Evaluating the Influence of Water Scarcity on the Host Response of Garlic to the Stem and Bulb Nematode Ditylenchus dipsaci.

Ditylenchus dipsaci is a plant-parasitic nematode with a great economic impact on bulbous crops, including garlic (Allium sativum L.), and is distributed worldwide, particularly in the Mediterranean region. Traditionally, garlic was a rainfed crop in Spain, but irrigated areas have increased during the last few decades. However, the expected climatic conditions, with longer and more intense droughts, will make it necessary to reduce the water supply to garlic crops. This poses the urgent need to select garlic cultivars more tolerant to water scarcity and that are also more resistant to plant pathogenic organisms. The aim of this work was to analyze the influence of water stress on the host response of garlic plants to D. dipsaci. The specific objectives were to evaluate the level of nematode infestation in plants from four garlic genotypes treated with a reduced irrigation regime and compare them with those of control plants not subjected to water stress. The observed results were correlated with changes in the bulb and root development, as well as in the physiological parameters (total chlorophyll concentration and proline accumulation). The effects were different depending on whether the plants were subjected to water stress before or after nematode inoculation, as well as whether the water stress was continuous or discontinuous. Garlic inter-cultivar variability also affected the obtained results.

Exogenously applied Casuarina equisetifolia leaf extracts act as an osmoprotectant on proline accumulation under drought stress in local rice from Indonesia.

The effects of exogenously supplied osmoprotectants in crops have not yet been extensively studied. In this study, an osmoprotectant containing a high concentration of proline (2.5 g mol-1 FW) was obtained from a Casuarina equisetifolia leaf extract. The effect of the extract was evaluated in local Indonesian rice cultivars Boawae Seratus Malam (BSM), Gogo Jak (GJ), Situ Bagendit (SB) (drought-tolerant), Kisol Manggarai (KM) and Ciherang (drought-susceptible) cultivars under drought at the morphological, physiological, and genetic levels. Under drought, the KM showed an increased level of OsWRKY, OsNAC, OsDREB1A, and OsDREB2A expression after application of the osmoprotectant, leading to the activation of proline synthesis genes including OsP5CS1, OsP5CR, and OsProDH, while the tolerant cultivars (BSM, GJ, and SB) showed no difference. The content of chlorophyll, carotenoids, anthocyanins, ascorbate peroxidase, catalase, and superoxide dismutase activities also increased in GJ and KM, during drought stress and applied osmoprotectants, but remained low in the BSM. We conclude that the foliar application of osmoprotectants derived from C.equisetifolia caused an accumulation of proline in susceptible plants. The existence of these extracts stabilizes leaf cells and supports photosynthetic compartments and carbon assimilation in plants, leading to growth.

The ubiquitin-binding protein MdRAD23D1 mediates drought response by regulating degradation of the proline-rich protein MdPRP6 in apple (Malus domestica).

RAD23 (RADIATION SENSITIVE23) proteins are a group of UBL-UBA (ubiquitin-like-ubiquitin-associated) proteins that shuttle ubiquitylated proteins to the 26S proteasome for breakdown. Drought stress is a major environmental constraint that limits plant growth and production, but whether RAD23 proteins are involved in this process is unclear. Here, we demonstrated that a shuttle protein, MdRAD23D1, mediated drought response in apple plants (Malus domestica). MdRAD23D1 levels increased under drought stress, and its suppression resulted in decreased stress tolerance in apple plants. Through in vitro and in vivo assays, we demonstrated that MdRAD23D1 interacted with a proline-rich protein MdPRP6, resulting in the degradation of MdPRP6 by the 26S proteasome. And MdRAD23D1 accelerated the degradation of MdPRP6 under drought stress. Suppression of MdPRP6 resulted in enhanced drought tolerance in apple plants, mainly because the free proline accumulation is changed. And the free proline is also involved in MdRAD23D1-mediated drought response. Taken together, these findings demonstrated that MdRAD23D1 and MdPRP6 oppositely regulated drought response. MdRAD23D1 levels increased under drought, accelerating the degradation of MdPRP6. MdPRP6 negatively regulated drought response, probably by regulating proline accumulation. Thus, "MdRAD23D1-MdPRP6" conferred drought stress tolerance in apple plants.

