NIR sensors combined with chemometric algorithms in intelligent quality evaluation of sweetpotato roots from 'Farm' to 'Table': Progresses, challenges, trends, and prospects.

NIR sensors, in conjunction with advanced chemometric algorithms, have proven to be a powerful and efficient tool for intelligent quality evaluation of sweetpotato roots throughout the entire supply chain. By leveraging NIR data in different wavelength ranges, the physicochemical, nutritional and antioxidant compositions, as well as variety classification of sweetpotato roots during the different stages were adequately evaluated, and all findings involving quantitative and qualitative investigations from the beginning to the present were summarized and analyzed comprehensively. All chemometric algorithms including both linear and nonlinear employed in NIR analysis of sweetpotato roots were introduced in detail and their calibration performances in terms of regression and classification were assessed and discussed. The challenges and limitations of current NIR application in quality evaluation of sweetpotato roots are emphasized. The prospects and trends covering the ongoing advancements in software and hardware are suggested to support the sustainable and efficient sweetpotato processing and utilization.

Environmental effects and their impact on yield in adjacent experimental plots of high-stem and short-stem wheat varieties.

BACKGROUND: In regional wheat trials, when short-stem wheat varieties and high-stem wheat varieties are planted adjacent to each other in small plots, changes in their marginal plot environment can lead to bias in yield evaluation. Currently, there is no relevant research revealing the degree of their mutual influence. RESULTS: In a regional wheat experiment, when high-stem wheat varieties and short-stem wheat varieties were planted adjacent to one another, there was no significant change in soil temperature or humidity in the high-stem wheat variety experimental plot from November to May compared to the control plot, while the soil humidity in the short-stem wheat variety experimental plot was greater than that in the control plot. In May, the soil temperature of the short-stem wheat varieties in the experimental plot was lower than that in the control plot. Illumination of the wheat canopy in the high-stem wheat variety experimental plot had a significant positive effect in April and May, while illumination of the wheat canopy in the short-stem wheat variety experimental plot had a negative effect. The chlorophyll fluorescence parameters of flag leaves in the high-stem wheat variety experimental plots showed an overall increasing trend, while the chlorophyll fluorescence parameters of flag leaves in the experimental plots of short-stem wheat varieties showed a decreasing trend. The analysis of the economic yield, biological yield, and yield factors in each experimental plot revealed that the marginal effects of the economic yield and 1000-grain weight were particularly significant and manifested as positive effects in the high-stem wheat variety experimental plot and as negative effects in the short-stem wheat variety experimental plot. The economic yield of the high-stem wheat variety experimental plot was significantly greater than that of the control plot, the economic yield of the short-stem wheat variety experimental plot was significantly lower than that of the control plot, and the economic yield of the high-stem experimental plot was significantly greater than that of the short-stem experimental plot. When the yield of the control plot of the high-stem wheat varieties was compared to that of the control plot of the short-stem wheat varieties, the yield of the control plot of the short-stem wheat varieties was significantly greater than that of the control plot of the high-stem wheat varieties. CONCLUSIONS: Based on these findings, it is concluded that plots with high-stem and short-stem wheat varieties are adjacent in regional wheat trials, the plots of high-stem wheat varieties are subject to marginal positive effects, resulting in a significant increase in economic yield; the plots of short-stem wheat varieties are subject to marginal negative effects, resulting in a decrease in economic yield. This study reveals the mutual influence mechanism of environment and yield with adjacent planting of high-stem and short-stem wheat varieties in regional wheat trials, providing a useful reference and guidance for optimizing the layout of regional wheat trials.

Advancements, limitations and challenges in hyperspectral imaging for comprehensive assessment of wheat quality: An up-to-date review.

The potential of hyperspectral imaging technology (HIT) for the determination of physicochemical and nutritional components, evaluation of fungal/mycotoxins contamination, wheat varieties classification, identification of non-mildew-damaged wheat kernels, as well as detection of flour adulteration is comprehensively illustrated and reviewed. The latest findings (2018-2023) of HIT in wheat quality evaluation through internal and external attributes are compared and summarized in detail. The limitations and challenges of HIT to improve assessment accuracy are clearly described. Additionally, various practical recommendations and strategies for the potential application of HIT are highlighted. The future trends and prospects of HIT in evaluating wheat quality are also mentioned. In conclusion, HIT stands as a cutting-edge technology with immense potential for revolutionizing wheat quality evaluation. As advancements in HIT continue, it will play a pivotal role in shaping the future of wheat quality assessment and contributing to a more sustainable and efficient food supply chain.

