Kentucky bluegrass (Poa pratensis L.) belong to Gramineae and is widely used in lawns, golf courses, landscapes, and sport fields as a prominent cool-season grass. Gene expression patterns during different stages of plant development can provide clues toward the understanding of its biological functions. The selection and validation of reference genes are the first steps in any real-time quantitative PCR gene expression study. Therefore, suitable reference genes are necessary for obtaining reliable results in real-time quantitative PCR analyses of Kentucky bluegrass. In the present study, 9 candidate reference genes were chosen, and their expression stability in the leaves and roots of Kentucky bluegrass under different stresses (drought, salt, heat, and cold) were evaluated using the GeNorm, NormFinder, BestKeeper, and RefFinder programs. The results showed that the expression stability of the candidate reference genes was dependent on the experimental conditions. The combination of SAM with GAPDH was the most stable in leaves under salt stress and cold stress, while TUB combined with ACT or GAPDH was stable in roots under salt or cold stress, respectively. ACT and SAM maintained stable expression in drought-treated leaves, and GAPDH combined with ACT was stable in drought-treated roots. SAM and TUB exhibited stable expression in heat-treated leaves. ACT and RPL were stable in heat-treated roots. In addition, the expression patterns of PpFEH in response to drought and cold stress were used to confirm the reliability of the selected reference genes, indicating that the use of an inappropriate reference gene as the internal control will cause erroneous results. This work is the first study on the expression stability of reference genes in Kentucky bluegrass and will be particularly useful in the selection of stress-tolerance genes and the identification of the molecular mechanisms conferring stress tolerance in this species.

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Drought stress occurs frequently and severely as a result of global climate change, and it exerts serious effects on plants. 5-Aminolevulinic acid (5-ALA) plays a crucial role in conferring abiotic stress tolerance in plants. To enhance the drought tolerance of turfgrass and investigate the effects of 5-ALA on antioxidant metabolism and gene expression under drought stress conditions, exogenous 5-ALA was applied by foliar spraying before Kentucky bluegrass (Poa pratensis L.) seedlings were exposed to drought [induced by 10% polyethylene glycol (PEG)] stress for 20 days. 5-ALA pretreatment increased turf quality (TQ) and leaf relative water content (RWC) while reducing reactive oxygen species (ROS) production including HO content and O generation rate, lipoxygenase (LOX) activity, and malondialdehyde (MDA) content under drought stress. 5-ALA pretreatment maintained ascorbate (AsA) and glutathione (GSH) contents and the ASA/DHA and GSH/GSSG ratios at high levels, and it enhanced the activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), glutathione peroxidase (GPX), dehydroascorbate reductase (DHAR), and glutathione reductase (GR), which are crucial for scavenging drought-induced ROS. In addition, 5-ALA upregulated the relative expression levels of Cu/ZnSOD, APX, GPX, and DHAR but downregulated those of CAT and GR under drought stress. These results indicated that the application of 5-ALA might improve turfgrass quality and promote drought tolerance in Kentucky bluegrass through reducing oxidative damage and increasing non-enzyme antioxidant levels and antioxidant enzyme activity at transcriptional and posttranscriptional levels.

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.

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