Understanding the responses of tillering to 2,4-D isooctyl ester in Setaria viridis L.

BACKGROUND: Green foxtail [Setaria viridis (L.)] is one of the most abundant and troublesome annual grass weeds in alfalfa fields in Northeast China. Synthetic auxin herbicide is widely used in agriculture, while how auxin herbicide affects tillering on perennial grass weeds is still unclear. A greenhouse experiment was conducted to examine the effects of auxin herbicide 2,4-D on green foxtail growth, especially on tillers. RESULTS: In the study, 2,4-D isooctyl ester was used. There was an inhibition of plant height and fresh weight on green foxtail after application. The photosynthetic rate of the leaves was dramatically reduced and there was an accumulation of malondialdehyde (MDA) content. Moreover, applying 2,4-D isooctyl ester significantly reduced the tillering buds at rates between 2100 and 8400 ga. i. /ha. Transcriptome results showed that applying 2,4-D isooctyl ester on leaves affected the phytohormone signal transduction pathways in plant tillers. Among them, there were significant effects on auxin, cytokinin, abscisic acid (ABA), gibberellin (GA), and brassinosteroid signaling. Indeed, external ABA and GA on leaves also limited tillering in green foxtail. CONCLUSIONS: These data will be helpful to further understand the responses of green foxtail to 2, 4-D isooctyl ester, which may provide a unique perspective for the development and identification of new target compounds that are effective against this weed species.

Effects of copper, and aluminium in ionic, and nanoparticulate form on growth rate and gene expression of Setaria italica seedlings.

This study aims to determine the effects of copper, copper oxide nanoparticles, aluminium, and aluminium oxide nanoparticles on the growth rate and expression of ACT-1, CDPK, LIP, NFC, P5CR, P5CS, GR, and SiZIP1 genes in five days old seedling of Setaria italica ssp. maxima, cultivated in hydroponic culture. Depending on their concentration (ranging from 0.1 to 1.8 mg L-1), all tested substances had both stimulating and inhibiting effects on the growth rate of the seedlings. Copper and copper oxide-NPs had generally a stimulating effect whereas aluminium and aluminium oxide-NPs at first had a positive effect but in higher concentrations they inhibited the growth. Treating the seedlings with 0.4 mg L-1 of each tested toxicant was mostly stimulating to the expression of the genes and reduced the differences between the transcript levels of the coleoptiles and roots. Increasing concentrations of the tested substances had both stimulating and inhibiting effects on the expression levels of the genes. The highest expression levels were usually noted at concentrations between 0.4 and 1.0 mg/L of each metal and metal nanoparticle, except for SiZIP1, which had the highest transcript amount at 1.6 mg L-1 of Cu2+ and at 0.1-0.8 mg L-1 of CuO-NPs, and LIP and GR from the seedling treated with Al2O3-NPs at concentrations of 0.1 and 1.6 mg L-1, respectively.

Genome-wide identification of the MED25 BINDING RING-H2 PROTEIN gene family in foxtail millet (Setaria italica L.) and the role of SiMBR2 in resistance to abiotic stress in Arabidopsis.

MAIN CONCLUSION: The SiMBR genes in foxtail millet were identified and studied. Heterologous expression of SiMBR2 in Arabidopsis can improve plant tolerance to drought stress by decreasing the level of reactive oxygen species. Foxtail millet (Setaria italica L.), a C4 crop recognized for its exceptional resistance to drought stress, presents an opportunity to improve the genetic resilience of other crops by examining its unique stress response genes and understanding the underlying molecular mechanisms of drought tolerance. In our previous study, we identified several genes linked to drought stress by transcriptome analysis, including SiMBR2 (Seita.7G226600), a member of the MED25 BINDING RING-H2 PROTEIN (MBR) gene family, which is related to protein ubiquitination. Here, we have identified ten SiMBR genes in foxtail millet and conducted analyses of their structural characteristics, chromosomal locations, cis-acting regulatory elements within their promoters, and predicted transcription patterns specific to various tissues or developmental stages using bioinformatic approaches. Further investigation of the stress response of SiMBR2 revealed that its transcription is induced by treatments with salicylic acid and gibberellic acid, as well as by salt and osmotic stresses, while exposure to high or low temperatures led to a decrease in its transcription levels. Heterologous expression of SiMBR2 in Arabidopsis thaliana enhanced the plant's tolerance to water deficit by reducing the accumulation of reactive oxygen species under drought stress. In summary, this study provides support for exploring the molecular mechanisms associated with drought resistance of SiMBR genes in foxtail millet and contributing to genetic improvement and molecular breeding in other crops.

