Asim Kadioglu

Salicylic acid pretreatment induces drought tolerance and delays leaf rolling by inducing antioxidant systems in maize genotypes

Salicylic acid (SA) acts as an endogenous signal molecule responsible for inducing abiotic stress tolerance in plants. In this study, the role of SA in improving drought tolerance in two maize cultivars (Zea mays L.) differing in their tolerance to drought was evaluated. The plants were regularly watered per pot and grown until the grain filling stage (R2) under a rainout shelter. At stage R2, parts of the plants were treated with SA, after which drought stress was applied. Leaf samples were harvested on the 10th and 17th days of the drought. Some antioxidant enzyme activity, such as the superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), glutathione reductase (GR), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), hydrogen peroxide (H2O2) and malondialdehyde (MDA) content, was measured during the drought period. Exogenous SA prevented water loss and delayed leaf rolling in comparison with control leaves in both cultivars. As a consequence of drought stress, lipid peroxidation, measured in terms of malondialdehyde content, was prevented by SA. SA pretreatment induced all antioxidant enzyme activities, and to a greater extent than the control leaves, during drought. SA also caused a reduction in the ascorbate (ASC) and glutathione (GSH) content in two maize cultivars. The H2O2 level was higher in SA pretreated plants than the controls in both cultivars. Pretreatment with SA further enhanced the activities of antioxidant enzymes and the concentrations of non-enzymatic antioxidants in the tolerant cultivar compared with the sensitive cultivar. Results suggested that exogenous SA could help reduce the adverse effects of drought stress and might have a key role in providing tolerance to stress by decreasing water loss and inducing the antioxidant system in plants with leaf rolling, an alternative drought protection mechanism.

A Dehydration Avoidance Mechanism: Leaf Rolling

Plants have several defense mechanisms against unfavorable environmental conditions. One of these mechanisms is leaf rolling. In this review, leaf rolling as a response to water deficit stress and biochemical changes during leaf rolling in higher plants are reported. For instance, the activities of some enzymes and osmotic substances change during leaf rolling. Leaf rolling increases drought resistance in numerous species in theGramineae as well as inCtenanthe setosa, a perennial herbaceous plant that is a suitable model for use in studies of leaf rolling.ZusammenfassungPflanzen besitzen Schutzmechanismen gegen ungünstige Umweltbedingungen. Einer dieser Mechanismen ist das Einrollen der Blätter. In diesem Übersichtsartikel wird erstmals über die Bedeutung des Einrollens der Blätter als Antwort auf Wassermangel und von biochemischen Änderungen während des Einrollens der Blätter bei höheren Pflanzen berichtet. Das Einrollen der Blätter ist nicht nur eine Reaktion der Pflanze auf Wassermangel, es treten auch biochemische Veränderungen zusammen mit dem Einrollen der Blätter auf. Zum Beispiel verändern sich einige Enzymaktivitäten und osmotisch-aktive Stoffe während des Einrollens der Blätter. Das Einrollen der Blätter erhöht die Widerstandsfähigkeit gegen Trockenheit bei vielen Gramineen wieCtenanthe setosa. C. setosa, eine mehrjährige krautige Pflanze, ist eine gute Modellpflanze für Untersuchungen des Einrollens der Blätter.

Exogenous salicylic acid alleviates effects of long term drought stress and delays leaf rolling by inducing antioxidant system

Salicylic acid (SA) is one of the important signal molecules modulating plant responses to environmental stress. In this study, the effects of exogenous SA on leaf rolling, one of drought avoidance mechanisms, and antioxidant system were investigated in Ctenanthe setosa during long term drought stress. The plants were subjected to 38-day drought period and they were treated with or without SA (10−6 M) on the 25th, 27th and 29th days of the period. Leaf samples were harvested on the 30th, 34th and 38th days. Some antioxidant enzyme activities (superoxide dismutase, catalase, ascorbate peroxidase, dehydroascorbate reductase, monodehydroascorbate reductase, glutathione reductase), reactive oxygen species (hydrogen peroxide and superoxide) and lipid peroxidation were determined during the drought period. Treatment with SA prevented water loss and delayed leaf rolling in comparison with control leaves. Exogenous SA induced all antioxidant enzyme activities more than control leaves during the drought. Ascorbate and glutathione, α-tocopherol, carotenoid and endogenous SA level were induced by the SA treatment. Levels of reactive oxygen species were higher in SA treated plants than control ones on the 34th day. Their levels on the 38th day, however, fastly decreased in SA treated plants. SA treatment prevented lipid peroxidation while the peroxidation increased in control plants. The results showed that exogenous SA can alleviate the damaging effect of long term drought stress by decreasing water loss and inducing the antioxidant system in the plant having leaf rolling, alternative protection mechanism to drought.

