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
Cong Li, Lie-Bao Han and Xunzhong Zhang
Drought stress is one of the major limiting factors for plant growth and development. The mechanism of drought tolerance has not been well understood. This study was designed to investigate proline and antioxidant metabolism associated with drought tolerance in transgenic tobacco (Nicotiana tabacum) plants overexpressing the OjERF gene relative to wild-type (WT) plants. The OjERF gene was isolated from mondo grass (Ophiopogon japonicus). The OjERF gene, driven by the CaMV35S promoter, was introduced into tobacco through agrobacterium (Agrobacterium tumefaciens)-mediated transformation. Five transgenic lines were regenerated, of which transgenic Line 5 (GT5) and Line 6 (GT6) were used to examine drought tolerance in comparison with WT plants in a growth chamber. Drought stress caused an increase in leaf malondialdehyde (MDA) and electrolyte leakage (EL), proline content, superoxide dismutase (SOD), and catalase (CAT) activity in both transgenic lines and WT plants. However, the transgenic lines had lower MDA content and EL and higher proline content, SOD and CAT activity relative to WT under drought stress. The activities of SOD and CAT were also greater in the transgenic lines relative to WT plants under well-watered conditions (Day 0). The OjERF activated the expression of stress-relative genes, including NtERD10B, NtERD10C, NtERF5, NtSOD, and NtCAT1 in tobacco plants. The results of this study suggest that the OjERF gene may confer drought stress tolerance through upregulating proline and antioxidant metabolism.
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.
Shanshan Sun, Mengying An, Liebao Han and Shuxia Yin
Perennial ryegrass (Lolium perenne L.) is a widely used turfgrass. In this study, the effect of exogenously applied 24-epibrassinolide (EBR) on salt stress tolerance of perennial ryegrass was investigated. The results indicated that pretreatment with four concentrations of EBR (0, 0.1, 10, 1000 nM) improved salt tolerance of perennial ryegrass. Exogenous EBR treatment decreased electrolyte leakage (EL), malondialdehyde (MDA), and H2O2 contents and enhanced the leaf relative water content (RWC), proline, soluble sugar, and soluble protein content under salt stress condition. Meanwhile, EBR reduced the accumulation of Na+ and increased K+, Ca2+, and Mg2+ contents in leaves after salt treatment. Moreover, EBR pretreatment also increased superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) activity, as well as ascorbic acid (AsA) and glutathione contents. These results suggested that EBR improved salt tolerance by enhancing osmotic adjustment and antioxidant defense systems in perennial ryegrass.
Lie-Bao Han, Gui-Long Song and Xunzhong Zhang
Traffic stress causes turfgrass injury and soil compaction but the underlying physiological mechanisms are not well documented. The objectives of this study were to investigate the physiological responses of kentucky bluegrass (Poa pratensis), tall fescue (Festuca arundinacea), and japanese zoysiagrass (Zoysia japonica) to three levels of traffic stress during the growing season under simulated soccer traffic conditions. Relative leaf water content (LWC), shoot density, leaf chlorophyll concentration (LCC), membrane permeability, and leaf antioxidant peroxidase (POD) activity were measured once per month. The traffic stress treatments caused a reduction in LWC, shoot density, LCC, and POD activity, and an increase in cell membrane permeability in all three species. Japanese zoysiagrass had less electrolyte leakage, and higher POD activity and shoot density than both kentucky bluegrass and tall fescue. The results suggest that turfgrass tolerance to traffic stress may be related to leaf antioxidant activity. Turfgrass species or cultivars with higher leaf antioxidant activity may be more tolerant to traffic stress than those with lower antioxidant activity.
Da Man, Yong-Xia Bao, Lie-Bao Han and Xunzhong Zhang
Drought stress is a major factor in turfgrass management; however, the underlying mechanisms of turfgrass drought tolerance are not well understood. This greenhouse study was designed to investigate proline and hormone responses to drought stress in two tall fescue [Festuca arundinacea (Schreb.)] cultivars differing in drought tolerance. The two cultivars, Van Gogh (relatively drought-tolerant) and AST7002 (relatively drought-sensitive), were established and grown under either well-watered (maintaining 90% container capacity) or drought stress (≈26% container capacity) and then re-watered. Drought stress reduced turfgrass quality, relative leaf water content (LWC), leaf indole-3-acetic acid (IAA) and cytokinin zeatin riboside (ZR) content, and increased proline and abscisic acid (ABA) content. ‘Van Gogh’ had greater turfgrass quality rating, LWC, proline, ABA, and ZR content relative to ‘AST7002’ under drought stress conditions. At the end of drought stress, leaf proline, ZR, and ABA content were 32%, 43%, and 50% higher in ‘Van Gogh’ relative to ‘AST7002’, respectively. No cultivar difference was observed under well-watered conditions. The results of this study suggest that the proline, ABA, and ZR content are associated with drought tolerance. Selection and use of the cultivars with higher proline, ABA, and ZR content under drought stress may be a practical approach to improve tall fescue drought tolerance.
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.
Nikolaos Ntoulas, Panayiotis A. Nektarios, Thomais-Evelina Kapsali, Maria-Pinelopi Kaltsidi, Liebao Han and Shuxia Yin
Several locally available materials were tested to create an optimized growth substrate for arid and semiarid Mediterranean extensive green roofs. The study involved a four-step screening procedure. At the first step, 10 different materials were tested including pumice (Pum), crushed tiles grade 1–2 mm (T1–2), 2–4 mm (T2–4), 5–8 mm (T5–8), 5–16 mm (T5–16), and 4–22 mm (T4–22); crushed bricks of either 2–4 mm (B2–4) or 2–8 mm (B2–8); a thermally treated clay (TC); and zeolite (Zeo). All materials were tested for their particle size distribution, pH, and electrical conductivity (EC). The results were compared for compliance with existing guidelines for extensive green roof construction. From the first step, the most promising materials were shown to include Pum, Zeo, T5–8, T5–16, and TC, which were then used at the second stage to develop mixtures between them. Tests at the second stage included particle size distribution and moisture potential curves. Pumice mixed with TC provided the best compliance with existing guidelines in relation to particle size distribution, and it significantly increased moisture content compared with the mixes of Pum with T5–8 and T5–16. As a result, from the second screening step, the best performing substrate was Pum mixed with TC and Zeo. The third stage involved the selection of the most appropriate organic amendment of the growing substrate. Three composts having different composition and sphagnum peat were analyzed for their chemical and physical characteristics. The composts were a) garden waste compost (GWC), b) olive (Olea europaea L.) mill waste compost (OMWC), and c) grape (Vitis vinifera L.) marc compost (GMC). It was found that the peat-amended substrate retained increased moisture content compared with the compost-amended substrates. The fourth and final stage involved the evaluation of the environmental impact of the final mix with the four different organic amendments based on their first flush nitrate nitrogen (NO3 −-N) leaching potential. It was found that GWC and OMWC exhibited increased NO3 −-N leaching that initially reached 160 and 92 mg·L−1 of NO3 −-N for OMWC and GWC, respectively. By contrast, peat and GMC exhibited minimal NO3 −-N leaching that was slightly above the maximum contaminant level of 10 mg·L−1 of NO3 −-N (17.3 and 14.6 mg·L−1 of NO3 −N for peat and GMC, respectively). The latter was very brief and lasted only for the first 100 and 50 mL of effluent volume for peat and GMC, respectively.
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.