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Root-knot nematodes were discovered in severely declining creeping bentgrass putting greens at a golf course in Indian Wells, Riverside County, California. The exhibited disease symptoms included chlorosis, stunted growth, and dieback. Based on morphological examination and measurements of J2 females and males, it was suggested that the causal pathogen was Meloidogyne marylandi. This identification was confirmed by analysis of the D2-D3 expansion segments of 28S rRNA and COI gene sequences. The host status of 28 plant species was evaluated in greenhouse trials. All tested monocots, except rye and Allium species, were found to be hosts, while no reproduction occurred on dicots. Temperature-tank experiments helped determine that the life cycle of M. marylandi was completed between 17–35 °C, with a base temperature of 8.3 °C and a required heat sum of 493 degree-days (DD). In greenhouse trials in pasteurized soil and near-ideal growing conditions, M. marylandi did not cause significant growth reduction of creeping bentgrass cv. Penn A-4, even at very high J2 inoculation densities. It is highly probable that other biotic and abiotic factors contributed to the observed putting green damage.

Abstract Background Isopentenyltransferases (IPT) serve as crucial rate-limiting enzyme in cytokinin synthesis, playing a vital role in plant growth, development, and resistance to abiotic stress. Results Compared to the wild type, transgenic creeping bentgrass exhibited a slower growth rate, heightened drought tolerance, and improved shade tolerance attributed to delayed leaf senescence. Additionally, transgenic plants showed significant increases in antioxidant enzyme levels, chlorophyll content, and soluble sugars. Importantly, this study uncovered that overexpression of the MtIPT gene not only significantly enhanced cytokinin and auxin content but also influenced brassinosteroid level. RNA-seq analysis revealed that differentially expressed genes (DEGs) between transgenic and wild type plants were closely associated with plant hormone signal transduction, steroid biosynthesis, photosynthesis, flavonoid biosynthesis, carotenoid biosynthesis, anthocyanin biosynthesis, oxidation-reduction process, cytokinin metabolism, and wax biosynthesis. And numerous DEGs related to growth, development, and stress tolerance were identified, including cytokinin signal transduction genes (CRE1, B-ARR), antioxidase-related genes (APX2, PEX11, PER1), Photosynthesis-related genes (ATPF1A, PSBQ, PETF), flavonoid synthesis genes (F3H, C12RT1, DFR), wax synthesis gene (MAH1), senescence-associated gene (SAG20), among others. Conclusion These findings suggest that the MtIPT gene acts as a negative regulator of plant growth and development, while also playing a crucial role in the plant’s response to abiotic stress.

Recently, there has been renewed interest in reducing excess inputs into turf, with a special emphasis on reducing water use. One potential mechanism to achieve this goal is the use of plant growth–promoting rhizobacteria (PGPR) applications. Plant growth–promoting rhizobacteria research in turfgrass is limited, but the few studies that have been conducted show that PGPR can reduce biotic and abiotic stress in turfgrass. Two creeping bentgrass (Agrostis stolonifera) cultivars (Penncross and 007) were treated with either PGPR Blend 20, PGPR DH44, water, or nitrogen fertilizer before being subjected to a dry-down period during which half of each treatment was either irrigated or drought stressed. Results indicate that PGPR DH44 may help maintain greater quality in drought-stressed ‘007’ creeping bentgrass compared with nitrogen-treated plots; however, quality did not differ between DH44- and water-treated plots. When drought stressed, PGPR did not help creeping bentgrass maintain clipping yield compared with nitrogen-fertilized or water-treated plots, nor did PGPR affect root biomass. More research is needed before recommendations can be made regarding PGPR applications to turfgrass.

