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Sanalkumar Krishnan and Emily B. Merewitz

Salinity stress is becoming more prevalent in turfgrass management with the increasing use of recycled water for irrigation. Creeping bentgrass (Agrostis stolonifera) is a cool-season turfgrass species that contains significant cultivar variation in salt stress tolerance, but the mechanism related to this cultivar variation is not well understood. Our objectives were to determine whether differential hormone content could play a role in cultivar variation of salt responses and to evaluate whether cell viability assays using dye techniques could differentiate salt stress damage levels in turfgrass species. Therefore, a growth chamber study with potted plants was conducted to evaluate salt ion concentrations, physiological responses, and hormone analysis [abscisic acid (ABA), indole-3-acetic acid (IAA), jasmonic acid (JA), salicylic acid (SA), zeatin riboside (ZR), and ethylene] at 4, 8, and 12 dS·m−1 in relatively salt-tolerant ‘Mariner’ compared with salt-sensitive ‘Penncross’ creeping bentgrass. A hydroponics-based growth chamber study was performed for evaluation of whether dead-cell stains coupled with image analysis could be a quick method for indicating cell viability variation between cultivars. Greater salt tolerance was evident in ‘Mariner’ at 12 dS·m−1, which showed significantly lower electrolyte leakage, higher leaf relative water content (RWC), osmotic potential, photochemical efficiency, and photochemical yield compared with ‘Penncross’. A higher K+ and lower Na+ content was maintained in leaves of ‘Mariner’ compared with ‘Penncross’ while roots of ‘Mariner’ maintained higher Ca2+ content under stressed and nonstressed conditions. Phytohormone levels showed a decline in salt-stressed roots compared with nonstressed plants but ‘Mariner’ roots were able to maintain levels higher than ‘Penncross’. ‘Mariner’ leaves showed an increased accumulation of ABA, JA, SA, and ZR while roots maintained higher IAA and SA compared with ‘Penncross’. The results suggest that ‘Mariner’ was able to mitigate salt stress by better ion regulation and differential regulation of hormones compared with ‘Penncross’. ‘Mariner’ leaves and roots showed significantly lower dead cells compared with ‘Penncross’ under salt stress. The results suggest that staining for cell viability could be a useful technique for studying turfgrass stress or other cellular responses.

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Sanalkumar Krishnan and Emily B. Merewitz

Polyamines (PAs), spermine (Spm), and spermidine (Spd) may enhance the abiotic stress tolerance and growth of creeping bentgrass (Agrostis stolonifera). Growth chamber studies were conducted to investigate the effect of PA application on the physiological response and hormone content in creeping bentgrass ‘Penn-G2’ under drought. Spm (1 mm) and Spd (5 mm) were applied exogenously under drought or well-watered conditions. PA-treated plants maintained significantly higher turf quality (TQ), relative water content (RWC), photochemical efficiency, and membrane health while maintaining lower canopy temperature. Spm at the 1-mm rate had a 2.46-fold higher osmotic adjustment (OA) at 10 d compared with control plants. A greater content of gibberellic acid (GA) isoforms (GA1, GA4, and GA20) were observed compared with controls during both studies for PA-treated plants under drought. After 7 days of drought stress in Expt.1, GA1 levels were 3.26 higher for Spm 1-mm-treated plants compared with drought controls. GA4 contents were 69% and 65% higher compared with drought-stressed-untreated plants for Spd 5-mm application after 9 and 11 days. Higher levels of GA20 were observed at 10 days (Spd 5 mm, 108.9% higher) due to PA treatment compared with drought controls. In addition to differential regulation of GA isoforms, we observed enhanced abscisic acid (ABA) due to PA application; however, not on a consistent basis. This study showed that PA application may play a role in GA1, GA4, and ABA accumulation in creeping bentgrass ‘Penn G-2’ under drought stress.

