Pb Stress Research Articles

Comparative efficiency of silica gel, biochar, and plant growth promoting bacteria on Cr and Pb availability to Solanum melongena L. in contaminated soil irrigated with wastewater

Crop irrigation with untreated wastewater is a routine practice in developing countries that causes multiple human health consequences. A comparative study was performed to regulate total Cr and Pb stress in soil and Solanum melongena L. plant. For this purpose, 0.2% chitosan polymerized silica gel (CP-silica gel), 1.5% zinc-enriched biochar (ZnBc), and three bacterial species such as Trichococcus sp. (B1), Pseudomonas alcaligenes (B2), and Bacillus subtilis (B3) were selected. Initially, a biosorption trial was conducted to test the heavy metal removal efficiency of three bacterial species B1, B2, and B3 for 24 h. Hence, B3 showed maximum Cr and Pb removal efficiency among the studied bacterial isolates. Then, a pot study was conducted with 12 different treatments having three replicates. After harvesting, different growth and biochemical parameters such as chlorophyll concentration, proteins, phenolics, reactive oxygen species, and antioxidant enzymes were analyzed. The results demonstrated that wastewater application significantly (p ≤ 0.01) reduced the fresh and dry weights of the root, stem, and leaves due to high total Cr and Pb toxicity. However, CP-silica gel and ZnBc treatments performed best when applied in combination with B3. The concentration of leaf total Cr was significantly decreased (91 and 85%) with the application of ZnBc + B3 and CP-Silica gel + B3, respectively, as compared to control. There was a reduction in stem hydrogen peroxide (87%) and malondialdehyde (81%) recorded with CP-silica gel + B3 treatment due to enhanced activities of antioxidant enzymes viz. ascorbate peroxidase (6-folds) and catalase (7-folds) relative to control. Similarly, leaf total phenolics (3-folds) and protein (6-folds) contents were enhanced with CP silica gel+B3 application relative to control. Overall, CP-silica gel and ZnBc with B3 application proved to be the most appropriate treatments and can be used in developing countries to limit the deleterious effects of total Cr and Pb pollution.

Combination of FOBIO biopesticide and Brassica rapa L. as remediator of heavy metal Pb in soil

Marginal soil is infertile soil caused by various factors, one of which is the excessive accumulation of heavy metals. FOBIO microorganism-based biopesticide formula containing phosphate solubilizing bacteria is expected to be used to reduce excess heavy metals in the soil. Brasica rapa L. is a variety resistant to Pb heavy metal stress and absorbs more heavy metals into root tissue than leaf tissue with the help of microorganisms. This study aimed to determine the combination of Brassica rapa L. and FOBIO biopesticide in absorbing Pb in the soil, determine the growth of Brassica rapa L. at each Pb concentration, and determine the population of phosphate-solubilizing bacteria in the soil at each level of Pb stress after remediation. This study used variations in the concentration of heavy metals, particularly Pb. Parameters observed included plant growth, Pb concentration in the soil before and after remediation, and the population of phosphate-solubilizing bacteria. The results showed that combining FOBIO biopesticides and Brassica rapa L. could reduce Pb concentration in the soil. The growth of Brassica rapa L. plants was significantly different after treatment. The findings confirmed that increasing, the concentration of Pb led to a decrease in the population of phosphate-solubilizing bacteria.

Priming with EDTA, IAA and Fe alleviates Pb toxicity in Trigonella Foneum graecum L. growth: Phytochemicals and secondary metabolites

