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Qingzhang Xu, Bingru Huang, and Zhaolong Wang

Turf quality of creeping bentgrass (Agrotis palustris L.) often declines during summer months. Reducing soil temperature alleviates bentgrass quality decline at supraoptimal air temperatures. The objective of this study was to investigate whether reducing soil temperature during the night is more effective than during the day in improving shoot and root growth when air temperature was supraoptimal for creeping bentgrass. The experiment was conducted in growth chambers using water baths to manipulate soil temperatures. Plants were exposed to the following temperature treatments: 1) optimal air and soil temperature during the day and night (20/20 °C, day/night, control); 2) high air and soil temperature during the day and night (35/35 °C, day/night); 3) lower soil temperatures during the day (20/35, 25/35, and 30/35 °C, day/night); and 4) lower soil temperature during the night (35/20, 35/25, and 35/30 °C) while air temperature was maintained at 35 °C during the day and night. Turf quality (on 1-9 scale) increased to the level of 6.5, 3.0, and 2.5 by reducing day soil temperature to 20, 25, or 30 °C, respectively, at 28 days of treatment, compared to the quality of 2.0 at 35/35 °C. Turf quality increased from 2.0 at 35/35 °C to 7.0, 6.0, and 4.5, respectively, by 28 days of exposure to night temperatures of 20, 25, and 30 °C. Chlorophyll content, root number, and root weight also were increased by reducing day or night soil temperature, and the increases were more pronounced for reduced night temperatures than day temperatures. These results demonstrated that reduced night soil temperature was more effective than reduced day soil temperature in improving shoot and root growth in creeping bentgrass under high air temperature conditions.

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Yali He, Xiaozhong Liu, and Bingru Huang

Various physiological processes may deteriorate in response to increasing temperatures, contributing to the decline in turf quality for cool-season turfgrasses during heat stress. This study was performed to investigate metabolic changes (membrane lipid peroxidation, total protein content, amino acid content, and protease activity) associated with turf quality decline for creeping bentgrass (Agrostis stolonifera Huds.) in response to gradually increasing temperatures for a short duration and prolonged exposure to lethally high temperature. Plants were subjected to increasing temperatures of 20, 25, 30, 35, and 40 °C for 7 days at each level of temperature [gradual heat stress (GHS)] or exposed to high temperature of 40 °C for 28 days [prolonged heat stress (PHS)] in growth chambers. During the GHS treatment, significant decline in turf quality occurred when plants were exposed to 30 °C for 7 days; simultaneously, malondialdehyde (MDA) content increased and total protein content in shoots decreased significantly compared to those at 20 °C. Protease activity increased at 25 °C and then decreased as temperature was elevated from 30 to 40 °C during the GHS treatment. Amino acid content decreased under GHS, beginning at 25 °C. Under the PHS treatment, turf quality declined and MDA content increased significantly, beginning at 14 days of PHS, while total protein content decreased at 7 days of PHS. Protease activity and amino acid content increased at 7 days of PHS, and then declined with longer stress duration. Our results indicated that protease activity, and amino acid and total protein content were more responsive to GHS or PHS than that of lipid peroxidation and turf quality. Changes in metabolic parameters of protease activity, amino acid and total protein content, and lipid peroxidation may contribute to leaf senescence and poor turf performance under severe or prolonged heat stress conditions for creeping bentgrass.

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Jinmin Fu, Bingru Huang, and Jack Fry

