Differential responses of two rice varieties to salt stress

Two rice varieties, viz. Nonabokra and Pokkali, have been evaluated for their responses to salinity in terms of some physiological and biochemical attributes. During the exposure to salinity (200 mM concentration of sodium chloride for 24, 48, and 72 h), a significant increase in sodium was recorded which was also concomitant with the changes of other metabolic profiles like proline, phenol, polyamine, etc. The protein oxidation was significantly increased and also varied between the two cultivars. The changes in activities of anti-oxidative enzymes under stress were significantly different to the control. The detrimental effects of salinity were also evident in terms of lipid peroxidation, chlorophyll content, protein profiles, and generation of free radicals; and these were more pronounced in Pokkali than in Nonabokra. The assessment and analysis of these physiological characters under salinity could unravel the mechanism of salt responses revealed in this present study and thus might be useful for selection of tolerant plant types under the above conditions of salinity.     

Differential responses of stress genes to low dose-rate gamma irradiation

In the past, most mechanistic studies of ionizing radiation response have employed very large doses, then extrapolated the results down to doses relevant to human exposure. It is becoming increasingly apparent, however, that this does not give an accurate or complete picture of the effects of most environmental exposures, which tend to be of low dose and protracted over time. We have initiated direct studies of low dose exposures, and using the relatively responsive ML-1 cell line, have shown that changes in gene expression can be triggered by doses of gamma-rays of 10 cGy and less in human cells. We have now extended these studies to investigate the effects on gene induction of reducing the rate of irradiation. In the ML-1 human myeloid leukemia cell line, we have found that reducing the dose rate over three orders of magnitude results in some protection against the induction of apoptosis, but still causes linear induction of the p53-regulated genes CDKN1A, GADD45A, and MDM2 between 2 and 50 cGy. Reducing the rate of exposure reduces the magnitude of induction of CDKN1A and GADD45A, but not the magnitude or duration of cell cycle delay. In contrast, MDM2 is induced to the same extent regardless of the rate of dose delivery. Microarray analysis has identified additional low dose-rate-inducible genes, and indicates the existence of two general classes of low dose-rate responders in ML-1. One group of genes is induced in a dose rate-dependent fashion, similar to GADD45A and CDKN1A. Functional annotation of this gene cluster indicates a preponderance of genes with known roles in apoptosis regulation. Similarly, a group of genes with dose rate-independent induction, such as seen for MDM2, was also identified. The majority of genes in this group are involved in cell cycle regulation. This apparent differential regulation of stress signaling pathways and outcomes in response to protracted radiation exposure has implications for carcinogenesis and risk assessment, and could not have been predicted from classical high dose studies.     

The influence of salt stress on ABA and auxin concentrations in two maize cultivars differing in salt resistance

The plant hormones abscisic acid (ABA) and auxin (IAA, IBA) play important roles in plant responses to environmental stresses such as salinity. Recent breeding improvements in terms of salt resistance of maize have lead to a genotype with improved growth under saline conditions. By comparing this salt-resistant hybrid with a sensitive hybrid, it was possible to show differences in hormone concentrations in expanding leaves and roots. In response to salinity, the salt-resistant maize significantly increased IBA concentrations in growing leaves and maintained IAA concentration in roots. These hormonal adaptations may help to establish favorable conditions for growth-promoting agents such as β-expansins and maintain growth of resistant maize hybrids under salt stress. Moreover, ABA concentrations significantly increased in resistant maize leaves under salt stress, which may contribute to acidifying the apoplast, which in turn is a prerequisite for growth.     