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Emily B. Merewitz and Sha Liu

Naturally derived products that may enhance the functionality of fertilizers or other agricultural inputs are needed to reduce inputs associated with stress damage and increase the sustainability of turfgrass management. Damage to high-value creeping bentgrass (Agrostis stolonifera) turf areas caused by heat stress is a widespread problem. This study aimed to evaluate multiple, diverse treatments that may illicit antioxidant responses in plants, melatonin, rutin, and Si, when applied as foliar pretreatments to heat stress. Creeping bentgrass plants were grown in growth chambers at optimal (23 °C) or heat stress conditions (35 °C). Turfgrass quality, chlorophyll content, leaf electrolyte leakage, photochemical efficiency, lipid peroxidation, antioxidant enzyme activity, and fatty acid content were measured to determine the effects of foliar treatments on heat stress responses. Melatonin, Si, and rutin were all found to improve some or all of the physiological parameters measured in the study, but only melatonin and Si reduced lipid peroxidation, increased antioxidant enzyme activity, and altered fatty acid contents. Melatonin- and Si-treated plants had greater superoxide dismutase and peroxidase activity and increased the content of the unsaturated fatty acid, linoleic acid, in creeping bentgrass leaves during heat stress compared with controls. Rutin improved turf quality and reduced electrolyte leakage during heat stress, but the mechanism associated with these changes is unclear because no changes were found in antioxidant enzyme activities or fatty acids. Melatonin and Si treatment promoted antioxidant enzyme activity and linoleic acid content of leaves, which have been associated with the improved heat tolerance of creeping bentgrass plants.

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Emily B. Merewitz, Thomas Gianfagna, and Bingru Huang

Drought stress is a widespread abiotic stress that causes a decline in plant growth. Drought injury symptoms have been associated with an inhibition in cytokinin (CK) synthesis. The objectives of this study were to investigate whether expression of a gene (ipt) encoding the enzyme adenine isopentenyl phosphotransferase for CK synthesis ligated to a senescence-activated promoter (SAG12) or a heat shock promoter (HSP18.2) would improve drought tolerance in creeping bentgrass (Agrostis stolonifera) and to examine shoot and root growth responses to drought stress associated with changes in endogenous production of CK, and the proportional change in CK and abscisic acid (ABA) due to ipt transformation. Most SAG12-ipt and HSP18.2-ipt transgenic lines exhibited significantly higher turf quality, photochemical efficiency, chlorophyll content, leaf relative water content, and root:shoot ratio under drought stress than the null transformant or the wild-type ‘Penncross’ plants. Transgenic lines that had better growth and turf performance generally had higher CK content and a higher CK-to-ABA ratio, although the direct correlation of CK and ABA content with individual physiological parameters in individual lines was not clear. Our results demonstrated that expressing ipt resulted in the improvement of turf performance under drought stress in creeping bentgrass in some of the transgenic plants with SAG12-ipt or HSP18.2-ipt, which could be associated with the suppression of leaf senescence and promoting root growth relative to shoot growth due to the maintenance of higher CK level and a higher ratio of CK to ABA.

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Sanalkumar Krishnan, Kevin Laskowski, Vijaya Shukla, and Emily B. Merewitz

Perennial ryegrass (Lolium perenne) is an important forage and turfgrass species that is sensitive to drought stress. The objective of this study was to investigate whether gamma aminobutyric acid (GABA) may play a role in promoting drought tolerance in grass species. GABA was exogenously applied as a foliar spray at the rate of 50 or 70 mm to perennial ryegrass ‘CSI’ under well-watered or drought-stressed conditions in a controlled-environment growth chamber. The effect of GABA on the growth physiology, drought stress response, antioxidant activity, and lipid peroxidation of perennial ryegrass exposed to drought stress was measured. GABA-treated perennial ryegrass exposed to drought stress had higher relative water content (RWC), turf quality, and peroxidase activity and lower wilt rating, canopy temperature depression, electrolyte leakage, and lipid peroxidation compared with untreated plants. GABA application had no significant effect on the activity of superoxide dismutase and catalase under well-watered and drought conditions. GABA application at 50 mm was found to be more effective in alleviating drought stress damage in perennial ryegrass. The results from this study suggest that GABA mitigated drought stress damage in perennial ryegrass by maintaining higher RWC and membrane stability.