Drought stress is one of the most important abiotic stresses limiting plant growth, while high recuperative capacity of plants from drought damages is critical for plant survival in periods of drought stress and rewatering. The objective of our study was to determine physiological and growth factors in association with drought tolerance and recuperative capacity of cool-season kentucky bluegrass (Poa pratensis cv. Excursion II) and warm-season zoysigrass (Zoysia matrella cv. Diomand), which were grown in controlled environment chambers and maintained well watered (control) or subjected to drought stress and subsequently rewatering. Compared with kentucky bluegrass, zoysiagrass maintained higher leaf hydration level during drought stress, as shown by greater relative water content (RWC), improved osmotic adjustment (OA), increased leaf thickness, and more extensive root system at deeper soil layers. Turf quality (TQ) and photosynthesis recovered to a greater level and sooner in response to rewatering for zoysiagrass, compared with kentucky bluegrass, which could be due to more rapid reopening of stomata [higher stomatal conductance (g S)] and leaf rehydration (higher RWC). The aforementioned physiological factors associated with leaf dehydration tolerance during drought and rapid resumption in turf growth and photosynthesis in zoysiagrass could be useful traits for improving drought tolerance in turfgrasses.
Jingjin Yu, Mengxian Liu, Zhimin Yang and Bingru Huang
Jianming Sun, Yiming Liu, Xianglin Li and Bingru Huang
Protein metabolism plays an important role in plant adaptation to drought stress. The objective of this study was to identify drought-responsive proteins associated with differential drought tolerance for a tolerant genotype (RU9) and a sensitive genotype (RU18) of tall fescue (Lolium arundinacea). Plants of both genotypes were grown under well-watered conditions or subjected to drought stress by withholding irrigation for 12 days in a growth chamber controlled at the optimal growth temperatures of 23/18 °C (day/night). Physiological analysis demonstrated that RU9 was relatively more drought tolerant than RU18, as shown by the higher leaf net photosynthetic rate (Pn) and photochemical efficiency at 12 days of drought treatment. Differentially expressed proteins between RU9 and RU18 exposed to drought stress were identified by two-dimensional electrophoresis and mass spectrometry (MS). Several proteins [photosystem I reaction center subunit II, Rubisco small subunit, and Glyceraldehyde-3-phosphate dehydrogenase (GADPH)] in photosynthesis, respiration, or oxidative regulation exhibited higher abundance in RU9 than RU18 under drought stress. These results suggested the critical importance of energy and oxidative metabolism in tall fescue adaptation to drought stress. Those abundant proteins in the drought-tolerant genotype could be used as biomarkers or developed to molecular markers to develop elite drought-tolerant germplasm in tall fescue and other cool-season perennial grass species.
Shaoyun Lu, Zhongcheng Wang, Yuejing Niu, Zhenfei Guo and Bingru Huang
Improving the drought tolerance of widely used bermudagrass [Cynodon dactylon (L.) Pers. var. dactylon] is important for water conservation and producing quality turf with limited irrigation. Mutants of bermudagrass were generated using gamma-ray irradiation with an aim toward developing dwarf and drought-resistant bermudagrass. The objectives of this study were to compare morphological characteristics between radiation-induced mutants and the wild-type of bermudagrass and to determine antioxidant responses associated with changes in drought resistance in the bermudagrass mutants. Three mutant lines (7-9, 10-5, and 10-12) that exhibit slow growth and good turf quality were chosen for this study. Plants were exposed to drought stress by withholding irrigation in a greenhouse. Mutant lines had lower canopy height, shorter internodes, and shorter leaves than the wild type under well-watered conditions. Under drought stress, all three dwarf mutant lines maintained higher relative water content and lower ion leakage and malondialdehyde content than the wild type. Antioxidant enzyme activities decreased in response to the drought stress in the mutant lines and the wild type, whereas nonenzymatic antioxidants increased under drought stress. Compared with the wild type, higher enzyme activities and antioxidant contents were maintained in mutant lines under drought stress. Our results indicated that bermudagrass mutants induced by gamma radiation exhibited dwarf characteristics and improved drought resistance, which was associated with maintenance of higher levels of antioxidant enzyme activities and nonenzymatic antioxidant contents.