Current advances in the investigation of leaf rolling caused by biotic and abiotic stress factors

Leaf rolling is known as a typical response to water deficit in numerous species such as rice, maize, wheat and sorghum. However, it results not only from the water deficit but also from other abiotic stress factors such as salt, temperature, heavy metals and UV radiation. In addition to the abiotic factors, herbivores, viruses, bacteria and fungi are biotic factors of leaf rolling. Leaf rolling is an effective protective mechanism from the effects of high light levels in agricultural fields and protects leaves of unirrigated plants from photodamage. The rolling reduces effective leaf area and transpiration, and thus is a potentially useful drought avoidance mechanism in dry areas. The current review focuses on the recent progress in understanding leaf rolling in relation to abiotic and biotic stress factors, the role of signal molecules, and the mechanisms of gene regulation.

Leaf rolling and photosystem II efficiency in Ctenanthe setosa exposed to drought stress

Photochemical efficiency of PSII of Ctenanthe setosa was investigated to understand the photosynthetic adaptation mechanism under drought stress causing leaf rolling. Stomatal conductance (gs), the levels of photosynthetic pigments and chlorophyll (Chl) fluorescence parameters were determined in leaves that had four different visual leaf rolling scores from 1 to 4, opened after re-watering and mechanically opened at score 4. gs value gradually decreased in adaxial and abaxial surfaces in relation to scores of leaf rolling. Pigment contents decreased until score 3 but approached score 1 level at score 4. No significant variations in effective quantum yield of PSII (ΦPSII), and photochemical quenching (qp) were found until score 3, while they significantly decreased at score 4. Non-photochemical quenching (NPQ) increased at score 2 but then decreased. After re-watering, the Chl fluorescence and other physiological parameters reached to approximately score 1 value, again. As for mechanically opened leaves, gs decreased during drought period. The decrease in adaxial surface was higher than that of the rolled leaves. NPQ was higher than that of the rolled leaves. ΦPSII and qp significantly declined and the decreases were more than those of the rolled leaves. In conclusion, the results indicate that leaf rolling protects PSII functionality from damage induced by drought stress.

The Relationship between Leaf Rolling and Ascorbate-Glutathione Cycle Enzymes in Apoplastic and Symplastic Areas of Ctenanthe setosa Subjected to Drought Stress

The ascorbate-glutathione (ASC-GSH) cycle has an important role in defensive processes against oxidative damage generated by drought stress. In this study, the changes that take place in apoplastic and symplastic ASC-GSH cycle enzymes of the leaf and petiole were investigated under drought stress causing leaf rolling in Ctenanthe setosa (Rose.) Eichler (Marantaceae). Apoplastic and symplastic extractions of leaf and petiole were performed at different visual leaf rolling scores from 1 to 4 (1 is unrolled, 4 is tightly rolled and the others are intermediate forms). Glutathione reductase (GR), a key enzyme in the GSH regeneration cycle, and ascorbate (ASC) were present in apoplastic spaces of the leaf and petiole, whereas dehydroascorbate reductase (DHAR), which uses glutathione as reductant, monodehydroascorbate reductase (MDHAR), which uses NAD(P)H as reductant, and glutathione were absent. GR, DHAR and MDHAR activities increased in the symplastic and apoplastic areas of the leaf. Apoplastic and symplastic ASC and dehydroascorbate (DHA), the oxidized form of ascorbate, rose at all scores except score 4 of symplastic ASC in the leaf. On the other hand, while reduced glutathione (GSH) content was enhanced, oxidized glutathione (GSSG) content decreased in the leaf during rolling. As for the petiole, GR activity increased in the apoplastic area but decreased in the symplastic area. DHAR and MDHAR activities increased throughout all scores, but decreased to the score 1 level at score 4. The ASC content of the apoplast increased during leaf rolling. Conversely, symplastic ASC content increased at score 2, however decreased at the later scores. While the apoplastic DHA content declined, symplastic DHA rose at score 2, but later was down to the level of score 1. While GSH content enhanced during leaf rolling, GSSG content did not change except at score 2. As well, there were good correlations between leaf rolling and ASC-GSH cycle enzyme activities in the leaf (GR and DHAR) and leaf rolling and GSSG. These results showed that in apoplastic and symplastic areas, ASC-GSH cycle enzymes leading ROS detoxification may have a role in controlling leaf rolling.

