DNA barcoding combined with high-resolution melting analysis to discriminate rhubarb species and its traditional Chinese patent medicines.

Introduction: Rhubarb is a frequently used and beneficial traditional Chinese medicine. Wild resources of these plants are constantly being depleted, meaning that rhubarb products have been subjected to an unparalleled level of adulteration. Consequentially, reliable technology is urgently required to verify the authenticity of rhubarb raw materials and commercial botanical drugs. Methods: In this study, the barcode-DNA high-resolution melting (Bar-HRM) method was applied to characterize 63 rhubarb samples (five Polygonaceae species: Rheum tanguticum, Rh. palmatum, Rh. officinale, Rumex japonicus and Ru. sp.) and distinguish the rhubarb contents of 24 traditional Chinese patent medicine (TCPM) samples. Three markers, namely ITS2, rbcL and psbA-trnH, were tested to assess the candidate DNA barcodes for their effectiveness in distinguishing rhubarb from its adulterants. A segment from ITS2 was selected as the most suitable mini-barcode to identify the botanical drug rhubarb in TCPMs. Then, rhubarbs and TCPM samples were subjected to HRM analysis based on the ITS2 barcode. Results: Among the tested barcoding loci, ITS2 displayed abundant sites of variation and was effective in identifying Polygonaceae species and their botanical origins. HRM analysis based on the ITS2 mini-barcode region successfully distinguished the authenticity of five Polygonaceae species and eight batches of TCPMs. Of the 18 TCPM samples, 66.7 % (12 samples) were identified as containing Rh. tanguticum or Rh. officinale. However, 33.3 % were shown to consist of adulterants. Conclusions: These results demonstrated that DNA barcoding combined with HRM is a specific, suitable and powerful approach for identifying rhubarb species and TCPMs, which is crucial to guaranteeing the security of medicinal plants being traded internationally.

Variations in Cold Resistance and Contents of Bioactive Compounds among Dendrobium officinale Kimura et Migo Strains.

Dendrobium officinale is a valuable traditional Chinese herbal plant that is both medicinal and edible. However, the yield of wild Dendrobium officinale is limited. Adverse stress affects the growth, development, and yield of plants, among which low temperature is the primary limiting factor for introducing Dendrobium officinale to high-latitude areas and expanding the planting area. Therefore, this study aims to explore the variations in growth ability, cold resistance, and contents of bioactive compounds among different Dendrobium officinale strains. Four strains of Dendrobium officinale were selected as experimental materials and were subjected to low-temperature stress (4 °C). The agronomic traits, physiological indices, as well as the expressions of cold resistance-related genes (HSP70, DcPP2C5, DoCDPK1, and DoCDPK6) in the roots and leaves of Dendrobium officinale, were determined. The contents of bioactive compounds, including polysaccharides, flavonoids, and phenols were also measured. Compared with the other strains, Xianju had the highest seed germination and transplantation-related survival rates. Under low-temperature stress, Xianju exhibited the strongest cold resistance ability, as revealed by the changes in water contents, chlorophyll levels, electrical conductivities, enzyme activities, and expressions of the cold resistance-related genes. Additionally, the polysaccharide content of Xianju increased the most, while the stem flavonoid and leaf phenol contents were elevated in all four strains under cold treatment. Therefore, selecting excellent performing strains is expected to expand the planting area, improve the yield, and increase the economic benefits of Dendrobium officinale in high latitude areas with lower temperatures.

Rice DST transcription factor negatively regulates heat tolerance through ROS-mediated stomatal movement and heat-responsive gene expression.

Plants are frequently subjected to a broad spectrum of abiotic stresses including drought, salinity and extreme temperatures and have evolved both common and stress-specific responses to promote fitness and survival. Understanding the components and mechanisms that underlie both common and stress-specific responses can enable development of crop plants tolerant to different stresses. Here, we report a rice heat stress-tolerant 1 (hst1) mutant with increased heat tolerance. HST1 encodes the DST transcription factor, which also regulates drought and salinity tolerance. Increased heat tolerance of hst1 was associated with suppressed expression of reactive oxygen species (ROS)-scavenging peroxidases and increased ROS levels, which reduced water loss by decreasing stomatal aperture under heat stress. In addition, increased ROS levels enhanced expression of genes encoding heat shock protein (HSPs) including HSP80, HSP74, HSP58 and small HSPs. HSPs promote stabilization of proteins and protein refolding under heat stress and accordingly mutation of HST1 also improved reproductive traits including pollen viability and seed setting under high temperature. These results broaden the negative roles of DST in abiotic stress tolerance and provide important new insights into DST-regulated tolerance to diverse abiotic stresses through both shared and stress-specific mechanisms.

