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Erick Amombo, Longxing Hu, Jibiao Fan, Zhengrong Hu, and Jinmin Fu

Clonal plants can consist of connected individual ramets that enhance resource sharing through physiological integration. This integration enables the whole clone to tolerate environmental stresses. The objective of this research was to investigate the effects of physical ramet connections on the integration of antioxidant enzymes in clonal common bermudagrass (Cynodon dactylon) growing under heterogeneously distributed water; i.e., nonuniform distribution of water due to 20% polyethylene glycol (PEG 6000) treatment on some ramets and not others. The bottom, middle, upper and three fragments of clonal common bermudagrass were subjected to 20% PEG 6000 with water potential of −1.8 MPa to induce heterogeneous and homogeneous drought stress. The control was not treated with 20% PEG 6000. Within the heterogeneous treatment, water stressed clonal fragments generally had higher leaf and root antioxidant enzyme activities with respect to superoxide dismutase, catalase, peroxidase (except for root peroxidase). There was no difference in antioxidant enzyme activity within the connected clonal ramets for homogeneous treatment; i.e., three connected ramets treated with 20% PEG 6000. Osmotically stressed clonal fragments under heterogeneous environments had a lower level of malonaldehyde (MDA) compared with those in homogeneous regimes. The antioxidant enzyme integration was affected by directionality and water availability contrast. This was indicated by significant decline in MDA levels within the heterogeneous treatments as compared with homogeneous treatment, which suggested reduced lipid peroxidation. These results suggested that ramet connections facilitate integration of antioxidant enzymes within clonal plants growing in heterogeneously available water. Enzymes were integrated from clonal fragments growing in water sufficient environment to those in water stressed regimes. This enhanced reactive oxygen species scavenging capacity of the entire clone hence improved drought tolerance.

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Longxing Hu, Tao Hu, Xunzhong Zhang, Huancheng Pang, and Jinmin Fu

Salinity stress may involve the accumulation of glycine betaine (GB). The objective of this study was to examine whether exogenous GB would ameliorate the detrimental effect of salinity stress on perennial ryegrass (Lolium perenne). The grass was subjected to two salinity levels (0 and 250 mm NaCl) and three GB levels (0, 20, and 50 mm). Salinity resulted in a remarkable decrease in vertical shoot growth rate (VSGR), shoot and root fresh weight, relative water content (RWC), relative transpiration rate (Tr), and chlorophyll (Chl) content, superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX) activities. Plants subjected to salt exhibited an increase in leaf electrolyte leakage (EL), lipid peroxidation (MDA), and proline content. Application of GB reduced EL, MDA, and proline content in salt-stressed plants. Perennial ryegrass subjected to salt stress plus GB had a greater level of VSGR, RWC, relative Tr, Chl content, and activities of SOD, CAT, and APX when compared with salt-stressed without GB. Salt stress increased Na+ and decreased K+ content, which resulted in a higher Na+/K+ ratio in perennial ryegrass. Application of 20 mm GB suppressed Na+ accumulation, whereas the K+ content was significantly increased in shoot, which led to a higher K+/Na+ ratio under saline conditions. These results suggested that GB-enhanced salt tolerance in perennial ryegrass was mainly related to the elevated SOD, CAT, and APX activity and alleviation of cell membrane damage by reducing oxidation of membrane lipid and improving the ion homeostasis under salt stress.

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Tao Hu, Haiying Yi, Longxing Hu, and Jinmin Fu