Fast quantitative analysis and chemical visualization of amylopectin and amylose in sweet potatoes via merging 1D spectra and 2D image.

The quantitative analysis and spatial chemical visualization of amylopectin and amylose in different varieties of sweet potatoes were studied by merging spectral and image information. Three-dimensional (3D) hyperspectral images carrying 1D spectra and 2D images of hundreds of the samples (amylopectin, n = 644; amylose, n = 665) in near-infrared (NIR) range of 950-1650 nm (426 wavelengths) were acquired. The NIR spectra were mined to correlate with the values of the two indexes using a linear algorithm, generating a best performance with correlation coefficients and root mean square error of prediction (rP and RMSEP) of 0.983 and 0.847 g/100 mg for amylopectin, and 0.975 and 0.500 g/100 mg for amylose, respectively. Then, 14 % of the wavelengths (60 for amylopectin, 61 for amylopectin) were selected to simplify the prediction with rP and RMSEP of 0.970 and 1.103 g/100 mg for amylopectin, and 0.952 and 0.684 g/100 mg for amylose, respectively, comparable to those of full-wavelength models. By transferring the simplified model to original images, the color chemical maps were created and the differences of the two indexes in spatial distribution were visualized. The integration of NIR spectra and 2D image could be used for the more comprehensive evaluation of amylopectin and amylose concentrations in sweet potatoes.

The 14-3-3 Protein BdGF14a Increases the Transcriptional Regulation Activity of BdbZIP62 to Confer Drought and Salt Resistance in Tobacco.

BdGF14a, a 14-3-3 gene from Brachypodium distachyon, induced by salt, H2O2, and abscisic acid (ABA), improved tolerance to drought and salt in tobacco, with a higher survival rate and longer roots under these stresses. Additionally, physiological index analyses showed that the heterologous expression of BdGF14a induced higher expression levels of antioxidant enzymes and their activities, leading to lighter DAB and NBT staining, denoting decreased H2O2 content. Additionally, the lower MDA content and ion leakage indicated enhanced cell membrane stability. Moreover, exogenous ABA resulted in shorter roots and a lower stomatal aperture in BdGF14a transgenic plants. BdGF14a interacted with NtABF2 and regulated the expression of stress-related genes. However, adding an ABA biosynthesis inhibitor suppressed most of these changes. Furthermore, similar salt and drought resistance phenotypes and physiological indicators were characterized in tobacco plants expressing BdbZIP62, an ABRE/ABF that interacts with BdGF14a. And Y1H and LUC assays showed that BdGF14a could enhance the transcription regulation activity of NtABF2 and BdbZIP62, targeting NtNECD1 by binding to the ABRE cis-element. Thus, BdGF14a confers resistance to drought and salinity through interaction with BdbZIP62 and enhances its transcriptional regulation activity via an ABA-mediated signaling pathway. Therefore, this work offers novel target genes for breeding salt- and drought-tolerant plants.

A 14-3-3 Protein-Encoding Gene, BdGF14g, Confers Better Drought Tolerance by Regulating ABA Biosynthesis and Signaling.

Abscisic acid (ABA), a phytohormone, enacts a cardinal function in coping with abiotic stress. 14-3-3 proteins can interact with ABA-responsive-element-binding transcription factors (ABFs), a chief constituent of ABA signaling, and play critical roles in the dehydration response involving ABA signaling. Meanwhile, whether and how 14-3-3 proteins regulate ABA signaling to respond to aridity stress is yet to be fully investigated. Herein, BdGF14g, a 14-3-3 gene induced by ABA, H2O2, and PEG treatments, was identified in Brachypodium distachyon (B. distachyon). Overexpression of BdGF14g improved drought stress tolerance in tobacco plants, with a higher survival rate, longer root length, enhanced cell membrane stability, and increased antioxidase activity compared with non-transgenic controls in coping with dehydration. Both drought and exogenous ABA treatments resulted in smaller stomatal apertures in BdGF14g-transgenic lines. Additionally, when an ABA biosynthesis inhibitor was added, the better growth statuses, less H2O2 accumulation, and higher activities of catalase and superoxide dismutase under mannitol stress disappeared. Moreover, BdGF14g interacted with NtABF2, upregulated the endogenous ABA content, and enhanced the transcription of ABA-related genes, including NtNCED1, a crucial ABA biosynthesis gene, under drought conditions. In conclusion, BdGF14g acts as a positive factor in the water deficiency response by affecting ABA biosynthesis and signaling in tobacco plants.

