Biochemical indicators of the water stress in maize seedlings

Free proline and chlorophyll contents, chlorophyllase, peroxidase and nitrate reductase activities were determined in maize seedlings grown under controlled conditions and submitted to water stress induced by 14 % PEG-4000 or NaCl (isotonic). All the parameters studied showed a significant response to the water stress, but the level of free proline and peroxidase activity together may preferentially be used for the diagnosis of osmotic stress or saline-osmotic stress.     

Biological seed priming mitigates the effects of water stress in sunflower seedlings

The sunflower (Helianthus annuus L. cv. PAC 36) seedlings were inoculated with plant growth promoting rhizobacteria (PGPR), viz. Azotobacter chroococcum (A+), Bacillus polymyxa (B+), separately and in combination of the two (AB+). Relative water content and seedling growth were maximum in AB+ seedlings under control. Water stress significantly decreased the RWC, growth and dry mass of non-inoculated seedlings. However, inoculated seedlings maintained higher growth even under water stress. Pigments and protein contents decreased under water stress, but higher amount of the same was observed in stressed AB+ seedlings. Enhanced activity of nitrate reductase was recorded in AB+ seedlings with maximum in control. Water stress significantly decreased the nitrate reductase activity. A significant increase in the activity of superoxide dismutase (SOD) in leaves was recorded under water stress except in B+ with maximum increase in non-inoculated seedlings. Catalase (CAT) activity decreased in stressed non-inoculated seedlings while increased in the leaves of A+ and AB+ seedlings. Almost similar trends were recorded for both leaves and cotyledons. PGPR improved the water status in stressed seedlings and thereby physiological and biochemical parameters and thus ameliorated the severe effects of water stress.     

Effects of bacterial single inoculation and co-inoculation on growth and phytohormone production of sunflower seedlings under water stress

The purpose of the study was to measure shoot and root dry matter (DM) and production of auxins, salicylic acid, abscisic acid, and jasmonic acid in sunflower (Helianthus annuus L.) seedlings cultivated under water stress and singly inoculated or co-inoculated with Achromobacter xylosoxidans (SF2) and Bacillus pumilus (SF3 and SF4) bacterial strains. Shoot DM was higher in non-stressed seedlings than in stressed seedlings for all inoculation treatments. Water stress resulted in decreased relative water content and reduction of shoot DM. Root DM was higher in stressed seedlings than in non-stressed seedlings. Salicylic acid was the most abundant phytohormone in shoots of stressed, singly inoculated and co-inoculated seedlings. High salicylic acid content in stressed seedlings suggests that this hormone plays a key role in abiotic stress. Abscisic acid was higher in stressed and co-inoculated seedlings than in non-stressed seedlings but was lower than that of salicylic acid. Auxin profile was similar to that of abscisic acid in co-inoculated seedlings. Shoot jasmonic acid content was increased in stressed seedlings co-inoculated with SF2/SF3 or SF2/SF4. Shoot hormonal profiles were different from those of root, suggesting a differential effect of bacterial inoculation on these plant organs. Our findings will be useful in future strategies to mitigate drought effects on crop plants through bacterial inoculation treatments.     

Biochemical indicators of thermal stress in Tilapia aurea (Steindachner)

Tilapia aurea muscle and liver adenylate nucleotides, the adenylate energy charge (EC), plasma glucose, cortisol and chloride were monitored during acute and chronic temperature stress. Muscle EC is unaffected during acute cold water exposure but decreases significantly when tilapia are exposed to chronic, sublethal, low temperature stress. The decrease in EC is primarily the result of a decrease in ATP concentration. Plasma glucose and cortisol increase when tilapia are exposed to 11–12° C for 60 min, 11 days, and a 5-week period. Incomplete compensation is evident in 5-week acclimated fish since glucose and cortisol levels are intermediate between controls and acutely stressed fish. Acclimation to 35° C does not significantly affect plasma glucose and cortisol compared to controls (22° C). Plasma chloride is relatively unaffected by acute and chronic temperature stress. Liver adenylates are not significantly affected when tilapia are subjected to a sudden drop in water temperature (22° down to 11° C). EC is a useful indicator of chronic low temperature stress in T. aurea , while plasma glucose and cortisol are sensitive to both acute and chronic temperature stress.     

