Assessment of the changes in growth, photosynthetic traits and gene expression in Cynodon dactylon against drought stress.

Drought stress considered a key restrictive factor for a warm-season bermudagrass growth during summers in China. Genotypic variation against drought stress exists among bermudagrass (Cynodon sp.), but the selection of highly drought-tolerant germplasm is important for its growth in limited water regions and for future breeding. Our study aimed to investigate the most tolerant bermudagrass germplasm among thirteen, along latitude and longitudinal gradient under a well-watered and drought stress condition. Current study included high drought-resistant germplasm, "Tianshui" and "Linxiang", and drought-sensitive cultivars; "Zhengzhou" and "Cixian" under drought treatments along longitude and latitudinal gradients, respectively. Under water deficit conditions, the tolerant genotypes showed over-expression of a dehydrin gene cdDHN4, antioxidant genes Cu/ZnSOD and APX which leads to higher antioxidant activities to scavenge the excessive reactive oxygen species and minimizing the membrane damage. It helps in maintenance of cell membrane permeability and osmotic adjustment by producing organic osmolytes. Proline an osmolyte has the ability to keep osmotic water potential and water use efficiency high via stomatal conductance and maintain transpiration rate. It leads to optimum CO2 assimilation rate, high chlorophyll contents for photosynthesis and elongation of leaf mesophyll, palisade and thick spongy cells. Consequently, it results in elongation of leaf length, stolon and internode length; plant height and deep rooting system. The CdDHN4 gene highly expressed in "Tianshui" and "Youxian", Cu/ZnSOD gene in "Tianshui" and "Linxiang" and APX gene in "Shanxian" and "Linxiang". The genotypes "Zhongshan" and "Xiaochang" showed no gene expression under water deficit conditions. Our results indicate that turfgrass show morphological modifications firstly when subjected to drought stress; however the gene expression is directly associated and crucial for drought tolerance in bermudagrass. Hence, current research has provided excellent germplasm of drought tolerant bermudagrass for physiological and molecular study and future breeding.

Comprehensive transcriptional analysis reveals salt stress-regulated key pathways, hub genes and time-specific responsive gene categories in common bermudagrass (Cynodon dactylon (L.) Pers.) roots.

BACKGROUND: Despite its good salt-tolerance level, key genes and pathways involved with temporal salt response of common bermudagrass (Cynodon dactylon (L.) Pers.) have not been explored. Therefore, in this study, to understand the underlying regulatory mechanism following the different period of salt exposure, a comprehensive transcriptome analysis of the bermudagrass roots was conducted. RESULTS: The transcripts regulated after 1 h, 6 h, or 24 h of hydroponic exposure to 200 mM NaCl in the roots of bermudagrass were investigated. Dataset series analysis revealed 16 distinct temporal salt-responsive expression profiles. Enrichment analysis identified potentially important salt responsive genes belonging to specific categories, such as hormonal metabolism, secondary metabolism, misc., cell wall, transcription factors and genes encoded a series of transporters. Weighted gene co-expression network analysis (WGCNA) revealed that lavenderblush2 and brown4 modules were significantly positively correlated with the proline content and peroxidase activity and hub genes within these two modules were further determined. Besides, after 1 h of salt treatment, genes belonging to categories such as signalling receptor kinase, transcription factors, tetrapyrrole synthesis and lipid metabolism were immediately and exclusively up-enriched compared to the subsequent time points, which indicated fast-acting and immediate physiological responses. Genes involved in secondary metabolite biosynthesis such as simple phenols, glucosinolates, isoflavones and tocopherol biosynthesis were exclusively up-regulated after 24 h of salt treatment, suggesting a slightly slower reaction of metabolic adjustment. CONCLUSION: Here, we revealed salt-responsive genes belonging to categories that were commonly or differentially expressed in short-term salt stress, suggesting possible adaptive salt response mechanisms in roots. Also, the distinctive salt-response pathways and potential salt-tolerant hub genes investigated can provide useful future references to explore the molecular mechanisms of bermudagrass.

Carbohydrate saving or biomass maintenance: which is the main determinant of the plant's long-term submergence tolerance?

