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Zoysia japonica is an important warm-season turfgrass used worldwide. The decreased aesthetic quality and functionality during leaf senescence hamper its further utilization. However, information about the transcriptional mechanism and genes involved in leaf senescence in Z. japonica needs to be more extensive. Therefore, to better understand leaf senescence in Z. japonica, we investigated the integrated analysis of chlorophyll fluorescence test (JIP-test) and RNA sequencing (RNA-seq) of mature and senescent leaves. First, we identified 22,049 genes, of which 4038 were differentially expressed genes (DEGs). The results for gene expression profiles were evaluated using quantitative real-time PCR. A total of 2515 genes have homologous genes in other plants. The matched known-function SAGs are mainly involved in chlorophyll degradation and plant hormone response. A total of 539 differentially expressed transcription factor genes, including AP2/ERF-ERF, NAC, WRKY, bHLH, and MYB, were identified to be associated with leaf senescence. Next, senescence represses chlorophyll biosynthesis while upregulating chlorophyll degradation. Senescence harms the integrity and functionality of PSII, PSI, and the intersystem electron transport chain. In addition, IAA biosynthesis was inhibited, whereas ABA and ET biosynthesis were activated in leaf senescence, and senescence activates signal transduction of IAA, ABA, and ET. These findings add to our understanding of the regulatory mechanism of leaf senescence. The senescence-associated genes are candidate targets for providing new insight into leaf senescence modeling in Z. japonica. They provided a theoretical foundation to reveal the functions of senescence-associated genes and chlorophyll catabolic genes involved in leaf senescence. [ABSTRACT FROM AUTHOR]

The degradation of chlorophyll is of great significance to plant growth. The chlorophyll b reductase NOL (NYC1-like) is in charge of catalyzing the degradation of chlorophyll b and maintaining the stability of the photosystem. However, the molecular mechanisms of NOL-mediated chlorophyll degradation, senescence, and photosynthesis and its functions in other metabolic pathways remain unclear, especially in warm-season turfgrass. In this study, ZjNOL was cloned from Zoysia japonica. It is highly expressed in senescent leaves. Subcellular localization investigation showed ZjNOL is localized in the chloroplast and the bimolecular fluorescence complementation (BiFC) results proved ZjNOL interacts with ZjNYC1 in vivo. ZjNOL promoted the accumulation of abscisic acid (ABA) and carbohydrates, and the increase of SAG14 at the transcriptional level. ZjNOL simultaneously led to the excessive accumulation of reactive oxygen species (ROS), the activation of antioxidant enzymes, and the generation of oxidative stress, which in turn accelerated senescence. Chlorophyll fluorescence assay (JIP-test) analysis showed that ZjNOL inhibited photosynthetic efficiency mainly through damage to the oxygen-evolving complex. In total, these results suggest that ZjNOL promotes chlorophyll degradation and senescence and negatively affects the integrity and functionality of the photosystem. It could be a valuable candidate gene for genome editing to cultivate Z. japonica germplasm with prolonged green period and improved photosynthesis efficiency. [ABSTRACT FROM AUTHOR]

Pheophytinase (PPH), the phytol hydrolase, plays important roles in chlorophyll degradation. Nevertheless, little attention has been paid to the PPHs in warm-season grass species; neither its detailed function in photosynthesis has been systematically explored to date. In this study, we isolated ZjPPH from Zoysia japonica , an excellent warm-season turfgrass species. Quantitative real-time PCR analysis and promoter activity characterization revealed that the expression of ZjPPH could be induced by senescence, ABA, and dark induction. Subcellular localization observation proved that ZjPPH was localized in the chloroplasts. Overexpression of ZjPPH accelerated the chlorophyll degradation and rescued the stay-green phenotype of the Arabidopsis pph mutant. Moreover, ZjPPH promoted senescence with the accumulation of ABA and soluble sugar contents, as well as the increased transcriptional level of SAG12 and SAG14. Transmission electron microscopy investigation revealed that ZjPPH caused the decomposition of chloroplasts ultrastructure in stable transformed Arabidopsis. Furthermore, chlorophyll a fluorescence transient measurement analysis suggested that ZjPPH suppressed photosynthesis efficiency by mainly suppressing both photosystem II (PSII) and photosystem I (PSI). In conclusion, ZjPPH plays an important role in chlorophyll degradation and senescence. It could be a valuable target for genetic editing to cultivate new germplasms with stay-green performance and improved photosynthetic efficiency. [ABSTRACT FROM AUTHOR]