Alternative Splicing Plays a Crucial Role in the Salt Tolerance of Foxtail Millet.

Foxtail millet is an important cereal crop that is relatively sensitive to salt stress, with its yield significantly affected by such stress. Alternative splicing (AS) widely affects plant growth, development, and adaptability to stressful environments. Through RNA-seq analysis of foxtail millet under different salt treatment periods, 2078 AS events were identified, and analyses were conducted on differential gene (DEG), differential alternative splicing gene (DASG), and overlapping gene. To investigate the regulatory mechanism of AS in response to salt stress in foxtail millet, the foxtail millet AS genes SiCYP19, with two AS variants (SiCYP19-a and SiCYP19-b), were identified and cloned. Yeast overexpression experiments indicated that SiCYP19 may be linked to the response to salt stress. Subsequently, we conducted overexpression experiments of both alternative splicing variants in foxtail millet roots to validate them experimentally. The results showed that, under salt stress, both SiCYP19-a and SiCYP19-b jointly regulated the salt tolerance of foxtail millet. Specifically, overexpression of SiCYP19-b significantly increased the proline content and reduced the accumulation of reactive oxygen species (ROS) in foxtail millet, compared to that in SiCYP19-a. This shows that SiCYP19-b plays an important role in increasing the content of proline and promoting the clearance of ROS, thus improving the salt tolerance of foxtail millet.

Potassium humate supplementation improves photosynthesis and agronomic and yield traits of foxtail millet.

Foxtail millet is a highly nutritious crop, which is widely cultivated in arid and semi-arid areas worldwide. Humic acid (HA), as a common plant growth regulator, is used as an organic fertilizer and feed additive in agricultural production. However, the impact of potassium humate KH on the photosynthetic rate and yield of foxtail millet has not yet been studied. We explored the effects of KH application on the morphology, photosynthetic ability, carbon and nitrogen metabolism, and yield of foxtail millet. A field experiment was performed using six concentrations of KH (0, 20, 40, 80, 160, and 320 kg ha-1) supplied foliarly at the booting stage in Zhangza 10 cultivar (a widely grown high-yield variety). The results showed that KH treatment increased growth, chlorophyll content (SPAD), photosynthetic rate (Pn), transpiration rate (Tr), and stomatal conductance (Gs). In addition, soluble protein content, sugar content, and nitrate reductase activity increased in KH-treated plants. With increased KH concentration, the effects became more evident and the peak values of each factor were achieved at 80 kg ha-1. Photosynthetic rate showed significant correlation with SPAD, Tr, Gs, and soluble protein content, but was negatively correlated with intercellular CO2 concentration. Compared to that of the control, the yield of foxtail millet under the T2, T3, T4, and T5 (40, 80, 160, and 320 kg ha-1 of KH) treatments significantly increased by 6.0%, 12.7%, 10.5%, and 8.6%, respectively. Yield exhibited a significant positive correlation with Tr, Pn, and Gs. Overall, KH enhances photosynthetic rate and yield of foxtail millet, therefore it may be conducive to stable millet production. These findings may provide a theoretical basis for the green and efficient production of millet fields.

New α-pyrone derivatives with herbicidal activity from the endophytic fungus Alternaria brassicicola.

Three pairs of undescribed enantiomeric α-pyrone derivatives (1a/1b-3a/3b) and six undescribed congeners (4-9), were obtained from the fungus Alternaria brassicicola that was isolated from the fresh leaves of Siegesbeckia pubescens Makino (Compositae). The structures of these new compounds were characterized by extensive NMR spectroscopic and HRESIMS data, and their absolute configurations were further elucidated by a modified Mosher's method, chemical conversion, single-crystal X-ray diffraction analysis, and ECD calculations. This is the first report of three pairs of enantiomeric α-pyrone derivatives from the fungus A. brassicicola, and these enantiomers were successfully acquired from scalemic mixtures via chiral HPLC. Compounds 1a/1b-3a/3b and 4-9 were evaluated for the herbicidal activity against Echinochloa crusgalli, Setaria viridis, Portulaca oleracea, and Taraxacum mongolicum. At a concentration of 100 µg/mL, compounds 1a and 1b could significantly inhibit the germination of monocotyledon weed seeds (E. crusgalli and S. viridis) with inhibitory ratios ranging from 68.6 ± 6.4% to 84.2 ± 5.1%, which was equivalent to that of the positive control (glyphosate). The potential structure-herbicidal activity relationship of these compounds was also discussed. To a certain extent, the results of this study will attract great interest for the potential practical application of promising fungal metabolites, α-pyrone derivatives, as ecofriendly herbicides.