Towards rapidly quantifying and visualizing starch content of sweet potato [Ipomoea batatas (L.) Lam] based on NIR spectral and image data fusion.

This study aimed to achieve the rapid quantification and visualization of the starch content in sweet potato via near-infrared (NIR) spectral and image data fusion. The hyperspectral images of the sweet potato samples containing 900-1700 nm spectral information within every pixel were collected. The spectra were preprocessed, analyzed and the 18 informative wavelengths were finally extracted to relate to the measured starch content using the multiple linear regression (MLR) algorithm, producing a good quantitative prediction accuracy with a correlation coefficient of prediction (rP) of 0.970 and a root-mean-square error of prediction (RMSEP) of 0.874 g/100 g by an external validation using a set of dependent samples. The MLR model was further verified in terms of soundness and predictive validity via F-test and t-test, and then transferred to each pixel of the original two dimensional images with the help of a developed algorithm, generating color distribution maps to achieve the vivid visualization of the starch distribution. The study demonstrated that the fusion of the NIR spectral and image data provided a good strategy for the rapidly and nondestructively monitoring the starch content of sweet potato. This technique can be applied to industrial use in the future.

Effect of fermentation on structural properties and antioxidant activity of wheat gluten by Bacillus subtilis.

Bacillus subtilis has been extensively studied for its ability to inhibit the growth of harmful microorganisms and its high protease activity. In this study, Bacillus subtilis was used to ferment gluten and assess the effects of the fermentation process on the physicochemical, microstructure and antioxidant properties of gluten. The results of Fourier infrared spectroscopy (FT-IR) and circular chromatography (CD) showed a significant decrease in the content of α-helix structures and a significant increase in the content of ß-sheet structures in gluten after fermentation (p < 0.05). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that glutenin was degraded into small molecular peptides with a molecular weight of less than 26 kDa after 24 h of fermentation; meanwhile, the fermentation process significantly increased the free amino acid content of the samples (p < 0.05), reaching 1923.38 µg/mL at 120 h of fermentation, which was 39.46 times higher than that at 24 h of fermentation (p < 0.05). In addition, the fermented back gluten has higher free radical scavenging activity and iron reduction capacity. Therefore, fermented gluten may be used as a functional food to alleviate oxidative stress. This study provides a reference for the high-value application of gluten.

Simultaneous quantifying and visualizing moisture, ash and protein distribution in sweet potato [Ipomoea batatas (L.) Lam] by NIR hyperspectral imaging.

This study aimed to achieve the rapid evaluation of moisture, ash and protein of sweet potato simultaneously by near-infrared (NIR) hyperspectral imaging (900-1700 nm). Hyperspectral images of 300 samples for each parameter were acquired and the spectra within images were extracted, averaged and preprocessed to relate to the three measured parameters, using partial least squares (PLS) algorithm, respectively, resulting in good performances. Nine, eleven and eleven informative wavelengths were selected to accelerate the prediction of the three parameters, generating a correlation coefficient of prediction (r P) of 0.984, 0.905, 0.935 and root mean square error of prediction (RMSEP) of 0.907%, 0.138%, 0.0941% for moisture, ash and protein, respectively. By transferring the best optimized PLS models to generate color chemical maps, the distributions and variations of the three parameters were visualized. NIR hyperspectral imaging is promising and can be applied to simultaneously evaluate multiple quality parameters of sweet potato.

Fermented Wheat Germ Alleviates Depression-like Behavior in Rats with Chronic and Unpredictable Mild Stress.

Depression is a chronic mental illness with devastating effects on a person's physical and mental health. Studies have reported that food fermentation with probiotics can enrich the nutritional values of food and produce functional microorganisms that can alleviate depression and anxiety. Wheat germ is an inexpensive raw material that is rich in bioactive ingredients. For example, gamma-aminobutyric acid (GABA) is reported to have antidepressant effects. Several studies concluded that Lactobacillus plantarum is a GABA-producing bacteria and can alleviate depression. Herein, fermented wheat germs (FWGs) were used to treat stress-induced depression. FWG was prepared by fermenting wheat germs with Lactobacillus plantarum. The chronic unpredictable mild stress (CUMS) model was established in rats, and these rats were treated with FWG for four weeks to evaluate the effects of FWG in relieving depression. In addition, the study also analyzed the potential anti-depressive mechanism of FWG based on behavioral changes, physiological and biochemical index changes, and intestinal flora changes in depressed rats. The results demonstrated that FWG improved depression-like behaviors and increased neurotransmitter levels in the hippocampus of CUMS model rats. In addition, FWG effectively altered the gut microbiota structure and remodeled the gut microbiota in CUMS rats, restored neurotransmitter levels in depressed rats through the brain-gut axis, and restored amino acid metabolic functions. In conclusion, we suggest that FWG has antidepressant effects, and its potential mechanism may act by restoring the disordered brain-gut axis.