Hydrogen Sulfide Mitigates Chilling Injury of Postharvest Banana Fruits by Regulating γ-Aminobutyric Acid Shunt Pathway and Ascorbate-Glutathione Cycle.

This study aimed to determine the effect of hydrogen sulfide on chilling injury (CI) of banana (Musa spp.) during cold storage (7°C). It was observed that hydrogen sulfide application (2 mmol L-1) markedly reduced the CI index and showed significantly higher chlorophyll contents, along with suppressed chlorophyll peroxidase and chlorophyllase enzyme activity. The treated banana fruits exhibited substantially higher peel lightness (L*), along with significantly a lower browning degree and soluble quinone content. The treated bananas had substantially a higher endogenous hydrogen sulfide content and higher activity of its biosynthesis-associated enzymes such as D-cysteine desulfhydrase (DCD) and L-cysteine desulfhydrase (LCD), along with significantly lower ion leakage, lipid peroxidation, hydrogen peroxide, and superoxide anion concentrations. Hydrogen sulfide-treated banana fruits showed an increased proline content and proline metabolism-associated enzymes including ornithine aminotransferase (OAT), Δ1-pyrroline-5-carboxylate synthetase (P5CS), and proline dehydrogenase (PDH). In the same way, hydrogen sulfide-fumigated banana fruits accumulated higher endogenous γ-aminobutyric acid (GABA) due to enhanced activity of glutamate decarboxylase (GAD) and GABA transaminase (GABA-T) enzymes. The hydrogen sulfide-treated fruits exhibited higher total phenolics owing to lower polyphenol oxidase (PPO) and peroxidase (POD) activity and stimulated phenylalanine ammonia lyase (PAL). The treated banana exhibited higher ascorbate peroxidase (APX), catalase (CAT), glutathione reductase (GR), dehydroascorbate reductase (DHAR), monodehydroascorbate reductase (MDHAR), and superoxide dismutase (SOD) activity, along with higher glutathione (GSH) and ascorbic acid (AsA) concentrations and a significantly lower dehydroascorbic acid (DHA) content. In conclusion, hydrogen sulfide treatment could be utilized for CI alleviation of banana fruits during cold storage.

Dehydrin responsive HVA1 driven inducible gene expression enhanced salt and drought tolerance in wheat.

Heterologous expression of plant genes is becoming an important strategy for the improvement of specific traits in existing cultivars. This study presents the response of a salt-sensitive high-yielding wheat variety under stress-inducible expression of barley HVA1 gene belonging to the Late embryogenesis abundance (LEA) gene family. Six homozygous transgenic wheat plants were developed and advanced for testing under various water regimes and salt stress conditions. Putative transgenic plants showed better germination and root shoot development at the early developmental stages under drought stress conditions. Moreover, transgenic plants illustrated higher values of physiological features as compared to non-transgenic plants under both drought and salinity stresses that indicate improved physiological processes in transgenic plants. Higher membrane stability index (MSI) and lower electrolyte leakage (EL) after exposure to abiotic stresses reveal improved cellular membrane stability (CMS) and reduced injury to chloroplast membrane. Interestingly, under salinity stress, transgenic wheat plants showed preference towards higher K+ accumulation in the shoot, which is not a well-understood HVA1 mediated Na + avoidance mechanism under excessive subsurface salts. The predisposition of K+/Na + under salt stress conditions on heterologous expression of the HVA1 gene in wheat needs to be studied in detail in further studies.

Plastic changes in cold and drought tolerance of Drosophila nepalensis correlate with sex-specific differences in body melanization, cuticular lipid mass, proline accumulation, and seasonal abundance.