Abstract Background Creeping bentgrass (Agrostis soionifera) is a perennial grass of Gramineae, belonging to cold season turfgrass, but has poor disease resistance. Up to now, little is known about the induced systemic resistance (ISR) mechanism, especially the relevant functional proteins, which is important to disease resistance of turfgrass. Achieving more information of proteins of infected creeping bentgrass is helpful to understand the ISR mechanism. Results With BDO treatment, creeping bentgrass seedlings were grown, and the ISR response was induced by infecting Rhizoctonia solani. High-quality protein sequences of creeping bentgrass seedlings were obtained. Some of protein sequences were functionally annotated according to the database alignment while a large part of the obtained protein sequences was left non-annotated. To treat the non-annotated sequences, a prediction model based on convolutional neural network was established with the dataset from Uniport database in three domains to acquire good performance, especially the higher false positive control rate. With established model, the non-annotated protein sequences of creeping bentgrass were analyzed to annotate proteins relevant to disease-resistance response and signal transduction. Conclusions The prediction model based on convolutional neural network was successfully applied to select good candidates of the proteins with functions relevant to the ISR mechanism from the protein sequences which cannot be annotated by database alignment. The waste of sequence data can be avoided, and research time and labor will be saved in further research of protein of creeping bentgrass by molecular biology technology. It also provides reference for other sequence analysis of turfgrass disease-resistance research.

Aeration and sand topdressing are important cultural practices for organic matter management on golf course putting greens. Many golf courses lack the budget for applications of new sand topdressing material. A 2-year study was conducted to investigate the effect of recycling sand from hollow-tine aerification cores on a sand-based creeping bentgrass (Agrostis stolonifera) putting green soil properties and playability. Treatments included traditional [T (cores removed and sand topdressed)], verticut [V (cores broken up with verticutter)], and recycle [R (cores recycled using a core recycler)]. There were no differences in root zone organic matter, bulk density, soil porosity, infiltration rates, percent sand recovered during mowing, surface firmness, and ball roll distance between treatments during the study. Immediately after aerification treatments, T had the highest percent green cover (PGC) (38.3%) compared with V (26.9%) and R (26.8%), indicating that T offered the least sand present on the surface. Seven days after treatments, there was no difference in PGC (85.3% to 90.1%), indicating all treatments recovered similarly. Alternative aerification treatments V and R could be useful techniques to minimize or reduce the amount of sand used for backfilling core aeration holes without compromising the putting green soil properties and playability.

Renovation is an opportune time for golf courses to address annual bluegrass (Poa annua L.) weed populations. Dazomet (tetrahydro-3,5-dimethyl-2H-1,3,5-thiadiazine-2-thione) is an effective fumigant, but without a tarp cover, it is only effective at the highest labeled rates. Fraise mowing cultivation could be used to help remove surface material and allow practitioners to effectively fumigate at lower rates. In Summer 2018 and Summer 2019, two cool-season fairway renovation experiments were conducted in East Lansing, MI. The objective of these experiments was to assess annual bluegrass control and creeping bentgrass establishment following dazomet applications to fraise mowed surfaces. In the first experiment (fraise mowing surface disturbance experiment), dazomet was applied at a fixed rate (294 kg·ha−1) to fraise mowed plots at varying levels of surface disturbance (0%, 15%, 50%, and 100%) to a depth of 1.9 cm. In the second experiment (dazomet rate experiment), fraise mowing removed 100% of surface material at a depth of 1.9 cm and dazomet was applied at five rates (0, 294, 588, 147 + 147, and 294 + 294 kg·ha−1). Both experiments were conducted on two soils (sand topdressed vs. native) and evaluated two methods of fumigant incorporation (solid-tine cultivation vs. tillage). Five days after treatments were applied, plots were seeded with ‘Pure Select’ creeping bentgrass (Agrostis stolonifera L.). The level of fraise mowing surface disturbance had no effect on annual bluegrass emergence, and creeping bentgrass cover was poorest in native soils at the highest levels of surface disturbance. In the dazomet rate experiment, dazomet applied twice at 294 kg·ha−1 provided the most consistent control of annual bluegrass. With the exception to single applications of 294 in 2018, all dazomet treatments allowed for greater creeping bentgrass establishment than the nontreated control. Fraise mowing cultivation may simplify the removal of surface material from large areas; however, even when combined with dazomet applied at the highest rates, it fails to provide complete annual bluegrass control.