It is hypothesized that plant submergence tolerance could be assessed from the decline of plant biomass due to submergence, as biomass integrates all eco-physiological processes leading to fitness. An alternative hypothesis stated that the consumption rate of carbohydrate is essential in differing tolerance to submergence. In the present study, the responses of biomass, biomass allocation, and carbohydrate content to simulated long-term winter submergence were assessed in four tolerant and four sensitive perennials. The four tolerant perennials occur in a newly established riparian ecosystem created by The Three Gorges Dam, China. They had 100% survival after 120 days' simulated submergence, and had full photosynthesis recovery after 30 days' re-aeration, and the photosynthetic rate was positively related to the growth during the recovery period. Tolerant perennials were characterized by higher carbohydrate levels, compared with the four sensitive perennials (0% survival) at the end of submergence. Additionally, by using a method which simulates posterior estimates, and bootstraps the confidence interval for the difference between strata means, it was found that the biomass response to post-hypoxia, rather than that to submergence, could be a reliable indicator to assess submergence tolerance. Interestingly, the differences of changes in carbohydrate content between tolerant and sensitive perennials during submergence were significant, which were distinct from the biomass response, supporting the hypothesis that tolerant perennials could sacrifice non-vital components of biomass to prioritize the saving of carbohydrates for later recovery. Our study provides some insight into the underlying mechanism(s) of perennials' tolerance to submergence in ecosystems such as temperate wetland and reservoir riparian.

Comparative proteomic analysis provides new insights into the specialization of shoots and stolons in bermudagrass (Cynodon dactylon L.).

BACKGROUND: Bermudagrass (Cynodon dactylon L.) is an important turfgrass species with two types of stems, shoots and stolons. Despite their importance in determining the morphological variance and plasticity of bermudagrass, the intrinsic differences between stolons and shoots are poorly understood. RESULTS: In this study, we compared the proteomes of internode sections of shoots and stolons in the bermudagrass cultivar Yangjiang. The results indicated that 376 protein species were differentially accumulated in the two types of stems. Pathway enrichment analysis revealed that five and nine biochemical pathways were significantly enriched in stolons and shoots, respectively. Specifically, enzymes participating in starch synthesis all preferentially accumulated in stolons, whereas proteins involved in glycolysis and diverse transport processes showed relatively higher abundance in shoots. ADP-glucose pyrophosphorylase (AGPase) and pyruvate kinase (PK), which catalyze rate-limiting steps of starch synthesis and glycolysis, showed high expression levels and enzyme activity in stolons and shoots, respectively, in accordance with the different starch and soluble sugar contents of the two types of stems. CONCLUSIONS: Our study revealed the differences between the shoots and stolons of bermudagrass at the proteome level. The results not only expand our understanding of the specialization of stolons and shoots but also provide clues for the breeding of bermudagrass and other turfgrasses with different plant architectures.

Characterization of Dehydrin protein, CdDHN4-L and CdDHN4-S, and their differential protective roles against abiotic stress in vitro.

BACKGROUND: Dehydrins play positive roles in regulating plant abiotic stress responses. The objective of this study was to characterize two dehydrin genes, CdDHN4-L and CdDHN4-S, generated by alternative splicing of CdDHN4 in bermudagrass. RESULTS: Overexpression of CdDHN4-L with φ-segment and CdDHN4-S lacking of φ-segment in Arabidopsis significantly increased tolerance against abiotic stresses. The growth phenotype of Arabidopsis exposed to NaCl at 100 mM was better in plants overexpressing CdDHN4-L than those overexpressing CdDHN4-S, as well as better in E.coli cells overexpressing CdDHN4-L than those overexpressing CdDHN4-S in 300 and 400 mM NaCl, and under extreme temperature conditions at - 20 °C and 50 °C. The CdDHN4-L had higher disordered characterization on structures than CdDHN4-S at temperatures from 10 to 90 °C. The recovery activities of lactic dehydrogenase (LDH) and alcohol dehydrogenase (ADH) in presence of CdDHN4-L and CdDHN4-S were higher than that of LDH and ADH alone under freeze-thaw damage and heat. Protein-binding and bimolecular fluorescence complementation showed that both proteins could bind to proteins with positive isoelectric point via electrostatic forces. CONCLUSIONS: These results indicate that CdDHN4-L has higher protective ability against abiotic stresses due to its higher flexible unfolded structure and thermostability in comparison with CdDHN4-S. These provided direct evidence of the function of the φ-segment in dehydrins for protecting plants against abiotic stress and to show the electrostatic interaction between dehydrins and client proteins.

Cholesterol accumulation by suppression of SMT1 leads to dwarfism and improved drought tolerance in herbaceous plants.

