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.

Autophagy and multivesicular body pathways cooperate to protect sulfur assimilation and chloroplast functions.

Autophagy and multivesicular bodies (MVBs) represent 2 closely related lysosomal/vacuolar degradation pathways. In Arabidopsis (Arabidopsis thaliana), autophagy is stress-induced, with deficiency in autophagy causing strong defects in stress responses but limited effects on growth. LYST-INTERACTING PROTEIN 5 (LIP5) is a key regulator of stress-induced MVB biogenesis, and mutation of LIP5 also strongly compromises stress responses with little effect on growth in Arabidopsis. To determine the functional interactions of these 2 pathways in Arabidopsis, we generated mutations in both the LIP5 and AUTOPHAGY-RELATED PROTEIN (ATG) genes. atg5/lip5 and atg7/lip5 double mutants displayed strong synergistic phenotypes in fitness characterized by stunted growth, early senescence, reduced survival, and greatly diminished seed production under normal growth conditions. Transcriptome and metabolite analysis revealed that chloroplast sulfate assimilation was specifically downregulated at early seedling stages in the atg7/lip5 double mutant prior to the onset of visible phenotypes. Overexpression of adenosine 5'-phosphosulfate reductase 1, a key enzyme in sulfate assimilation, substantially improved the growth and fitness of the atg7/lip5 double mutant. Comparative multi-omic analysis further revealed that the atg7/lip5 double mutant was strongly compromised in other chloroplast functions including photosynthesis and primary carbon metabolism. Premature senescence and reduced survival of atg/lip5 double mutants were associated with increased accumulation of reactive oxygen species and overactivation of stress-associated programs. Blocking PHYTOALEXIN DEFICIENT 4 and salicylic acid signaling prevented early senescence and death of the atg7/lip5 double mutant. Thus, stress-responsive autophagy and MVB pathways play an important cooperative role in protecting essential chloroplast functions including sulfur assimilation under normal growth conditions to suppress salicylic-acid-dependent premature cell-death and promote plant growth and fitness.

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.

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".

[Effects of intercropping on cadmium uptake by maize and tomato].

In order to explore the effect of intercropping on the uptake of heavy metal cadmium (Cd), pot experiments were undertaken using three different planting methods: monoculture, restrictive intercropping and intercropping. The effects of Cd accumulation in different plant parts, and their causes, were examined using a plant species regarded as a relatively high heavy metal accumulator (tomato: Lycopersicon esculentum var. Zhongshu 4) and a species regarded as a relatively low heavy metal accumulator (maize: Zea mays L. var. Jinzhumi). Cd levels for all experiments were 3.70 mg/kg. Results indicate that restricted intercropping and intercropping of tomato and maize increased the accumulation of Cd (from 13.52 mg/kg to 24.94 mg/kg and 27.30 mg/kg in tomato leaf, respectively). Compared with the control group, pH levels in soil surrounding tomato roots in the intercropped samples decreased and the activity of acid phosphatase increased, while the activity of urease decreased. Intercropping can also change the structure of the crop root microorganism population, increase the abundance of microbiological species that promote the uptake of heavy metals, and finally achieve high accumulation of Cd in tomatoes. Our research results provide reference for controlling soil heavy metal pollution and ensuring food safety by using an intercropping model.

Soil cadmium extraction in Chinese cabbage and cabbage intercropping / Extração de cádmio no solo em repolho chinês e consorciação de repolho

ABSTRACT: Toxic metals contamination of soil has become a serious problem in recent years. In this study, Chinese cabbage (a relatively high-accumulator of cadmium (Cd)) and cabbage (a relatively low-accumulator of Cd) were cultured in monoculture and in intercropping in the Cd-contaminated soil, to evaluate the effect of intercropping on the alteration of Cd extraction. Both the pot experiments and field experiments indicated that intercropping increased the Cd extraction by Chinese cabbage and decreased the Cd extraction by cabbage. Thus, Cd extraction was advanced while safe production was obtained. Further pot experiment was conducted to investigate the alterations of soil Cd fractions, soil pH, and soil enzyme activities to reveal their possible relationship with Cd extraction between different planting patterns. Results revealed that three individual Chinese cabbages in one intercropping pot played the same effect on alteration of these factors as six individual Chinese cabbages in one monoculture pot. The intercropping increased Cd extraction by Chinese cabbage and decreased Cd extraction by cabbage, probably by influencing mechanisms such as soil enzyme activities (especially the urease activity) in the cultivation system. Effect of intercropping on Cd accumulation is an important issue in cultivation of vegetables in potentially contaminated land.