Biochemical indicators of salt stress in Plantago maritima: Implications for environmental stress assessment

Our study is focused on native spontaneous species of saline ecosystems Plantago maritima. Plants were cultivated at several salt concentrations (0, 50, 100, 200, 300, 400 and 500 mM NaCl) in a glass greenhouse under semi-controlled conditions. Growth parameters, water parameters and ionic status were determined and they were used as criteria to assess the response of P. maritima under a salinity gradient. Catalase, guaiacaol and ascobate peroxidase activities, total protein and proline were also determined. Our results show that P. maritima is a facultative halophyte capable of expressing its maximum growth potential at relatively low concentrations of salt (less than 3 g l⁻¹ NaCl). At high doses of salt (concentrations > 200 mM), the decrease in the growth of P. maritima is associated to a decrease in the uptake of K⁺. There is a disruption of the water intake of their organs and therefore results an invasion of the cytoplasm by Na⁺ toxic ion. However, stressed plants use K⁺ more sparingly. They invest especially in the production of biomass expressed by the dry weight of the shoots, and they use Na⁺ and proline for osmotic adjustment. The halophyte studied is able to accumulate high levels of proline in response to increasing salt concentration. The accumulation of the amino compound, mainly in roots, is interpreted as an indicator of salt tolerance. Additionally, a significant correlation between the tolerance of the plants to salinity and the activity of several antioxidant enzymes has been observed. Hence, we suggest the possibility of using these activities as a biochemical indicator for salt tolerance in P. maritima. Our study points out two types of biomarkers of salt exposure: enzymatic biomarkers in the leaves and proline content in the roots. Both did show very good correlation with salt exposure, and thus may be considered good biomarkers of exposure with a very good dose–response relationship.     

Lipase activity in germinating sunflower seedlings

Studying lipase in germinating sunflower seedlings, we looked for an activator of the lipolytic activity. In the presence of 1.25 mM ATP, the enzyme activity increased 2-fold. Lipid-body lipase solubilization was realized using two detergents: Tween 80 and CHAPS. Lipolytic activity was increased 10-fold in the presence of 2% (w/v) CHAPS, showing the probable 'complexity' of the enzyme. Looking for the possible lipolytic activity of the 10000 g pellet we detected the presence of the enzyme. The pellet extract was mixed, in a range of concentrations, with the oil-body fraction. The resulting lipolytic activity was 4-fold higher. These results give clues as to the subcellular distribution of lipase and its intracellular transport.     

Pigments formed in etiolated sunflower seedlings

Etiolated plants must be irradiated before chlorophyll can form in them. Chlorophyll develops most readily in young irradiated plants. More carotene develops in seedlings grown from young seeds than in seedlings grown from older ones. There may be a relationship between the viability of seeds and the potential power to produce carotene. Carotinoid-pigment formation precedes chlorophyll formation. Probably some necessary substance is translocated in the plant for the formation of pigment. Carotene and xanthophyll may be precursors of chlorophyll. They probably form a respiratory mechanism for the plant. Carotene may be a precursor of auxin.     