Effects of deficit irrigation applied to home lawns, used as means of water conservation, are an important issue. However, the impact of deficit irrigation on sucrose metabolism in tall fescue (Festuca arundinacea) is unknown and important because sucrose is the dominant form of carbohydrate transported to developing plant organs. The objectives of this study were to investigate the effects of deficit irrigation on leaf water content, osmotic potential (ψS), sucrose level, and the activity of sucrose phosphate synthase (SPS; EC 2.4.1.14), sucrose synthase (SS; EC 2.4.1.13), and acid invertase (AI; EC 3.2.1.26) in tall fescue leaves. Sods of ‘Falcon II’ tall fescue were established in polyvinylchloride (PVC) tubes (10 cm diameter × 40 cm long) filled with a mixture of sand and fritted clay [9:1 (v:v)] and then placed in growth chambers. Reference evapotranspiration rate [ETo (millimeters of water per day)] was determined by weighing the PVC tubes containing well-watered turfgrass every 3 days to determine water loss on a daily basis as ETo. Deficit irrigation treatments were applied as follows: well-watered control, mild drought stress (60% ETo), and severe drought stress (20% ETo). Leaf water content was lower at 6, 12, and 20 days of treatment for the 20% ETo treatment and 20 days after treatment began for the 60% ETo treatment. Compared with the well-watered control, ψS was lower in the 60% ETo treatment on all three measurement dates. Sucrose was higher at 8 and 14 days after treatment began in the 60% ETo treatment and on all three measurement dates in the 20% ETo treatment relative to the well-watered control. No difference in sucrose level was observed between the 20% ETo and 60% ETo irrigation regimes at 8 and 14 days of treatment. Beginning 14 days after treatment, tall fescue had a higher level of SPS in the 60% ETo and 20% ETo treatments compared with the well-watered treatment. Tall fescue receiving 60% or 20% ETo had a lower level of AI activity on all measurement dates. Results suggest that the decrease in ψS was accompanied by higher sucrose levels, which were the result of the increased level of SPS and SS activity and a decline in AI activity.

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Lixin Xu, Liebao Han, and Bingru Huang

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.

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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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Nanqing Liu, Yixin Shen, and Bingru Huang

Compatible solute accumulation regulating osmotic adjustment (OA) is associated with drought tolerance. The objectives of this study were to examine genetic variations in OA among a diverse group of bentgrass (Agrostis sp.) genotypes or lines with differential drought tolerance, and determine major types of organic osmoregulants contributing to OA and accounting for the genetic variations in drought tolerance. A wild type cultivar of creeping bentgrass [Agrostis stolonifera (Penncross)], a transgenic line of creeping bentgrass (SAGIPT41), and four hybrid bentgrass lines [Agrostis capillaris × Agrostis stolonifera (ColxCr14, ColxCr190, ColxCr481, and ColxCr679)] were exposed to drought stress by withholding irrigation for 17 days in growth chambers. Among genotypes, ColxCr14, ColxCr190, and SAGIPT41 showed superior drought tolerance, as manifested by higher turf quality (TQ) and leaf relative water content (RWC), as well as OA than ‘Penncross’, ColxCr679, and ColxCr481 under drought stress. SAGIPT41 leaves accumulated greater content of soluble sugars (glucose, sucrose, and fructose), proline, glycine betaine (GB), and spermine; ColxCr190 had higher content of soluble sugars and spermidine; and ColxCr14 accumulated more soluble sugars and GB, compared with the three drought-sensitive genotypes. Soluble sugars were predominant contributors to OA, followed by GB and proline, with all three forms of polyamine (PA) as minor contributors in bentgrass genotypes. The osmolytes highly correlated to OA and superior drought tolerance could be used as biomarkers to select for drought-tolerant germplasm of bentgrass and other cool-season turfgrass species.

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Zhongchun Jiang, Chenping Xu, and Bingru Huang

Low nitrogen (N) rates are recommended for creeping bentgrass (Agrostis stolonifera) putting greens to prevent excessive shoot growth and potential nitrate leaching, but low N rates could lead to N deficiency, which induces leaf senescence. This study was conducted to examine the effects of N deficiency on two enzymes involved in organic N metabolism as well as amino acid (AA) and soluble protein (SP) contents in both young and old leaves and roots of creeping bentgrass. Creeping bentgrass plants (cv. Penncross) were grown in a nutrient solution containing either 6 mm nitrate (+N plants) or zero N (−N plants), and each of the two treatments had four replicate pots. Young leaves on upper portions of the stolons and old leaves on lower portions of the stolons were separated and sampled at 14, 21, and 28 days of treatment, and roots were sampled at 28 days. Nitrogen deficiency increased glutamine synthetase (GS) transferase activity in all three tissues and at all three dates and GS biosynthetic activity in young leaves at all three dates. Prolonged N deficiency at 21 and 28 days increased glutamate dehydrogenase (GDH) deamination and amination activities in old leaves. In the roots, N deficiency at 28 days increased GS transferase activity but decreased GDH deamination activity. The N deficiency decreased AA content in all three tissues and at all three dates and SP content in young leaves at all three dates and in old leaves at 21 and 28 days. Decreasing organic N reserves in AA and SP and increasing GS and GDH activities in senescing leaves may be adaptive responses to N deficiency.