Plants possess abiotic stress responses that alter photosynthetic metabolism under salinity stress. The objective of this study was to identify the stomatal and metabolic changes associated with photosynthetic responses to NaCl stress in perennial ryegrass (Lolium perenne). Five-week-old seedlings of two perennial ryegrass genotypes, PI 516605 (salt-sensitive) and BARLP 4317 (salt-tolerant), were subjected to 0 and 250 mm NaCl for 8 days. The salt tolerance in perennial ryegrass was significantly associated with leaf relative water content (RWC) and photosynthetic capacity through the maintenance of greater metabolic activities under prolonged salt stress. BARLP 4317 maintained greater turf quality, RWC, and stomatal limitations but a lower level of lipid peroxidation [malondialdehyde (MDA)] and intercellular CO2 concentration than PI 516605 at 8 days after treatment (DAT). Ribulose-1, 5-bisphosphate carboxylase:oxygenase (Rubisco) activity and activation state, transcriptional level of rbcL gene, and expression level of Rubisco large subunit (LSU) declined in stressed perennial ryegrass but were higher in salt-tolerant genotype at 8 DAT. Furthermore, photosynthetic rate increased linearly with increasing Rubisco activity, Rubisco activation state, and RWC in both genotypes. The same linear relationship was found between RWC and Rubisco activity. However, MDA content decreased linearly with increasing RWC in both genotypes. Salinity-induced inhibition of photosynthesis in perennial ryegrass was mainly the result of stomatal limitation during early salt stress and metabolic limitation associated with the inhibition of RWC, activity of Rubisco, expression level of rbcL gene, and LSU under a prolonged period of severe salinity.

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Longxing Hu, Zehui Huang, Shuqian Liu, and Jinmin Fu

Plant adaptation to salt stress may be associated with morphological, physiological, and gene expression alterations. The objective of this study was to investigate the effect of salt stress on morphological and antioxidant enzyme changes and its gene expressions in bermudagrass (Cynodon dactylon). Salt-tolerant ‘C43’ and salt-sensitive ‘C198’, previously determined in our preliminary study, were subjected to four salinity levels: 0 mm (control), 100 mm (low), 200 mm (moderate), and 400 mm (high) NaCl for 21 days. Salt stress decreased turf quality and canopy height, especially in ‘C198’. Salt stress increased root length, root number, root fresh weight, and root/shoot length ratio, to a greater extent in salt-tolerant genotype. Salt stress increased Na+ and decreased K+ content, which resulted in a higher Na+/K+ ratio in bermudagrass, to a great extent in shoot and root of ‘C198’. Moderate (200 mm) and high (400 mm) salt concentration increased malondialdehyde and hydrogen peroxide content in old leaves of ‘C198’. ‘C43’ exhibited a greater activity of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and dehydro-ascorbate reductase (DHAR) than ‘C198’ in old leaves subjected to 200 and 400 mm NaCl. Antioxidant gene expressions were upregulated in new leaves and downregulated in old leaves with increasing salinity levels for both genotypes. Salt-tolerant genotypes exhibited a relatively greater antioxidant gene expression than salt-sensitive ones when exposed to the same level of salt stress. These results suggested that SOD, CAT, APX, and DHAR might be involved in scavenging salt stress-induced reactive oxygen species in bermudagrass at the level of gene expression. Salt tolerance might be attributed to the development and maintenance of a more extensive root system under saline conditions and induced antioxidant gene expressions, leading to more efficient enzyme stimulation and protection in bermudagrass.

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Zhuangjun Zhao, Margaret Mukami Gitau, Tao Hu, Yan Xie, Longxing Hu, and Jinmin Fu

Plants growing in salt-affected soils may have retarded growth and inhibited or altered metabolic processes. This study aims at investigating the impact of subsurface soil salinity on root growth and metabolic processes in perennial ryegrass (Lolium perenne). The seeds of perennial ryegrass (cv. Quick Start II) were planted in polyvinyl chloride (PVC) tubes (10 cm diameter × 42 cm long) for 2 months. The experiment consisted of three treatments: 1) control, 40 cm filled with sand–peat mixture (7 sand : 3 peat wt/wt); 2) T20, a 20-cm-deep layer of saline soil covered with a 20-cm-deep layer of sand–peat mixture; and 3) T30, a 30-cm-deep layer of saline soil covered with a 10-cm-deep layer of sand–peat mixture. Our study showed that soil salinity at the subsurface inhibited the growth of perennial ryegrass roots. Compared with the control, the root activity in saline soil layer decreased, whereas it remained high in the mixture-soil zone. The content of amino acids in the roots obtained from the surface soil (0–10 cm) in T30 was greater than that in both the T20 and the control regimes. The content of soluble sugars in the roots went up with the decrease of the depth of sand–peat mixture. The increased root activity and free amino acids content in the roots sampled from the upper soil layers coupled with the increased soluble sugars in the roots subjected to soil salinity stress in the bottom soil layer represents some adaptive responses and regulative mechanisms in perennial ryegrass.