Molecular responses reveal that two glutathione S-transferase CsGSTU8s contribute to detoxification of glyphosate in tea plants (Camellia sinensis).

Tea plant (Camellia sinensis) is an important economical crop that frequently suffers from various herbicides, especially glyphosate. However, the molecular responses and regulatory mechanisms of glyphosate stress in tea plants remain poorly understood. Here, we reported a transcriptome dataset and identified large number of differentially expressed genes (DEGs) under glyphosate exposure. Next, two glutathione S-transferase genes (CsGSTU8-1 and CsGSTU8-2) that upregulated significantly were screened as candidate genes. Tissue-specific expression patterns showed that both CsGSTU8-1 and CsGSTU8-2 had extremely high expression levels in the roots and were predominantly localized in the nucleus and plasma membrane based on subcellular localization. Both were significantly upregulated at different time points under various stressors, including drought, cold, salt, pathogen infections, and SA treatments. An enzymatic activity assay showed that CsGSTU8-1 catalyzes the conjugation of glutathione with 2,4-dinitrochlorobenzene (CDNB). Functional analysis in yeast verified that the two genes significantly contributed to the detoxification of glyphosate, and CsGSTU8-1 had a stronger role in detoxification than CsGSTU8-2. Taken together, these findings provide insights into the molecular responses of tea plants to glyphosate and the functions of CsGSTU8s in glyphosate detoxification, which can be used as a promising genetic resource for improving herbicide resistance in tea cultivars.

The Laccase Family Gene CsLAC37 Participates in Resistance to Colletotrichum gloeosporioides Infection in Tea Plants.

Fungal attacks have become a major obstacle in tea plantations. Colletotrichum gloeosporioides is one of the most devastating fungal pathogens in tea plantations that can severely affect tea yield and quality. However, the molecular mechanism of resistance genes involved in anthracnose is still largely unknown in tea plants. Here, we found that the laccase gene CsLAC37 was involved in the response to fungal infection based on a transcriptome analysis. The full-length CDS of CsLAC37 was cloned, and its protein sequence had the closest relationship with the Arabidopsis AtLAC15 protein compared to other AtLACs. Tissue-specific expression analysis showed that CsLAC37 had higher expression levels in mature leaves and stems than in the other tissues. Subcellular localization showed that the CsLAC37 protein was predominantly localized in the cell membrane. The expression levels of CsLAC37 were upregulated at different time points under cold, salt, SA, and ABA treatments. qRT-PCR confirmed that CsLAC37 responded to both Pestalotiopsis-like species and C. gloeosporioides infections. Functional validation showed that the hydrogen peroxide (H2O2) content increased significantly, and POD activity decreased in leaves after antisense oligonucleotide (AsODN) treatment compared to the controls. The results demonstrated that CsLAC37 may play an important role in resistance to anthracnose, and the findings provide a theoretical foundation for molecular breeding of tea varieties with resistance to fungal diseases.

Integrated physiological, metabolite and proteomic analysis reveal the glyphosate stress response mechanism in tea plant (Camellia sinensis).

Glyphosate residues can tremendously impact the physiological mechanisms of tea plants, thus threatening tea security and human health. Herein, integrated physiological, metabolite, and proteomic analyses were performed to reveal the glyphosate stress response mechanism in tea plant. After exposure to glyphosate (≥1.25 kg ae/ha), the leaf ultrastructure was damaged, and chlorophyll content and relative fluorescence intensity decreased significantly. The characteristic metabolites catechins and theanine decreased significantly, and the 18 volatile compounds content varied significantly under glyphosate treatments. Subsequently, tandem mass tags (TMT)-based quantitative proteomics was employed to identify the differentially expressed proteins (DEPs) and to validate their biological functions at the proteome level. A total of 6287 proteins were identified and 326 DEPs were screened. These DEPs were mainly catalytic, binding, transporter and antioxidant active proteins, involved in photosynthesis and chlorophyll biosynthesis, phenylpropanoid and flavonoid biosynthesis, sugar and energy metabolism, amino acid metabolism, and stress/defense/detoxification pathway, etc. A total of 22 DEPs were validated by parallel reaction monitoring (PRM), demonstrating that the protein abundances were consistent between TMT and PRM data. These findings contribute to our understanding of the damage of glyphosate to tea leaves and molecular mechanism underlying the response of tea plants to glyphosate.