RESUMO: A contaminação do solo por metais tóxicos tornou-se um problema grave nos últimos anos. Neste estudo, dois tipos de repolhos, o repolho chinês (maior acumulador de cádmio (Cd)) e o repolho comum (menor acumulador de Cd) foram cultivados em monocultivo e em consórcio, em solo contaminado com esse metal, para avaliar o efeito do consórcio na extração de Cd do solo. Os experimentos em vasos de campo indicaram que o consórcio aumentou a extração desse metal pelo repolho chinês e diminuiu a extração pelo repolho comum. Assim, aumentou a extração de Cd do solo, proporcionando segurança alimentar. Outro experimento em vaso foi conduzido para investigar as alterações das frações de Cd do solo, o pH do solo e atividades enzimáticas do solo, para revelar possíveis efeitos na extração desse metal entre os sistemas de plantio. Os resultados do experimento em vaso revelaram que três repolhos da china, em cultivo consorciado, proporcionaram o mesmo efeito nessas variáveis, que os observados com seis plantas desse tipo de repolho, em monocultura. O consórcio influenciou as frações de Cd do solo devido a maior influência das hortaliças no pH do solo e nas atividades enzimáticas (especialmente a urease), resultando maior extração de Cd pelo repolho chinês e menor pelo repolho comum.

Biogenesis and Function of Multivesicular Bodies in Plant Immunity.

Multivesicular bodies (MVBs) are specialized endosomes that contain intraluminal vesicles generated from invagination and budding of the limiting membrane. In the endocytic pathway, MVBs are late endosomes whose content can be degraded through fusion with lysosomes/vacuoles or released into the extracellular space after fusion with the plasma membrane (PM). The proteins retained on the limiting membrane of MVBs are translocated to the membrane of lysosomes/vacuoles or delivered back to the PM. It has been long suspected that MVBs might fuse with the PM to form paramural bodies in plant cells, possibly leading to release of building blocks for deposition of papillae and antimicrobial molecules against invading pathogens. Over the past decade or so, major progress has been made in establishing the critical roles of MVBs and associated membrane trafficking in pathogen recognition, defense signaling, and deployment of defense-related molecules during plant immune responses. Regulatory proteins and signaling pathways associated with induced biogenesis and trafficking of MVBs during plant immune responses have also been identified and characterized. Recent successful isolation of plant extracellular vesicles and proteomic profiling of their content have provided additional support for the roles of MVBs in plant-pathogen interactions. In this review, we summarize the important progress and discuss how MVBs, particularly through routing of cellular components to different destinations, contribute to the complex network of plant immune system.

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.

Comparative transcriptome combined with morpho-physiological analyses revealed key factors for differential cadmium accumulation in two contrasting sweet sorghum genotypes.

Cadmium (Cd) is a widespread soil contaminant threatening human health. As an ideal energy plant, sweet sorghum (Sorghum bicolor (L.) Moench) has great potential in phytoremediation of Cd-polluted soils, although the molecular mechanisms are largely unknown. In this study, key factors responsible for differential Cd accumulation between two contrasting sweet sorghum genotypes (high-Cd accumulation one H18, and low-Cd accumulation one L69) were investigated. H18 exhibited a much higher ability of Cd uptake and translocation than L69. Furthermore, Cd uptake through symplasmic pathway and Cd concentrations in xylem sap were both higher in H18 than those in L69. Root anatomy observation found the endodermal apoplasmic barriers were much stronger in L69, which may restrict the Cd loading into xylem. The molecular mechanisms underlying these morpho-physiological traits were further dissected by comparative transcriptome analysis. Many genes involved in cell wall modification and heavy metal transport were found to be Cd-responsive DEGs and/or DEGs between these two genotypes. KEGG pathway analysis found phenylpropanoid biosynthesis pathway was over-represented, indicating this pathway may play important roles in differential Cd accumulation between two genotypes. Based on these results, a schematic representation of main processes involved in differential Cd uptake and translocation in H18 and L69 is proposed, which suggests that higher Cd accumulation in H18 depends on a multilevel coordination of efficient Cd uptake and transport, including efficient root uptake and xylem loading, less root cell wall binding, and weaker endodermal apoplasmic barriers.