Autumn-collected flies of Himalayan Drosophila nepalensis differ in body color phenotypes (males more melanized relative to females) and in their behavior (males abundant in the open sites vs. shelters-seeking females). In contrast, winter-collected flies of both sexes are equally melanized and abundant in the open sites. We tested developmental and adult plasticity changes in cold or drought tolerance in D. nepalensis flies reared under winter or autumn simulated conditions. In D. nepalensis flies reared at 21 °C, male flies were more cold tolerant (as shown by shorter chill-coma recovery time and lower cold-shock mortality). Further, male flies also exhibited greater drought tolerance (increased levels of desiccation resistance, cuticular lipid mass, melanization, hydration level, and dehydration tolerance) as compared to females. We observed sex-specific differences in the adult plasticity responses due to rapid cold or drought hardening (RCH or RDH); and for the persistence of cold acclimation effects. RCH or RDH-induced changes in the level of proline accumulations are negatively correlated with a decrease in the chill-coma recovery time. Therefore, cold or drought hardening treatments are likely to influence cold tolerance through proline accumulation. Developmental acclimation and adult hardening responses revealed significant interaction effects between sexes and thermal treatments. Thus, sex-specific differences in morphological traits (body melanization and cuticular lipid mass) and physiological traits (adult plasticity changes in cold tolerance and proline accumulation) correlate with behavioral divergence (habitat usage) across sexes.

A Wild Allele of Pyrroline-5-Carboxylate Synthase1 Leads to Proline Accumulation in Spikes and Leaves of Barley Contributing to Improved Performance Under Reduced Water Availability.

Water stress (WS) during spike development strongly affects final grain yield and grain quality in cereals. Proline, an osmoprotectant amino-acid, may contribute to alleviating the effects of cell and tissue dehydration. We studied five spring barley genotypes contrasting in their drought response, including two introgression lines, S42IL-143 and S42IL-141, harboring a Pyrroline-5-carboxylate synthase1- P5cs1 allele originating from the wild barley accession ISR42-8. We tested the hypothesis that barley genotypes harboring a wild allele at P5cs1 locus are comparatively more drought-tolerant at the reproductive stage by inducing proline accumulation in their immature spikes. At the booting stage, we subjected plants to well-watered and WS treatments until physiological maturity. Several morpho-physiological traits had significant genotype by treatment interaction and reduction under WS. Varying levels of genotypic proline accumulation and differences in WS tolerance were observed. Spike proline accumulation was higher than leaf proline accumulation for all genotypes under WS. Also, introgression lines carrying a wild allele at P5cs1 locus had a markedly higher spike and leaf proline content compared with the other genotypes. These introgression lines showed milder drought symptoms compared with elite genotypes, remained photosynthetically active under WS, and maintained their intrinsic water use efficiency. These combined responses contributed to the achievement of higher final seed productivity. Magnetic resonance imaging (MRI) of whole spikes at the soft dough stage showed an increase in seed abortion among the elite genotypes compared with the introgression lines 15 days after WS treatment. Our results suggest that proline accumulation at the reproductive stage contributes to the maintenance of grain formation under water shortage.

Physiological and molecular responses of Setaria viridis to osmotic stress.

Drought-tolerant species, such as Setaria viridis, a C4 model plant, make physiological and biochemical adjustments water limitation and recover from the stress upon its release. We investigated S. viridis (A10.1 accession) responses to continuing osmotic stress. The osmotic stress was imposed using polyethylene glycol (PEG) 8000 (7.5%) for 10 days. Morphological traits and stomatal conductance were measured daily for the 10 days. On days 6 and 10, the following traits were measured separately for root and shoot: relative water content (RWC), osmotic potential (OP), electrolytic leakage (EL), and proline content. qPCR analysis was used to evaluate the expression of five selected genes in roots (SvLEA, SvDREB1C, SvPIP2-1, SvHSP20, and SvP5CS2), and chlorophyll a fluorescence was measured on three key days. The morphological data demonstrated a drastic reduction in shoot biomass as an effect of water deficit caused by the osmotic stress. Shoot biomass reduction could be associated with putative ABA-dependent signaling involved in SvDREB1C expression. Stomatal conductance and photosynthesis were severely affected up until day 6, however, stomatal conductance and some photosynthetic parameters such as FV/FM, ABS/RC, and DI0/RC showed total or slight recovery on day 10. Root EL decreased in treated plants suggesting an investment in membrane protection by osmoregulator expression such as dehydrin (SvLEA) and proline (SvP5CS2) genes. Our data suggest that S. viridis exhibited a partial recovery from an imposed and constant osmotic stress within 10 days.