Differential molecular responses to abscisic acid and osmotic stress in viviparous maize embryos

Substantial quantities of mRNA encoding the abundant Em polypeptide accumulate, in planta, in developing embryos of maize (Zea mays L.). By contrast, accumulation of Em mRNA is only barely detectable in embryos with the vp-5/vp-5 genotype [an abscisic acid (ABA)-deficient viviparous phenotype]. Em mRNA is not detectable within viviparous embryos of the vp-1/vp-1 genotype that are non-responsive to ABA. Culture of immature wild-type and vp-5/vp-5 embryos in the presence of exogenous ABA or of an osmotically active agent prevents precocious germination and results in expression of the Em genes. When vp-1/vp-1 embryos are cultured under similar conditions, only the application of osmotic stress prevents precocious germination. However, Em mRNA does not accumulate either in ABA-treated or stressed, arrested embryos, indicating a requirement for ABA perception through a VP-1-mediated mechanism for Em gene expression. Nevertheless, vp-1/vp-1 embryos do show both ABA and stress responses at the molecular level. Treatment with ABA causes the accumulation of mRNA encoding a polypeptide of approx. 30 kDa, whilst osmotic stress induces the accumulation both of a 30-kDa polypeptide and a set of approx. 20-kDa polypeptides. This indicates the existence of discrete, parallel ABA and stress response pathways in developing maize embryos.Abbreviations ABA abscisic acid - cDNA copy-DNA - DAP days after pollination - kDa kilodaltons - MS Murashige and Skoog medium - LEA late embryogenesis abundant - NEpHGE non-equilibrium pH gradient gel electrophoresis - SDS-PAGE sodium dodecyl sulphate-polyacrylamide gel electrophoresis     

Isolation and characterization of maize PMP3 genes involved in salt stress tolerance

Plasma membrane protein 3 (PMP3), a class of small hydrophobic polypeptides with high sequence similarity, is responsible for salt, drought, cold, and abscisic acid. These small hydrophobic ploypeptides play important roles in maintenance of ion homeostasis. In this study, eight ZmPMP3 genes were cloned from maize and responsive to salt, drought, cold and abscisic acid. The eight ZmPMP3s were membrane proteins and their sequences in trans-membrane regions were highly conserved. Phylogenetic analysis showed that they were categorized into three groups. All members of group II were responsive to ABA. Functional complementation showed that with the exception of ZmPMP3-6, all were capable of maintaining membrane potential, which in turn allows for regulation of intracellular ion homeostasis. This process was independent of the presence of Ca(2+). Lastly, over-expression of ZmPMP3-1 enhanced growth of transgenic Arabidopsis under salt condition. Through expression analysis of deduced downstream genes in transgenic plants, expression levels of three ion transporter genes and four important antioxidant genes in ROS scavenging system were increased significantly in transgenic plants during salt stress. This tolerance was likely achieved through diminishing oxidative stress due to the possibility of ZmPMP3-1's involvement in regulation of ion homeostasis, and suggests that the modulation of these conserved small hydrophobic polypeptides could be an effective way to improve salt tolerance in plants.     

Differential stress responses of early salt-stress responding genes in common wheat

Four early salt-stress responding genes (WESR1-4) in common wheat (Triticum aestivum L.) were analyzed for their temporal accumulation of mRNA during salt stress, osmotic stress and abscisic acid (ABA) treatment. All genes showed transient stimulation by 0.15 M NaCl treatment. WESR1 and WESR2 were induced by both osmotic stress and exogenous ABA treatment. WESR3 responded to exogenous ABA, but not to osmotic stress. WESR4 did not show significant response to either osmotic stress or exogenous ABA treatment. These results suggest that wheat has at least two salt stress signal transduction pathways, an ABA-dependent and ABA-independent pathway.     

Electrophysiological responses of maize roots to low water potentials: relationship to growth and ABA accumulation