This study evaluated the effects of the exogenous application of ethylenediaminetetraacetic acid (EDTA), indole-3-acetic acid (IAA) and iron sulfate (FeSO4) upon the phytochemical mechanisms of fenugreek grown under Pb-excess (2000 mg L−1 PbCl2). The results showed that chemical additives of EDTA and IAA as well as FeSO4 improved fenugreek germination parameters. The radicle length and the amylase activity were significantly improved under IAA treatment compared to EDTA and FeSO4. Exogenous FeSO4 was more effective to improving growth parameters. Moreover, the decrease in hydrogen peroxide (H2O2) and malondialdehyde (MDA) levels was noted under all chemical additives especially under IAA application. In addition, it was more effective than EDTA and Fe in increasing catalase, glutathione (GSH), ascorbate peroxidase (APX), flavonoids and phenols while the increment superoxide dismutase (SOD) production was more pronounced under EDTA addition to Pb than other chelators. HPLC analysis revealed that the gallic was the major phenol produced under all chelators addition especially with IAA. In addition, the syringic acid was only produced with exogenous IAA while the quercetin was only detected under EDTA addition. Our results exhibited a higher IAA efficiency than EDTA and FeSO4 in mitigating Pb stress in fenugreek through up-regulated mechanisms of the antioxidant system for reducing reactive oxygen species (ROS) activities and enhancing special phenols.

Phytochemical composition, cytotoxicity, antioxidant and antimicrobial responses of Lavandula dentata L. grown under different levels of heavy metals stress condition

In order to know if the heavy metals stress condition is boon or bane for the plants growth, Lavandula dentata species was planted in pots under different levels of heavy metals stress condition and the phytochemical composition, cytotoxicity, antioxidant and antimicrobial responses of their leaf ethanolic extracts toward this stress condition were investigated compared to the control samples. Our findings showed significant differences in heavy metals bioaccumulation, photosynthetic pigments and total phenolic/flavonoids contents among L. dentata leafs ethanolic extracts, grown under different levels of heavy metals stress condition. The L. dentata leafs extracts, grown under Zn and Cu stress condition, showed the highest antioxidant and antimicrobial activities than those grown under Cd and Pb stress condition. Comparatively, the L. dentata leafs extracts, grown under Zn stress condition, showed higher antioxidant activity, and those, grown under Cu stress condition, showed higher antimicrobial activity. The highest cytotoxicity was showed by L. dentata leaf extracts, grown under Cd and Pb stress condition, which lead to conclude that these extracts could be served as a novel scaffold in search for new drugs against cancer. In conclusion, the highlighted variability reflects the high impact of heavy metals stress condition on phytochemical composition and consequently on the biological activities of medicinal plants. Such impact led to conclude that we should select medicinal plants extracts to be investigated carefully depending on this stress condition, in order to isolate the bioactive components or to have the best quality of extracts in terms of biological activities.

Exogenous Gamma-Aminobutyric Acid Application Induced Modulations in the Performance of Aromatic Rice Under Lead Toxicity.

Gamma-aminobutyric acid (GABA) is a non-protein amino acid and has a multi-functional role in abiotic stress tolerance. A pot experiment was conducted to assess the role of exogenous gamma-aminobutyric acid (GABA) application to modulate the growth, yield, and related physio-biochemical mechanisms in two aromatic rice cultivars, that is, Guixiangzhan (GXZ) and Nongxiang 18 (NX-18), under Pb toxic and normal conditions. The experimental treatments were comprised of Ck: without Pb and GABA (control), GABA: 1 mM GABA is applied under normal conditions (without Pb), Pb + GABA: 1 mM GABA is applied under Pb toxicity (800 mg kg−1 of soil), and Pb= only Pb (800 mg kg−1 of soil) is applied (no GABA). The required concentrations of GABA were applied as a foliar spray. Results revealed that Pb stress induced oxidative damage in terms of enhanced malondialdehyde (MDA), electrolyte leakage (EL), and H2O2 contents, while exogenous GABA application improved leaf chlorophyll, proline, protein and GABA contents, photosynthesis and gas exchange, and antioxidant defense under Pb toxicity in both rice cultivars. Moreover, glutamine synthetase (GS) and nitrate reductase (NR) activities were variably affected due to GABA application under Pb stress. The yield and related traits, that is, productive tillers/pot, grains/panicle, filled grain %, 1,000-grain weight, and grain yield were 13.64 and 10.29, 0.37% and 2.26%, 3.89 and 19.06%, 7.35 and 12.84%, and 17.92 and 40.56 lower under Pb treatment than Pb + GABA for GXZ and NX-18, respectively. Furthermore, exogenous GABA application in rice reduced Pb contents in shoot, leaves, panicle, and grains compared with Pb-exposed plants without GABA. Overall, GXZ performed better than NX-18 under Pb toxic conditions.