Dwarfism and drought tolerance are 2 valuable traits in breeding of many crops. In this study, we report the novel physiological roles of cholesterol in regulation of plant growth and drought tolerance. Compared with the wild type, sterol-C24-methyltransferase 1 (SMT1) gene transcript was greatly reduced in a bermudagrass mutant with dwarfism and enhanced drought tolerance, accompanied with cholesterol accumulation, elevated transcript levels of a small group of genes including SAMDC, and increased concentrations of putrescine (Put), spermidine (Spd), and spermine (Spm). Knock-down of OsSMT1 expression by RNA interference resulted in similar phenotypic changes in transgenic rice. Moreover, exogenously applied cholesterol also led to elevated transcripts of a similar set of genes, higher levels of Put, Spd, and Spm, improved drought tolerance, and reduced plant height in both bermudagrass and rice. We revealed that it is Spm, but not Spd, that is responsible for the height reduction in bermudagrass and rice. In conclusion, we suggest that cholesterol induces expression of SAMDC and leads to dwarfism and elevated drought tolerance in plants as a result of the promoted Spd and Spm synthesis.

Reflectance, absorbance and transmittance spectra of bermudagrass and manilagrass turfgrass canopies.

Leaves act as a primary organ for the interception of solar radiation and their spatial arrangement determines how the plant canopy interacts with light. Many studies have been carried out on the penetration of radiation into crops however to date, few results are available on turfgrasses, mainly due to the difficulties of introducing sensors into the turf without disturbing the natural position of the leaves. In the present research two warm season turfgrasses, hybrid bermudagrass (Cynodon dactylon × transvaalensis) 'Patriot' and manilagrass (Zoysia matrella) 'Zeon', were studied. The aim was to describe their canopy architecture grown with minimal disturbance to the natural arrangement of the leaves and stems, and to determine the potential effects of canopy architecture on light penetration and reflectance. Radiometric measurements were carried out at eight different profile levels of turfgrasses that were up to 12 cm tall. A LI-COR 1800 spectroradiometer with an optical fiber cable and a 7 mm diameter sensor was used. Measurements were carried out in the 390-1100 nm region at 5 nm intervals. The LAI value was higher for the manilagrass (9.0) than for the hybrid bermudagrass (5.6). The transmitted radiation was found to be closely dependent on downward cumulative LAI. Despite a more upright habit (mean insertion angle of 22.4° ±3.4), Zoysia matrella showed a higher NIR reflectance compared to Cdxt, which has a horizontal leaf arrangement (mean insertion angle 62.1° ± 9.6). The species studied showed substantial differences both in terms of phytometric characteristics and in the capacity to attenuate solar radiation.

Amelioration of Salt Stress on Bermudagrass by the Fungus Aspergillus aculeatus.

There is considerable evidence that plant abiotic-stress tolerance can be evoked by the exploitation of a globally abundant microbe. A. aculeatus, which was initially isolated from the rhizosphere of bermudagrass, has been shown to increase heavy metal tolerance in turfgrasses. Here, we report on the potential of A. aculeatus to induce tolerance to salt stress in bermudagrass. Physiological markers for salt stress, such as plant growth rate, lipid peroxidation, photosynthesis, and ionic homeostasis were assessed. Results indicated that strain A. aculeatus produced indole-3-acetic acid (IAA) and siderophores and exhibited a greater capacity for Na+ absorption under salt stress. The plant inoculation by A. aculeatus increased plant growth and attenuated the NaCl-induced lipid peroxidation in roots and leaves of bermudagrass. The fungus significantly elevated the amount of IAA and glutathione and slightly enhanced photosynthetic efficiency of salt-treated bermudagrass. Tissues of inoculated plants had significantly increased concentrations of K+ but lower Na+ concentrations than those of uninoculated regimes. It appears that the role of A. aculeatus in alleviating bermudagrass salt stress is partly to produce IAA, to increase the activity of antioxidases, to absorb Na+ by fungal hyphae, and to prevent the plant from ionic homeostasis disruption.

Photosynthesis, antioxidant system and gene expression of bermudagrass in response to low temperature and salt stress.