Phytotoxic Activity of Alkaloids in the Desert Plant Sophora alopecuroides.

Sophora alopecuroides is known to produce relatively large amounts of alkaloids; however, their ecological consequences remain unclear. In this study, we evaluated the allelopathic potential of the main alkaloids, including aloperine, matrine, oxymatrine, oxysophocarpine, sophocarpine, sophoridine, as well as their mixture both in distilled H2O and in the soil matrix. Our results revealed that all the alkaloids possessed inhibitory activity on four receiver species, i.e., Amaranthus retroflexus, Medicago sativa, Lolium perenne and Setaria viridis. The strength of the phytotoxicity of the alkaloids was in the following order: sophocarpine > aloperine > mixture > sophoridine > matrine > oxysophocarpine > oxymatrine (in Petri dish assays), and matrine > mixture > sophocarpine > oxymatrine > oxysophocarpine > sophoridine > aloperine (in pot experiments). In addition, the mixture of the alkaloids was found to significantly increase the IAA content, MDA content and POD activity of M. sativa seedlings, whereas CTK content, ABA content, SOD activity and CAT activity of M. sativa seedlings decreased markedly. Our results suggest S. alopecuroides might produce allelopathic alkaloids to improve its competitiveness and thus facilitate the establishment of its dominance; the potential value of these alkaloids as environmentally friendly herbicides is also discussed.

Potassium supply promotes the mitigation of NaCl-induced effects on leaf photochemistry, metabolism and morphology of Setaria viridis.

Soil salinity has the potential to severely affect crop performance. To maintain cell functioning and improve salt tolerance, the maintenance of K+ homeostasis is crucial in several plant metabolism processes. Besides, potassium fertilization can efficiently alleviate the perilous effects of salinity. We characterized impacts in Setaria viridis exposed to NaCl and KCl to underlying photochemistry mechanisms, K+ and Na+ shoot contents, enzymatic activity, electrolytic leakage, and morphological responses focusing on non-stomatal limitation of photosynthesis. Plants were exposed to sodium chloride (NaCl; 0, 150 and 250 mM) and potassium chloride (KCl; 0, 5, 9 mM). The exposure to NaCl affected S. viridis leaves morphological and physiologically. Plants submitted to 150 mM showed reductions in performance indexes (PIabs and PItotal; JIP-test), and the presence of positive K- and L-bands. Plants exposed to 250 mM exhibited blockage in electron flow further than QA within 48h and permanent photoinhibition at 96 h. The presence of 9 and 5 mM of KCl counteracted the effects of NaCl on plants submitted to 150 mM, concomitant with increases in K+ accumulation and cell turgidity conservation, causing positive effects in plant growth and metabolism. Neither KCl concentrations were effective in reducing NaCl-induced effects on plants exposed to 250 mM of NaCl. Our results support the conclusion that greater availability of K+ alleviates the harmful effects of salinity in S. viridis under moderate stress and that application of KCl as means of lightning saline stress has a concentration and a salt level limit that must be experimentally determined.

Transcriptome analysis identified the mechanism of synergy between sethoxydim herbicide and a mycoherbicide on green foxtail.

