The objectives of this study were to examine antioxidant enzyme responses to drought stress and rewatering at both enzymatic activity and transcript levels and to determine the major antioxidant processes associated with drought tolerance and post-drought recovery for a perennial grass species, kentucky bluegrass (Poa pratensis). Antioxidant enzyme responses to drought and rewatering in a drought-tolerant cultivar (Midnight) and a drought-sensitive cultivar (Brilliant) were compared in a growth chamber. Plants were exposed to 22 days of drought stress for ‘Midnight’ and 18 days for ‘Brilliant’ before rewatering to allow the leaf relative water content (RWC) of both cultivars to drop to the same level. ‘Midnight’ exhibited higher photochemical efficiency (Fv/Fm) and lower electrolyte leakage compared with ‘Brilliant’ when at the same water deficit status (26% to 28% RWC). After 6 days of rewatering, all physiological parameters returned to the control level for ‘Midnight’, but only Fv/Fm fully recovered for ‘Brilliant’. The transcript level of cytosolic copper/zinc superoxide dismutase (cyt Cu/Zn SOD) and ascorbate peroxidase (APX) was significantly higher in ‘Midnight’ than in ‘Brilliant’ when exposed to the same level of water deficit (26% to 28% RWC), suggesting that SOD and APX could be involved in scavenging oxidative stress-induced reactive oxygen species in kentucky bluegrass through changes in the level of gene expression. Significantly higher activities of APX, monodehydroascorbate reductase, glutathione reductase, and dehydroascorbate reductase as well as lower lipid peroxidation levels were observed in ‘Midnight’ versus ‘Brilliant’ when exposed to drought. However, the activities of SOD, catalase (CAT), and guaiacol peroxidase (POD) did not differ between the two cultivars. After 6 days of rewatering, ‘Midnight’ displayed significantly higher activity levels of CAT, POD, and APX compared with ‘Brilliant’. The enzyme activity results indicate that enzymes involved in the ascorbate–glutathine cycle may play important roles in antioxidant protection to drought damage, whereas CAT, POD, and APX could be associated with better post-drought recovery in kentucky bluegrass.
Lixin Xu, Liebao Han, and Bingru Huang
Feifei Li, Da Zhan, Lixin Xu, Liebao Han, and Xunzhong Zhang
Heat stress is a major limiting factor for growth of cool-season perennial grass species, and mechanisms of heat tolerance have not been well understood. This study was designed to investigate antioxidant enzyme and hormone metabolism responses to heat stress in two kentucky bluegrass (Poa pratensis L.) cultivars contrasting in heat tolerance. The plants were subjected to 20/20 °C [day/night (control)] or 38/30 °C [day/night (heat stress)] for 28 days in growth chambers. Heat stress increased leaf electrolyte leakage (EL) and malondialdehyde (MDA) with heat-tolerant cultivar EverGlade exhibiting lower levels of EL and MDA relative to heat-sensitive cultivar Kenblue under heat stress. Superoxide dismutase (SOD) and catalase (CAT) activity increased and then declined during 28 days of heat stress. Peroxidase (POD) and ascorbate peroxidase (APX) activity declined and then increased during heat stress. ‘EverGlade’ had greater activities of SOD, CAT, POD, and APX relative to ‘Kenblue’ under heat stress. In addition, ‘EverGlade’ had two additional SOD isozymes and three additional POD isozymes relative to ‘Kenblue’ under heat stress. Leaf abscisic acid (ABA) increased in response to heat stress. Leaf indole-3-acetic acid (IAA) increased and then declined during heat stress. ‘OverGlade’ had higher ABA and IAA content relative to ‘Kenblue’. At the end of heat stress, leaf IAA and ABA content were 27.8% and 73% higher in ‘EverGlade’ relative to ‘Kenblue’, respectively. The results indicated that antioxidant enzymes and the hormones (ABA and IAA) were associated with kentucky bluegrass heat tolerance. Selection and use of cultivars with higher IAA and ABA content and greater antioxidant enzyme activities may improve kentucky bluegrass growth and quality under heat stress.
Lixin Xu, Mili Zhang, Xunzhong Zhang, and Lie-Bao Han
Zoysiagrass (Zoysia spp.), a warm-season turfgrass species, experiences freezing damage in many regions. The mechanisms of its cold acclimation and freezing tolerance have not been well understood. This study was designed to investigate effects of cold acclimation treatment on leaf abscisic acid (ABA), cytokinin (transzeatin riboside (t-ZR), and antioxidant metabolism associated with freezing tolerance in zoysiagrass (Zoysia japonica). ‘Chinese Common’ zoysiagrass was subjected to either cold acclimation treatment with temperature at 8/2 °C (day/night) and a photosynthetically active radiation (PAR) of 250 µmol·m−2·s−1 over a 10-hour photoperiod or normal environments (temperature at 28/24 °C (day/night), PAR at 400 µmol·m−2·s−1 and 14-hour photoperiod) for 21 days in growth chambers. Cold treatment caused cell membrane injury as indicated by increased leaf cell membrane electrolyte leakage (EL) and malondialdehyde (MDA) content after 7 days of cold treatment. Cold treatment increased leaf ABA and hydrogen peroxide content and reduced t-ZR content. Leaf superoxide dismutase (SOD), ascorbate peroxidase (APX) activity, and proline content increased, whereas catalase (CAT) and peroxidase (POD) activity declined in response to cold treatment. Cold treatment increased freezing tolerance as LT50 declined from −4.8 to −12.5 °C. The results of this study indicated that cold acclimation treatment might result in increases in ABA and H2O2, which induce antioxidant metabolism responses and improved freezing tolerance in zoysiagrass.