Alcaligenes faecalis Juj3 alleviates Plasmodiophora brassicae stress to cabbage via promoting growth and inducing resistance

Clubroot is a devastating disease threatening global cruciferous vegetable production caused by Plasmodiophora brassicae (Pb). We have evaluated the positive effects of the Alcaligenes faecalis Juj3 on cabbage growth promotion and Pb stress alleviation through pot and field experiments. The Juj3 strain was isolated from a healthy cabbage rhizosphere with growth-promoting characteristics and was identified as A. faecalis based on morphological traits and phylogeny. Seed germination assays revealed that Juj3 inoculation enhances cabbage bud shoot and root growth. In pot experiments, inoculation with Juj3 fermentation powder at cabbage sowing dates significantly improved the seedling biomass. Combining seed treatments with root irrigation after transplanting considerably reduced the clubroot disease index and resulted in appreciable biocontrol efficacy (83.7%). Gene expression analyses of cabbage after Juj3 inoculation showed that PR2 and EIN3 expression were significantly up-regulated. Physiologically, Juj3 inoculation enhanced cabbage chlorophyll content and root activity in a normal environment. Irrespective of whether plants were under normal environment or Pb stresses, Juj3 improved photosynthesis. Field trial analyses revealed that Juj3 exhibits satisfactory biocontrol efficacy in cabbage (51.4%) and Chinese cabbage (37.7%). Moreover, Juj3 could also enhance cabbage and Chinese cabbage biomass to improve the yield quality. These findings pave the way for future use of A. faecalis as biocontrol agents for clubroot and reveal the great potential of the rhizobacterium for plant growth-promoting applications in agriculture and horticulture.

Comparative proteomic analysis reveals insights into the response of Cryptolaemus montrouzieri to bottom-up transfer of cadmium and lead across a multi-trophic food chain

Contamination of agro-ecosystems with heavy metals can affect the development and reproduction of insect natural enemies. This study reports a detailed Tandem Mass Tag based quantitative proteomic analysis of underlying mechanisms responsible for stress response of Cryptolaemus montrouzieri against heavy metals (cadmium (Cd) and lead (Pb)) transported across a multi-trophic food chain. A total of 6639 proteins were detected under Cd as well as Pb stress. In Pb versus the control cluster, 69 proteins (28 up-regulated and 41 down-regulated) were differentially expressed whereas 268 proteins were differentially expressed under Cd versus the control cluster, having 198 proteins up-regulated and 70 down-regulated proteins. The analysis of differentially expressed proteins showed that 27 proteins overlapped in both clusters representing the core proteome to Pb and Cd stress. The bioinformatics analysis demonstrated that these proteins were mapped to 57 and 99 pathways in Pb versus control and Cd versus control clusters, respectively. The functional classification by COG, GO and KEGG databases showed significant changes in protein expression by C. montrouzieri under Pb and Cd stress. The heavy metal stress (Pb and Cd) induced significant changes in expression of proteins like hexokinase (HK), succinyl-CoA, trypsin like proteins, cysteine proteases, cell division cycle proteins, and yellow gene proteins. The results provide detailed information on the protein expression levels of C. montrouzieri and will serve as basic information for future proteomic studies on heavy metal responses of insect predators within a multi-trophic food chain.

Genetic diversity and growth responses of Indonesian tomato (Solanum lycopersicum L.) genotypes under lead stress.

Tomato (Solanum lycopersicum L.) is cultivated and consumed worldwide, including in Indonesia. It is used in the food, cosmetic, and pharmaceutical industries, due to its high content of carotenoid (lycopene) compounds that have antioxidant and anticancer activities. In Indonesia, although several cultivars of tomato are cultivated, including Opal, Permata, Mutiara, and Rewako, studies on their genetic information are limited. Unpredicted climate change as well as heavy metal contamination, especially Pb pollution, has threatened Indonesian food security. Therefore, our study aimed to analyze the genetic diversity of the four local tomatoes using random amplified polymorphic DNA (RAPD) markers and to determine the growth responses of several local tomato genotypes under Pb stress. In this study, morphological responses to Pb, including plant height and root length were observed. The RAPD analysis showed that Rewako and Permata were distinct, whereas Opal and Mutiara were closely related, possessing 81.8% similarity. Pb stress influenced plant height and root length in the four tomato genotypes, with each genotype exhibiting different morphological responses than others. However, the closely related Mutiara and Opal genotypes demonstrated similar responses to Pb stress to Permata and Rewako. Our study demonstrates that RAPD are sensitive and efficient for elucidating the genomic profile of the tomato genotypes. In addition, our results suggest that genetic variation among tomato genotypes might influence the morphological responses against Pb stress.