Light Control of Salt-Induced Proline Accumulation Is Mediated by ELONGATED HYPOCOTYL 5 in Arabidopsis.

Plants have to adapt their metabolism to constantly changing environmental conditions, among which the availability of light and water is crucial in determining growth and development. Proline accumulation is one of the sensitive metabolic responses to extreme conditions; it is triggered by salinity or drought and is regulated by light. Here we show that red and blue but not far-red light is essential for salt-induced proline accumulation, upregulation of Δ1-PYRROLINE-5-CARBOXYLATE SYNTHASE 1 (P5CS1) and downregulation of PROLINE DEHYDROGENASE 1 (PDH1) genes, which control proline biosynthetic and catabolic pathways, respectively. Chromatin immunoprecipitation and electrophoretic mobility shift assays demonstrated that the transcription factor ELONGATED HYPOCOTYL 5 (HY5) binds to G-box and C-box elements of P5CS1 and a C-box motif of PDH1. Salt-induced proline accumulation and P5CS1 expression were reduced in the hy5hyh double mutant, suggesting that HY5 promotes proline biosynthesis through connecting light and stress signals. Our results improve our understanding on interactions between stress and light signals, confirming HY5 as a key regulator in proline metabolism.

Proline Accumulation Influenced by Osmotic Stress in Arbuscular Mycorrhizal Symbiotic Plants.

Salinity and drought are the major osmotic stress limitations that affect plant growth and crop yield in agriculture worldwide. The alternative response mediated by plants in response to salinity and drought are principally proline accumulation which regulates stress combat strategies owing to sustainable production in the realm of agricultural production even under severe stress. Symbiotic and soil associated arbuscular mycorrhizal fungi (AMF) are regarded as efficient biofertilizers in several crops under these stresses. Summarily AMF is renowned for effective scavengers of free radicals in soil thereby increasing soil parameters optimal for plant growth. AMF contribute to augment host plant tolerance to stress specifically salinity and drought. Mycorrhizal colonization positively regulates root uptake of available nutrients and enhance growth even when bestowed by water constraints which has contributory roles due to proline accumulation providing several intriguing researches on AMF symbiosis pertaining to plant productivity and yield. Mycorrhizal plants and their non-mycorrhizal counterparts show varied expression pattern regarding proline amass. Hence, the precise role of proline with respect to stress tolerance and equivocal mechanisms involved in evasion of osmotic stress has not been extensively reviewed earlier. Further molecular forecasting in this arena is still an underexploited research field. This review comprehensively addresses the observable facts pertaining to proline accumulation upon AMF association and adherence relevant to stress tolerance and host plant efficiency and efficacy.

A differential tolerance to mild salt stress conditions among six Italian rice genotypes does not rely on Na+ exclusion from shoots.

Rice is very sensitive to salt stress at the seedling level, with consequent poor crop establishment. A natural variability in susceptibility to moderate saline environments was found in a group of six Italian temperate japonica rice cultivars, and the physiological determinants for salt tolerance were investigated. Cation (Na+, K+ and Mg++) levels were determined in shoots from individual rice plantlets grown in the absence or in the presence of inhibitory, yet sublethal salt levels, and at increasing time after salt treatments. Significant variations were found among genotypes, but these were unrelated to the relative tolerance, which seems to result from neither mechanism(s) for reduced Na+ translocation to the aerial part, nor its increased retrieval from the xylem mediating Na+ exclusion from leaves. Accordingly, thiobarbituric acid reactive substance levels raised in leaf tissues of salt-treated seedlings, and osmo-induced proline accumulation was found in all genotypes. Data suggest that the difference in salt tolerance most likely depends on mechanisms for osmotic adjustment and/or antioxidative defence.

Kunitz Proteinase Inhibitors Limit Water Stress Responses in White Clover (Trifolium repens L.) Plants.