There is widespread distribution of salinized lands in northern China. Harnessing such land is essential to environmental health. Bermudagrass [Cynodon dactylon (L.) Pers.] has the potential to improve the salinized lands. However, low temperature remarkably limits the growth of bermudagrass in winter. Currently, there is no information about the interaction of cold and salt in this plant. Hence, the objectives of this study were to figure out the effects of combined cold and salinity stress on bermudagrass. In this study, 4 °C and 200 mM salt solution was used as cold and salt treatments respectively while 4 °C along with 200 mM salt solution were applied as combined stress. After 5 days treatment, bermudagrass displayed a dramatic decline in the turf quality and chlorophyll content, but higher malonaldehyde, electrolyte leakage, hydrogen peroxide content, antioxidant enzyme activity in the combined stress regime as compared to cold or salt treated alone. Analysis of chlorophyll a revealed that the combined stress aggravated stress-induced inhibition of photosystem II. In addition, the expressions of stress-related genes were up-regulated with a lower expression level when cold and salt applied together. In summary, the grass exposed to combined stress presented a relatively lower stress tolerance and suffered a more severe damage than grass grown in the other regimes. These findings are crucial for elucidating the molecular mechanisms of cold and salt combined stress in bermudagrass, and provide information for breeding programs to select and develop bermudagrass cultivars that are suitable for improvement of the northern China salinized land.

Effects of ethylene on photosystem II and antioxidant enzyme activity in Bermuda grass under low temperature.

The phytohormone ethylene has been reported to mediate plant response to cold stress. However, it is still debated whether the effect of ethylene on plant response to cold stress is negative or positive. The objective of the present study was to explore the role of ethylene in the cold resistance of Bermuda grass (Cynodon dactylon (L).Pers.). Under control (warm) condition, there was no obvious effect of the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC) or the antagonist Ag(+) of ethylene signaling on electrolyte leakage (EL) and malondialdehyde (MDA) content. Under cold stress conditions, ACC-treated plant leaves had a greater level of EL and MDA than the untreated leaves. However, the EL and MDA values were lower in the Ag(+) regime versus the untreated. In addition, after 3 days of cold treatment, ACC remarkably reduced the content of soluble protein and also altered antioxidant enzyme activity. Under control (warm) condition, there was no significant effect of ACC on the performance of photosystem II (PS II) as monitored by chlorophyll α fluorescence transients. However, under cold stress, ACC inhibited the performance of PS II. Under cold condition, ACC remarkably reduced the performance index for energy conservation from excitation to the reduction of intersystem electron acceptors (PI(ABS)), the maximum quantum yield of primary photochemistry (φP0), the quantum yield of electron transport flux from Q(A) to Q(B) (φE0), and the efficiency/probability of electron transport (ΨE0). Simultaneously, ACC increased the values of specific energy fluxes for absorption (ABS/RC) and dissipation (DI0/RC) after 3 days of cold treatment. Additionally, under cold condition, exogenous ACC altered the expressions of several related genes implicated in the induction of cold tolerance (LEA, SOD, POD-1 and CBF1, EIN3-1, and EIN3-2). The present study thus suggests that ethylene affects the cold tolerance of Bermuda grass by impacting the antioxidant system, photosystem II, as well as the CBF transcriptional regulatory cascade.

Overwintering, Oviposition, and Larval Survival of Hunting Billbugs (Coleoptera: Curculionidae) and Implications for Adult Damage in North Carolina Turfgrass.

The hunting billbug, Sphenophorus venatus vestitus Chittenden, is one of the most widely recognized billbug turfgrass pests. Since 2000, damage to warm-season turfgrass caused by hunting bill bugs has increased and a need for information on hunting billbug biology is necessary for the development of management plans. Field and laboratory studies were conducted to collect data on overwintering, oviposition behavior, larval survival at various levels of soil moisture, and adult damage. Turfgrass samples from 'Tifway 419' bermudagrass(Cynodon dactylon (L.) Pers x Cynodon transvaalensis Burtt Davy) on golf courses were collected to determine overwintering behavior, and 10 female adult billbugs were collected weekly to determine oviposition behavior.Survival of medium-sized larvae (head capsule width: 1.0 and 1.7 mm) was evaluated in containers with 20, 40,60, or 80% of the total pore space occupied by water. Zero, two, four, or six adult billbugs were placed in bermudagrass, zoysiagrass, or tall fescue containers and images were collected for 4 weeks to determine adult damage. We observed that hunting billbugs overwinter as adults and all larval sizes. Adults became active in March and began to oviposit, which continued through October. Larval mortality was lowest with 20% of the total pores pace occupied by water, while increases in moisture caused significant mortality. Adults caused a greater reduction in warm-season turfgrass cover than cool-season turfgrass cover. This research builds on the existing biological information for the hunting billbug biology in transition zones and will be pivotal in developing practical and sustainable management plans.