Synthesis and application of starch-stablized Fe–Mn/biochar composites for the removal of lead from water and soil

Starch-stablized and Fe/Mn bimetals modified biochar derived from corn straw (SFM@CBC and SFM@CBC-350) were firstly prepared, characterized (FTIR, XRD, SEM, EDS, BET and XPS), and applied in Pb removal from water and soil. SFM@CBC and SFM@CBC-350 displayed highly effective adsorption performance of Pb2+ from wastewater with the maximum adsorption capacity of 170.91 mg g−1 and 190.17 mg g−1, respectively, which were much greater than that of FM@CBC (149.25 mg g−1) and CBC (101.10 mg g−1). Studies of adsorption kinetics, isotherms and thermodynamics indicated that the absorption of Pb2+ by SFM@CBC and SFM@CBC-350 was spontaneous and endothermic reaction, and it was controlled by monolayer chemisorption. The mechanism studies indicated that Pb2+ removal involved with multiple mechanism, including complexation (dominant process confirmed by XPS analysis), physical adsorption, electrostatic attraction, and cation exchange. The reusability test demonstrated that SFM@CBC and SFM@CBC-350 had very good stability and reusability. In addition, in order to further explore Pb removal performance of the modified biochar, SFM@CBC-350 was used in soil-ryegrass pot systems. Compared with the controls, the addition of SFM@CBC-350 reduced Pb content in soil and ryegrass, increased the biomass and total chlorophyll content, reduced the activity of antioxidant enzymes (CAT, SOD, MDA and POD) and ROS fluorescence intensity of ryegrass, thus alleviating Pb stress of ryegrass. Besides, the addition of SFM@CBC-350 could increase the richness and diversity of soil microorganisms, which was beneficial to the growth of ryegrass. Hence, SFM@CBC-350 has the potential of being used as a green, efficient and promising adsorbent in Pb removal from wastewater and soil.

Transcriptome analysis reveals candidate genes involved in multiple heavy metal tolerance in hyperaccumulator Sedum alfredii

Sedum alfredii Hance is a perennial herb native to China that can particularly be found in regions with abandoned Pb/Zn mines. It is a Cd/Zn hyperaccumulator that is highly tolerant to Pb, Cu, Ni, and Mn, showing potential for phytoremediation of soils contaminated with multiple heavy metals. A better understanding of how this species responds to different heavy metals would advance the phytoremediation efficiency. In this study, transcriptomic regulation of S. alfredii roots after Cd, Zn, Pb, and Cu exposure was analyzed to explore the candidate genes involved in multi-heavy metal tolerance. Although Zn and Cd, Pb and Cu had similar distribution patterns in S. alfredii, distinct expression patterns were exhibited among these four metal treatments, especially about half of the differentially expressed genes were upregulated under Cu treatment, suggesting that it utilizes distinctive and flexible strategies to cope with specific metal stress. Most unigenes regulated by Cu were enriched in catalytic activity, whereas the majority of unigenes regulated by Pb had unknown functions, implying that S. alfredii may have a unique strategy coping with Pb stress different from previous cognition. The unigenes that were co-regulated by multiple heavy metals exhibited functions of antioxidant substances, antioxidant enzymes, transporters, transcription factors, and cell wall components. These metal-induced responses at the transcriptional level in S. alfredii were highly consistent with those at the physiological level. Some of these genes have been confirmed to be related to heavy metal absorption and detoxification, and some were found to be related to heavy metal tolerance for the first time in this study, like Metacaspase-1 and EDR6. These results provide a theoretical basis for the use of genetic engineering technology to modify plants by enhancing multi-metal tolerance to promote phytoremediation efficiency.