The response of plants to water deficiency or drought is a complex process, the perception of which is triggered at the molecular level before any visible morphological responses are detected. It was found that different groups of plant proteinase inhibitors (PIs) are induced and play an active role during abiotic stress conditions such as drought. Our previous work with the white clover (Trifolium repens L.) Kunitz Proteinase Inhibitor (Tr-KPI) gene family showed that Tr-KPIs are differentially regulated to ontogenetic and biotic stress associated cues and that, at least some members of this gene family may be required to maintain cellular homeostasis. Altered cellular homeostasis may also affect abiotic stress responses and therefore, we aimed to understand if distinct Tr-PKI members function during drought stress. First, the expression level of three Tr-KPI genes, Tr-KPI1, Tr-KPI2, and Tr-KPI5, was measured in two cultivars and one white clover ecotype with differing capacity to tolerate drought. The expression of Tr-KPI1 and Tr-KPI5 increased in response to water deficiency and this was exaggerated when the plants were treated with a previous period of water deficiency. In contrast, proline accumulation and increased expression of Tr-NCED1, a gene encoding a protein involved in ABA biosynthesis, was delayed in plants that experienced a previous drought period. RNAi knock-down of Tr-KPI1 and Tr-KPI5 resulted in increased proline accumulation in leaf tissue of plants grown under both well-watered and water-deficit conditions. In addition, increased expression of genes involved in ethylene biosynthesis was found. The data suggests that Tr-KPIs, particularly Tr-KPI5, have an explicit function during water limitation. The results also imply that the Tr-KPI family has different in planta proteinase targets and that the functions of this protein family are not solely restricted to one of storage proteins or in response to biotic stress.

Identification of Δ1-pyrroline 5-carboxylate synthase (P5CS) genes involved in the synthesis of proline in Lotus japonicus.

Proline accumulation is a common response of plants to different biotic and abiotic stresses. In the model legume Lotus japonicus, osmotic stress-induced proline accumulation is one of the first responses of the plant, converting proline in a reliable stress marker. The main biosynthetic pathway of proline is from glutamate and the reaction catalyzed by the enzyme Δ1-pyrroline 5-carboxylate synthase (P5CS) is the rate limiting step. L. japonicus has been suggested to have three different P5CS genes. Here the predicted P5CS genes of L. japonicus were analyzed in silico and their expression under osmotic stress was determined. Contrary to previous suggestions this study demonstrated that L. japonicus has two different P5CS genes, as most dicotyledonous plants do. The gene that is inducible by osmotic stress and is located on chromosome 1, was called LjP5CS1, and the one located on chromosome 2 and not inducible by osmotic stress was called LjP5CS2.

Exogenous Application of GABA Improves PEG-Induced Drought Tolerance Positively Associated with GABA-Shunt, Polyamines, and Proline Metabolism in White Clover.

In order to investigate the physiological effects of exogenous γ-aminobutyric acid (GABA) on drought tolerance in white clover (Trifolium repens), GABA shunt, polyamines (PAs), and proline (Pro) metabolism were examined after plants pretreated with or without GABA (8 mM) and then exposed to water or 15% PEG-induced drought stress in growth chamber. In this study, exogenous application of GABA effectively alleviated drought-induced damage in leaves, as reflected by significantly higher relative water content, lower electrolyte leakage, lipid peroxidation, and leaf wilt. Exogenous GABA further promoted drought-induced increases in GABA transaminase and alpha ketone glutarate dehydrogenase activities, but inhibited glutamate decarboxylase activity under both control and drought conditions, resulting in an increase in endogenous glutamate (Glu) and GABA content. Besides, exogenous GABA could well accelerated PAs synthesis and suppressed PAs catabolism, which lead to the extremely enhanced different types of PAs content (free Put and Spd, insoluble bound Spd and Spm, soluble conjugated Spd and Spm, and total Put, Spd and Spm) under drought stress. In addition, exogenous GABA application further activated drought-induced Δ1-pyrroline-5-carboxylate synthetase and proline dehydrogenase activities, but suppressed drought-facilitated ornithine -δ-amino transferase activities, leading to a higher Pro accumulation and metabolism in GABA-pretreated plants in the middle and last period of drought. The results suggested that increased endogenous GABA by exogenous GABA treatment could improve drought tolerance of white clover associated with a positive regulation in the GABA-shunt, PAs and Pro metabolism.

Arabidopsis AMINO ACID PERMEASE1 Contributes to Salt Stress-Induced Proline Uptake from Exogenous Sources.

