Increased dehydrin level decreases leaf rolling grade by altering the reactive oxygen species homeostasis and abscisic acid content in maize subjected to osmotic stress.

Dehydrins (DHNs) are stress proteins involved in the development of protective reactions in plants against dehydration. The relationship between DHNs and morphological responses such as leaf rolling in plants exposed to water deficit is not well known. In this study, we detected how variations in DHN levels affect the leaf rolling response in maize exposed to osmotic stress in relation to the antioxidant system and ABA level. In this context, we altered the DHN levels in maize seedlings by treatment with bio-regulators (salicylic acid and abscisic acid) under PEG6000-free and PEG6000-induced osmotic stress. When the DHN levels were increased by the bio-regulators (25 µM SA and 100 µM ABA), the relative expression level of the Zea mays dehydrin COR410 gene increased in the seedlings, while reactive oxygen species (ROS) and leaf rolling grade decreased. Moreover, induction of DHNs caused increases in the antioxidant enzyme activity and content of antioxidant substances, and very high amounts of endogenous abscisic acid. When DHN level was suppressed by a bio-regulator (200 µM SA) in the maize seedlings, dehydrin COR410 expression level decreased, while ROS and the leaf rolling grade increased. Moreover, the antioxidant enzyme activity and content of antioxidant substances decreased in the seedlings, while the amount of abscisic acid increased. Taken all together, an increase in DHN level by bio-regulator treatment can stimulate the antioxidant system, enable abscisic acid regulation, and thus reduce leaf rolling through decreased ROS levels. The results also indicated that DHNs may be involved in the signal pathways inducing expression of some genes related to leaf rolling response, possibly by modulating ROS levels, in maize seedlings exposed to osmotic stress.

Proline-stimulated signaling primarily targets the chlorophyll degradation pathway and photosynthesis associated processes to cope with short-term water deficit in maize.

Increased photosynthetic efficiencies in genotypes with greater proline level and in crops treated with proline under water deficit have been reported in recent years, but the biochemical and molecular mechanisms of this process are still not known. We examined photosystem II (PSII) activity, photosynthetic enzymes, ribulose 1,5-bisphosphate carboxylase/oxygenase (rubisco), phosphoenolpyruvate carboxylase (PEPc), rubisco activase (RCA), and chlorophyll metabolic enzymes, magnesium chelatase (Mg-CHLI), and chlorophyllase (Chlase), which would be the primary targets of exogenous proline to provide photosynthetic protection to plants under PEG-induced short-term water deficit. Two maize genotypes W23/M14 with greater proline content and Safak with low proline content were hydroponically grown for 21-23 days, and then the seedlings were subjected to water deficit (- 0.75 MPa) induced by PEG6000 for 0, 4, and 8 h. Before the seedlings were exposed to the water deficit, proline (1 mM) was applied to the rooting medium of the Safak genotype for 2 days. The time course effects of the applications showed that exogenous proline significantly enhanced PSII efficiency, PEPc activity, rubisco activity, and the relative expression levels of PEPc, rubisco large subunit, rubisco small subunit, and RCA genes at 0, 4, and 8 h. The W23/M14 genotype had higher rubisco quantity than the Safak genotype at all time periods. Proline pre-treatment under the stress-free and PEG conditions reduced the activity of Chlase and the gene expressions of Chlase, while it enhanced Mg-CHLI gene expression at 0, 4, and 8 h. Taken together, the results indicated that the primary target of proline-stimulated signaling in maize seedlings exposed to short-term severe water deficit may be to induce PSII efficiency, activities of carbon dioxide fixation enzymes and chlorophyll metabolism and mitigate chlorophyll degradation.

Exogenous alpha lipoic acid can stimulate photosystem II activity and the gene expressions of carbon fixation and chlorophyll metabolism enzymes in maize seedlings under drought.