Certain synthetic herbicides can act synergistically with specific bioherbicides. In this study, a sethoxydim herbicide at 0.1× label rate improved biocontrol of herbicide-sensitive green foxtail (Setaria viridis, GFT) by Pyricularia setariae (a fungal bioherbicide agent), but did not change the efficacy on a herbicide-resistant GFT biotype. Reference transcriptomes were constructed for both GFT biotypes via de novo assembly of RNA-seq data. GFT plants treated with herbicide alone, fungus alone and herbicide + fungus were compared for weed-control efficacy and differences in transcriptomes. On herbicide-sensitive GFT, sethoxydim at the reduced rate induced ABA-activated signaling pathways and a bZIP transcription factor 60 (TF bZIP60), while improved the efficacy of biocontrol. The herbicide treatment did not increase these activities or improve biocontrol efficacy on herbicide-resistant plants. An exogenous application of ABA to herbicide-sensitive plants also enhanced bZIP60 expression and improved biocontrol efficacy, which supported the results of transcriptome analysis that identified the involvement of ABA and bZIP60 in impaired plant defense against P. setariae. It is novel to use transcriptome analysis to decipher the molecular basis for synergy between a synthetic herbicide and a bioherbicide agent. A better understanding of the mechanism underlining the synergy may facilitate the development of weed biocontrol.

Expression of PHT1 family transporter genes contributes for low phosphate stress tolerance in foxtail millet (Setaria italica) genotypes.

MAIN CONCLUSION: This is a first comprehensive study to analyze the 12 PHT1 family phosphate transporter genes in 20 foxtail millet genotypes for the improvement of millets and other crops for phosphate use efficiency. Phosphorus (P), absorbed from soil solutions as inorganic phosphate (Pi), is a limiting nutrient for plant growth and yield. Twenty genotypes of foxtail millet (Setaria italica) with contrasting degree of growth and Pi uptake responses under low Pi (LP) and high Pi (HP) supply were chosen based on a previous study. To gain molecular insights, expression dynamics of 12 PHosphate Transporter 1 (PHT1) family (SiPHT1;1 to 1;12) genes were analyzed in these 20 genotypes and compared with their Pi and total P (TP) contents. SiPHT1;1, 1;2, 1;3 and 1;8 genes were expressed in shoot tissues of three (ISe 1209, ISe 1305 and Co-6) of the LP best performing genotypes (LPBG); however, they were expressed in only one of the LP worst performing genotype (LPWG) (ISe 748). More importantly, this is correlating with higher shoot Pi and TP contents of the LPBG compared to LPWG. Apart from this condition, expression of SiPHT1 genes and their Pi and TP contents do not correlate directly for many genotypes in other conditions; genotypes with low Pi and TP contents induced more SiPHT1 genes and vice versa. Promoter analysis revealed that genotype ISe 1888 with a high level of SiPHT1;8 expression possesses two additional root box motifs compared to other genotypes. The PHT1 family genes seem to play a key role for LP stress tolerance in foxtail millet and further studies will help to improve the P-use efficiency in foxtail millet and other cereals.

Simple, Effective, and Ecofriendly Strategy to Inhibit Droplet Bouncing on Hydrophobic Weed Leaves.

Despite small-molecule surfactants and polymers being widely used as pesticide adjuvants to inhibit droplet bouncing and splashing, they still have intrinsic drawbacks either in the easy wind drift and evaporation, the unfavorable wettability, or the usage of nonrenewable resources. In this paper, we found that upon droplet impacting, 1D nanofibers assembled from natural glycyrrhizic acid (GL) could pin on the rough hydrophobic surface and delay the retraction rate of droplets effectively. Using GL as a tank-mixed adjuvant, the efficiency of glyphosate to control the weed growth was improved significantly in the field experiment, which addressed the dilemmas of current adjuvants elegantly. Our work not only provides a constructive way to overcome droplet bouncing but also prompted us to verify in future if all 1D nanofibers assembled from different small molecules can display similar control efficiencies.

High throughput deep sequencing elucidates the important role of lncRNAs in Foxtail millet response to herbicides.

Long non-coding RNAs (lncRNAs) play an important function in plant growth and development as well as response to stresses. However, little information was known in foxtail millet; no study was reported on lncRNAs in plant response to herbicide treatment. In this study, by using deep sequencing and advanced bioinformatic analysis, a total of 2547 lncRNAs were identified, including 787 known and 1760 novel lncRNAs. These lncRNAs are distributed across all 9 chromosomes, and the majority were located in the intergenic region with 1-2 exons. These lncRNAs were differentially expressed between different genotypes under different herbicide treatments. lncRNAs regulate plant growth and development as well as response to herbicide treatments through targeting protein-coding genes that directly relate to chemical metabolism and defense system. Multiple potential target genes and lncRNA-mRNA-miRNA gene networks were discovered. These results elucidate the potential roles of lncRNAs in plant response to herbicides.

An investigation into the beneficial effects and molecular mechanisms of humic acid on foxtail millet under drought conditions.