Abstract Background Small heat shock proteins (sHSPs) are critical for plant response to biotic and abiotic stresses, especially heat stress. They have also been implicated in various aspects of plant development. However, the acting mechanisms of the sHSPs in plants, especially in perennial grass species, remain largely elusive. Results In this study, AsHSP26.8a, a novel chloroplast-localized sHSP gene from creeping bentgrass (Agrostis stolonifera L.) was cloned and its role in plant response to environmental stress was studied. AsHSP26.8a encodes a protein of 26.8 kDa. Its expression was strongly induced in both leaf and root tissues by heat stress. Transgenic Arabidopsis plants overexpressing AsHSP26.8a displayed reduced tolerance to heat stress. Furthermore, overexpression of AsHSP26.8a resulted in hypersensitivity to hormone ABA and salinity stress. Global gene expression analysis revealed AsHSP26.8a-modulated expression of heat-shock transcription factor gene, and the involvement of AsHSP26.8a in ABA-dependent and -independent as well as other stress signaling pathways. Conclusions Our results suggest that AsHSP26.8a may negatively regulate plant response to various abiotic stresses through modulating ABA and other stress signaling pathways.

Heat stress is a major concern for the growth of cool-season creeping bentgrass (Agrostis stolonifera L.). Nonetheless, there is a lack in a clear and systematic understanding of thermotolerance mechanisms for this species. This study aimed to assess heat tolerance in experimental lines and cultivars to determine important physiological and biochemical traits responsible for improved tolerance, including the use of OJIP fluorescence. Ten creeping bentgrass lines were exposed to either control (20/15 °C day/night) or high temperature (38/33 °C day/night) conditions for 35 d via growth chambers at Griffin, GA. Principal component analysis and clustering analysis were performed to rank stress performance and divide lines into different groups according to their tolerance similarities, respectively. At the end of the trial, S11 729-10 and BTC032 were in the most thermotolerant group, followed by a group containing BTC011, AU Victory and Penncross. Crenshaw belonged to the most heat-sensitive group while S11 675-02 and Pure Eclipse were in the second most heat-sensitive group. The exceptional thermotolerance in S11 729-10 and BTC032 was associated with their abilities to maintain cell membrane stability and protein metabolism, plus minimize oxidative damages. Additionally, among various light-harvesting steps, energy trapping, dissipation and electron transport from QA to PQ were more heat-sensitive than electron transport from QA to final PSI acceptors. Along with the strong correlations between multiple OJIP parameters and other traits, it reveals that OJIP fluorescence could be a valuable tool for dissection of photosynthetic processes and identification of the critical steps responsible for photosynthetic declines, enabling a more targeted heat-stress screening. Our results indicated that variability in the level of heat tolerance and associated mechanisms in creeping bentgrass germplasm could be utilized to develop new cultivars with improved thermotolerance.

Results suggest that sand topdressing was more consistent at reducing dollar spot (Clarireedia jacksonii) in fairway turfgrass more so than rolling. This practice could be an effective cost-saving alternative to reduce frequent fungicide applications. Research was conducted from 2011 to 2014 on a simulated golf fairway and examined dollar spot severity responses in a mixed-stand of creeping bentgrass (Agrostis stolonifera) and annual bluegrass (Poa annua ssp. reptans) to sand topdressing and rolling. Treatments consisted of biweekly sand topdressing, rolling at three frequencies (one, three, or five times weekly), a control, and three replications. Infection was visually estimated. Sand topdressing significantly (P < 0.05) reduced disease up to 50% at the peak of the dollar spot activity in 2011, 2013, and 2014. Results on the effects of rolling on dollar spot were inconsistent.

Extensive stolon development and growth are superior traits for rapid establishment as well as post-stress regeneration in stoloniferous grass species. Despite the importance of those stoloniferous traits, the regulation mechanisms of stolon growth and development are largely unknown. The objectives of this research were to elucidate the effects of the reallocation of soluble sugars for energy reserves and endogenous hormone levels for cell differentiation and regeneration in regulating stolon growth of a perennial turfgrass species, creeping bentgrass (Agrostis stolonifera L.). Plants were grown in growth chambers with two CO2 concentrations: ambient CO2 concentration (400 ± 10 µmol mol−1) and elevated CO2 concentration (800 ± 10 µmol mol−1). Elevated CO2 enhanced stolon growth through increasing stolon internode number and internode length in creeping bentgrass, as manifested by the longer total stolon length and greater shoot biomass. The content of glucose, sucrose, and fructose as well as endogenous IAA were accumulated in stolon nodes and internodes but not in leaves or roots under elevated CO2 concentration. These results illustrated that the production and reallocation of soluble sugars to stolons as well as the increased level of IAA in stolon nodes and internodes could contribute to the enhancement of stolon growth under elevated CO2 in creeping bentgrass.

