Aykut Sağlam

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.

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.

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.

Scavenging of reactive oxygen species in apoplastic and symplastic areas of rolled leaves in Ctenanthe setosa under drought stress.

The correspondence among apoplastic and symplastic antioxidant status, stomatal conductance and water potential was investigated during leaf rolling in Ctenanthe setosa (Rosc.) Eichler (Marantaceae) under drought stress. 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 form). In the leaf symplast, the highest changes were found in catalase (CAT) and guaiacol peroxidase (GPX) activities when compared to score 1 during leaf rolling. No significant change was observed in superoxide dismutase (SOD) and ascorbate peroxidase (APX) activities in the symplast of leaf during the rolling. The same phenomenon was also present in the symplast of petiole except APX activity. In the leaf apoplast, the highest increase occurred in APX and GPX activities, whilst a slight increase in CAT and SOD activities. In the apoplast of petiole, the highest increment was found only in GPX activity, while there were small increases in SOD, APX and CAT activities. Hydrogen peroxide content increased up to score 3 in the apoplast and symplast of leaf and petiole but then slightly decreased. Also, superoxide production increased in the leaf and petiole apoplast but its quantity in the apoplast was much more than that of the symplast. On the other hand, NAD(P)H oxidase activity increased in the leaf but no change was observed in the petiole. In conclusion, as a result of water deficit during leaf rolling antioxidant enzymes are induced to scavenging of ROS produced in symplast and apoplast.

Leaf Rolling Reduces Photosynthetic Loss in Maize Under Severe Drought

Abstract Effects of leaf rolling (LR) on maize photosynthesis under severe drought stress were studied in two cultivars with opposite drought responses, Batem 56-55 (drought tolerant) and Batem 51-52 (drought sensitive). Drought stress and artificial prevention of leaf rolling (PLR) were applied at grain filling stage for 30 days. LR in Batem 56-55 occurred later than in Batem 51-52. Leaf water potential (Ψleaf) did not change in Batem 56-55 but decreased in Batem 51-52 at LR. Maximum quantum yield of photosystem II (Fv/Fm), effective quantum yield of photosystem II (ΦPSII) and electron transport rate (ETR) of the cultivars decreased during LR more significantly in Batem 56-55 in comparison to Batem 51-52. The same was observed for the decrease in net photosynthetic rate (Pn), stomatal conductance (gs), transpiration (E) and intracellular level of CO2 (C). Rubis-co activity and content were reduced at LR, but were less affected in Batem 56-55 than in Batem 51-52. Ear and kernel weights also decreased at LR. All parameters at PLR were more reduced than those of LR. These results implied that LR was an important and necessary mechanism protecting photosynthesis and reducing yield loss under drought stress by maintaining the leaf hydration, preventing loss of the photosynthetic pigments, sustaining the activity of PSU, keeping the stornata open, and conserving the activity of Rubisco.

The relations between antioxidant enzymes and chlorophyll fluorescence parameters in common bean cultivars differing in sensitivity to drought stress

Effects of the antioxidant system and chlorophyll fluorescence on drought tolerance of four common bean (Phaseolus vulgaris L.) cultivars were studied. The cultivars were positioned in the order of a decrease in their drought tolerance: Yakutiye, Pinto Villa, Ozayse, and Zulbiye on the basis of changes in the water potential, stomatal conductance, photosynthetic pigment content, and lipid peroxidation. Under drought conditions, the level of H2O2 was not changed in cv. Pinto Villa but decreased in other cultivars. Antioxidant enzymes (superothide dismutase (SOD), guaiacol peroxidase (GPX), catalase (CAT), ascorbate peroxidase (APX), and glutathione reductase (GR)) were generally activated in all cultivars. Interestingly, CAT, APX, and GR activities were not changed in cv. Pinto Villa, APX activity decreased in cv. Yakutiye, and CAT activity was not changed in cv. Zulbiye. The increases in SOD and GPX activities in cv. Ozayse were higher than in other cultivars. Drought stress reduced the effective quantum yield of PS2 (ΦPS2) and the photochemical quenching (qp), while it increased nonphotochemical quenching (NPQ) in all cultivars. The reduction or increase was more pronounced in cv. Zulbiye. There were generally significant correlations between qp, NPQ, and ROS scavenging by SOD and APX. Also, there were significant correlations between SOD and qp in tolerant cultivars and APX and qp in sensitive ones. The results indicate that activation of SOD and APX was closely related to the efficiency of PS2 in common bean cultivars. This interaction was essential for protection of photosystems and plant survival under drought.

Physiological changes in them in post-stress emerging Ctenanthe setosa plants under drought conditions

The capability of Ctenanthe setosa (Rosc.) Eichler (Marantaceae), pre-exposed to severe drought stress, of adaptation to a new drought conditions was investigated. C. setosa unstressed plants were exposed to drought period of 70 days. These plants were named as primary-stress plants (PS). They were trimmed, and new plants were produced from their rhizomes. New growing plants were named as post-stress emerging (PSE) plants. It was observed that leaf area and petiole length of PSE plants were reduced, as compared to unstressed plants. When PSE plants were exposed to a second drought-stress cycle, it was determined that the degree of their leaf rolling was higher but their relative water content (RWC) was less than these parameters of PS plants during drought period. However, at the 81% rolling degree, RWC in PSE plants was higher than in PS plants. Consequently, it can be said that the PSE plant were more resistant to a new drought stress than unstressed and PS plants. In the leaves of PSE plants, which were exposed to a second drought-stress period, the content of soluble protein decreased in the early period of drought, but increased in the later periods. Also, the content of reducing sugar in PSE plants gradually decreased during the stress period. Proline content increased markedly during drought period, whereas peroxidase activity increased up to the 60th day and then decreased.