Research Progress of Soil Microorganisms in Response to Heavy Metals in Rice.

Soil heavy-metal pollution leads to excessive heavy metals in rice and other food crops, which has caused serious impacts on the ecological environment and on human health. In recent years, environmental friendly treatment methods that reduce the bioavailability of heavy metals in soil by soil microorganisms improving the tolerance of heavy metals in rice and reducing the transfer of heavy metals from the roots to the above-ground parts of rice have attracted much attention. This paper reviews the role and mechanism of soil microorganisms in alleviating heavy-metal stress in rice at home and abroad in recent years. At present, microorganisms tolerant to heavy metals mainly include bacteria and fungi, and their mechanisms include the adsorption of heavy metals by microorganisms, the secretion of growth-promoting substances (growth hormone, ACC deaminase, IAA), changing the physical and chemical properties of the soil and the composition of the microbial community, changing the transport mode of heavy metals in soil, the improvement of the antioxidant capacity of rice, etc. Hence, soil microorganisms have good application value and prospects in rice and other crops. However, the vast majority of current research focuses on a single strain, the screening principles of strains are limited, the pathogenicities of the strains have not been evaluated, and there are still few field experiments under natural conditions. In the future, we should strengthen the action of soil microorganisms on rice in response to the above problems in heavy metals, to better promote the microbial remediation technology.

[Screening of tomato cultivars in cadmium-polluted areas and study on their antioxidant capacity].

To screen the available tomato pollution-safe cultivar varieties and reduce the potential food safety risks in Cd-polluted areas, the differences of Cd accumulation in different tomato (Solanum lycopersicum) varieties in southern China were studied by soil culture and hydroponic experiments. Firstly, the high and low accumulation varieties were selected from 25 tomato varieties under 2.94 mg/kg Cd stress by soil culture test, and then the responses of high and low accumulation tomato varieties to Cd stress were determined by hydroponic experiments. The results of soil culture test show that under 2.94 mg/kg Cd stress, there were significant differences in plant height, total biomass and yield among 25 tomato cultivars, and the Cd contents of fruits of all 25 tomato cultivars exceeded the highest limit value (0.05 mg/kg) of CAC (Codex alimentarius commission). Through cluster analysis, 7, 4 and 14 varieties accumulating relatively high, medium, and low concentrations of Cd in the fruits were screened, among which the highest, the lowest, and the average Cd contents in the fruits were 3.06 mg/kg DW, 1.47 mg/kg DW, and 2.21 mg/kg DW, respectively. The results of hydroponic experiment show that under the same concentration of Cd stress, Qiantangxuri F1, a high Cd accumulating variety, absorbed Cd faster, accumulated more Cd, used shorter oxidative stress response time and had stronger tolerance to Cd than Zhefen 3053, a low Cd accumulating variety. The typical high and low Cd accumulating varieties can provide a reference for agricultural production in heavy metal polluted areas and the development of molecular-assisted breeding methods of PSC. At present, cultivating low Cd accumulating PSC varieties and dynamic monitoring of Cd contents in tomato fruits are feasible methods in medium and light Cd-polluted areas.

High-Resolution Melting Analysis of COI Sequences Distinguishes Pufferfish Species (Takifugu spp.) in China.

Pufferfish is a traditional, delicious dish in Asia. However, eating wild or improperly processed pufferfish causes serious poisoning. This study aimed to exploit the high-resolution melting (HRM) method for authenticating four species of Takifugu pufferfish (Takifugu xanthopterus, T. fasciatus, T. flavidus, and T. rubripes). Candidate DNA barcodes, including the cytochrome c oxidase subunit I (COI), cytochrome oxidase b (Cytb), and the control region (D-loop), were analyzed, with COI selected as the optimal DNA barcode. An HRM method was developed to identify 57 commercial fish samples in China, including 33 commercial pufferfish products and 24 unlabeled fish products. The findings revealed that the pufferfish products were T. rubripes or T. fasciatus, and four T. xanthopterus samples were detected in unlabeled fish products. These results showed that DNA barcode coupled with HRM analysis was a rapid and efficient tool to identify pufferfish, which might aid in the prevention of consumer fraud or mislabeling of fish products.

Application of exogenous salicylic acid reduces Cd toxicity and Cd accumulation in rice.