The V-ATPase subunit A is essential for salt tolerance through participating in vacuolar Na+ compartmentalization in Salicornia europaea.

MAIN CONCLUSION: The V-ATPase subunit A participates in vacuolar Na + compartmentalization in Salicornia europaea regulating V-ATPase and V-PPase activities. Na+ sequestration into the vacuole is an efficient strategy in response to salinity in many halophytes. However, it is not yet fully understood how this process is achieved. Particularly, the role of vacuolar H+-ATPase (V-ATPase) in this process is controversial. Our previous proteomic investigation in the euhalophyte Salicornia europaea L. found a significant increase of the abundance of V-ATPase subunit A under salinity. Here, the gene encoding this subunit named SeVHA-A was characterized, and its role in salt tolerance was demonstrated by RNAi directed downregulation in suspension-cultured cells of S. europaea. The transcripts of genes encoding vacuolar H+-PPase (V-PPase) and vacuolar Na+/H+ antiporter (SeNHX1) also decreased significantly in the RNAi cells. Knockdown of SeVHA-A resulted in a reduction in both V-ATPase and vacuolar H+-PPase (V-PPase) activities. Accordingly, the SeVHA-A-RNAi cells showed increased vacuolar pH and decreased cell viability under different NaCl concentrations. Further Na+ staining showed the reduced vacuolar Na+ sequestration in RNAi cells. Taken together, our results evidenced that SeVHA-A participates in vacuolar Na+ sequestration regulating V-ATPase and V-PPase activities and thereby vacuolar pH in S. europaea. The possible mechanisms underlying the reduction of vacuolar V-PPase activity in SeVHA-A-RNAi cells were also discussed.

Genome-Wide Identification of Sorghum bicolor Laccases Reveals Potential Targets for Lignin Modification.

Laccase is a key enzyme in plant lignin biosynthesis as it catalyzes the final step of monolignols polymerization. Sweet sorghum [Sorghum bicolor (L.) Moench] is considered as an ideal feedstock for ethanol production, but lignin greatly limits the production efficiency. No comprehensive analysis on laccase has ever been conducted in S. bicolor, although it appears as the most promising target for engineering lignocellulosic feedstock. The aim of our work is to systematically characterize S. bicolor laccase gene family and to identify the lignin-specific candidates. A total of twenty-seven laccase candidates (SbLAC1-SbLAC27) were identified in S. bicolor. All SbLACs comprised the equivalent L1-L4 signature sequences and three typical Cu-oxidase domains, but exhibited diverse intron-exon patterns and relatively low sequence identity. They were divided into six groups by phylogenetic clustering, revealing potential distinct functions, while SbLAC5 was considered as the closest lignin-specific candidate. qRT-PCR analysis deciphered that SbLAC genes were expressed preferentially in roots and young internodes of sweet sorghum, and SbLAC5 showed high expression, adding the evidence that SbLAC5 was bona fide involved in lignin biosynthesis. Besides, high abundance of SbLAC6 transcripts was detected, correlating it a potential role in lignin biosynthesis. Diverse cis regulatory elements were recognized in SbLACs promoters, indicating putative interaction with transcription factors. Seven SbLACs were found to be potential targets of sbi-miRNAs. Moreover, putative phosphorylation sites in SbLAC sequences were identified. Our research adds to the knowledge for lignin profile modification in sweet sorghum.