Isolation, Structural, Functional, and Bioactive Properties of Cereal Arabinoxylan─A Critical Review.

Arabinoxylans (AXs) are widely distributed in various cereal grains, such as wheat, corn, rye, barley, rice, and oat. The AX molecule contains a linear (1,4)-ß-D-xylp backbone substituted by α-L-araf units and occasionally t-xylp and t-glcpA through α-(1,2) and/or α-(1,3) glycosidic linkages. Arabinoxylan shows diversified functional and bioactive properties, influenced by their molecular mass, branching degree, ferulic acid (FA) content, and the substitution position and chain length of the side chains. This Review summarizes the extraction methods for various cereal sources, compares their structural features and functional/bioactive properties, and highlights the established structure-function/bioactivity relationships, intending to explore the potential functions of AXs and their industrial applications.

Genome-wide identification of alcohol dehydrogenase (ADH) gene family under waterlogging stress in wheat (Triticum aestivum).

BACKGROUND: Alcohol dehydrogenase (ADH) plays an important role in plant survival under anaerobic conditions. Although some research about ADH in many plants have been carried out, the bioinformatics analysis of the ADH gene family from Triticum aestivum and their response to abiotic stress is unclear. METHODS: A total of 22 ADH genes were identified from the wheat genome, and these genes could be divided into two subfamilies (subfamily I and subfamily II). All TaADH genes belonged to the Medium-chain ADH subfamily. Sequence alignment analysis showed that all TaADH proteins contained a conservative GroES-like domain and Zinc-binding domain. A total of 64 duplicated gene pairs were found, and the Ka/Ks value of these gene pairs was less than 1, which indicated that these genes were relatively conservative and did not change greatly in the process of duplication. RESULTS: The organizational analysis showed that nine TaADH genes were highly expressed in all organs, and the rest of TaADH genes had tissue specificity. Cis-acting element analysis showed that almost all of the TaADH genes contained an anaerobic response element. The expression levels of ADH gene in waterlogging tolerant and waterlogging sensitive wheat seeds were analyzed by quantitative real-time PCR (qRT-PCR). This showed that some key ADH genes were significantly responsive to waterlogging stress at the seed germination stage, and the response of waterlogging tolerant and waterlogging sensitive wheat seeds to waterlogging stress was regulated by different ADH genes. The results may be helpful to further study the function of TaADH genes and to determine the candidate gene for wheat stress resistance breeding.

Wheat heat tolerance is impaired by heightened deletions in the distal end of 4AL chromosomal arm.

Heat stress (HS) causes substantial damages to worldwide crop production. As a cool season crop, wheat (Triticum aestivum) is sensitive to HS-induced damages. To support the genetic improvement of wheat HS tolerance (HST), we conducted fine mapping of TaHST1, a locus required for maintaining wheat vegetative and reproductive growth under elevated temperatures. TaHST1 was mapped to the distal terminus of 4AL chromosome arm using genetic populations derived from two BC6 F6 breeding lines showing tolerance (E6015-4T) or sensitivity (E6015-3S) to HS. The 4AL region carrying TaHST1 locus was approximately 0.949 Mbp and contained the last 19 high confidence genes of 4AL according to wheat reference genome sequence. Resequencing of E6015-3S and E6015-4T and haplotype analysis of 3087 worldwide wheat accessions revealed heightened deletion polymorphisms in the distal 0.949 Mbp region of 4AL, which was confirmed by the finding of frequent gene losses in this region in eight genome-sequenced hexaploid wheat cultivars. The great majority (86.36%) of the 3087 lines displayed different degrees of nucleotide sequence deletions, with only 13.64% of them resembling E6015-4T in this region. These deletions can impair the presence and/or function of TaHST1 and surrounding genes, thus rendering global wheat germplasm vulnerable to HS or other environmental adversities. Therefore, conscientious and urgent efforts are needed in global wheat breeding programmes to optimize the structure and function of 4AL distal terminus by ensuring the presence of TaHST1 and surrounding genes. The new information reported here will help to accelerate the ongoing global efforts in improving wheat HST.