The aim of this study was to determine the effects and underlying molecular mechanisms of humic acid (HA) on foxtail millet (Setaria italica Beauv.) under drought conditions. The rainless climate of the Shanxi Province (37°42'N, 112°58'E) in China provides a natural simulation of drought conditions. Two foxtail millet cultivars, Jingu21 and Zhangza10, were cultivated in Shanxi for two consecutive years (2017-2018) based on a split-plot design. Plant growth, grain quality, and mineral elements were analyzed in foxtail millet treated with HA (50, 100, 200, 300, and 400 mg L-1) and those treated with clear water. Transcriptome sequencing followed by bioinformatics analysis was performed on plants in the normal control (CK), drought treatment (D), and drought + HA treatment (DHA) groups. Results were verified using real-time quantitative PCR (RT-qPCR). HA at a concentration of 100-200 mg L-1 caused a significant increase in the yield of foxtail millet and had a positive effect on dry weight and root-shoot ratio. HA also significantly increased P, Fe, Cu, Zn, and Mg content in grains. Moreover, a total of 1098 and 409 differentially expressed genes (DEGs) were identified in group D vs. CK and D vs. DHA, respectively. A protein-protein interaction network and two modules were constructed based on DEGs (such as SETIT_016654mg) between groups D and DHA. These DEGs were mainly enriched in the metabolic pathway. In conclusion, HA (100 mg L-1) was found to promote the growth of foxtail millet under drought conditions. Furthermore, SETIT_016654mg may play a role in the effect of HA on foxtail millet via control of the metabolic pathway. This study lays the foundation for research into the molecular mechanisms that underlie the alleviating effects of HA on foxtail millet under drought conditions.

Sensitivity of yellow foxtail (Setaria glauca L.) and barnyardgrass (Echinochloa crus-galli L.) to aqueous extracts or dry biomass of cover crops.

BACKGROUND: The common weeds Echinochloa crus-galli L. and Setaria glauca L. were studied for their sensitivity to aqueous extracts or dry biomass of the following cover crops (CCs): Brassicaceae (Sinapis alba, Raphanus sativus var. Oleiformis, Camellina sativa), Fagopyrum esculentum and Guizotia abyssinica. RESULTS: Treating E. crus-galli with aqueous extracts of mixed CCs or individual brassica CC significantly reduced germination. Treating S. glauca with aqueous extracts of C. sativa or G. abyssinicia reduced germination. Aqueous extracts of all CCs significantly reduced radicle length of E. crus-galli and S. glauca, with C. sativa and mixed CCs showing the greatest effect. Aqueous extracts of nearly all CCs delayed start and middle germination of E. crus-galli and S. glauca, with S. alba and R. sativus showing the strongest effects. Aqueous extracts of Brassicaceae leaf and flower significantly reduced germination, coleoptile length, radicle length and seedling biomass of E. crus-galli and S. glauca. Brassicaceae leaves and flowers contained higher phenolics than other tissues. Adding 4 or 8% S. alba and R. sativus dry powder to soil significantly reduced growth of E. crus-galli and S. glauca; even concentrations of 1% measurably slowed growth of E. crus-galli. CONCLUSIONS: Brassicaceae may be allelopathic to S. glauca and E. crus-galli. Aqueous extracts of leaves and flowers showed greater phytotoxic activity than other tissues and also contained more phenolics. Therefore Brassicaceae CCs may be most effective against S. glauca and E. crus-galli if incorporated into soil during their flowering stage. © 2020 Society of Chemical Industry.

Genomic dissection and expression analysis of stress-responsive genes in C4 panicoid models, Setaria italica and Setaria viridis.