Comparative proteomics of root plasma membrane proteins reveals the involvement of calcium signalling in NaCl-facilitated nitrate uptake in Salicornia europaea.

Improving crop nitrogen (N) use efficiency under salinity is essential for the development of sustainable agriculture in marginal lands. Salicornia europaea is a succulent euhalophyte that can survive under high salinity and N-deficient habitat conditions, implying that a special N assimilation mechanism may exist in this plant. In this study, phenotypic and physiological changes of S. europaea were investigated under different nitrate and NaCl levels. The results showed that NaCl had a synergetic effect with nitrate on the growth of S. europaea. In addition, the shoot nitrate concentration and nitrate uptake rate of S. europaea were increased by NaCl treatment under both low N and high N conditions, suggesting that nitrate uptake in S. europaea was NaCl facilitated. Comparative proteomic analysis of root plasma membrane (PM) proteins revealed 81 proteins, whose abundance changed significantly in response to NaCl and nitrate. These proteins are involved in metabolism, cell signalling, transport, protein folding, membrane trafficking, and cell structure. Among them, eight proteins were calcium signalling components, and the accumulation of seven of the above-mentioned proteins was significantly elevated by NaCl treatment. Furthermore, cytosolic Ca(2+) concentration ([Ca(2+)]cyt) was significantly elevated in S. europaea under NaCl treatment. The application of the Ca(2+) channel blocker LaCl3 not only caused a decrease in nitrate uptake rate, but also attenuated the promoting effects of NaCl on nitrate uptake rates. Based on these results, a possible regulatory network of NaCl-facilitated nitrate uptake in S. europaea focusing on the involvement of Ca(2+) signalling was proposed.

H(+) -pyrophosphatase from Salicornia europaea confers tolerance to simultaneously occurring salt stress and nitrogen deficiency in Arabidopsis and wheat.

High salinity and nitrogen (N) deficiency in soil are two key factors limiting crop productivity, and they usually occur simultaneously. Here we firstly found that H(+) -PPase is involved in salt-stimulated NO3 (-) uptake in the euhalophyte Salicornia europaea. Then, two genes (named SeVP1 and SeVP2) encoding H(+) -PPase from S. europaea were characterized. The expression of SeVP1 and SeVP2 was induced by salt stress and N starvation. Both SeVP1 or SeVP2 transgenic Arabidopsis and wheat plants outperformed the wild types (WTs) when high salt and low N occur simultaneously. The transgenic Arabidopsis plants maintained higher K(+) /Na(+) ratio in leaves and exhibited increased NO3 (-) uptake, inorganic pyrophosphate-dependent vacuolar nitrate efflux and assimilation capacity under this double stresses. Furthermore, they had more soluble sugars in shoots and roots and less starch accumulation in shoots than WT. These performances can be explained by the up-regulated expression of ion, nitrate and sugar transporter genes in transgenic plants. Taken together, our results suggest that up-regulation of H(+) -PPase favours the transport of photosynthates to root, which could promote root growth and integrate N and carbon metabolism in plant. This work provides potential strategies for improving crop yields challenged by increasing soil salinization and shrinking farmland.

Overexpression of SeNHX1 improves both salt tolerance and disease resistance in tobacco.

Recently, we found NHX1, the gene encoding a Na(+)/H(+) exchanger, participated in plant disease defense. Although NHX1 has been confirmed to be involved in plant salt tolerance, whether the NHX1 transgenic plants exhibit both salt tolerance and disease resistance has not been investigated. The T1 progenies of Nicotiana tabacum L. lines expressing SeNHX1 (from Salicornia europaea) were generated for the present study. Compared with PBI-type control plants, SeNHX1 transgenic tobaccos exhibited more biomass, longer root length, and higher K(+)/Na(+) ratio at post germination or seedling stage under NaCl treatment, indicating enhanced salt tolerance. The vacuolar H(+) efflux in SeNHX1 transgenic tobacco was increased after treatment of NaCl with different concentration. Meanwhile, the SeNHX1 transgenic tobaccos showed smaller wilted spot area, less H2O2 accumulation in leaves after infection of Phytophthora parasitica var. nicotianae. Further investigation demonstrated a larger NAD(P)(H) pool in SeNHX1 transgenic tobacco. These evidences revealed that overexpression of SeNHX1 intensified the compartmentation of Na(+) into vacuole under salt stress and improved the ability of eliminating ROS after pathogen attack, which then enhanced salt tolerance and disease resistance simultaneously in tobacco. Our findings indicate NHX1 has potential value in creating crops with both improved salt tolerance and disease resistance.