Transcriptomic and anatomic profiling reveal the germination process of different wheat varieties in response to waterlogging stress.

BACKGROUND: Waterlogging is one of the most serious abiotic stresses affecting wheat-growing regions in China. Considerable differences in waterlogging tolerance have been found among different wheat varieties, and the mechanisms governing the waterlogging tolerance of wheat seeds during germination have not been elucidated. RESULTS: The results showed no significant difference between the germination rate of 'Bainong 207' (BN207) (after 72 h of waterlogging treatment) and that of the control seeds. However, the degree of emulsification and the degradation rate of endosperm cells under waterlogging stress were higher than those obtained with the control treatment, and the number of amyloplasts in the endosperm was significantly reduced by waterlogging. Transcriptomic data were obtained from seed samples (a total of 18 samples) of three wheat varieties, 'Zhoumai 22' (ZM22), BN207 and 'Bainong 607' (BN607), subjected to the waterlogging and control treatments. A comprehensive analysis identified a total of 2775 differentially expressed genes (DEGs). In addition, an analysis of the correlations among the expression difference levels of DEGs and the seed germination rates of the three wheat varieties under waterlogging stress revealed that the relative expression levels of 563 and 398 genes were positively and negatively correlated with the germination rate of the wheat seeds, respectively. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses showed that the difference in the waterlogging tolerance among the three wheat varieties was related to the abundance of key genes involved in the glycolysis pathway, the starch and sucrose metabolism pathway, and the lactose metabolism pathway. The alcohol dehydrogenase (ADH) gene in the endosperm of BN607 was induced immediately after short-term waterlogging, and the energy provided by the glycolysis pathway enabled the BN607 seeds to germinate as early as possible; in addition, the expression of the AP2/ERF transcription factor was upregulated to further enhance the waterlogging tolerance of this cultivar. CONCLUSIONS: Taken together, the results of this study help elucidate the mechanisms through which different wheat varieties respond to waterlogging stress during germination.

PDIL1-2 can indirectly and negatively regulate expression of the AGPL1 gene in bread wheat.

BACKGROUND: ADP-glucose pyrophosphorylase (AGPase), the key enzyme in plant starch biosynthesis, is a heterotetramer composed of two identical large subunits and two identical small subunits. AGPase has plastidial and cytosolic isoforms in higher plants, whereas it is mainly detected in the cytosol of grain endosperms in cereal crops. Our previous results have shown that the expression of the TaAGPL1 gene, encoding the cytosolic large subunit of wheat AGPase, temporally coincides with the rate of starch accumulation and that its overexpression dramatically increases wheat AGPase activity and the rate of starch accumulation, suggesting an important role. METHODS: In this study, we performed yeast one-hybrid screening using the promoter of the TaAGPL1 gene as bait and a wheat grain cDNA library as prey to screen out the upstream regulators of TaAGPL1 gene. And the barley stripe mosaic virus-induced gene-silencing (BSMV-VIGS) method was used to verify the functional characterization of the identified regulators in starch biosynthesis. RESULTS: Disulfide isomerase 1-2 protein (TaPDIL1-2) was screened out, and its binding to the TaAGPL1-1D promoter was further verified using another yeast one-hybrid screen. Transiently silenced wheat plants of the TaPDIL1-2 gene were obtained by using BSMV-VIGS method under field conditions. In grains of BSMV-VIGS-TaPDIL1-2-silenced wheat plants, the TaAGPL1 gene transcription levels, grain starch contents, and 1000-kernel weight also significantly increased. CONCLUSIONS: As important chaperones involved in oxidative protein folding, PDIL proteins have been reported to form hetero-dimers with some transcription factors, and thus, our results suggested that TaPDIL1-2 protein could indirectly and negatively regulate the expression of the TaAGPL1 gene and function in starch biosynthesis.