Effect of fruit maturation on sugar and organic acid composition in two blueberries (Vaccinium arctostaphylos and V. myrtillus) native to Turkey

Abstract Some organic acids and sugars in two blueberry species from north‐east Anatolia, Turkey, were studied. The fruits of the blueberry species (Vaccinium arctostaphylos L. and V. myrtillus L.) were collected at three stages of maturity (immature, mid ripe, and ripe), and the organic acids and sugars were studied by gas liquid chromatography (GLC) and verified by gas liquid chromatography‐mass spectrometry (GLC‐MS). The soluble sugars fructose, glucose, and sucrose, and the sugar alcohol inositol were identified and quantified. Fructose and glucose represented the major sugars in the fruits. Quinic and citric acids were found to be the major organic acids in both blueberry species. The levels of these acids were significantly different (P = 0.05) at all three stages of fruit maturation. The level of malic acid increased gradually in both species, whereas the levels of citric and quinic acids decreased rapidly. The total acid level in the fruits of the two species was also significantly different. The results of this study can be used to compare the two species with other Vaccinium species.

Hydrogen peroxide pretreatment induces osmotic stress tolerance by influencing osmolyte and abscisic acid levels in maize leaves

Hydrogen peroxide (H2O2) functions as a signal molecule in plants under abiotic and biotic stresses. Leaves of detached maize (Zea mays L.) seedlings were used to study the function of H2O2 pretreatment in osmotic stress resistance. Low H2O2 concentration (10 mM) which did not cause a visual symptom of water deficit (leaf rolling) was applied to the seedlings. Exogenous H2O2 alone increased leaf water potential, endogenous H2O2 content, abscisic acid (ABA) concentration, and metabolite levels including soluble sugars, proline, and polyamines while it decreased lipid peroxidation and stomatal conductance. Osmotic stress induced by polyethylene glycol (PEG 6000) decreased leaf water potential and stomatal conductance but enhanced lipid peroxidation, endogenous H2O2 content, the metabolite levels, and ABA content. H2O2 pretreatment also induced the metabolite accumulation and improved water status, stomatal conductance, lipid peroxidation, ABA, and H2O2 levels under osmotic stress. These results indicated that H2O2 pretreatment may alleviate water loss and induce osmotic stress resistance by increasing the levels of soluble sugars, proline, and polyamines thus ABA and H2O2 production slightly decrease in maize seedlings under osmotic stress.

Spermine and putrescine enhance oxidative stress tolerance in maize leaves

The protective effects of spermine (SPM) and putrescine (PUT) against paraquat (PQ), a herbicide in agriculture and oxidative stress inducer, were investigated in the leaves of maize. Maize leaves were pretreated to SPM and PUT at concentrations of 0.2 and 1 mM and treated with PQ afterwards. Pretreatment with 1 mM of SPM and PUT significantly prevented the losses in chlorophyll and carotenoid levels induced by PQ. Ascorbic acid content in the leaves pretreated with both polyamines was found to be higher than those of the leaves pretreated with water. Also, pretreatment with SPM and PUT was determined to have some effects on the activities of superoxide dismutase (SOD) and peroxidase (POD). 1 mM of SPM increased SOD activity, but PUT has no significant effect on SOD activity. On the other hand, POD activity was recorded to increase slightly in response to both concentrations of SPM and 1 mM of PUT. The results showed that such polyamine pretreated plants may become more tolerant to oxidative stress due to increases in the antioxidative enzymes and antioxidants.

Co-overexpressing a Plasma Membrane and a Vacuolar Membrane Sodium/Proton Antiporter Significantly Improves Salt Tolerance in Transgenic Arabidopsis Plants

The Arabidopsis gene AtNHX1 encodes a vacuolar membrane-bound sodium/proton (Na+/H+) antiporter that transports Na+ into the vacuole and exports H+ into the cytoplasm. The Arabidopsis gene SOS1 encodes a plasma membrane-bound Na+/H+ antiporter that exports Na+ to the extracellular space and imports H+ into the plant cell. Plants rely on these enzymes either to keep Na+ out of the cell or to sequester Na+ into vacuoles to avoid the toxic level of Na+ in the cytoplasm. Overexpression of AtNHX1 or SOS1 could improve salt tolerance in transgenic plants, but the improved salt tolerance is limited. NaCl at concentration >200 mM would kill AtNHX1-overexpressing or SOS1-overexpressing plants. Here it is shown that co-overexpressing AtNHX1 and SOS1 could further improve salt tolerance in transgenic Arabidopsis plants, making transgenic Arabidopsis able to tolerate up to 250 mM NaCl treatment. Furthermore, co-overexpression of AtNHX1 and SOS1 could significantly reduce yield loss caused by the combined stresses of heat and salt, confirming the hypothesis that stacked overexpression of two genes could substantially improve tolerance against multiple stresses. This research serves as a proof of concept for improving salt tolerance in other plants including crops.