Antioxidant response by alfalfa (Medicago sativa L.) to Pb pollution - A study to value the feasibility of soil phytoremediation

With the surrounding environment of Inner Mongolia lead (Pb) ore as the research background, the germination and physio-biochemical effects of Pb stress on alfalfa were discussed to employ this species for the remediation of Pb contaminated soil. Research has shown that a low Pb stress concentration could improve the biological resistance of alfalfa seeds, while a high Pb stress concentration cannot be tolerated. Interestingly, when the Pb concentration was 5 mg/L, the germination rate of the seed was promoted, and the chlorophyll content was especially increased. As the Pb content and stress increased, the amount of malondialdehyde (MDA), H2O2, catalase (CAT) increased; while the root cell viability, chlorophyll and soluble protein content decreased. In consequence, alfalfa was tolerant to Pb stress of 5 mg/L, inversely, its growth was inhibited at levels higher than 5 mg/L, and it was poisoned at 500 mg/L. Based on the principal component analysis (PCA), the H2O2, O2–, chlorophyll total, chlorophyll a, CAT and proline content explicitly reflected the change in the physiology on the alfalfa and its tolerance under Pb stress.

Mitigation of lead toxicity in Vigna radiata genotypes by silver nanoparticles

Heavy metal (HM) contamination of the soil through anthropogenic activities influences the living systems and drastically impacts food chain. This study examined the application of silver nanoparticles (AgNPs) in two genotypes (G1 and G2) of Mung bean (Vigna radiata) for ameliorating the Pb toxicity. Different doses of Pb (0, 25, 50 μM) were differentially tackled by AgNPs with the aim of ameliorating the plant attributes. Both genotypes displayed statistically significant quantitative and qualitative modulations for Pb tolerance. In G2, the most prominent increase in plant height (43.79%), fresh biomass (49.56%) and total chlorophyll (20%) was observed at L2 (AgNPs 10 mg/L) in comparison with the control. Overall, photosynthetic rate was increased by 26% in G2 at L6 (AgNPs 25 mg/L + Pb 25 μM). In addition, the results presented 78.5% increase in water use efficiency of G2 while G1 experienced a maximum internal CO2 concentration (209.8%) at L8 (Pb 50 μM). AgNPs triggered balanced uptake of minerals and improved growth of Vigna genotypes. 50 μM Pb was most hazardous and caused maximum reduction in growth of Vigna plants along with a significant suppression in photosynthetic activity, increase in MDA (199.7%) in G1 and H2O2 (292.8%) in G2. In comparison to control, maximum superoxide dismutase (376%), peroxidase (659.8%) and catalase (9.3%) activity was observed in G2 at L11. The application of AgNPs substantially enhanced plant growth and helped them in surviving well in absence as well as presence of Pb. G2 genotype exhibited substantial tolerance capability and revealed less impairment in the studied attributes than G1 and treatment of AgNPs i.e. 25 mg/L was the best level that yielded best results in both genotypes. The results demonstrate that AgNPs mediate response(s) of plants under Pb stress and particularly contributed to HM tolerance of plants and thus showing great promise for use in phytoremediation.

Low-molecular-weight organic acid-mediated tolerance and Pb accumulation in centipedegrass under Pb stress