Dollar spot is caused by the fungus Clarireedia jacksonii and is the most common disease of golf course turfgrass in temperate climates. Oxalic acid (OA) is an important pathogenicity factor in other fungal plant pathogens, such as the dicot pathogen Sclerotinia sclerotiorum, but its role in C. jacksonii pathogenicity on monocot hosts remains unclear. Herein, we assess fungal growth, OA concentration, and pH change in potato dextrose broth (PDB) following incubation of C. jacksonii. In addition, OA production by C. jacksonii and S. sclerotiorum was compared in PDB amended with creeping bentgrass or common plant cell wall components (cellulose, lignin, pectin, or xylan). Our results show that OA production is highly dependent on the environmental pH, with twice as much OA produced at pH 7 than pH 4 and a corresponding decrease in PDB pH from 7 to 5 following 96 h of C. jacksonii incubation. In contrast, no OA was produced or changes in pH observed when C. jacksonii was incubated in PDB at a pH of 4. Interestingly, C. jacksonii increased OA production in response to PDB amended with creeping bentgrass tissue and the cell wall component xylan, a major component of grass cell walls. S. sclerotiorum produced large amounts of OA relative to C. jacksonii regardless of treatment, and no treatment increased OA production by this fungus, though pectin suppressed S. sclerotiorum’s OA production. These results suggest that OA production by C. jacksonii is reliant on host specific components within the infection court, as well as the ambient pH of the foliar environment during its pathogenic development.

Low cost and favorable handling characteristics make urea (46-0-0) a leading nitrogen source for frequent, foliar N fertilization of golf course putting greens in season. Yet few field investigations of resulting NH3 volatilization from putting greens have been directed. Meanwhile, NH3 emissions degrade air and surface water quality. Our objective was to quantify NH3 volatilization following practical, low-N rate, and foliar application of commercial urea-N fertilizers. Over the 2019 and 2020 growing seasons in University Park, PA, USA, an industrial vacuum pump, H3BO3 scrubbing flasks, and sixteen dynamic flux chambers were employed in four unique experiments to measure NH3 volatilization from creeping bentgrass putting greens (Agrostis stolonifera L. ‘Penn G2’) in the 24 h period ensuing foliar application of urea based-N at a 7.32 or 9.76 kg/ha rate. Simultaneous and replicated flux chamber trapping efficiency trials showing 35% mean NH3 recovery were used to adjust NH3 volatilization rates from treated plots. Under the duration and conditions described, 3.1 to 8.0% of conventional urea N volatilized from the putting greens as NH3. Conversely, 0.7 to 1.1% of methylol urea liquid fertilizer (60% short-chain methylene urea) or 0.7 to 2.2% of urea complimented with dicyandiamide (DCD) and N-(n-butyl) thiophosphoric triamide (NBPT) volatilized as NH3.

Cadmium (Cd) toxicity seriously affects the growth and development of plants, so studies on uptake, translocation, and accumulation of Cd in plants are crucial for phytoremediation. However, the molecular mechanism of the plant response to Cd stress remains poorly understood. The main objective of this study was to reveal differentially expressed genes (DEGs) under lower (BT2_5) and higher (BT43) Cd concentration treatments in creeping bentgrass. A total of 463,184 unigenes were obtained from creeping bentgrass leaves using RNA sequencing technology. Observation of leaf tissue morphology showed that the higher Cd concentration damages leaf tissues. Four key transcription factor (TF) families, WRKY, bZIP, ERF, and MYB, are associated with Cd stress in creeping bentgrass. Our findings revealed that these four TFs play crucial roles during the creeping bentgrass response to Cd stress. This study is mainly focused on the molecular characteristics of DEGs under Cd stress using transcriptomic analysis in creeping bentgrass. These results provide novel insight into the regulatory mechanisms of respond to Cd stress and enrich information for phytoremediation.

