Understanding antioxidant mechanisms for heat stress is important for improving heat tolerance in cool-season plant species. The objective of this study was to identify antioxidant enzymes associated with cultivar variations in heat tolerance in kentucky bluegrass (Poa pratensis) by comparing heat responses of activity and isoforms of antioxidant enzymes in two cultivars contrasting in heat tolerance. Plants of heat-tolerant ‘Eagleton’ and heat-sensitive ‘Brilliant’ were exposed to 20 °C (control) or 40 °C (heat stress) for 28 days in growth chambers. Chlorophyll (Chl) a content remained unchanged and Chl b content increased in ‘Eagleton’, while both of them decreased in ‘Brilliant’, and by 28 days, ‘Eagleton’ had significantly higher Chl a and b content than ‘Brilliant’. The activities of superoxide dismutase (SOD) were significantly higher in ‘Eagleton’ than in ‘Brilliant’ by 28 days of heat stress. An isozyme SOD2 was induced early during heat stress in ‘Eagleton’, while isozyme SOD3 degraded, to a lesser extent in ‘Eagleton’ than in ‘Brilliant’. Catalase (CAT) activity significantly increased in ‘Brilliant’ but remained constant in ‘Eagleton’, and ‘Brilliant’ had a significantly higher CAT activity and isozyme CAT1 than ‘Eagleton’ during heat stress. Significant increases in ascorbate peroxidase (APX) activities occurred under heat stress, to a greater extent in ‘Eagleton’, whereas isozymes did not exhibit difference between cultivars. Guaiacol-peroxidase (POD) activity declined during heat stress in both cultivars. The intensity of POD isozymes in ‘Brilliant’ remained constant, while ‘Eagleton’ showed a transient increases in POD1 at 7 days of heat stress. Our results indicated that antioxidant defense mechanisms for heat tolerance in kentucky bluegrass could be mainly associated with changes in activity and forms of isozymes of SOD for O2 scavenging and APX activity for H2O2 scavenging under heat stress.
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.
Yali He, Xiaozhong Liu, and Bingru Huang
The acclimation of plants to moderately high temperature plays an important role in inducing plant tolerance to subsequent lethal high temperatures. This study was performed to investigate the effects of heat acclimation and sudden heat stress on protein synthesis and degradation in creeping bentgrass (Agrostis palustris Huds.). Plants of the cultivar Penncross were subjected to two temperature regimes in growth chambers: 1) heat acclimation—plants were exposed to a gradual increase in temperatures from 20 to 25, 30, and 35 °C for 7 days at each temperature level before being exposed to 40 °C for 28 days; and 2) sudden heat stress (nonacclimation)—plants were directly exposed to 40 °C for 28 days from 20 °C without acclimation through the gradual increase in temperatures. Heat acclimation increased plant tolerance to subsequent heat stress, as demonstrated by lower electrolyte leakage (relative EL) in leaves of heat-acclimated plants compared to nonacclimated plants at 40 °C. Heat acclimation induced expression of some heat shock proteins (HSPs), 57 and 54 kDa, detected in a salt-soluble form (cystoplasmic proteins), which were not present in unacclimated plants under heat stress. However, HSPs of 23, 36, and 66 kDa were induced by both sudden and gradual exposure to heat stress. In general, total protein content decreased under both heat acclimation and sudden heat stress. Cystoplasmic proteins was more sensitive to increasing temperatures, with a significant decline initiated at 25 °C, while sodium dodecyl sulphate (SDS)-soluble (membrane) protein content did not decrease significantly until temperature was elevated to 30 °C. The results demonstrated that both a gradual increase in temperature and sudden heat stress caused protein degradation and also induced expression of newly synthesized HSPs. Our results suggested that the induction of new HSPs during heat acclimation might be associated with the enhanced thermotolerance of creeping bentgrass, although direct correlation of these two factors is yet to be determined. This study also indicated that protein degradation could be associated with heat injury during either gradual increases in temperature or sudden heat stress.
Mahalaxmi Veerasamy, Yali He, and Bingru Huang
Heat stress induces leaf senescence and causes changes in protein metabolism. The objective of this study was to investigate effects of exogenous application of a synthetic form of cytokinin, zeatin riboside (ZR), on protein metabolism associated with leaf senescence under heat stress for a cool-season grass species. Creeping bentgrass (Agrostis stolonifera L.) (cv. Penncross) plants were exposed to optimum temperature control (20/15 °C, day/night) and heat stress (35/30 °C) in growth chambers. Before heat stress treatments, foliage was sprayed with 10 μmol ZR or water (untreated) for 3 days and then once per week during 35 days of heat stress. Leaf chlorophyll content, photochemical efficiency (Fv/Fm), and soluble protein content declined, whereas protease activity increased during heat stress. Treatments with ZR helped maintain higher leaf chlorophyll content, Fv/Fm, and soluble protein content under heat stress. Protease activity in ZR-treated plants was lower than that of untreated plants. Zeatin riboside-treated plants had less severe degradation of ribulose-1,5-bisphosphate carboxylase proteins than untreated plants exposed to heat stress. In addition, ZR treatment upregulated the expression of 32- and 57-kDa proteins under heat stress conditions. These results demonstrated that the exogenous application of ZR ameliorated the negative effects of heat stress, as manifested by suppression or delay of leaf senescence. Cytokinins may have helped to alleviate heat stress injury, probably by slowing down the action of protease and by induction or upregulation of heat-shock proteins.