The maintenance of root elongation is an important adaptive response to low water potentials (psi(w)), but little is known about its regulation. An important component may be changes in root cell electrophysiology, which both signal and maintain growth maintenance processes. As a first test of this hypothesis, membrane potentials (E(m)) were measured within the cell elongation zone of maize (Zea mays L.) primary roots. Seedlings were grown in oxygenated solution culture, and low psi(w) was imposed by the gradual addition of polyethylene glycol. Cells hyperpolarized approximately 25 mV in response to low psi(w), and after 48 h resting potentials remained significantly hyperpolarized at psi(w) lower than -0.3 MPa compared with roots at high psi(w). Inhibitor experiments showed that the hyperpolarization was dependent on plasma membrane H(+)-ATPase activity. Previous work showed that accumulation of abscisic acid (ABA) is required for the maintenance of maize primary root elongation at low psi(w). To determine if the mechanism of action of ABA involves changes in root electrophysiology, E(m) measurements were made during long-term exposure to low psi(w). Steady-state resting E(m) were measured in regions in which maintenance of cell elongation was dependent on ABA accumulation (2-3 mm from the apex), or in which elongation was inhibited regardless of ABA status (6-8 mm from the apex). E(m) was substantially more negative in ABA-deficient roots specifically in the 2-3 mm region. The results suggest that set-points for ion homeostasis shifted in association with the maintenance of root cell elongation at low psi(w), and that ABA accumulation plays a role in regulating the ion transport processes involved in this response.     

镉胁迫下玉米幼苗生理生态的变化

用不同浓度Cd2 + 处理玉米种子 ,在室内常规培养 ,研究了种子萌发和幼苗生理生态的变化。结果表明 ,镉影响玉米种子的萌发和幼苗的生长。当Cd2 + 浓度高于 5 0mg·L-1时 ,显著抑制种子的发芽率 ;随Cd2 + 浓度的增加 ,根系长度和侧根数减小 ;在Cd2 + 浓度小于 5mg·L-1时 ,镉刺激苗高和根系及地上部干物质量增加 ,当Cd2 + 浓度超过相应浓度时 ,苗高和根系及地上部的生长量随Cd2 + 浓度提高而降低。镉胁迫下幼苗根系活力和叶绿素含量明显降低 ,根内丙二醛含量增加。镉影响玉米幼苗对矿质元素的吸收。根系和茎叶中Ca、Mg、Fe、Cu的吸收量随Cd2 + 浓度提高而增加 ,K、Zn的吸收量随Cd2 + 浓度提高而减少。     

Photosynthetic and stomatal responses of spinach leaves to salt stress

The gas exchange of spinach plants, salt-stressed by adding NaCl to the nutrient solution in increments of 25 millimolar per day to a final concentration of 200 millimolar, was studied 3 weeks after starting NaCl treatment. Photosynthesis became light saturated at 1100 to 1400 micromoles per square meter per second in salt-treated plants and at approximately 2000 micromoles per square meter per second in control plants. Photosynthetic capacity of the mesophyll measured as a function of intercellular partial pressure of CO₂ at the light intensity prevailing during growth and at light saturation were both decreased in the salttreated plants. The CO₂ compensation points and relative enhancements of photosynthesis at low O₂ were not affected by salinity. The lower photosynthetic rates in salt-treated leaves at 450 micromoles per square meter per second were associated with a 70% reduction in stomatal conductance and low intercellular CO₂ (219 microbars; cf. 285 microbars for controls). Increasing photon flux density to light saturation extended the linear portions of the CO₂ response curves, increased stomatal conductances, increased intercellular CO₂ in the salt-treated plants, but lowered it in controls, and accentuated differences in photosynthetic rate (area basis) between the treatments.

Leaves from salt-treated plants were thicker but contained about 73% of the chlorophyll per unit area of control plants. When photosynthetic rates were expressed on a chlorophyll basis there was no difference in initial slope of assimilation versus intercellular CO₂ between treatments. Photosynthetic rates (chlorophyll basis) at light saturation differed only by 20% which was also observed earlier with isolated, intact chloroplasts (Robinson et al. 1983 Plant Physiol 73: 238-242).

Measurement of carbon isotope ratio revealed less discrimination against ¹³C with salt treatment and confirmed the persistence of low intercellular partial pressures of CO₂ during plant growth. The development of a thicker leaf with less chlorophyll per unit area during salt treatment permitted stomatal conductance and intercellular partial pressure of CO₂ to decline without restricting photosynthesis and had the benefit of greatly increasing water use efficiency.