The study reports the identification and expression profiling of five major classes of C4 pathway-specific genes, namely, carbonic anhydrase (CaH), phosphoenolpyruvate carboxylase (PEPC), pyruvate orthophosphate dikinase (PPDK), NADP-dependent malate dehydrogenase (MDH) and NADP-dependent malic enzyme (NADP-ME), in the model species, Setaria italica and Setaria viridis. A total of 42 and 41 genes were identified in S. italica and S. viridis, respectively. Further analysis revealed that segmental and tandem duplications have contributed to the expansion of these gene families. RNA-Seq derived expression profiles of the gene family members showed their differential expression pattern in tissues and dehydration stress. Comparative genome mapping and Ks dating provided insights into their duplication and divergence in the course of evolution. Expression profiling of candidate genes in contrasting S. italica cultivars subjected to abiotic stresses and hormone treatments showed distinct stress-specific upregulation of SiαCaH1, SißCaH5, SiPEPC2, SiPPDK2, SiMDH8, and SiNADP-ME5 in the tolerant cultivar. Overexpression of SiNADP-ME5 in heterologous yeast system enabled the transgenic cells to survive and grow in dehydration stress conditions, which highlights the putative role of SiNADP-ME5 in conferring tolerance to dehydration stress. Altogether, the study highlights key genes that could be potential candidates for elucidating their functional roles in abiotic stress response.

Foxtail millet (Setaria italica (L.) P. Beauv) CIPKs are responsive to ABA and abiotic stresses.

CBL-interacting protein kinases (CIPKs) have been shown to regulate a variety of environmental stress-related signalling pathways in plants. Foxtail millet (Setaria italica (L.) P. Beauv) is known worldwide as a relatively stress-tolerant C4 crop species. Although the foxtail millet genome sequence has been released, little is known about the functions of CIPKs in foxtail millet. Therefore, a systematic genome-wide analysis of CIPK genes in foxtail millet was performed. In total, 35 CIPK members were identified in foxtail millet and divided into four subgroups (I to IV) on the basis of their phylogenetic relationships. Phylogenetic and gene structure analyses clearly divided all SiCIPKs into intron-poor and intron-rich clades. Cis-element analysis subsequently indicated that these SiCIPKs may be involved in responses to abiotic stimuli, hormones, and light signalling during plant growth and development, and stress-induced expression profile analysis revealed that all the SiCIPKs are involved in various stress signalling pathways. These results suggest that the CIPK genes in foxtail millet exhibit the basic characteristics of CIPK family members and play important roles in response to abiotic stresses. The results of this study will contribute to future functional characterization of abiotic stress responses mediated by CIPKs in foxtail millet.

Photosynthesis and yield response to elevated CO2, C4 plant foxtail millet behaves similarly to C3 species.

Foxtail millet (Setaria italica) is a nutrient-rich food source traditionally grown in arid and semi-arid areas, as it is well adapted to drought climate. Yet there is limited information as how the crop responses to the changing climate. In order to investigate the response of foxtail millet to elevated [CO2] and the underlying mechanism, the crop was grown at ambient [CO2] (400 µmol mol-1) and elevated [CO2] (600 µmol mol-1) in an open-top chamber (OTC) experimental facility in North China. The changes in leaf photosynthesis, chlorophyll fluorescence, biomass, yield and global gene expression in response to elevated [CO2] were determined. Despite foxtail millet being a C4 photosynthetic crop, photosynthetic rates (PN) and intrinsic water-use efficiency (WUEi), were increased under elevated [CO2]. Similarly, grain yield and above-ground biomass also significantly increased (P <  0.05) for the two years of experimentation under elevated [CO2]. Increases in seeds and tiller number, spike and stem weight were the main contributors to the increased grain yield and biomass. Using transcriptomic analyses, this study further identified some genes which play a role in cell wall reinforcement, shoot initiation, stomatal conductance, carbon fixation, glycolysis / gluconeogenesis responsive to elevated [CO2]. Changes in these genes reduced plant height, increased stem diameters, and promote CO2 fixation. Higher photosynthetic rates at elevated [CO2] demonstrated that foxtail millet was not photosynthetically saturated at elevated [CO2] and its photosynthesis response to elevated [CO2] were analogous to C3 plants.

Enhanced drought tolerance of foxtail millet seedlings by sulfur dioxide fumigation.