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Hongmei Du, Zhaolong Wang, and Bingru Huang

Heat stress may limit the growth of turfgrasses through the induction of oxidative stress, causing cellular and physiological damage. The objective of the study was to examine the association of heat and oxidative stresses between warm-season (C4) and cool-season (C3) turfgrasses. Plants of zoysiagrass (Zoysia matrella L. Merr. cv. Manila) (C4) and tall fescue (Festuca arundinacea Shreber cv. Barlexus) (C3) were exposed to optimal temperature conditions (24 °C for tall fescue and 34 °C for zoysiagrass) or heat stress (10 °C above the respective optimal temperature for each species) in growth chambers. Zoysiagrass exhibited less severe decline in turf quality and photochemical efficiency and less severe oxidative damage in cellular membranes as demonstrated by lower membrane electrolyte leakage and lipid peroxidation compared with tall fescue when both were exposed to heat stress. The activities of superoxide dismutase (SOD) and peroxidase (POD) declined with heat stress for both species, but to a lesser extent in zoysiagrass than in tall fescue, whereas catalase activity did not change significantly under heat stress and did not exhibit species variation. Our results demonstrate that the superior heat tolerance in zoysiagrass in comparison with tall fescue was associated with greater oxidative scavenging capacity as a result of the maintenance of higher SOD and POD activities.

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Liang Cheng, Ning Zhang, and Bingru Huang

The accumulation of 1-aminocyclopropane-1-carboxylate (ACC), which is a precursor for ethylene production, in plant roots exposed to salinity stress can be detrimental to plant growth. The objectives of this study were to determine whether inoculating roots with bacteria containing deaminase enzymes that break down ACC (ACC-deaminase) could improve plant tolerance to salinity in perennial ryegrass (Lolium perenne) and to examine growth and physiological factors, as well as nutrition status of plants affected by the ACC-deaminase bacteria inoculation under salinity stress. Plants of perennial ryegrass (cv. Pangea) were inoculated with either Burkholderia phytofirmans PsJN or Burkholderia gladioli RU1 and irrigated with either fresh water (control) or a 250 mm NaCl solution to induce salinity stress. The bacterium-inoculated plants had less ACC content in shoots and roots under both nonstressed and salinity conditions. Salinity stress inhibited root and shoot growth, but the bacterium-inoculated plants exhibited higher visual turf quality (TQ), tiller number, root biomass, shoot biomass, leaf water content, and photochemical efficiency, as well as lower cellular electrolyte leakage (EL) under salinity stress. Plants inoculated with bacteria had lower sodium content and higher potassium to sodium ratios in shoots under salinity stress. Shoot and root nitrogen content and shoot potassium content increased, whereas shoot and root calcium, magnesium, iron, and aluminum content all decreased due to bacterial inoculation under salinity treatment. ACC-deaminase bacteria inoculation of roots was effective in improving salinity tolerance of perennial ryegrass and could be incorporated into turfgrass maintenance programs in salt-affected soils.

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Xiuju Bian, Emily Merewitz, and Bingru Huang

Understanding factors influencing drought resistance traits is important for improving turfgrass growth in water-limited environments. The objectives of this study were to examine effects of a plant growth regulator, trinexapac-ethyl (TE), on turf growth and water use for creeping bentgrass (Agrostis stolonifera L.) exposed to drought stress, and to determine changes in the accumulation of solutes involved in osmotic adjustment associated with TE application. Plant foliage of cultivar L-93 was sprayed with 1.95 mL·L−1 of TE at 0.113% a.i. 14 days before and at the initiation of drought stress. TE-treated and untreated plants were exposed to well-watered or drought stress conditions for 28 days in a growth chamber. TE-treated plants exhibited a reduced rate of water depletion from the soil as demonstrated by higher soil water content, lower evapotranspiration rates, and higher leaf relative water content during 28 days of drought stress compared with non-TE-treated plants. During the later phase of drought stress, TE-treated plants had a greater reduction in leaf ψS at full turgor or greater osmotic adjustment, which was associated with increased accumulation of soluble sugars and inorganic ions (Ca and K) in leaves of TE-treated plants. Proline content increased in response to drought stress, but was unaffected by TE application, suggesting that it may not contribute to the effects of TE on osmotic adjustment. TE-treated plants maintained significantly higher turf quality and leaf photochemical efficiency under drought stress. The results suggest that the promotive effects of TE application on turf growth during drought stress were associated with the reduction in water depletion or lower water use and increases in osmotic adjustment due to the accumulation of inorganic solutes and soluble sugars.