Cd pollution in farmland is becoming a serious problem because it affects the safety of rice production and human health. Salicylic acid (SA) plays crucial roles in plant development and mediates plant responses to biotic and abiotic stress. This study assessed the molecular and physiological mechanisms of SA spraying effects on Cd tolerance and Cd accumulation in rice. Spraying of 0.1 mM SA had no great effect on the agronomic traits of rice, but significantly decreased Cd accumulation in rice grains, and SA spraying increased the Cd contents in leaves (only at the mature stage) and decreased the Cd contents in panicles (only at the filling and mature stage), but had no evident impact on the Cd content of other tissues and other growth stages. SA spraying reduced Cd accumulation in rice grains by promoting the deposition and fixation of Cd in the cell wall of leaves, thus preventing Cd being transferred from leaves to rice grains at the filling stage. SA spraying also decreased Cd toxicity by reducing H2O2 and MDA accumulation and increasing the chlorophyll content in rice leaves. Furthermore, SA spraying remarkably decreased Cd accumulation in rice grains by modulating the expression level of the genes associated with Cd translocation and accumulation to control the Cd accumulation in rice. Hence, SA spraying reduced the inhibition of Cd on the plant height caused by Cd and increased the dry weight of shoots in the vegetative growth period of rice seedlings, and it reduced Cd transport from leaves to grains, thus reducing Cd content in rice. These findings provide a novel perspective and a new method for reducing Cd accumulation in rice.

Salicylic acid application alleviates cadmium accumulation in brown rice by modulating its shoot to grain translocation in rice.

Cadmium (Cd) contamination, which poses a serious threat to human health, has been recognized as a major threat to the agricultural system and crop production. Salicylic acid (SA) is a signaling molecule that plays an important role in against Cd toxicity. Previously, we found that spraying rice with SA could reduce the Cd accumulation in rice grains grown in Cd-contaminated soil. In this study, we studied the specific mechanism of SA spray on reducing Cd accumulation in rice grain. The results showed that treatment with SA could alleviate Cd toxicity in rice by increasing the activities of antioxidant enzymes that reduce hydrogen peroxide (H2O2) accumulation, but not by changing the pH, or total or available Cd of the soil. The key factor by which SA treatment reduced Cd accumulation in rice grains was by decreasing the Cd content in rice leaves at the flowering stage. This indicated that SA could modulate the Cd accumulation in shoots, reducing the Cd translocation to rice grains. Furthermore, SA could increase the H2O2 content, activating the SA-signaling pathway and modulating the expression levels of Cd transporters (OsLCT1 and OsLCD) in rice leaves to increase Cd tolerance and reduce Cd accumulation in the rice grain. Thus, spraying rice with SA may be effective measure to cope with Cd contamination in paddy soils.

[Effects of Kochia scoparia-Brassica rapa rotation on Cd uptake by Brassica rapa].

In this study, pot and field experiments were conducted to study the enrichment of soil cadmium by Kochia scoparia. Further, rotations in pot experiments were carried out with four varieties of Brassica rapa to verify the remediation effect of Kochia scoparia on cadmium contamination in soil. The enrichment capacity of Kochia scoparia was leaf > root > stem with bioconcentration factors (BCFCd) of 15.07, 5.44 and 2.96, respectively. The total cadmium in soil decreased by 6.02% to 13.60% after planting Kochia scoparia, and the activities of soil urease and acid phosphatase also increased. The results of pot cultivation shows that the above-ground cadmium content of Brassica rapa in Kochia scoparia-Brassica rapa rotation system decreased by 17.21% on average compared with the control group without rotation, whereas the biomass increased slightly, and the above-ground translocation factors (TFCd) did not change significantly. These results suggest that the rotation of Brassica rapa with Kochia scoparia could increase the yield of Brassica rapa, and effectively reduce the cadmium content in edible parts of Brassica rapa, toward the purpose of realizing the green agricultural concept of "harnessing while producing".

Emerging Roles of microRNAs in Plant Heavy Metal Tolerance and Homeostasis.