PDIL1-2 can indirectly and negatively regulate expression of the AGPL1 gene in bread wheat

BACKGROUND: ADP-glucose pyrophosphorylase (AGPase), the key enzyme in plant starch biosynthesis, is a heterotetramer composed of two identical large subunits and two identical small subunits. AGPase has plastidial and cytosolic isoforms in higher plants, whereas it is mainly detected in the cytosol of grain endosperms in cereal crops. Our previous results have shown that the expression of the TaAGPL1 gene, encoding the cytosolic large subunit of wheat AGPase, temporally coincides with the rate of starch accumulation and that its overexpression dramatically increases wheat AGPase activity and the rate of starch accumulation, suggesting an important role. METHODS: In this study, we performed yeast one-hybrid screening using the promoter of the TaAGPL1 gene as bait and a wheat grain cDNA library as prey to screen out the upstream regulators of TaAGPL1 gene. And the barley stripe mosaic virus-induced gene-silencing (BSMV-VIGS) method was used to verify the functional characterization of the identified regulators in starch biosynthesis. RESULTS: Disulfide isomerase 1-2 protein (TaPDIL1-2) was screened out, and its binding to the TaAGPL1-1D promoter was further verified using another yeast one-hybrid screen. Transiently silenced wheat plants of the TaPDIL1-2 gene were obtained by using BSMV-VIGS method under field conditions. In grains of BSMV-VIGS-TaPDIL1-2-silenced wheat plants, the TaAGPL1 gene transcription levels, grain starch contents, and 1000-kernel weight also significantly increased. CONCLUSIONS: As important chaperones involved in oxidative protein folding, PDIL proteins have been reported to form hetero-dimers with some transcription factors, and thus, our results suggested that TaPDIL1-2 protein could indirectly and negatively regulate the expression of the TaAGPL1 gene and function in starch biosynthesis.

Low pH stress responsive transcriptome of seedling roots in wheat (Triticum aestivum L.).

Soil acidification is one of major problems limiting crop growth and especially becoming increasingly serious in China owing to excessive use of nitrogen fertilizer. Only the STOP1 of Arabidopsis was identified clearly sensitive to proton rhizotoxicity and the molecular mechanism for proton toxicity tolerance of plants is still poorly understood. The main objective of this study was to investigate the transcriptomic change in plants under the low pH stress. The low pH as a single factor was employed to induce the response of the wheat seedling roots. Wheat cDNA microarray was used to identify differentially expressed genes (DEGs). A total of 1057 DEGs were identified, of which 761 genes were up-regulated and 296 were down-regulated. The greater percentage of up-regulated genes involved in developmental processes, immune system processes, multi-organism processes, positive regulation of biological processes and metabolic processes of the biological processes. The more proportion of down-regulation genes belong to the molecular function category including transporter activity, antioxidant activity and molecular transducer activity and to the extracellular region of the cellular components category. Moreover, most genes among 41 genes involved in ion binding, 17 WAKY transcription factor genes and 17 genes related to transport activity were up-regulated. KEGG analysis showed that the jasmonate signal transduction and flavonoid biosynthesis might play important roles in response to the low pH stress in wheat seedling roots. Based on the data, it is can be deduced that WRKY transcription factors might play a critical role in the transcriptional regulation, and the alkalifying of the rhizosphere might be the earliest response process to low pH stress in wheat seedling roots. These results provide a basis to reveal the molecular mechanism of proton toxicity tolerance in plants.

The roles of H2S and H2O2 in regulating AsA-GSH cycle in the leaves of wheat seedlings under drought stress.

This paper investigated the roles of hydrogen sulfide (H2S) and hydrogen peroxide (H2O2) and the possible relationship between them in regulating the AsA-GSH cycle in wheat leaves under drought stress (DS). Results showed that DS markedly increased the production of H2S and H2O2, the transcript levels and activities of ascorbate peroxidase (APX), glutathione reductase (GR), monodehydroascorbate reductase (MDHAR), and dehydroascorbate reductase (DHAR); malondialdehyde (MDA) content; and electrolyte leakage (EL). Meanwhile, DS markedly reduced plant height and biomass. Above increases induced by drought stress except MDA content and EL were all suppressed by pretreatments with H2S synthesis inhibitor aminooxyaceticacid (AOA) and H2O2 synthesis inhibitor diphenylene iodonium (DPI). Besides, pretreatments with AOA and DPI further significantly increased MDA content and EL and significantly reduced plant height and biomass under DS. DPI reduced the production of H2O2 and H2S induced by DS. AOA also reduced the production of H2S and H2O2 induced by DS. Pretreatments with NaHS + AOA and H2O2 + DPI reversed above effects of AOA and DPI. Our results suggested that H2S and H2O2 all participated in the up-regulation of AsA-GSH cycle in wheat leaves by DS and possibly affected each other.