Lead (Pb) is one of the most harmful, toxic pollutants to the ecological environment and humans. Centipedegrass, a fast-growing warm-season turfgrass, is excellent for Pb pollution remediation. Exogenous low-molecular-weight organic acid (LMWOA) treatment is a promising approach for assisted phytoremediation. However, the effects of this treatment on the tolerance and Pb accumulation of centipedegrass are unclear. This study investigated these effects on the physiological growth response and Pb accumulation distribution characteristics of centipedegrass. Applications of 400 μM citric acid (CA), malic acid (MA) and tartaric acid (TA) significantly reduced membrane lipid peroxidation levels of leaves and improved biomass production of Pb-stressed plants. These treatments mainly increased peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX) activities and enhanced free protein (Pro), ascorbic acid (AsA) and phytochelatins (PCs) contents, ultimately improving the Pb tolerance of centipedegrass. Their promoting effects decreased as follows: TA>CA>MA. All the treatments decreased root Pb concentrations and increased stem and leaf Pb concentrations, thus increasing total Pb accumulation and TF values. MA had the best and worst effects on Pb accumulation and Pb transportation, respectively. CA had the best and worst effects on Pb transportation and Pb accumulation, respectively. TA exhibited strong effects on both Pb accumulation and transport. Furthermore, all treatments changed the subcellular Pb distribution patterns and distribution models of the chemical forms of Pb in each tissue. The root Pb concentration was more highly correlated with the Pb subcellular fraction distribution pattern, while the stem and leaf Pb concentrations were more highly correlated with the distribution models of the chemical forms of Pb. Overall, TA improved plant Pb tolerance best and promoted both Pb absorption and transportation well and is considered the best candidate for Pb-contaminated soil remediation with centipedegrass. This study provides a new idea for Pb-contaminated soil remediation with centipedegrass combined with LMWOAs.

Integrated physiological, transcriptomic and metabolomic analysis of the response of Trifolium pratense L. to Pb toxicity

Lead (Pb) interferes with plant gene expression, alters metabolite contents and affects plant growth. However, the molecular mechanism underlying the plant response to Pb is not completely understood. In the present study, Trifolium pratense L. was exposed to Pb concentrations of 0 (Pb0), 500 (Pb500), 1000 (Pb1000), 2000 (Pb2000) and 3000 (Pb3000) mg/kg in soils. Pb stress affected the ability of T. pratense to accumulate and transport Pb, increased the activity of peroxidase (POD) and the contents of malondialdehyde (MDA) and proline, decreased the amount of photosynthetic pigments and soluble proteins, and led to changes in growth and biomass. Transcriptomic and metabolomic analyses showed that Pb mainly affected eight pathways, and LHC, flavonoids, organic acids, amino acids and carbohydrates were upregulated or downregulated. Moreover, Pb500 induced the upregulation of serA, promoted the synthesis of citric acid, maintained photosynthetic pigment levels, and ultimately promoted an increase in stem length. Pb3000 induced the upregulation of ARF, GH3 and SAUR genes, but the saccharide contents and stem length decreased in response to Pb stress. We used a variety of methods to provide a molecular perspective on the mechanism underlying the response of T. pratense to Pb stress.

Physiological, nutritional, and biochemical indicators of lead tolerance in sunflower genotypes

This study aimed to select and classify sunflower genotypes tolerant to lead (Pb) stress and evaluate their capacity of phytoextraction based on physiological, nutritional, and biochemical responses. Two experiments were carried out under lead stress. In the first experiment, out of 21 genotypes studied three showed higher relative biomass yield and were characterized as Pb-tolerant and five showed lower relative biomass production and were considered Pb-sensitive. In the second experiment, one Pb-tolerant (BRS-G27) and two Pb-sensitive (H251 and AG963) genotypes were studied. In this experiment, Pb stress reduced the growth and contents photosynthetic pigments in all genotypes, but more pronouncedly in sensitive genotypes. There were no substantial changes in micronutrient levels in the leaves and stem, but the levels of Cu and Mn in the stressed roots of sensitive genotypes were much lower than in BRS-G27. The contents of organic solutes in the roots suggest that sensitive genotypes have higher energy costs for osmoregulation by carbohydrates and amino acids synthesis. However, the accumulation of proline may be related to a greater Pb tolerance. Considering the results of dry mass yield, transfer coefficient, translocation factor, and tolerance index, the BRS-G27 genotype can be recommended for use in phytoremediation of Pb-contaminated soils.