Stress-induced proline accumulation in plants is thought to result primarily from enhanced proline biosynthesis and decreased proline degradation. To identify regulatory components involved in proline transport, we screened for Arabidopsis thaliana T-DNA mutants with enhanced tolerance to toxic levels of exogenous proline (45 mM). We isolated the proline resistant 1-1 (pre1-1) mutant and map-based cloning identified PRE1 as AMINO ACID PERMEASE1 (AAP1, At1g58360), which encodes a plasma membrane-localized amino acid permease. AAP1 expression is induced by salt stress and abscisic acid, but not by proline. In pre1-1 mutants, a 19-nucleotide deletion in the AAP1 coding region produced a premature stop codon. When grown on proline-containing medium, pre1-1 mutants accumulated significantly less proline than did the wild type. Under salt stress, proline uptake decreased significantly in pre1-1 mutants. By contrast, proline uptake increased significantly in the wild type. These results suggest that AAP1 functions in the increase of proline uptake during salt stress. In addition, proline uptake promotes salt tolerance in Arabidopsis seedlings. We conclude that plants can increase proline accumulation by AtAAP1-mediated proline uptake from exogenous source, which help to improve the salt tolerance of seedlings.

Screening of Bread Wheat Genotypes for Drought Tolerance Using Phenotypic and Proline Analyses.

Drought stress is one of the leading constraints to wheat (Triticum aestivum L.) production globally. Breeding for drought tolerance using novel genetic resources is an important mitigation strategy. This study aimed to determine the level of drought tolerance among diverse bread wheat genotypes using agronomic traits and proline analyses and to establish correlation of proline content and agronomic traits under drought-stress conditions in order to select promising wheat lines for breeding. Ninety-six diverse genotypes including 88 lines from the International Maize and Wheat Improvement Center (CIMMYT)'s heat and drought nurseries, and eight local checks were evaluated under greenhouse and field conditions during 2014/15 and 2015/16 making four testing environments. The following phenotypic traits were collected after stress imposed during the heading to anthesis period: the number of days to heading (DTH), days to maturity (DTM), productive tiller number (TN), plant height (PH), spike length (SL), spikelet per spike (SPS), kernels per spike (KPS), thousand kernel weight (TKW) and grain yield (GY) and proline content (PC). Analysis of variance, Pearson's correlation coefficient, principal component and stress tolerance index were calculated. Genotypes with high yield performance under stressed and optimum conditions maintained high values for yield components. Proline content significantly increased under stress, but weakly correlated with agronomic traits under both optimal and water limited conditions. The positive correlation observed between grain yield and proline content under-drought stress conditions provides evidence that proline accumulation might ultimately be considered as a tool for effective selection of drought tolerant genotypes. The study selected 12 genotypes with high grain yields under drought stressed conditions and favorable adaptive traits useful for breeding.

Effect of progressive drought stress on growth, leaf gas exchange, and antioxidant production in two maize cultivars.

Drought stress is one of the major environmental factors responsible for reduction in crop productivity. In the present study, responses of two maize cultivars (Rung Nong 35 and Dong Dan 80) were examined to explicate the growth, yield, leaf gas exchange, leaf water contents, osmolyte accumulation, membrane lipid peroxidation, and antioxidant activity under progressive drought stress. Maize cultivars were subjected to varying field capacities (FC) viz., well-watered (80 % FC) and drought-stressed (35 % FC) at 45 days after sowing. The effects of drought stress were analyzed at 5, 10, 15, 20, ad 25 days after drought stress (DAS) imposition. Under prolonged drought stress, Rung Nong 35 exhibited higher reduction in growth and yield as compared to Dong Dan 80. Maize cultivar Dong Dan 80 showed higher leaf relative water content (RWC), free proline, and total carbohydrate accumulation than Run Nong 35. Malondialdehyde (MDA) and superoxide anion were increased with prolongation of drought stress, with higher rates in cultivar Run Nong 35 than cultivar Dong Dan 80. Higher production of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) and glutathione reductase (GR) resulted in improved growth and yield in Dong Dan 80. Overall, the cultivar Dong Dan 80 was better able to resist the detrimental effects of progressive drought stress as indicated by better growth and yield due to higher antioxidant enzymes, reduced lipid peroxidation, better accumulation of osmolytes, and maintenance of tissue water contents.