The effect of decreasing soil moisture on the water relations of sunflower seedlings

Автор исследовал инт енсивность корневог о плача, наличный водн ый дефицит семядолей, конечный водный дефи цит семядолей (после 24-часового пребывания растений в среде, насыщенной во дяным паром), сырой вес растений, сухой вес се мядолей и ход завядания (график ? 1) в з ависимости от постеп енного падения почве нной влаги вследствие транспир ации и испарения. Опыт ы проводились в лабор аторных условиях, на сеянцах подсолнеч ника, выращиваемых в п робирках. Предельная капилляр ная влагоемкость исп ользованной почвы со ставляла 29,5% от сухого веса. Точка д лительного завядани я находилась около 8%, т очка конечного (нулевого) дефицита ок оло 9% и точка нулевого корневого плача окол о 13% влажности использованной почв ы. Из приведенных данны х вытекает, что почти ч етвертая часть из общ его количества почвенной влаги межд у точкой длительного завядания и предельн ой капиллярной влагоемкостью почвы оказывается недосту пной для корневого пл ача сеянцев под-солнечника. Сравнивая разные физ иологические индика торы доступности поч венной влаги для сеянцев подсолнечни ка, автор показывает, ч то корневой плач явля ется чувствительным индикатором, примени мым при почвенной вла жности выше точки нул евого корневого плача, когда различии в водном дефиците явл яются относительно н ебольшими.     

Biochemical stress indicators of greater wax moth exposure to organophosphorus insecticides

Although acetylcholinesterase (AChE) is the primary target of organophosphorus insecticides (OPs), increasing evidence regarding their secondary effects suggests that OPs disturb homeostasis of insects by generating free radical intermediates that trigger lipid peroxidation. We therefore investigated alterations in lipid peroxidation product, malondialdehyde (MDA) content, and alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, in conjunction with AChE activity as biochemical stress indicators in greater wax moth, Galleria mellonella (L.) larvae for OPs methyl parathion (MP) and ethyl parathion (EP). The effects of MP and EP were first investigated by rearing the young larvae on an artificial diet containing 0.01, 0.1, 1, 10, and 100 ppm of each insecticide. Second, the mature larvae were injected with 0.05, 0.5, 5, 50, and 500 ng of insecticides for determining the changes in biochemical stress responses. The diet with lowest level of MP significantly decreased the activities of all measured enzymes, whereas it increased MDA content. However ALT and AST were significantly higher in the larvae reared with the diet with high levels of MP than in control larvae. All tested levels of MP resulted in a decrease in AChE activity. The lowest level of EP in diet (0.01 ppm) significantly increased ALT activity, whereas it reduced that of AChE. This insecticide at 0.1 ppm resulted in reduced AST activity, but 1 ppm in diet elevated AST activity and MDA content. EP at 0.1 ppm and higher levels in the diet reduced ALT activity. All dietary EP levels significantly decreased AChE activity. ALT, AST, and AChE were lower in larvae fed with the diet containing 100 ppm ethyl parathion compared with larvae on control diet. MP at 50 ng per larva increased ALT and AST activities from 35.42 +/- 0.74 and 26.34 +/- 0.83 to 203.57 +/- 1.09, and 122.90 +/- 1.21 U/g, respectively, when the mature larvae were injected. All injected doses of EP dramatically reduced both ALT and AST activities, but only the lowest and highest levels of this insecticide decreased AChE activity. The lowest level of this insecticide also significantly increased MDA content in larvae. High levels of both insecticides increased MDA content. We observed a significant higher increase in MDA content in the larvae reared with 10 ppm EP (102.16 +/- 1.57 nmol/g protein) than the control group (30.28 +/- 1.42 nmol/g protein). These results suggest that OPs caused the metabolic and synaptic dysfunctions in greater wax moth and alter its biochemical physiology in response to oxidative stress.     

Tissue-specific expression of sunflower heat shock proteins in response to water stress

Accumulation of mRNA and synthesis of low-molecular-weight heat shock proteins (LMW HSPs) was investigated in water-stressed sunflower, under experimental conditions resulting in little or no thermal stress. Using probes and antibodies derived from developmentally expressed LMW HSPs, it was shown that homologous mRNAs and proteins accumulate in the stem and root of water-stressed plants. This expression is quantitatively comparable with the response to heat shock: protein and mRNA accumulate to similar, high, levels and persist for comparable times during recovery from either environmental stress. However, it is shown that LMW HSPs with different molecular weights and isoelectric points are expressed in response to heat shock or water stress. Furthermore in situ localizations show a differential tissue-specificity for the water-stress- and heat-shock-induced LMW HSPs. Whereas the latter are localized mostly around the xylem vessels in the stem, the water-stress-induced proteins accumulate in the fascicular and interfascicular cambium. The possible functional implications for this specific expression are discussed.     