Heavy metal stress is a major growth- and yield-limiting factor for plants. Heavy metals include essential metals (copper, iron, zinc, and manganese) and non-essential metals (cadmium, mercury, aluminum, arsenic, and lead). Plants use complex mechanisms of gene regulation under heavy metal stress. MicroRNAs are 21-nucleotide non-coding small RNAs as important modulators of gene expression post-transcriptionally. Recently, high-throughput sequencing has led to the identification of an increasing number of heavy-metal-responsive microRNAs in plants. Metal-regulated microRNAs and their target genes are part of a complex regulatory network that controls various biological processes, including heavy metal uptake and transport, protein folding and assembly, metal chelation, scavenging of reactive oxygen species, hormone signaling, and microRNA biogenesis. In this review, we summarize the recent molecular studies that identify heavy-metal-regulated microRNAs and their roles in the regulation of target genes as part of the microRNA-associated regulatory network in response to heavy metal stress in plants.

MicroRNA166 Modulates Cadmium Tolerance and Accumulation in Rice.

MicroRNAs (miRNAs) are 20- to 24-nucleotide small noncoding RNAs that regulate gene expression in eukaryotic organisms. Several plant miRNAs, such as miR166, have vital roles in plant growth, development and responses to environmental stresses. One such environmental stress encountered by crop plants is exposure to cadmium (Cd), an element highly toxic to most organisms, including humans and plants. In this study, we analyzed the role of miR166 in Cd accumulation and tolerance in rice (Oryza sativa). The expression levels of miR166 in both root and leaf tissues were significantly higher in the reproductive stage than in the seedling stage in rice. The expression of miR166 in the roots of rice seedlings was reduced after Cd treatment. Overexpression of miR166 in rice improved Cd tolerance, a result associated with the reduction of Cd-induced oxidative stress in transgenic rice plants. Furthermore, overexpression of miR166 reduced both Cd translocation from roots to shoots and Cd accumulation in the grains. miR166 targets genes encoding the class-III homeodomain-Leu zipper (HD-Zip) family proteins in plants. In rice, HOMEODOMAIN CONTAINING PROTEIN4 (OsHB4) gene (Os03g43930), which encodes an HD-Zip protein, was up-regulated by Cd treatment but down-regulated by overexpression of miR166 in transgenic rice plants. Overexpression of OsHB4 increased Cd sensitivity and Cd accumulation in the leaves and grains of transgenic rice plants. By contrast, silencing OsHB4 by RNA interference enhanced Cd tolerance in transgenic rice plants. These results indicate a critical role for miR166 in Cd accumulation and tolerance through regulation of its target gene, OsHB4, in rice.

MicroRNA268 Overexpression Affects Rice Seedling Growth under Cadmium Stress.

MicroRNAs (miRNAs) are 21-24-nucleotide-long RNAs that function as ubiquitous post-transcriptional regulators of gene expression in plants and animals. Increasing evidence points to the important role of miRNAs in plant responses to abiotic and biotic stresses. Cadmium (Cd) is a nonessential heavy metal highly toxic to plants. Although many genes encoding metal transporters have been characterized, the mechanisms for the regulation of the expression of the heavy-metal transporter genes are largely unknown. In this study, we found that the expression of miR268 in rice was significantly induced under Cd stress. By contrast, expression of natural resistance-associated macrophage protein 3 (NRAMP3), a target gene of miR268, was dramatically decreased by Cd treatment. Overexpression of miR268 inhibited rice seedling growth under Cd stress. The transgenic miR268-overexpressing plant leaves contained increased levels of hydrogen peroxide and malondialdehyde, and their seedlings accumulated increased levels of Cd when compared to those in wild-type plants. These results indicate that miR268 acts as a negative regulator of rice's tolerance to Cd stress. Thus, miRNA-guided regulation of gene expression plays an important role in plant responses to heavy-metal stress.

The role of receptor-like protein kinases (RLKs) in abiotic stress response in plants.

KEY MESSAGE: We review and introduce recent studies on RLK s involved in the abiotic stress response and provide insights into potential regulatory mechanisms for alleviating abiotic stress. Abiotic stresses are important factors affecting plant growth and development, resulting in crop production reduction and even plant death. To survive, plants utilize different mechanisms to respond and adapt to continuously changing environmental factors. Understanding of the molecular mechanisms of plant response to various stresses will aid in improving tolerance of plants to abiotic stress through genetic engineering, which would greatly promote the development of modern agriculture. RLKs, the largest gene family in plants, play critical roles in the regulation of plant developmental processes, signaling networks and disease resistance. Many RLKs have been shown to be involved in abiotic stress responses, including the abscisic acid response, calcium signaling and antioxidant defense. This review summarizes recent studies on RLKs involved in plant responses to abiotic stress, including drought, salt, cold, toxic metals/metalloids and other stresses, and emphasizes the upstream and downstream factors in RLK signal transduction pathways under abiotic stress.