Hexaconazole-Cu complex improves the salt tolerance of Triticum aestivum seedlings.

Hexaconazole is one of the triazole complexes that are broadly used as systemic fungicides with non-traditional plant growth regulator properties. Hexaconazole-Cu complex (Hex-Cu) is a new triazole derivative, and the biological effect of Hex-Cu has been rarely studied. In this work, we investigated the functions of Hex-Cu in regulating growth and the response to salt stress in the seedlings of Triticum aestivum. Pretreated with 60µmolL(-1) Hex-Cu, the seedling plants got increased root/shoot ratio by 42.0%, and the contents of chlorophyll and soluble protein were also increased by 38.1% and 27.9%, respectively. Furthermore, Hex-Cu alleviated the growth inhibition caused by salt stress, enabled the seedlings to maintain a higher proline content and lower malondialdehyde accumulation. The functions of Hex-Cu in regulating the expression of proline synthetase (P5CS and P5CR) genes were investigated by quantitative real-time PCR (qPCR). Under 100mmolL(-1) NaCl stress, the expression of P5CS and P5CR in the seedlings by Hex-Cu pretreatment were significantly up-regulated. It attributed to the enhanced salt tolerance in plants.

Proline accumulation and metabolism-related genes expression profiles in Kosteletzkya virginica seedlings under salt stress.

Proline accumulation is a common response to salt stress in many plants. Salt stress also increased proline concentration in roots, stems, and leaves of Kosteletzkya virginica seedling treated with 300 mM NaCl for 24 h and reached 3.75-, 4.76-, and 6.83-fold higher than controls. Further study on proline content in leaves under salt stress showed that proline content increased with increasing NaCl concentrations or time. The proline level peaked at 300 mM NaCl for 24 h and reached more than sixfold higher than control, but at 400 mM NaCl for 24 h proline content fell back slightly along with wilting symptom. To explore the cause behind proline accumulation, we first cloned full length genes related to proline metabolism including KvP5CS1, KvOAT, KvPDH, and KvProT from K. virginica and investigated their expression profiles. The results revealed that the expressions of KvP5CS1 and KvProT were sharply up-regulated by salt stress and the expression of KvOAT showed a slight increase with increasing salt concentrations or time, while the expression of KvPDH was not changed much and slightly decreased before 12 h and then returned to the original level. As the key enzyme genes for proline biosynthesis, the up-regulated expression of KvP5CS1 played a more important role than KvOAT for proline accumulation in leaves under salt stress. The low expression of KvPDH for proline catabolism also made a contribution to proline accumulation before 12 h.

Mechanisms of salt tolerance in habanero pepper plants (Capsicum chinense Jacq.): Proline accumulation, ions dynamics and sodium root-shoot partition and compartmentation.

Despite its economic relevance, little is known about salt tolerance mechanisms in pepper plants. To address this question, we compared differences in responses to NaCl in two Capsicum chinense varieties: Rex (tolerant) and Chichen-Itza (sensitive). Under salt stress (150 mM NaCl over 7 days) roots of Rex variety accumulated 50 times more compatible solutes such as proline compared to Chichen-Itza. Mineral analysis indicated that Na(+) is restricted to roots by preventing its transport to leaves. Fluorescence analysis suggested an efficient Na(+) compartmentalization in vacuole-like structures and in small intracellular compartments in roots of Rex variety. At the same time, Na(+) in Chichen-Itza plants was compartmentalized in the apoplast, suggesting substantial Na(+) extrusion. Rex variety was found to retain more K(+) in its roots under salt stress according to a mineral analysis and microelectrode ion flux estimation (MIFE). Vanadate-sensitive H(+) efflux was higher in Chichen-Itza variety plants, suggesting a higher activity of the plasma membrane H(+)-ATPase, which fuels the extrusion of Na(+), and, possibly, also the re-uptake of K(+). Our results suggest a combination of stress tolerance mechanisms, in order to alleviate the salt-induced injury. Furthermore, Na(+) extrusion to apoplast does not appear to be an efficient strategy for salt tolerance in pepper plants.