Secondary metabolites production combined with lead bioremediation by Halamphora sp. marine diatom microalgae and their physiological response

This study was designed to investigate the physiological and biochemical response of the diatom microalgae Halamphora sp. (SB1 MK575516.1) to the toxicity of lead (Pb) as well as its ability as phytoremediation. Four different concentrations of Pb (50, 100, 150, and 200 mg L-1) were applied for 10 days. Fatty acid profile, mineral composition, secondary metabolite contents, and physiological responses have been determined in Halamphora biomass. We found that this metal was mainly removed by bio adsorption on cell surfaces and that Halamphora sp. could acclimatize upon long-term exposure to Pb stress. A decrease in the cell’s number and size, polyunsaturated fatty acids as well as mineral content in Halamphora sp were observed under Pb stress. However, an increase in polyphenol, flavonoid, and carotenoid contents has been recorded at 100 mg Pb L-1, with stimulation of the antioxidant capacity as measured by DPPH and ABTS radical scavenging activities. An increase in MDA, proline, and H2O2 levels were also observed. On the other hand, the deleterious effect of Pb resulting from the cellular oxidative state can be alleviated by the enzymatic system such as Superoxide dismutase (SOD), Glutathione peroxidase (GPx), and catalase (CAT). The present study indicates the ability of Halamphora sp. to remove heavy metals from the aquatic environment and produce antioxidant biomolecules.

Genome-Wide Characterization Analysis of CCT Genes in Raphanus sativus and Their Potential Role in Flowering and Abiotic Stress Response

CCT genes play vital roles in flowering, plant growth, development, and response to abiotic stresses. Although they have been reported in many plants, the characterization and expression pattern of CCT genes is still limited in R. sativus. In this study, a total of 58 CCT genes were identified in R. sativus. Phylogenetic tree, gene structure, and conserved domains revealed that all CCT genes were classified into three groups: COL, CMF, and PRR. Genome-wide identification and evolutionary analysis showed that segmental duplication expanded the CCT gene families considerably, with the LF subgenome retaining more CCT genes. We observed strong purifying selection pressure for CCT genes. RsCCT genes showed tissue specificity, and some genes (such as RsCCT22, RsCCT36, RsCCT42 and RsCCT51) were highly expressed in flowers. Promoter cis-elements and RNA-seq data analysis showed that RsCCT genes could play roles in controlling flowering through the photoperiodic pathway and vernalization pathway. The expression profiles of RsCCT genes under Cd, Cr, Pb, and heat and salt stresses revealed that many RsCCT genes could respond to one or more abiotic stresses. Our findings could provide essential information for further studies on the function of RsCCT genes.

Arbuscular Mycorrhizal Fungi and Glomalin Play a Crucial Role in Soil Aggregate Stability in Pb-Contaminated Soil.

With the rapid development of industrialization and urbanization, soil contamination with heavy metal (HM) has gradually become a global environmental problem. Lead (Pb) is one of the most abundant toxic metals in soil and high concentrations of Pb can inhibit plant growth, harm human health, and damage soil properties, including quality and stability. Arbuscular mycorrhizal fungi (AMF) are a type of obligate symbiotic soil microorganism forming symbiotic associations with most terrestrial plants, which play an essential role in the remediation of HM-polluted soils. In this study, we investigated the effects of AMF on the stability of soil aggregates under Pb stress in a pot experiment. The results showed that the hyphal density (HLD) and spore density (SPD) of the AMF in the soil were significantly reduced at Pb stress levels of 1000 mg kg−1 and 2000 mg kg−1. AMF inoculation strongly improved the concentration of glomalin-related soil protein (GRSP). The percentage of soil particles >2 mm and 2–1 mm in the AMF-inoculation treatment was higher than that in the non-AMF-inoculation treatment, while the Pb stress increased the percentage of soil particles <0.053 mm and 0.25–0.53 mm. HLD, total glomalin-related soil protein (T-GRSP), and easily extractable glomalin-related soil protein (EE-GRSP) were the three dominant factors regulating the stability of the soil aggregates, based on the random forest model analysis. Furthermore, the structural equation modeling analysis indicated that the Pb stress exerted an indirect effect on the soil-aggregate stability by regulating the HLD or the GRSP, while only the GRSP had a direct effect on the mean weight diameter (MWD) and geometric mean diameter (GMD). The current study increases the understanding of the mechanism through which soil degradation is caused by Pb stress, and emphasizes the crucial importance of glomalin in maintaining the soil-aggregate stability in HM-contaminated ecosystems.