Changes in southern Piedmont grassland community structure and nutritive quality with future climate scenarios of elevated tropospheric ozone and altered rainfall patterns.

Forage species common to the southern USA Piedmont region, Lolium arundinacea, Paspalum dilatatum, Cynodon dactylon and Trifolium repens, were established in a model pasture system to test the future climate change scenario of increasing ozone exposure in combination with varying rainfall amounts on community structure and nutritive quality. Forages were exposed to two levels of ozone [ambient (non-filtered; NF) and twice ambient (2×) concentrations] with three levels of precipitation (average or ±20% of average) in modified open-top chambers (OTCs) from June to September 2009. Dry matter (DM) yield did not differ over the growing season between forage types, except in primary growth grasses where DM yield was higher in 2× than NF treatment. Primary growth clover decreased in nutritive quality in 2× ozone because of increased concentrations of neutral detergent fibre (NDF), acid detergent fibre (ADF) and acid detergent lignin (ADL). Re-growth clover exhibited the largest decrease in nutritive quality, whereas grasses were not adversely affected in 2× ozone. Re-growth grasses responded positively to 2× ozone exposure, as indicated in increased relative food value (RFV) and percentage crude protein (CP) than NF-exposed re-growth grasses. Effects of precipitation were not significant over the growing season for primary or re-growth forage, except in primary growth grasses where DM yield was higher in chambers with above average (+20%) precipitation. Total canopy cover was significantly higher over the growing season in chambers receiving above average precipitation, but no significant effects were observed with ozone. Results indicate shifts in plant community structure and functioning related to mammalian herbivore herbivory in future climate change scenarios.

Competition between a Lawn-Forming Cynodon dactylon and a Tufted Grass Species Hyparrhenia hirta on a South-African Dystrophic Savanna.

South African savanna grasslands are often characterised by indigestible tufted grass species whereas lawn grasses are far more desirable in terms of herbivore sustenance. We aimed to investigate the role of nutrients and/or the disturbance (grazing, trampling) by herbivores on the formation of grazing lawns. We conducted a series of common garden experiments to test the effect of nutrients on interspecific competition between a typical lawn-forming grass species (Cynodon dactylon) and a species that is frequently found outside grazing lawns (Hyparrhenia hirta), and tested for the effect of herbivore disturbance in the form of trampling and clipping. We also performed a vegetation and herbivore survey to apply experimentally derived insights to field observations. Our results showed that interspecific competition was not affected by soil nutrient concentrations. C. dactylon did show much more resilience to disturbance than H. hirta, presumably due to the regenerative capacity of its rhizomes. Results from the field survey were in line with these findings, describing a correlation between herbivore pressure and C. dactylon abundance. We conclude that herbivore disturbance, and not soil nutrients, provide C. dactylon with a competitive advantage over H. hirta, due to vegetative regeneration from its rhizomes. This provides evidence for the importance of concentrated, high herbivore densities for the creation and maintenance of grazing lawns.

Suitability of Creeping Bentgrass and Bermudagrass Cultivars for Black Cutworms and Fall Armyworms (Lepidoptera: Noctuidae).

The black cutworm, Agrotis ipsilon Hufnagel, and fall armyworm, Spodoptera frugiperda Smith (Lepidoptera: Noctuidae), are common turfgrass pests of golf courses in the southeastern United States. Heat-tolerant bentgrass (Agrostis stolonifera L.) cultivars are expanding the range of bentgrass further south, but these cultivars have not been studied for their potential host plant resistance to black cutworm or fall armyworm. The goals of the study were to investigate feeding response of black cutworm and fall armyworm to these newer heat-tolerant creeping bentgrass cultivars, as well as commonly used cultivars of bermudagrass [Cynodon dactylon (Loppers.)]. Choice and no-choice feeding assays and fecundity tests were conducted in the laboratory and greenhouse to evaluate performance and preference of the two insects. When given a choice, neither black cutworm nor fall armyworm showed a preference for the majority of new cultivars tested. There were no differences in leaf area consumption or insect development for either pest in no-choice feeding assays. Black cutworm females preferred laying eggs in bentgrass compared with bermudagrass, but will oviposit onto bermudagrass, suggesting that both turf species are suitable hosts of this pest. The broad host ranges of generalist caterpillar pests of turfgrass hinder the application of host plant resistance in integrated pest management on golf courses.