Role of salicylic acid in resistance to cadmium stress in plants.

KEY MESSAGE: We review and introduce the importance of salicylic acid in plants under cadmium stress, and provide insights into potential regulatory mechanisms for alleviating cadmium toxicity. Cadmium (Cd) is a widespread and potentially toxic environmental pollutant, originating mainly from rapid industrial processes, the application of fertilizers, manures and sewage sludge, and urban activities. It is easily taken up by plants, resulting in obvious toxicity symptoms, including growth retardation, leaf chlorosis, leaf and root necrosis, altered structures and ultrastructures, inhibition of photosynthesis, and cell death. Therefore, alleviating Cd toxicity in plants is a major aim of plant research. Salicylic acid (SA) is a ubiquitous plant phenolic compound that has been used in many plant species to alleviate Cd toxicity by regulating plant growth, reducing Cd uptake and distribution in plants, protecting membrane integrity and stability, scavenging reactive oxygen species and enhancing antioxidant defense system, improving photosynthetic capacity. Furthermore, SA functions as a signaling molecule involved in the expression of several important genes. Significant amounts of research have focused on understanding SA functions and signaling in plants under Cd stress, but several questions still remain unanswered. In this article, the influence of SA on Cd-induced stress in plants and the potential regulation mechanism for alleviating Cd toxicity are reviewed.

Heavy Metal Pollution in a Soil-Rice System in the Yangtze River Region of China.

Heavy metals are regarded as toxic trace elements in the environment. Heavy metal pollution in soil or rice grains is of increasing concern. In this study, 101 pairs of soil and rice samples were collected from the major rice-producing areas along the Yangtze River in China. The soil properties and heavy metal (i.e., Cd, Hg, Pb and Cr) concentrations in the soil and rice grains were analyzed to evaluate the heavy metal accumulation characteristics of the soil-rice systems. The results showed that the Cd, Hg, Pb and Cr concentrations in the soil ranged from 0.10 to 4.64, 0.01 to 1.46, 7.64 to 127.56, and 13.52 to 231.02 mg·kg(-)¹, respectively. Approximately 37%, 16%, 60% and 70% of the rice grain samples were polluted by Cd, Hg, Pb, and Cr, respectively. The degree of heavy metal contamination in the soil-rice systems exhibited a regional variation. The interactions among the heavy metal elements may also influence the migration and accumulation of heavy metals in soil or paddy rice. The accumulation of heavy metals in soil and rice grains is related to a certain extent to the pH and soil organic matter (SOM). This study provides useful information regarding heavy metal accumulation in soil to support the safe production of rice in China. The findings from this study also provide a robust scientific basis for risk assessments regarding ecological protection and food safety.

Different responses of low grain-Cd-accumulating and high grain-Cd-accumulating rice cultivars to Cd stress.

Cadmium (Cd) is a major heavy metal pollutant which is highly toxic to plants and animals. The accumulation of Cd in rice grains is a major agricultural problem in regions with Cd pollution. A hydroponics experiment using low grain-Cd-accumulating rice (xiushui 11) and high grain-Cd-accumulating rice (xiushui 110) was carried out to characterize the different responses of rice cultivars to Cd stress. We found that xiushui 11 was more tolerant to Cd than xiushui 110, and xiushui 11 suffered less oxidative damage. Cell walls played an important role in limiting the amount of Cd that entered the protoplast, especially in xiushui 11. Cd stored in organelles as soluble fractions, leading to greater physiological stress of Cd detoxification. We found that Cd can disturb the ion homeostasis in rice roots because Cd(2+) and Ca(2+) may have a similar uptake route. Xiushui 11 had a faster root-to-shoot transport of Cd, and the expression level of OsPCR1 gene which was predicted related with Cd accumulation in rice was consist with the Cd transport of root-to-shoot in rice and maintain the greater Cd tolerance of xiushui 11. These results suggest there are different Cd detoxification and accumulation mechanisms in rice cultivars.

Heteroexpression of the wheat phytochelatin synthase gene (TaPCS1) in rice enhances cadmium sensitivity.