Recently, sulfur dioxide (SO2) has been considered to be a beneficial bio-regulator in animals. However, the positive roles of SO2 in plant adaptation to drought stress are still unclear. In this study, we investigated the physiological and molecular changes that are induced by SO2 fumigation to improve the drought tolerance of foxtail millet seedlings. The relative water content in the leaves of drought-stressed seedlings was significantly improved by pre-exposure to 30 mg/m3 SO2. These responses might be related to decreased stomatal apertures and a reduced leaf transpiration rate, which were induced by SO2 under drought conditions. In addition, the SO2 pretreatment markedly enhanced proline accumulation in the leaves of drought-stressed seedlings, which was supported by increased Δ1-pyrroline-5-carboxylate synthetase (P5CS) activity, decreased proline dehydrogenase (ProDH) activity, and the corresponding transcripts. Moreover, the SO2 application upregulated the enzyme activity of catalase (CAT) and peroxidase (POD) in the leaves of drought-stressed plants, as well as their transcripts, which contributed to the scavenging of hydrogen peroxide (H2O2) and alleviated drought-induced oxidative damage, as indicated by the decreased malondialdehyde (MDA) level in SO2-pretreated plants. Together, these results indicate that the application of SO2 might enhance drought tolerance by reducing stomatal apertures, increasing proline accumulation, and promoting antioxidant defence in foxtail millet seedlings. This study presents new insight into the beneficial roles of SO2 in plant responses to drought stress.

Effect of irrigation with detergent-containing water on foxtail millet shoot biomass and ion accumulation.

Water shortage leads farmers to use sewages for irrigation. Sewages contain a large amount of laundry detergent. In this study the impact of irrigation by contaminated water on shoot biomass and seed germination of foxtail millet (Setaria italica) was investigated. The research was conducted as laboratory and pot experiments. Iso-potentials (- 0.042, - 0.077, and - 0.415 MPa) of polyethylene glycol (PEG, water deficit treatment) and laundry detergent (contamination treatment) made the laboratory experiment treatments. The pot experiment included contamination factor (0, 0.1, 1, and 10 g L-1 of laundry detergent) and deficit irrigation factor (irrigation interval of 1, 2, and 3 days). Results of this study showed that at the iso-potential, laundry detergent had more negative effect on seed germination traits when compared with PEG. There was no germination at - 0.415 MPa of laundry detergent. Both drought and contamination reduced dry forage yield, plant height, leaf number, leaf area, leaf dry and fresh weight, stem dry, and fresh weight. Detergent concentration of 10 g L-1 with irrigation interval of 3 days had a forage yield reduction of 63% compared to control (laundry detergent concentration of 0 g L-1 with irrigation interval of 1 day). Detergent concentration of 10 g L-1 with irrigation interval of 1 day had a sodium increase of 1847% compared to control. Based on the results of this study, it is recommended not to irrigate foxtail millet farm by contaminated water with laundry detergent higher than 1 g L-1.

Sulfur dioxide derivatives alleviate cadmium toxicity by enhancing antioxidant defence and reducing Cd2+ uptake and translocation in foxtail millet seedlings.

Sulfur dioxide (SO2) was recently proposed as a novel bio-regulator in mammals. However, the possible advantageous effects of SO2 in plant adaptation to heavy metal-contaminated environments are largely unknown. In the present study, using Na2SO3/NaHSO3 derivatives as SO2 donors, we investigated the possible roles and regulation mechanisms of SO2 in alleviating Cd2+ toxicity in foxtail millet seedlings. Exogenous SO2 derivatives (0.5 mM) application significantly reduced the seedling growth inhibition caused by Cd2+ stress. Cd2+-induced oxidative damage was also alleviated by SO2 derivatives, which was supported by the decreased malondialdehyde (MDA) level in the leaves of seedlings pretreated with SO2 derivatives. These responses were related to the enhanced activities of representative antioxidant enzymes, including catalase and superoxide dismutase, as well as the up-regulation of ascorbate-glutathione cycle, which contributed to the scavenging of Cd2+-elicited O2•－ and H2O2 within the leaves of foxtail millet seedlings. Also, SO2 derivative application promoted sulfur assimilation and increased the content of glutathione and phytochelatins, which may help to enhance Cd2+ detoxification capacity in foxtail millet seedlings. Moreover, application of SO2 derivatives caused down-regulation of the transcript expression levels of several genes involved in Cd2+ uptake and translocation, such as NRAMP1, NRAMP6, IRT1, IRT2, HMA2, and HMA4, thus resulting in reduced Cd2+ accumulation in the shoots and roots of Cd2+-stressed seedlings. Collectively, these results suggest that exogenous SO2 derivative application can alleviate oxidative damage and restrict Cd2+ buildup, thereby reducing Cd2+-induced growth inhibition in foxtail millet seedlings upon Cd2+ exposure. This novel finding indicates that the usage of SO2 derivatives may be an effective approach for enhancing Cd2+ tolerance in foxtail millet and other crops.