     

Differential expression of candidate genes for lignin biosynthesis under drought stress in maize leaves

In order to provide information for the development of molecular selection markers for drought tolerance improvement, the methods of prometric analysis, quantitative real-time PCR and field evaluation were employed for the identification of the differential expression of candidate genes under drought stress in maize. At seventeen, twenty-four and forty-eight hours of polyethylene glycol-simulated drought stress at the seventh leaf stage, leaf samples were collected from two drought-tolerant inbred lines for prometric analysis by two-dimensional electrophoresis and peptide mass fingerprinting. Fifty-eight proteins out of more than 500 were found in response to drought stress. Three drought-induced spots 2506, 3507 and 4506 showed sequence similarity with cinnamyl alcohol dehydrogenase, cytochrome protein 96A8 and S-adenosyl-L-methionine synthase, respectively. The expression of two key enzymes to lignin biosynthesis was quantified by quantitative real-time PCR among three drought-tolerant and one drought-sensitive inbred lines under drought stress and well-watered control conditions. After a decrease at the beginning of drought stress, the expression of cinnamyl alcohol dehydrogenase and caffeateO-methyltransferase recovered at twenty-four hours of the drought stress in the three drought-tolerant lines, but not in the drought-sensitive lines. Leaf lignin content, anthesis-silking interval and grain weight per plant were investigated with six inbred lines of varying drought tolerance under drought stress and well-watered control. Drought tolerance coefficients of these three characters were calculated and the correlation coefficients among these drought tolerance coefficients were estimated. Significant difference in leaf lignin content was found among the inbred lines and in response to drought stress. Close correlations were observed between the drought tolerant coefficients for leaf lignin content and grain weight per plant, and between the drought tolerant coefficients for leaf lignin content and anthesis-silking interval. These results indicate that leaf lignin content is a useful index for evaluation of drought tolerance in maize. Molecular selection markers can be developed on the basis of differential expression of the candidate genes and applied to maize improvement for drought tolerance.     

Stevia rebaudiana Bertoni responses to salt stress and chitosan elicitor

This study examined the effect of chitosan elicitor with four different concentrations (0, 0.2, 0.4 and 0.6 g/l) on physiological and biochemical properties of stevia under four levels of salinity stress (0, 50, 100, 150 mM level of NaCl). Salt stress caused reduction of chlorophyll a (Chl a), chlorophyll b (Chl b), total chlorophyll, carotenoid and total protein content. The increment of malondialdehyde (MDA) content was not significant in all NaCl levels, while the CAT and POX activities were increased as well as stevioside and rebaudioside A under salinity stress. On one side, chitosan treatments could compensate the reduction of physiological traits such as photosynthetic pigments and protein content. On the other side, chitosan caused multiple increases in malondialdehyde content, antioxidant enzymes activity (catalase and peroxidase), steviol glycosides (stevioside and rebaudioside A) under salt stress. We report for the first time, the potential of chitosan to enhance salinity-tolerant abilities in stevia through increment of the salt-adaptive factors and to diminish harmful damages caused by NaCl stress.     

Differential responses of antioxidative enzymes and lipid peroxidation to salt stress in salt-tolerant Plantago maritima and salt-sensitive Plantago media

The changes in plant growth, relative water content (RWC), stomatal conductance, lipid peroxidation and antioxidant system in relation to the tolerance to salt stress were investigated in salt-tolerant Plantago maritima and salt-sensitive Plantago media. The 60 days old P. maritima and P. media seedlings were subjected to 0, 100 and 200 mM NaCl for 7 days. Reduction in shoot length was higher in P. media than in P. maritima after exposure to 200 mM NaCl, but 100 mM NaCl treatment did not show any effect on shoot length of P. maritima. Shoot dry weight decreased in P. media and did not change in P. maritima. Two hundred millimolar NaCl treatment had no effect on leaf RWC in P. maritima, but it was reduced in P. media. Salt stress caused reduction in stomatal conductance being more pronounced in P. media than in P. maritima. Activities of superoxide dismutase (SOD; EC 1.15.1.1), catalase (CAT; EC 1.11.1.6), glutathione reductase (GR; EC 1.6.4.2) decreased in P. media with increasing salinity. Ascorbate peroxidase (APX; EC 1.11.1.11) activity in leaves of P. media was increased and showed no change under 100 and 200 mM NaCl, respectively. However, activities of CAT, APX and GR increased under 200 mM NaCl while their activities did not change under 100 mM NaCl in P. maritima. SOD activity in leaves of P. maritima increased with increasing salinity. Concomitant with this, four SOD activity bands were identified in leaves of P. maritima, two bands only were observed in P. media. Peroxidase (POX; EC 1.11.1.7) activity increased under both salt concentrations in P. maritima, but only under 200 mM NaCl in P. media. Confirming this, five POX activity bands were identified in leaves of P. maritima, but only two bands were determined in P. media. Malondialdehyde levels in the leaves increased under salt stress in P. media but showed no change and decreased in P. maritima at 100 and 200 mM NaCl, respectively. These results suggest that the salt-tolerant P. maritima showed a better protection mechanism against oxidative damage caused by salt stress by its higher induced activities of antioxidant enzymes than the salt-sensitive P. media.     