Carbendazim alleviates effects of water stress on chickpea seedlings

Carbendazim (methyl-2-benzimidazole carbamate) promoted root growth of chickpea (Cicer arietinum L.) seedlings subjected to polyethylene glycol (PEG, osmotic potential −0.5 MPa) induced water stress. The relative water content, membrane stability index, 2,3,5-triphenyltetrazolium chloride reduction and contents of some osmolytes (proline, sucrose, glucose and fructose) enhanced significantly while the contents of lipid peroxides and hydrogen peroxide diminished effectively by addition of 0.05 % carbendazim into PEG solution. This revised version was published online in September 2005 with the corrected author information.     

四种灌木幼苗对水分胁迫的生理响应

柠条锦鸡儿(Caragana korshinskii)、白皮锦鸡儿(C.leucophloea)、刺叶锦鸡儿(C.acanthophylla)和长枝木蓼(Atraphaxis virgata)是乌鲁木齐周边植被组成中的重要植物种,在植被恢复中具有潜在价值。研究通过人工控制水分比较这4种灌木对干旱胁迫的生理响应,结果表明:叶片的组织含水量和叶绿素含量随着干旱胁迫的加剧而降低;脯氨酸在干旱胁迫下的积累程度较可溶性糖大;除刺叶锦鸡儿外其它植物幼苗的丙二醛在干旱胁迫下均无明显的积累(P>0.05)。4种灌木在干旱胁迫下通过渗透调节、保持膜系统稳定等途径维持正常的生理活动,对干旱具有一定的适应能力。4种灌木抗旱能力的排序为白皮锦鸡儿>柠条锦鸡儿>长枝木蓼>刺叶锦鸡儿。     

Cytokinin oxidase activity and cytokinin content in roots of sunflower under water stress

Contents of trans-zeatin riboside (ZR), dihydrozeatin riboside (DZR) and N6-(delta2-isopentenyl) adenosine (iPA) was quantified by an indirect ELISA using polyclonal antibodies, in the roots, xylem sap and leaves of pot grown sunflower plants subjected to water stress (RWC of leaves approximately 65 per cent). The delivery rates of all three cytokinins decreased significantly under stress. Cytokinin levels also decreased in roots and in leaves of stressed plants. Three-fold increase in cytokinin oxidase activity was observed in stressed roots after polymin P-ammonium sulphate fractionation. Further purification using Con A agarose resulted in elution of protein with cytokinin oxidase activity and was found to be 30 kDa protein on SDS-PAGE.     

Biochemical identification of proteasome-associated endonuclease activity in sunflower

Proteasomes have been purified from sunflower hypocotyles. They elute with a molecular mass of 600 kDa from gel filtration columns and two-dimensional gel electrophoresis indicates that the complex contains at least 20 different protein subunits. Peptide microsequencing revealed the presence of four subunits homologous to subunits Beta2, Beta6, Alpha5 and Alpha6 of plant proteasomes. These proteasomes have chymotrypsin-like activity and the highly purified fraction of this complex is associated with an endonuclease activity hydrolyzing Tobacco mosaic virus RNA and Lettuce mosaic virus RNA with a cleavage pattern showing fragments of well-defined size. This is the first evidence of a RNA endonuclease activity associated with plant proteasomes.     