Virus-induced gene silencing reveals control of reactive oxygen species accumulation and salt tolerance in tomato by γ-aminobutyric acid metabolic pathway.

γ-Aminobutyric acid (GABA) accumulates in many plant species in response to environmental stress. However, the physiological function of GABA or its metabolic pathway (GABA shunt) in plants remains largely unclear. Here, the genes, including glutamate decarboxylases (SlGADs), GABA transaminases (SlGABA-Ts) and succinic semialdehyde dehydrogenase (SlSSADH), controlling three steps of the metabolic pathway of GABA, were studied through virus-induced gene silencing approach in tomato. Silencing of SlGADs (GABA biosynthetic genes) and SlGABA-Ts (GABA catabolic genes) led to increased accumulation of reactive oxygen species (ROS) as well as salt sensitivity under 200 mm NaCl treatment. Targeted quantitative analysis of metabolites revealed that GABA decreased and increased in the SlGADs- and SlGABA-Ts-silenced plants, respectively, whereas succinate (the final product of GABA metabolism) decreased in both silenced plants. Contrarily, SlSSADH-silenced plants, also defective in GABA degradation process, showed dwarf phenotype, curled leaves and enhanced accumulation of ROS in normal conditions, suggesting the involvement of a bypath for succinic semialdehyde catabolism to γ-hydroxybutyrate as reported previously in Arabidopsis, were less sensitive to salt stress. These results suggest that GABA shunt is involved in salt tolerance of tomato, probably by affecting the homeostasis of metabolites such as succinate and γ-hydroxybutyrate and subsequent ROS accumulation under salt stress.

Na⁺/H⁺ exchanger 1 participates in tobacco disease defence against Phytophthora parasitica var. nicotianae by affecting vacuolar pH and priming the antioxidative system.

Despite the importance of NHX1 (Na(+)/H(+) exchanger 1) in plant salt tolerance, little is known about its other functions. In this study, intriguingly, it was found that NHX1 participated in plant disease defence against Phytophthora parasitica var. nicotianae (Ppn) in Nicotiana benthamiana. NbNHX1 was originally isolated from N. benthamiana, and characterized. The subcellular localization of NbNHX1 with its C-terminus fused with green fluorescent protein indicated that NbNHX1 localized primarily to the tonoplast. Tobacco rattle virus-induced NbNHX1 silencing led to reduced H(+) efflux from the vacuole to cytoplasts, and decreased Ppn resistance in N. benthamiana. After attack by Ppn, NbNHX1-silenced plants exhibited impaired ability to scavenge reactive oxidative species (ROS) induced by the pathogen. Pea early browning virus-mediated ectopic expression of SeNHX1 (from Salicornia europaea) or AtNHX1 (from Arabidopsis thaliana) both conferred enhanced Ppn resistance to N. benthamiana, with a lower H2O2 concentration after Ppn inoculation. Further investigation of the role of NHX1 demonstrated that transient overexpression of NbNHX1 improved the vacuolar pH and cellular ROS level in N. benthamiana, which was coupled with an enlarged NAD(P) (H) pool and higher expression of ROS-responsive genes. In contrast, NbNHX1 silencing led to a lower pH in the vacuole and a lower cellular ROS level in N. benthamiana, which was coupled with a decreased NAD(P) (H) pool and decreased expression of ROS-responsive genes. These results suggest that NHX1 is involved in plant disease defence; and regulation of vacuolar pH by NHX1, affecting the cellular oxidation state, primes the antioxidative system which is associated with Ppn resistance in tobacco.