Effect of Exogenous Melatonin Application on the Grain Yield and Antioxidant Capacity in Aromatic Rice under Combined Lead-Cadmium Stress.

This study aimed to determine the mechanism of exogenous melatonin application in alleviating the combined Pb and Cd (Pb-Cd) toxicity on aromatic rice (Oryza sativa L.). In this study, a pot experiment was conducted; two aromatic rice varieties, Yuxiangyouzhan and Xiangyaxiangzhan, were selected, and sprays using 50, 100, 200, and 400 μmol L−1 melatonin (denoted as S50, S100, S200, and S400) and irrigation using 100, 300, and 500 μmol L−1 melatonin (denoted as R100, R300, and R500) were also selected. The results showed that, under the S50, S100, and S200 treatments, the Pb content of aromatic rice grain decreased, and the grain yield increased significantly. Moreover, the application of exogenous melatonin significantly reduced the accumulation of H2O2 in rice leaves at maturity under Cd–Pb stress and reduced the MDA content in Xiangyaxiangzhan leaves. In addition, the microbial community structure changed significantly under S50 and R300 treatments. Some pathways, such as the synthesis of various amino acids and alanine, aspartate, and glutamate metabolism, were regulated by S50 treatment. Overall, melatonin application improved aromatic rice grain yield while reducing heavy metal accumulation by regulating the antioxidant capacity and metabolites in aromatic rice plants and altering the physicochemical properties and microbial community structures of the soil.

Transcriptome Analysis on Key Metabolic Pathways in Rhodotorula mucilaginosa Under Pb(II) Stress

Rhodotorula mucilaginosa shows adaption to a broad range of Pb2+ stress. In this study, three key pathways, i.e., glycolysis (EMP), the tricarboxylic acid (TCA) cycle, and oxidative phosphorylation (OXPHOS), were investigated under 0-2,500 mg · L-1 Pb stress, primarily based on biochemical analysis and RNA sequencing. R. mucilaginosa cells showed similar metabolic response to low/medium (500/1000 mg · L-1) Pb2+ stress. High (2,500 mg · L-1) Pb2+ stress exerted severe cytotoxicity to R. mucilaginosa. The downregulation of HK under low-medium Pb2+ suggested a correlation with the low hexokinase enzymatic activity in vivo. However, IDH3, regulating a key step of circulation in TCA, was upregulated to promote ATP feedstock for downstream OXPHOS. Then, through activation of complex I & IV in the electron transport chain (ETC) and ATP synthase, ATP production was finally enhanced. This mechanism enabled fungal cells to compensate for ATP consumption under low-medium Pb2+ toxicity. Hence, R. mucilaginosa tolerance to such a broad range of Pb2+ concentrations can be attributed to energy adaption. In contrast, high Pb2+ stress caused ATP deficiency. Then, the subsequent degradation of intracellular defense systems further intensified Pb toxicity. This study correlated responses of EMP, TCA, and OXPHOS pathways in R. mucilaginosa under Pb stress, hence providing new insights into the fungal resistance to heavy metal stress. IMPORTANCE Glycolysis (EMP), the tricarboxylic acid (TCA) cycle, and oxidative phosphorylation (OXPHOS) are critical metabolism pathways for microorganisms to obtain energy during the resistance to heavy metal (HM) stress. However, these pathways at the genetic level have not been elucidated to evaluate their cytoprotective functions for Rhodotorula mucilaginosa under Pb stress. In this study, we investigated these three pathways based on biochemical analysis and RNA sequencing. Under low-medium (500-1,000 mg · L-1) Pb2+ stress, ATP production was stimulated mainly due to the upregulation of genes associated with the TCA cycle and the electron transport chain (ETC). Such an energy compensatory mechanism could allow R. mucilaginosa acclimation to a broad range of Pb2+ concentrations (up to 1000 mg · L-1). In contrast, high (2500 mg · L-1) Pb2+ stress exerted its excessive toxicity by provoking ATP deficiency and damage to intracellular resistance systems. This study provided new insights into R. mucilaginosa resistance to HM stress from the perspective of metabolism.