A transcriptomic analysis of bermudagrass (Cynodon dactylon) provides novel insights into the basis of low temperature tolerance.

BACKGROUND: Cold stress is regarded as a key factor limiting widespread use for bermudagrass (Cynodon dactylon). Therefore, to improve cold tolerance for bermudagrass, it is urgent to understand molecular mechanisms of bermudagrass response to cold stress. However, our knowledge about the molecular responses of this species to cold stress is largely unknown. The objective of this study was to characterize the transcriptomic response to low temperature in bermudagrass by using RNA-Seq platform. RESULTS: Ten cDNA libraries were generated from RNA samples of leaves from five different treatments in the cold-resistant (R) and the cold-sensitive (S) genotypes, including 4 °C cold acclimation (CA) for 24 h and 48 h, freezing (-5 °C) treatments for 4 h with or without prior CA, and controls. When subjected to cold acclimation, global gene expressions were initiated more quickly in the R genotype than those in the S genotype. The R genotype activated gene expression more effectively in response to freezing temperature after 48 h CA than the S genotype. The differentially expressed genes were identified as low temperature sensing and signaling-related genes, functional proteins and transcription factors, many of which were specifically or predominantly expressed in the R genotype under cold treatments, implying that these genes play important roles in the enhanced cold hardiness of bermudagrass. KEGG pathway enrichment analysis for DEGs revealed that photosynthesis, nitrogen metabolism and carbon fixation pathways play key roles in bermudagrass response to cold stress. CONCLUSIONS: The results of this study may contribute to our understanding the molecular mechanism underlying the responses of bermudagrass to cold stress, and also provide important clues for further study and in-depth characterization of cold-resistance breeding candidate genes in bermudagrass.

Physiological and Molecular Mechanism of Nitric Oxide (NO) Involved in Bermudagrass Response to Cold Stress.

Bermudagrass is widely utilized in parks, lawns, and golf courses. However, cold is a key factor limiting resource use in bermudagrass. Therefore, it is meaningful to study the mechanism of bermudagrass response to cold. Nitric oxide (NO) is a crucial signal molecule with multiple biological functions. Thus, the objective of this study was to investigate whether NO play roles in bermudagrass response to cold. Sodium nitroprusside (SNP) was used as NO donor, while 2-phenyl-4,4,5,5-tetramentylimidazoline-l-oxyl-3-xide (PTIO) plus NG-nitro-L-arginine methyl ester (L-NAME) were applied as NO inhibitor. Wild bermudagrass was subjected to 4 °C in a growth chamber under different treatments (Control, SNP, PTIO + L-NAME). The results indicated lower levels of malondialdehyde (MDA) content and electrolyte leakage (EL), higher value for chlorophyll content, superoxide dismutase (SOD) and peroxidase (POD) activities after SNP treatment than that of PTIO plus L-NAME treatments under cold stress. Analysis of Chlorophyll (Chl) a fluorescence transient displayed that the OJIP transient curve was higher after treatment with SNP than that of treated with PTIO plus L-NAME under cold stress. The values of photosynthetic fluorescence parameters were higher after treatment with SNP than that of treated with PTIO plus L-NAME under cold stress. Expression of cold-responsive genes was altered under cold stress after treated with SNP or PTIO plus L-NAME. In summary, our findings indicated that, as an important strategy to protect bermudagrass against cold stress, NO could maintain the stability of cell membrane, up-regulate the antioxidant enzymes activities, recover process of photosystem II (PSII) and induce the expression of cold-responsive genes.

Performance of a system with full- and pilot-scale sludge drying reed bed units treating septic tank sludge in Brazil.

This study investigated the performance of sludge drying reed beds (SDRB) at full- and pilot-scale treating sludge from septic tanks in the city of Belo Horizonte, Brazil. The treatment units, planted with Cynodon spp., were based on an adaptation of the first-stage of the French vertical-flow constructed wetland, originally developed for treating sewage. Two different operational phases were investigated; in the first one, the full-scale unit was used together with six pilot-scale columns in order to test different feeding strategies. For the second phase, only the full-scale unit was used, including a recirculation of the filtered effluent (percolate) to one of the units of the French vertical wetland. Sludge application was done once a week emptying a full truck, during 25 weeks. The sludge was predominantly diluted, leading to low solids loading rates (median values of 18 kgTS m(-2) year(-1)). Chemical oxygen demand removal efficiency in the full-scale unit was reasonable (median of 71%), but the total solids removal was only moderate (median of 44%) in the full-scale unit without recirculation. Recirculation did not bring substantial improvements in the overall performance. The other loading conditions implemented in the pilot columns also did not show statistically different performances.