S‐nitrosylation/denitrosylation as a regulatory mechanism of salt stress sensing in sunflower seedlings

Nitric oxide (NO) and various reactive nitrogen species produced in cells in normal growth conditions, and their enhanced production under stress conditions are responsible for a variety of biochemical aberrations. The present findings demonstrate that sunflower seedling roots exhibit high sensitivity to salt stress in terms of nitrite accumulation. A significant reduction in S‐nitrosoglutathione reductase (GSNOR) activity is evident in response to salt stress. Restoration of GSNOR activity with dithioerythritol shows that the enzyme is reversibly inhibited under conditions of 120 mM NaCl. Salt stress‐mediated S‐nitrosylation of cytosolic proteins was analyzed in roots and cotyledons using biotin‐switch assay. LC‐MS/MS analysis revealed opposite patterns of S‐nitrosylation in seedling cotyledons and roots. Salt stress enhances S‐nitrosylation of proteins in cotyledons, whereas roots exhibit denitrosylation of proteins. Highest number of proteins having undergone S‐nitrosylation belonged to the category of carbohydrate metabolism followed by other metabolic proteins. Of the total 61 proteins observed to be regulated by S‐nitrosylation, 17 are unique to cotyledons, 4 are unique to roots whereas 40 are common to both. Eighteen S‐nitrosylated proteins are being reported for the first time in plant systems, including pectinesterase, phospholipase d ‐alpha and calmodulin. Further physiological analysis of glyceraldehyde‐3‐phosphate dehydrogenase and monodehydroascorbate reductase showed that salt stress leads to a reversible inhibition of both these enzymes in cotyledons. However, seedling roots exhibit enhanced enzyme activity under salinity stress. These observations implicate the role of S‐nitrosylation and denitrosylation in NO signaling thereby regulating various enzyme activities under salinity stress in sunflower seedlings.     

Root respiration during grain filling in sunflower: The effects of water stress

Instantaneous rates of (soil + root) respiration were measured periodically during grain filling in sunflower crops that were i) irrigated at weekly intervals and ii) subjected to water stress for the last 25 days of the 40-day grain filling period. Daily (soil + root) respiration was calculated using instantaneous respiration rates, an empirically determined temperature response function, and diurnal records of soil temperature. Daily soil respiration was estimated using empirically determined functions linking soil respiration to soil temperature and water content. Between anthesis and maturity, daily root respiration of the irrigated crop dropped by about one half from ca. 1.8 g C m^-2 d^-1, exhibiting a strong association with daily crop gross photosynthesis. Water stress brought about a rapid decrease in root respiration, which fell to about 0.1 g C m^-2 d^-1 at maturity. Root respiration during grain filling was 46 and 30 g C m^-2 for irrigated and stressed crops, respectively.     

Effect of chlorsulfuron on phenylpropanoid metabolism in sunflower seedlings

The effect of the herbicide chlorsulfuron on phenylpropanoid titer and metabolism and the role of endogenous ethylene in this response was examined in light-grown sunflower (Helianthus annuus L.) seedlings. Application of chlorsulfuron to the apex resulted in large increases in both total phenolic and hydroxycinnamic acid content in hypocotyls isolated from the treated seedlings. Both of these parameters were increased within 24 h of herbicide treatment, and both reached a maximum level 3–4 days post-treatment. An increase in ethylene evolution was found to proceed in parallel with the alterations of phenolic content. The extractable activities of phenylalanine ammonia lyase,trans-cinnamic-4-hydroxylase, and soluble peroxidase were increased by chlorsulfuron treatment. Chlorsulfuron had little effect on total phenolic content when administered directly to isolated hypocotyl segments. Exogenous ethylene had no effect on the endogenous titer of phenolic compounds. Root-fed cobalt chloride (5 × 10^?4 M) inhibited chlorsulfuron-induced ethylene production by 92% and also inhibited the accumulation of phenolic materials by 56%. Exogenous ethylene was unable to reverse the inhibition caused by cobalt chloride. It was concluded that chlorsulfuron-induced increases in phenolic compounds were not mediated solely by endogenous ethylene.