Differential stress responses to NaCl salt application in early- and late-maturing diploid potato (Solanum sp.) clones

Cultivated tetraploid potatoes (Solanum tuberosum L.) are moderately salt sensitive but greater stress tolerance exists in diploid wild types. However, little work has been published on salt-tolerance in diploid potato. This study utilized sensitive and tolerant diploid potatoes as well as a commercially cultivated potato to investigate mechanisms of stress tolerance. Stem cuttings from salt-tolerant (T) and sensitive (S) clones of early-maturing (EM) and late-maturing (LM) diploid potato clones were stressed for 5 days at the tuber initiation stage with 150 mmol NaCl in a hydroponic sand culture under greenhouse conditions. The stress responses of the early- and late-maturing potato clones were distinctly different. Under stress, early-maturing clones accumulated Na⁺ in the leaf tissues while late-maturing clones generally excluded Na⁺ from the leaf tissues. Salt tolerant clones of both maturity types were able to tolerate high levels of Na⁺ in the leaf tissues. The lower leaves accumulated more Na⁺ than the upper leaves in both maturity types. The potassium to sodium ratio was significantly greater in the leaves of the late-maturing types, reflecting differences in Na⁺ accumulation rather than alterations in K⁺ levels. Proline levels increased upon salt exposure but were not clearly associated with salinity tolerance. Tolerance was manifested in maintenance of vegetative growth, tuber yield, and reduced leaf necrosis. These responses require efficient uptake of water and source–sink translocation. Maintenance of stomatal conductance under stress was not associated with these responses but tuber yield was related to lower-leaf osmotic potential (OP) in both early- and late-maturity types. Salt tolerant clones of both maturity types also had less negative tuber OP under salt stress than sensitive types. High yielding EMT and LMT clones either minimized tuber yield loss or even increased yield after exposure to salt stress. Mechanistic studies and screening experiments for salt tolerant clones should consider maturity type, leaf position and source–sink relationships enhancing tuber yield.     

Tissue-specific regulation of rice molybdenum cofactor sulfurase gene in response to salt stress and ABA

Molybdenum-containing aldehyde oxidase is a key enzyme for catalyzing the final step of abscisic acid (ABA) biosynthesis in plants. Sulfuration of the molybdenum cofactor (MoCo) is an essential step for activating aldehyde oxidase. The molybdenum cofactor sulfurase (MCSU) that transfers the sulfur ligand to aldehyde oxidase-bound MoCo is thus considered an important factor in regulating the ABA levels in plant tissues. In this study, we identified the rice MCSU cDNA (OsMCSU), which is the first MCSU gene cloned in monocot species. According to the functional domain analysis of the predicted amino acid sequence, the OsMCSU protein contains a Nifs domain at its N-terminus and a MOSC domain at the C-terminus. Expression of the OsMCSU gene was up-regulated by salt stress in root tissues of rice seedlings, but this effect was not observed in leaf tissues. In roots, regulations of OsMCSU expressions could be mediated by both ABA-dependent and ABA-independent signaling pathways under salt stress condition.