June Liu, Zhimin Yang, Weiling Li, Jingjin Yu and Bingru Huang
Cold stress is a major factor limiting the growth of warm-season turfgrass species. Cold tolerance in warm-season turfgrass species could be improved through in vitro selection for somaclonal variations. The objectives of this study were to establish an effective in vitro culture protocol for generating plants from calli using mature seeds of seashore paspalum (Paspalum vaginatum) and to determine whether in vitro cold selection of somaclonal variations would lead to improved cold tolerance in seashore paspalum. The optimal concentrations of supplemental compounds in the culture medium for callus induction, embryogenic callus formation, and plant regeneration were determined. The supplemental compounds included 2,4–dichlorophenoxy acetic acid (2,4-D), 6-benzylaminopurine (6-BA), kinetin (KT), naphthalene-1-acetic acid (NAA), CuSO4, and acidic hydrolysis casein (AHC). The highest rates of callus induction (97.50%), embryogenic callus formation (66.88%), and regeneration (55.94%) were obtained with the supplemental compounds of 3.0 mg·L−1 2,4-D and 10.0 mg·L−1 CuSO4 for callus induction; with 3.0 mg·L−1 2,4-D, 15 mg·L−1 CuSO4, and 1.0 g·L−1 AHC for embryogenic callus formation; and with 8.0 mg·L−1 6-BA, 0.2 mg·L−1 KT, 0.5 mg·L−1 NAA, and 10 mg·L−1 CuSO4 for plant regeneration. Embryogenic calli were subjected to 2 or 6 °C treatment for 90 days for in vitro cold selection of somaclonal variation. Plants regenerated from calli surviving cold treatment (cold-selected) for 45 or 60 days were then exposed to low temperatures [15/10 or 5/3 °C (day/night)]. Plant variants derived from cold-selected calli exhibited significant improvement in their tolerance to low temperature of either 15/10 or 5/3 °C (day/night), as manifested by higher turf quality, leaf chlorophyll content, and membrane stability as well as lower levels of lipid peroxidation compared with the control plants. This study demonstrated the feasibility of in vitro selection for cold tolerance in seashore paspalum. The cold-tolerant variants could be useful germplasm for breeding programs and further molecular characterization of cold tolerance mechanisms.
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.
Jinyu Wang, Bo Yuan, Yi Xu and Bingru Huang
Amino acid and protein metabolism are interrelated and both play important roles in plant adaptation to heat stress. The objective of this study was to identify amino acids and soluble proteins associated with genetic variation in heat tolerance of hard fescue (Festuca trachyphylla). According to a previous screening experiment, the hard fescue cultivars Reliant IV and Predator were selected as heat-tolerant and heat-sensitive cultivars, respectively. Plants of these two hard fescue cultivars were exposed to heat stress at 38/33 °C (day/night) or optimal temperature at 21/18 °C in growth chambers. Each cultivar had four replications under each temperature, and the experimental design was a split-plot design, temperature as the main plots and cultivars as the subplots. Under heat stress, ‘Reliant IV’ exhibited higher turf quality (TQ) and greater membrane stability than ‘Predator’. In response to heat stress, total amino acid content increased, whereas total soluble protein content decreased in both cultivars. The greater accumulation of amino acids in ‘Reliant IV’ was contributed by the greater increase of proteins involved in the glycolysis and the tricarboxylic acid (TCA) cycle that provided carbon skeleton for amino acid synthesis. ‘Reliant IV’ leaves exhibited greater extent of increases in the content of six individual amino acids (histidine, glutamine, proline, threonine, aspartate, and tryptophan) than ‘Predator’ under heat stress. Several soluble proteins were upregulated in response to heat stress, to a greater extent in ‘Reliant IV’ than ‘Predator’, including the proteins involved in photosynthesis, protein folding, redox hemostasis, stress signaling, stress defense, cell organization, and metabolism. These differentially accumulated free amino acids and soluble proteins could be associated with the genetic variation in heat tolerance of hard fescue.
Diheng Zhong, Hongmei Du, Zhaolong Wang and Bingru Huang
Fatty acid metabolism may be involved in plant adaptation to drought stress. The objective of this study was to identify saturated and unsaturated fatty acids associated with leaf dehydration tolerance by comparing fatty acid composition and unsaturation levels at equivalent leaf water status of two bermudagrass genotypes contrasting in drought resistance. A drought-resistant hybrid bermudagrass (Cynodon dactylon × C. transvaalensis) genotype (‘Tifway’) and a drought-sensitive bermudagrass (C. dactylon) genotype (‘C299’) were maintained under well-watered (control) or water-withheld (drought) conditions. Drought treatment was imposed until soil water content decreased to 5% or leaf relative water content (RWC) dropped to 28% to 29%. ‘Tifway’ maintained higher RWC and lower electrolyte leakage (EL) at 5 and 10 days of drought stress. Leaves of ‘Tifway’ maintained lower EL when RWC of both genotypes declined to the same level of water deficit (28% to 29%) by the end of drought periods. The degree of fatty acid unsaturation, expressed as the double bond index, decreased in both genotypes during drought stress, which was mainly associated with the decline in linoleic (C18:2) and linolenic acids (C18:3) and an increase in palmitic (C16:0) and stearic acids (C18:0). A lipid composition characterized by a greater amount of unsaturated fatty acids was detected in ‘Tifway’ relative to ‘C299’ exposed to the same level of water deficit, mainly as a result of a greater content of C18:2 and a lower content of C16:0 and C18:0. Our results suggest that the ability to maintain a greater composition of unsaturated fatty acids in membrane lipids may contribute to superior leaf dehydration tolerance in bermudagrass.