Creeping bentgrass (Agrostis stolonifera) is the preferred green lawn grass, with excellent turf characteristics but poor disease resistance. At present, the mechanisms of disease resistance in creeping bentgrass are poorly understood, especially the ethylene signal transduction pathway under the induced systemic resistance (ISR) response. In this study, butanediol (BDO), as a new type of disease-resistance compound, was applied to creeping bentgrass seedlings to induce the ISR response. Then, we measured ethylene production and related enzyme activities. Additionally, transcript profiling and gene identification were performed in association to ethylene signal transduction pathways. The changes of ethylene production and related enzyme 1-aminocyclopropane-1-carboxylic acid oxidase (ACO) and 1-aminocyclopropane-1-carboxylic acid synthases (ACS) activities showed significant difference at 24 h after Rhizoctonia solani inoculation among five treatments of various BDO concentrations. After 100 µmol L−1 BDO treatment, ethylene production and related enzyme activities reached their peak levels. Additionally, 208,672 unigenes of creeping bentgrass were obtained by de novo assembly. In total, 15,903 annotated unigenes were grouped into 33 canonical pathways in the KEGG (Kyoto Encyclopedia of Genes and Genomes) analysis. Among those, 1803 unigenes were classified as ‘signal transduction’. There were 6766 differentially expressed genes (DEGs) among B24 (inoculated-rhizobacteria in MS medium with 100 µmol L−1 BDO for 24 h), NB24, B72 and NB24 (no rhizobacteria in MS medium with 100 µmol L−1 BDO for 24 h) libraries, and 4,639 DEGs between B24 and B72 (inoculated-rhizobacteria in MS medium with 100 µmol L−1 BDO for 72 h) libraries, with 4489 DEGs in all three libraries. As suggested by the RT-PCR assay, the expression levels of ethylene-responsive and defense-related genes were variable among treated samples during the BDO-induced ISR responses. The expression levels of EIN, ERF, NPR1, PR3 and PR4 genes increased and reached their peaks in the first 24 h after R. solani infection in the BDO-induced ISR reaction compared with NB24 treatments. This results is consistent with the changes of important ethylene biosynthetic enzymes and ethylene concentrations during the BDO-induced ISR responses. We further found the intermediate substances for the signaling pathway, and the relationships between the expression levels of BDO-induced ISR disease-resistance genes and those of the response genes for ethylene signal pathway. Our findings present a genetic basis for systemic resistance of creeping bentgrass through transcriptomic analysis and our study provides a theoretical and practical basis for the improvement of turfgrass disease resistance and quality.

Bacterial volatile compounds (BVCs) have been reported to enhance plant growth and elicit plant defenses against fungal infection and insect damage. The objective of this study was to determine transcriptomic changes in response to synthetic BVC that could be associated with plant resistance to Rhizoctonia solani in creeping bentgrass. The 2,3-butanediol (BD) (250 µM) was sprayed on creeping bentgrass leaves grown in jam jars. The result showed that synthetic BD induced plant defense against R. solani for creeping bentgrass. Transcriptomic analysis demonstrated that more genes were repressed by BD while less showed up-regulation. BD suppressed the expression of some regular stress-related genes in creeping bentgrass, such as pheromone activity, calcium channel activity, photosystem II oxygen evolving complex, and hydrolase activity, while up-regulated defense related transcription factors (TFs), such as basic helix-loop-helix (bHLH) TFs, cysteine2-cysteine2-contans-like (C2C2-CO) and no apical meristem TFs (NAC). Other genes related to disease resistance, such as jasmonic acid (JA) signaling, leucine rich repeats (LRR)-transmembrane protein kinase, pathogen-related (PR) gene 5 receptor kinase and nucleotide binding site-leucine rich repeats (NBS-LRR) domain containing plant resistance gene (R-gene) were also significantly up-regulated. These results suggest that BD may induce changes to the plant transcriptome in induced systemic resistance (ISR) pathways.