Phytochelatin synthase (PCS) (EC 2.3.2.15) catalyzes the final step of phytochelatins (PCs) biosynthesis. PCs are a family of cysteine-rich thiol-reactive and heavy metal-binding peptides that play an important role in sequestration and detoxification of heavy metals in plants. Previous studies have indicated that plants that overexpressed PCS displayed contrasting phenotypes, ranging from enhanced cadmium (Cd) tolerance to Cd hypersensitivity in Arabidopsis thaliana. In this study, the wheat phytochelatin synthase gene, TaPCS1, was heteroexpressed in wild-type rice (Oryza sativa L., cv. Zhonghua 11) to evaluate the relationship between synthesis of PCs and Cd tolerance in rice. Data showed that the heteroexpression of TaPCS1 in rice enhanced Cd sensitivity and significantly increased Cd accumulation in shoots, but not in roots. Additionally, the PCS line exhibited a much higher content of PCs and non-protein thiols (NPTs) in shoots. Prominent changes in NPT composition led to reduced glutathione pool depletion and higher Cd content in cell organelles in shoots, followed by higher oxidative stress, which might result in Cd sensitivity. Therefore, the heteroexpression of TaPCS1 in rice is capable of increasing Cd accumulation in rice shoots and enhancing Cd sensitivity.

Comparative proteomic study and functional analysis of translationally controlled tumor protein in rice roots under Hg2+ stress.

So far, very little is known about mercury stress-induced intercellular metabolic changes in rice roots at the proteome level. To investigate the response of rice roots to mercury stress, changes in protein expression in rice roots were analyzed using a comparative proteomics approach. Six-leaf stage rice seedlings were treated with 50 micromol/L HgCl2 for 3 hr; 29 protein spots showed a significant changes in abundance under stress when compared with the Hg2+ -tolerant rice mutant and wild type (Zhonghua 11). Furthermore, all these protein spots were identified by mass spectrometry to match 27 diverse protein species. The identified proteins were involved in several processes, including stress response, redox homeostasis, signal transduction, regulation and metabolism; some were found to be cellular structure proteins and a few were unknown. Among the up-regulated proteins, OsTCTP (translationally controlled tumor protein) was chosen to perform hetereologous expression in yeast which was presumed to participate in the Hg2+ tolerance of rice, providing evidence for its role in alleviating Hg2+ damage. Among the many tests, we found that OsTCTP-overexpressed yeast strains were more resistant to Hg2+ than wild-type yeast. Thus, we propose that OsTCTP contributes to Hg2+ resistance. Here we present, for the first time, the functional characterization of OsTCTP in connection with Hg2+ stress in plants.

[Heterologous expression of a rice syntaxin-related protein KNOLLE gene (OsKNOLLE) in yeast and its functional analysis in the role of abiotic stress].

Syntaxin-related protein KNOLLE, a multifunctional protein family belonging to the SNARE superfamily, plays an important role in many physiological processes in plants. In order to understand the function of the syntaxin-related protein KNOLLE (OsKNOLLE) in rice (Oryza sativa), the CDS sequence of OsKNOLLE gene isolated from a japonica rice cultivar "Zhonghua 11" was fused into an expression vector pYX212 and transformed into the S. cerevisiae strain BY4741 by lithium acetate/single-stranded carrier DNA/polyethylene glycol method. The transformants with OsKNOLLE showed better survival abilities than the transformants with the empty vectors based on their phenotypes in responses to different abiotic stresses such as salt, Cu2+, H2O2, Cd2+, and Hg2+. These data suggests that OsKNOLLE plays a crucial role in re-sponses to abiotic stresses. This experimental system sets up a method for studying functions of the OsKNOLLE gene in the future and clarifies the relationship between OsKNOLLE and abiotic stresses.

Relationship between proline and Hg2+-induced oxidative stress in a tolerant rice mutant.

There has been little agreement regarding the mechanism by which proline reduces heavy metal stress. The present work examines the relationship between Hg(2+)-induced oxidative stress and proline accumulation in rice and explores the possible mechanisms through which proline protects against Hg(2+) stress. The effect of proline on alleviation of Hg(2+) toxicity was studied by spectrophotography and enzymatic methods. Hg(2+) induced oxidative stress in rice by increasing lipid peroxidation. Pretreatment of the rice with 2 mM proline for 12 h profoundly alleviated Hg(2+)-induced lipid peroxidation and minimized H(2)O(2) accumulation. Proline pretreatment significantly reduced (p < 0.01) the Hg(2+) content in rice leaves. A comparison of the effects of proline pretreatment on H(2)O(2) accumulation by Hg(2+) and aminotrazole suggested that proline protected cells from Hg(2+)-induced oxidative stress by scavenging reactive oxygen species. The present work demonstrates a protective effect of proline on Hg(2+) toxicity through detoxifying reactive oxygen species, rather than chelating metal ions or maintaining the water balance under Hg(2+) stress.