Zhaolong Wang, Bingru Huang, Stacy A. Bonos and William A. Meyer
Drought is a major factor limiting plant growth, which has been associated with the accumulation of absicsic acid (ABA) in various species. The objective of the study was to determine the relationship between ABA accumulation and drought tolerance for kentucky bluegrass (Poa pratensis L.) during short-term drought stress. Eight kentucky bluegrass cultivars (`Midnight', `A82-204', `RSP', `Alpine', `Moonlight', `Brilliant', `Washington', and `Baruzo') were subjected to drought stress in a growth chamber. Water relations, gas exchange rate, and ABA content of leaves were determined at various times during drought stress. Turf quality decreased with drought duration for all eight cultivars. Leaf ABA content increased linearly with drought stress within 11 days of treatment; the rate of the increase was negatively related to the rate of turf quality decline. The rate of ABA accumulation during drought stress was positively correlated with the rates of decrease in turf quality (r 2 = 0.6346), increase in electrolyte leakage (r 2 = 0.7128), and decrease in relative water content (r 2 = 0.5913). There were highly significant negative correlations between ABA content and leaf water potential (r 2 = 0.9074), stomatal conductance (r 2 = 0.6088), transpiration rate (r 2 = 0.6581), net photosynthesis rate (r 2 = 0.6956), and a positive correlation between ABA content and electrolyte leakage (r 2 = 0.7287). The results indicate that drought tolerance is negatively related to ABA accumulation during shortterm drought stress. ABA accumulation in response to drought stress could be used as a metabolic factor to select for drought tolerance in kentucky bluegrass.
Yiming Liu, Hongmei Du, Kai Wang, Bingru Huang and Zhaolong Wang
Salinity is a detrimental abiotic stress for plant growth in salt-affected soils. The objective of this study was to examine photosynthetic responses to salinity stress in two warm-season turfgrasses differing in salinity tolerance. Salt-tolerant species seashore paspalum (Paspalum vaginatum) and salt-sensitive species centipedegrass (Eremochloa ophiuroides) were exposed to salinity at three NaCl concentrations (0, 300, and 500 mm) in a growth chamber. Turf quality, relative water content (RWC), and leaf photochemical efficiency (Fv/Fm) declined, whereas electrolyte leakage (EL) increased under the two NaCl regimes for both grass species, and the changes were more dramatic in centipedegrass than that in seashore paspalum as well as in the higher salinity concentration. Two grass species showed different phytosynthetic responses to salinity stress. The earlier inhibition of photosynthesis in seashore paspalum was mainly associated with stomatal closure. As salinity increased and salinity stress prolonged, the inhibition of photosynthesis in seashore paspalum was mainly associated with non-stomatal factors. The inhibition of photosynthesis in centipedegrass was associated with both stomatal closure and non-stomatal factors at both salinity levels. The sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) analysis demonstrated the Rubisco large subunit had no obvious decrease during the whole stress period under the 300-mm and 500-mm treatments in seashore paspalum, whereas it significantly decreased in centipedegrass under both the 300-mm and 500-mm treatments. The results indicated that the superior salinity tolerance in seashore paspalum, compared with centipedegrass, could be attributed to its maintenance of Rubisco stability, chlorophyll content, photochemical efficiency as well as photosynthetic rate (Pn) capacity under salinity stress.
Hua Shen, Hongmei Du, Zhaolong Wang and Bingru Huang
The objective of this study was to compare differential nutrient responses to heat stress in relation to heat tolerance for warm-season (C4) common bermudagrass [Cynodon dactylon (L.) Pers.] and cool-season (C3) kentucky bluegrass (Poa pratensis L.). Both species were exposed to two temperature regimes in growth chambers: optimal day/night temperature conditions (24/20 °C for kentucky bluegrass and 34/30 °C for bermudagrass) or heat stress (10 °C above the respective optimal temperature for each species). Heat injury in leaves was evaluated and the concentrations of several major macronutrients [nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), and magnesium (Mg)] in both grass species were measured at 0, 7, 14, 21, and 28 days of treatment. Heat stress reduced leaf photochemical efficiency and cellular membrane stability in both species, but bermudagrass leaves exhibited less damage in these parameters than kentucky bluegrass. Heat stress caused a significant decline in N, P, and K concentration, beginning at 7 days in kentucky bluegrass, but had no significant effects on N, P, and K concentration in bermudagrass during the 28-day treatment period. The concentration of Ca and Mg increased under heat stress in both kentucky bluegrass and bermudagrass, but there were no significant differences between the species under optimal or high-temperature conditions, suggesting they were not involved in heat responses in either species. The differential responses of N, P, and K to heat stress could at least partially account for the differences in heat tolerance between the two species and demonstrate the importance of sufficient N, P, and K in turfgrass adaptation to heat stress.