Protective compounds such as non-enzymatic antioxidants, osmolytes and signal molecules have been applied to plants exposed to various environmental stresses to increase their stress tolerance. However, there are not enough records about the response of plants to alpha lipoic acid (ALA) application with antioxidant properties. Therefore, this study was designed to evaluate the function of exogenous ALA on the photosynthetic performance of maize seedlings grown in hydroponic conditions under drought stress. Three weeks old seedlings were treated with or without ALA (12 µM) and they were subjected to drought stress induced by 10% polyethylene glycol (PEG6000) for 24 h. Chlorophyll content, gas exchange parameters, chlorophyll fluorescence and the expression levels of genes involved in CO2 fixation (ribulose-1,5-bisphosphate carboxylase (rubisco), phosphoenolpyruvate carboxylase (PEPc), Rubisco activase (RCA)) and chlorophyll metabolism (magnesium chelatase (Mg-CHLI) and chlorophyllase (Chlase)) were determined. The application of ALA increased chlorophyll content and the activity of photosystem II in comparison to the untreated seedlings under drought stress. The relative expression levels of Rubisco, PEPc, RCA and Mg-CHLI significantly increased while the Chlase gene expression decreased in seedlings to which ALA was applied in comparison those to which it was not applied under the stress. These results suggest that exogenous ALA can enhance the photosynthetic performance of maize seedlings exposed to drought by inducing photosystem II activity and the gene expressions of carbon fixation and chlorophyll metabolism enzymes.

Exogenous hydrogen peroxide increases dry matter production, mineral content and level of osmotic solutes in young maize leaves and alleviates deleterious effects of copper stress.

BACKGROUND: The effects of exogenously applied H2O2 on growth, water status, the mineral ion content (Na+, K+, Ca2+, Mg2+ and Cu2+), proline, total sugars and soluble proteins were assessed in leaves of maize (Zea mays L.) cultivars, Akpinar and Pegaso exposed to excess copper (0.5 mM). Seedlings were grown in equal-sizes plastic pots and irrigated with Hoagland nutrient solution containing H2O2 or/and copper. Different treatments taken for pot experiments were named as the control (C), H2O2 treatment only (H2O2), excess Cu (Cu) and, Cu stress combined with H2O2 pretreatment (Cu + H2O2). RESULTS: Treatment of H2O2 caused the increases in growth, water content, mineral concentration, proline, total sugar and soluble protein contents compared to the control groups in the leaves of both cultivars. Yet excess copper caused reductions in the growth, leaf water potential, Na+, K+, Ca+, Mg2+ concentrations and soluble protein levels but increases in proline, total soluble sugars and Cu2+ contents compared to the control group. Dry matter, leaf water potential and mineral content of Cu + H2O2 group revealed a lower decrease than Cu group ones. A higher increase was also observed in proline and total sugar contents of Cu + H2O2 group than Cu group ones in both cultivars. CONCLUSIONS: These data revealed that exogenous H2O2 might increase the dry matter production and the mineral ion distribution in maize seedlings. Moreover, osmotic regulation might be involved in alleviation of copper toxicity of maize leaves by pretreatment of H2O2.

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.

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.

Photosystem II functionality and antioxidant system changes during leaf rolling in post-stress emerging Ctenanthe setosa exposed to drought.

We studied the changes in antioxidant system and chlorophyll fluorescence parameters in post-stress emerging Ctenanthe setosa (Rosc.) Eichler (Marantaceae) plants (PSE plants) having reduced leaf area under drought stress causing leaf rolling and re-watering. PSE plants were compared to primary stressed plants (PS) in previous studies. The parameters were measured at different visual leaf rolling scores from 1 to 4 (1 is unrolled, 4 is tightly rolled and the others is intermediate form). Water potentials and stomatal conductance of leaves were gradually decreased during leaf rolling. Similarly, maximum quantum efficiency of open PS II center and quantum yield of PS II decreased during the rolling period. Non-photochemical quenching of chlorophyll fluorescence decreased at score 2 then increased while photochemical quenching did not change during leaf rolling. Electron transport rate decreased only at score 4 but approximately reached to score 1 level after re-watering. Superoxide dismutase activity was not constant at all leaf rolling scores. Ascorbate peroxidase, catalase and glutathione reductase activities generally tended to increase during leaf rolling. Lipid peroxidation and H 2 O 2 content increased at score 2 but decreased at the later scores. On the other hand, O 2 .- production increased during the rolling period. After re-watering of the plants having score 4 of leaf rolling, antioxidant enzyme activities were lower than those of score 1. Other physiological parameters also tended to reach the value of score 1. The results indicated that PSE plants gained drought tolerance by reducing leaf area effectively induced their antioxidant systems and protected the photosynthesis under drought stress similar to PS plants.

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.

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 ...