Physiological integration enhanced the tolerance of Cynodon dactylon to flooding.

Many flooding-tolerant species are clonal plants; however, the effects of physiological integration on plant responses to flooding have received limited attention. We hypothesise that flooding can trigger changes in metabolism of carbohydrates and ROS (reactive oxygen species) in clonal plants, and that physiological integration can ameliorate the adverse effects of stress, subsequently restoring the growth of flooded ramets. In the present study, we conducted a factorial experiment combining flooding to apical ramets and stolon severing (preventing physiological integration) between apical and basal ramets of Cynodon dactylon, which is a stoloniferous perennial grass with considerable flooding tolerance. Flooding-induced responses including decreased root biomass, accumulation of soluble sugar and starch, as well as increased activity of superoxide dismutase (SOD) and ascorbate peroxidase (APX) in apical ramets. Physiological integration relieved growth inhibition, carbohydrate accumulation and induction of antioxidant enzyme activity in stressed ramets, as expected, without any observable cost in unstressed ramets. We speculate that relief of flooding stress in clonal plants may rely on oxidising power and electron acceptors transferred between ramets through physiological integration.

Comparative physiological, metabolomic, and transcriptomic analyses reveal mechanisms of improved abiotic stress resistance in bermudagrass [Cynodon dactylon (L). Pers.] by exogenous melatonin.

Melatonin (N-acetyl-5-methoxytryptamine), a well-known animal hormone, is also involved in plant development and abiotic stress responses. In this study, it is shown that exogenous application of melatonin conferred improved salt, drought, and cold stress resistances in bermudagrass. Moreover, exogenous melatonin treatment alleviated reactive oxygen species (ROS) burst and cell damage induced by abiotic stress; this involved activation of several antioxidants. Additionally, melatonin-pre-treated plants exhibited higher concentrations of 54 metabolites, including amino acids, organic acids, sugars, and sugar alcohols, than non-treated plants under abiotic stress conditions. Genome-wide transcriptomic profiling identified 3933 transcripts (2361 up-regulated and 1572 down-regulated) that were differentially expressed in melatonin-treated plants versus controls. Pathway and gene ontology (GO) term enrichment analyses revealed that genes involved in nitrogen metabolism, major carbohydrate metabolism, tricarboxylic acid (TCA)/org transformation, transport, hormone metabolism, metal handling, redox, and secondary metabolism were over-represented after melatonin pre-treatment. Taken together, this study provides the first evidence of the protective roles of exogenous melatonin in the bermudagrass response to abiotic stresses, partially via activation of antioxidants and modulation of metabolic homeostasis. Notably, metabolic and transcriptomic analyses showed that the underlying mechanisms of melatonin could involve major reorientation of photorespiratory and carbohydrate and nitrogen metabolism.

Comparative proteomic responses of two bermudagrass (Cynodon dactylon (L). Pers.) varieties contrasting in drought stress resistance.

Drought (water-deficit) stress is a serious environmental problem in plant growth and cultivation. As one of widely cultivated warm-season turfgrass, bermudagrass (Cynodon dactylon (L). Pers.) exhibits drastic natural variation in the drought stress resistance in leaves and stems of different varieties. In this study, proteomic analysis was performed to identify drought-responsive proteins in both leaves and stems of two bermudagrass varieties contrasting in drought stress resistance, including drought sensitive variety (Yukon) and drought tolerant variety (Tifgreen). Through comparative proteomic analysis, 39 proteins with significantly changed abundance were identified, including 3 commonly increased and 2 decreased proteins by drought stress in leaves and stems of Yukon and Tifgreen varieties, 2 differentially regulated proteins in leaves and stems of two varieties after drought treatment, 23 proteins increased by drought stress in Yukon variety and constitutively expressed in Tifgreen variety, and other 3 differentially expressed proteins under control and drought stress conditions. Among them, proteins involved in photosynthesis (PS), glycolysis, N-metabolism, tricarboxylicacid (TCA) and redox pathways were largely enriched, which might be contributed to the natural variation of drought resistance between Yukon and Tifgreen varieties. These studies provide new insights to understand the molecular mechanism underlying bermudagrass response to drought stress.