A Survey of Enhanced Cold Tolerance and Low-Temperature-Induced Anthocyanin Accumulation in a Novel Zoysia japonica Biotype

Yang

Plant Biotechnology Journal

et al. 2023

SummaryLow temperature is the main environmental factor affecting the yield, quality and geographical distribution of crops, which significantly restricts development of the fruit industry. The NAC (NAM, ATAF1/2 and CUC2) transcription factor (TF) family is involved in regulating plant cold tolerance, but the mechanisms underlying these regulatory processes remain unclear. Here, the NAC TF MdNAC104 played a positive role in modulating apple cold tolerance. Under cold stress, MdNAC104‐overexpressing transgenic plants exhibited less ion leakage and lower ROS (reactive oxygen species) accumulation, but higher contents of osmoregulatory substances and activities of antioxidant enzymes. Transcriptional regulation analysis showed that MdNAC104 directly bound to the MdCBF1 and MdCBF3 promoters to promote expression. In addition, based on combined transcriptomic and metabolomic analyses, as well as promoter binding and transcriptional regulation analyses, we found that MdNAC104 stimulated the accumulation of anthocyanin under cold conditions by upregulating the expression of anthocyanin synthesis‐related genes, including MdCHS‐b, MdCHI‐a, MdF3H‐a and MdANS‐b, and increased the activities of the antioxidant enzymes by promoting the expression of the antioxidant enzyme‐encoding genes MdFSD2 and MdPRXR1.1. In conclusion, this study revealed the MdNAC104 regulatory mechanism of cold tolerance in apple via CBF‐dependent and CBF‐independent pathways.

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

MdNAC104 positively regulates apple cold tolerance via CBF‐dependent and CBF‐independent pathways

Yang

Plant Biotechnology Journal

et al. 2023

“…Zoysiagrass is primarily used in golf courses and lawns because of its fine texture, fast spreading and high density. Efforts are ongoing to breed for and select cultivars that have excellent turf quality, late winter dormancy, early spring green‐up and cold hardiness (Jin et al, 2022; Sun et al, 2020). The stay‐green ( SGR ) phenotype is a highly desirable trait in zoysiagrasses; however, the trait is lacking in these genetic resources.…”

Section: Discussionmentioning

confidence: 99%

Agrobacterium‐mediated transformation via establishment of stable tissue culture system inZoysia matrella(L.) Merrill ‘Wakaba’

Gondo

Ushiyama

et al. 2022

Grassland Science

Zoysia matrella (L.) Merrill is a perennial C4 warm-season turfgrass grown for landscapes, golf courses, sports fields and recreation parks. To create a new breeding method by genome editing, an efficient genetic transformation system is essential. In this study, we report the efficient protocol of Agrobacterium-mediated transformation through the establishment of a stable tissue culture system for Z. matrella 'Wakaba'.The embryogenic callus was induced from shoot apices of nodal segments incubated in Murashige and Skoog (MS) medium containing 2 mg/l 2,4-dichlorophnoxyacetic acid (2,4-D), 0.1 mg/l 6-benzylaminopurine (BAP) and 5 μM CuSO 4 (MS-DBC).Repeated subculture of compact high-quality callus in MS-DBC medium produced a highly regenerative callus with dense pre-embryogenic clusters, and it was used as a transformation target. Agrobacterium strain EHA105 harboring pANIC8B vector containing the β-glucuronidase gene (GUS) and hygromycin B phosphotransferase gene was used. Agrobacterium-infected calli were cocultured for 5 days with 100 μM acetosyringone and then subjected to selection pressure of 50 mg/l hygromycin. This optimized protocol yielded transformation efficiencies of up to 6.6%. Southern blot analysis verified one to three copies of the GUS gene in different independent transgenic plants. All transgenic plants were morphologically normal, and the GUS expressions were stable. Our optimized in vitro and transgenic system will facilitate the new breeding technology of genome editing in zoysiagrass.

“…Several genomic and transcriptomic studies have been published on Z. japonica along with their datasets (Ahn et al, 2015; Wei et al, 2015; Xie et al, 2015; Li et al, 2018; Wang et al, 2020; Guan et al, 2022; Jin et al, 2022). These studies aim to explore the genetic makeup and gene expression patterns of Z. japonica under different conditions.…”

Section: Introductionmentioning

confidence: 99%

Unveiling unique alternative splicing responses to low temperature in Zoysia japonica through ZjRTD1.0, a high‐quality reference transcript dataset

Wu,

He,

Wang

et al. 2024

Physiologia Plantarum

Self Cite

Inadequate reference databases in RNA‐seq analysis can hinder data utilization and interpretation. In this study, we have successfully constructed a high‐quality reference transcript dataset, ZjRTD1.0, for Zoysia japonica, a widely‐used turfgrass with exceptional tolerance to various abiotic stress, including low temperatures and salinity. This dataset comprises 113,089 transcripts from 57,143 genes. BUSCO analysis demonstrates exceptional completeness (92.4%) in ZjRTD1.0, with reduced proportions of fragmented (3.3%) and missing (4.3%) orthologs compared to prior datasets. ZjRTD1.0 enables more precise analyses, including transcript quantification and alternative splicing assessments using public datasets, which identified a substantial number of differentially expressed transcripts (DETs) and differential alternative splicing (DAS) events, leading to several novel findings on Z. japonica's responses to abiotic stresses. First, spliceosome gene expression influenced alternative splicing significantly under abiotic stress, with a greater impact observed during low‐temperature stress. Then, a significant positive correlation was found between the number of differentially expressed genes (DEGs) encoding protein kinases and the frequency of DAS events, suggesting the role of protein phosphorylation in regulating alternative splicing. Additionally, our results suggest possible involvement of serine/arginine‐rich (SR) proteins and heterogeneous nuclear ribonucleoproteins (hnRNPs) in generating inclusion/exclusion isoforms under low‐temperature stress. Furthermore, our investigation revealed a significantly enhanced overlap between DEGs and differentially alternatively spliced genes (DASGs) in response to low‐temperature stress, suggesting a unique co‐regulatory mechanism governing transcription and splicing in the context of low‐temperature response. In conclusion, we have proven that ZjRTD1.0 will serve as a reliable and useful resource for future transcriptomic analyses in Z. japonica.