Na Zhang, Lu Han, Lixin Xu, and Xunzhong Zhang
Ethephon [ETH (2-chloroethylphosphonic acid, an ethylene-releasing compound)] has been used as a plant growth regulator in turfgrass management. The aim of the study was to assess the effects of ETH seed treatment on drought tolerance of kentucky bluegrass (Poa pratensis) seedlings. Seeds of two kentucky bluegrass cultivars, Midnight and Nuglade, were exposed to ETH treatment or untreated as controls. Seedlings were then exposed to two water regimes: well-watered conditions and polyethylene glycol (PEG)–induced drought conditions. ETH-treated plants exhibited better turf performance relative to the untreated control under PEG-stressed conditions illustrated by higher relative water content (RWC) and lower lipid peroxidation and lower electrolyte leakage (EL). In both cultivars, ETH treatment increased enzyme activity of ascorbate peroxidase (APX), peroxidase (POD), and catalase (CAT); proline content; and soluble protein content under PEG-induced drought conditions. The results suggest that ETH seed treatment can improve drought tolerance in kentucky bluegrass seedlings.
Zhimin Yang, Lixin Xu, Jingjin Yu, Michelle DaCosta, and Bingru Huang
Carbohydrate metabolism is important for plant adaptation to drought stress. The objective of this study was to examine major forms of carbohydrates associated with superior drought tolerance and post-drought recovery in kentucky bluegrass (Poa pratensis) by comparing responses of different forms of carbohydrates with drought stress and re-watering in two cultivars contrasting in drought tolerance. Plants of drought-tolerant ‘Midnight’ and drought-sensitive ‘Brilliant’ were maintained well watered or subjected to drought stress for 10 days by withholding irrigation, and drought-stressed plants were re-watered for 3 days. Physiological analysis (turf quality, relative water content, and electrolyte leakage) confirmed the genetic variability of the two cultivars in drought tolerance. The two cultivars exhibited differential responses to drought stress and re-watering for the content of water-soluble sugars (sucrose, fructose, and glucose) and storage carbohydrates (starch and fructan), and ‘Midnight’ maintained higher sucrose content at 10 days of drought stress and more fructan at 3 days of re-watering. The greater accumulation of sucrose in ‘Midnight’ under drought stress corresponded with higher activities of two sucrose-synthesizing enzymes (sucrose phosphate synthase and sucrose synthase) but was not related to the sucrose-degrading enzyme activity (acid invertase). These results suggested that increased sucrose accumulation resulting from the maintenance of active sucrose synthesis could be associated with superior turf performance during drought stress, whereas increased fructan accumulation could contribute to rapid re-growth and post-drought recovery on re-watering in kentucky bluegrass.
Li-Juan Zhang, Tian-Xiu Zhong, Li-Xin Xu, Lie-bao Han, and Xunzhong Zhang
Soil water deficit impacts cold acclimation and freezing tolerance in creeping bentgrass (Agrostis stolonifera L.), but the mechanisms underlying have not been well understood. The objectives of this study were to investigate the effects of deficit irrigation before and during cold acclimation on osmoprotectants, antioxidant metabolism, and freezing tolerance in creeping bentgrass. The grass was subjected to three-soil moisture levels: well-watered [100% container capacity (CC)], deficit irrigation induced-mild drought stress (60% CC), and severe drought stress (30% CC) for 35 days including 14 days at 24/20 °C (day/night) and then 21 days under cold acclimation treatment (2 °C) in growth chambers. Leaf proline and total soluble sugar (TSS) levels were higher in the grass under mild drought stress relative to that under severe drought stress. Superoxide (O2 −·), hydrogen peroxide (H2O2), and malondialdehyde (MDA) content were higher in the grass under severe drought relative to that under well-watered and mild drought stress at day 35. Mild drought stress increased catalase (CAT) and guaiacol peroxidase (POD) activity, induced new isoforms and increased band intensities of superoxide dismutase (SOD), CAT, and POD during cold acclimation (days 14 to 35). No differences in osmoprotectants, antioxidant metabolism, and freezing tolerance were found between mild drought and well-watered treatments. The results of this study suggest deficit irrigation-induced mild drought stress in late fall and winter could induce accumulation of osmoprotectants and improve antioxidant metabolism, and freezing tolerance, but severe drought stress could reduce freezing tolerance of creeping bentgrass in the region with limited precipitation.