g·g−1 and 2.79 g·g−1, respectively. In particular, compared with other reported COF-based adsorbents, iCOF-AB-50 exhibited the highest I2 adsorption capacity under both static and dynamic conditions. Additionally, the authors also investigated the I2 capture performance in the presence of water. This groundbreaking work provided a new design strategy for the development of high-performance iodine adsorption materials.Another notable strategy effectively promoting Ification . The so Cl) and 2,5-dimethoxyterephthalaldehyde (A3), which can efficiently capture I2 under industrial conditions [2 through coulombic interactions to trap I2. The saturated I2 uptake capacity of TGDM was determined to be 29.24 wt%, which is higher than that of the well-known iCOF-AB-50 (8.27 wt%). This may be due to the different ionic sites (quaternary ammonium groups for iCOF-AB-50), and also the fact that the ionic site density of iCOF-AB-50 is lower than that of TGDM (number of ionic sites per gram: 1.09 × 1021 vs. 1.59 × 1021). Remarkably, TGDM can be easily recycled and reused without losing its adsorption capacity. The characterization and theoretical calculations indicate that iodine adsorption follows different mechanisms at low and high temperatures  through the reaction of thiamine-guanidine hydrochloride  for the first time [2-Th) bearing both amine and hydrazide functionalities was designed to react with benzene-1,3,5-tricarbaldehyde (Bta). The different activities of the amine and hydrazide groups toward aldehyde underpinned the synthesis of the unprecedented NH2-Th-Bta COF via a simple and straightforward route  in comparison to that of the nonfunctionalized Th-Bta COF counterpart (0.68 g·g−1), despite its specific low surface area (10 m2·g−1). This work proved that the introduced free amine group plays an essential role in enhancing iodine adsorption capacity, which gives rise to a charge-transfer complex with the iodine molecule. Moreover, the NH2-Th-Bta COF possessed exceptional cycling capability and retained high iodine uptake, even after six cycles. This strategy of increasing the density of active sites improves the iodine adsorption capacity, which sheds light on the future design of advanced porous materials. It is reasonable to speculate that if such a strategy can be used to obtain COFs with large enough surface areas, the iodine adsorption capacity will inevitably increase significantly.Recently, Zhang et al. developed a site-selective synthetic strategy for the facile preparation of an amine-functionalized hydrazone-linked COF  COFs have attracted much attention for their excellent iodine adsorption performance. According to the work of Fang’s group . The rearactions .2 adsorption capacity and adsorption rate due to their strong Coulomb interactions between ionic sites and polyiodide anions. However, in practice, it is very challenging to incorporate ionic sites into COFs while maintaining high crystallinity and accessible porosity [2 concentration, pressure, static/dynamic conditions, and other different test conditions, will also significantly affect the iodine adsorption performance of COFs.In principle, COFs can be functionalized with the desired adsorption ionic sites through the design of appropriate monomers or post-synthesis modification; through this, they can achieve a high Iporosity ,76. In a2 adsorption capacity is determined by the collective effects of multiple factors: the textural properties  of the adsorbent, the affinity between the binding site and the I2 molecule, and the density of binding sites. Therefore, an optimal balance between these factors is essential for achieving high adsorption capacity. Based on the above theory, knowledge of how to construct COFs with regular channels and improve the density of adsorption sites on the materials—while effectively ensuring high activity and efficiency of these adsorption sites, as well as high affinity for iodine—has become the key to preparing high-performance iodine capture materials.In short, IAs a sustainable and low-carbon energy supply, nuclear energy is expected to play a more important role in the future. Radioactive iodine in the waste of the nuclear industry is one of the important factors hindering its popularization. COFs exhibit unique properties including large surface areas, ultrahigh porosity, tunable pore sizes, and relatively high chemical/thermal stability combined with tailorable architecture, which are not seen in other porous materials. Because of these appealing properties, vast research has been conducted on COF-based materials for vapor iodine capture in recent years.In general, an outstanding COF-based material for radioiodine capture that can be used in industry should satisfy the following requirements: (I) high adsorption capacity and adsorption kinetics for vapor iodine under industrial conditions; (II) high selectivity within a mixture of competitive gases; (III) long retention time of the adsorbed iodine; (IV) and excellent recyclability and low-cost.2 at high temperatures and under other harsh industrial conditions; (III) design and synthesize 3D COFs; and (IV) evaluate the I2 adsorption capacities of COFs under dynamic conditions. These strategies may yet lead to progress in the construction of COFs with enhanced adsorption capacity and adsorption kinetics under industrial conditions.According to recently published works, there is still much room for further improvement in the iodine capture performance of COF materials, which also provides great opportunities for researchers in this field to: (I) pay attention to the selective adsorption of COFs on vapor iodine under industrial conditions; (II) develop high-performance adsorbent COFs that can efficiently capture low-concentration IIn summary, we provide an overview of the research progress in the iodine capture application of COF materials, which is a very active subject. We hope this work will provide guidance for the design and synthesis of COFs with excellent iodine capture performance, and inspire innovations in this emerging field. Additionally, we believe that in the near future, this research area will experience rapid growth followed by industrial application.