Transcriptome analysis reveals the roles of phytohormone signaling in tea plant (Camellia sinensis L.) flower development.

BACKGROUND: Tea plant (Camellia sinensis (L.) O. Kuntze) is an important economic tea crop, but flowering will consume a lot of nutrients of C. sinensis, which will seriously affect the nutritional growth of C. sinensis. However, there are few studies on the development mechanism of C. sinensis flower, and most studies focus on a single C. sinensis cultivar. RESULTS: Here, we identified a 92-genes' C. sinensis flower development core transcriptome from the transcriptome of three C. sinensis cultivars ('BaiYe1', 'HuangJinYa' and 'SuChaZao') in three developmental stages (bud stage, white bud stage and blooming stage). In addition, we also reveal the changes in endogenous hormone contents and the expression of genes related to synthesis and signal transduction during the development of C. sinensis flower. The results showed that most genes of the core transcriptome were involved in circadian rhythm and autonomous pathways. Moreover, there were only a few flowering time integrators, only 1 HD3A, 1 SOC1 and 1 LFY, and SOC1 played a dominant role in the development of C. sinensis flower. Furthermore, we screened out 217 differentially expressed genes related to plant hormone synthesis and 199 differentially expressed genes related to plant hormone signal transduction in C. sinensis flower development stage. CONCLUSIONS: By constructing a complex hormone regulation network of C. sinensis flowering, we speculate that MYC, FT, SOC1 and LFY play key roles in the process of endogenous hormones regulating C. sinensis flowering development. The results of this study can a provide reference for the further study of C. sinensis flowering mechanism.

Multiomic Analyses Reveal the Effects of Supplementing Phytosterols on the Metabolic Function of the Rumen Microbiota in Perinatal Cows.

Phytosterols are natural steroids in plants, possessing bioactivities that could modify gut microbes. This experiment aimed to evaluate the effects of feeding phytosterols on the community structures and metabolic functions of the rumen microbiota in perinatal cows. Perinatal cows were supplied with 0 mg (control) or 200 mg (treatment) phytosterols per day. Multiomic analyses were used to analyze the community structures and metabolic functions of rumen microbiota. Results showed that dietary phytosterols increased the copy number of total ruminal bacteria, the concentration of microbial crude protein, and the molar percentage of propionate in the rumen of perinatal cows but had no effects on the alpha diversity of ruminal bacteria. However, they enriched three genera (i.e., Fibrobacter) and seven species (i.e., Fibrobacter succinogenes) within active ruminal bacteria. Metatranscriptomic and metabolomic analyses revealed that dietary phytosterols enhanced the pathway of glycolysis and the family of glycoside hydrolase 13 but depressed the citrate cycle and pyruvate metabolism and several pathways of amino acid biosynthesis. In conclusion, dietary addition of phytosterols improved the growth of ruminal bacteria and changed rumen fermentation by modifying the rumen microbiome and the energy metabolism pathways, which would be beneficial for the energy utilization of perinatal cows. IMPORTANCE Perinatal cows suffer serious physiological stress and energy deficiency. Phytosterols have bioactive functions for gut microbes. However, little knowledge is available on their effects on rumen microbiota and rumen fermentation. Results of the present experiment revealed that dietary supplementation of phytosterols could improve the growth of ruminal bacteria and changed the rumen fermentation to provide more glycogenetic precursors for the perinatal cows by modifying the ruminal bacteria community and altering the energy metabolism pathways of the rumen microbiota. These findings suggest that dietary supplementation of phytosterols would be beneficial for perinatal cows suffering from a negative energy balance.

Camellia sinensis small GTPase gene (CsRAC1) involves in response to salt stress, drought stress and ABA signaling pathway.

RAC/ROP gene (RACs) is a plant-specific small GTPases. RACs play an irreplaceable role in the tissue dynamics of cytoskeleton, vesicle transport and hormone signal transmission in plants. In the present study, a novel gene from RACs family, CsRAC1, was identified from tea [Camellia sinensis (L.) O. Kuntze]. CsRAC1 contained a 591-bp open reading frame and encoded a putative protein of 197 amino acids. Subcellular localization analysis in leaves of transgenic tobacco and root tips of Arabidopsis thaliana showed that CsRAC1 targeted the nucleus and cell membrane. The expression of CsRAC1 induced by abiotic stresses such as cold, heat, drought, salt and abscisic acid has also been verified by RT-qPCR. Further verification of biological function of CsRAC1 showed that overexpression of CsRAC1 increased the sensitivity of A. thaliana to salt stress, improved the tolerance of mature A. thaliana to drought stress, and enhanced the inhibition of ABA on seed germination of A. thaliana. In addition, the antioxidant system regulated by CsRAC1 mainly worked in mature A. thaliana. The results indicate that CsRAC1 is involved in the response of C. sinensis to salt, drought stress and ABA signaling pathway.

The performance of water-soluble fluoride transformation in soil-tea-tea infusion chain system and the potential health risk assessment.

BACKGROUND: Water-soluble fluoride (WS-F) can be absorbed directly by tea plants from soil and comprises a major source of dietary F in tea consumers. To reveal the WS-F accumulation in tea leaves and assess WS-F health risks, 70 sets of samples including tea leaves at three maturity stages and corresponding topsoil were collected from Xinyang, China. The WS-F contents in tea samples and pH values in soil samples were determined. RESULTS: The contents of WS-F in tea leaves exhibited a positive correlation with leaf maturity. The contents of WS-F in tea leaves showed a positive correlation with WS-F contents in the soil as the soil pH value exceeds 5. All the bud with two leaves samples, 84.29% of the third to sixth leaves samples, and 78.57% mature leaves samples in 5-min infusion tend to be no health threat. The leaching characteristics of WS-F from tea leaves were influenced by the leaf maturity and soaking time. CONCLUSION: Taking measures to control pH and WS-F concentration of plantations soil, as well as drinking tea infusion made from young leaves or reducing soaking time could decrease the WS-F health risk. © 2021 Society of Chemical Industry.

Identification and response analysis of xyloglucan endotransglycosylase/hydrolases (XTH) family to fluoride and aluminum treatment in Camellia sinensis.

BACKGROUND: Xyloglucan endotransglycosylase/hydrolases (XTH) can disrupt and reconnect the xyloglucan chains, modify the cellulose-xyloglucan complex structure in the cell wall to reconstruct the cell wall. Previous studies have reported that XTH plays a key role in the aluminum (Al) tolerance of tea plants (Camellia sinensis), which is a typical plant that accumulates Al and fluoride (F), but its role in F resistance has not been reported. RESULTS: Here, 14 CsXTH genes were identified from C. sinensis and named as CsXTH1-14. The phylogenetic analysis revealed that CsXTH members were divided into 3 subclasses, and conserved motif analysis showed that all these members included catalytic active region. Furthermore, the expressions of all CsXTH genes showed tissue-specific and were regulated by Al3+ and F- treatments. CsXTH1, CsXTH4, CsXTH6-8 and CsXTH11-14 were up-regulated under Al3+ treatments; CsXTH1-10 and CsXTH12-14 responded to different concentrations of F- treatments. The content of xyloglucan oligosaccharide determined by immunofluorescence labeling increased to the highest level at low concentrations of Al3+ or F- treatments (0.4 mM Al3+ or 8 mg/L F-), accompanying by the activity of XET (Xyloglucan endotransglucosylase) peaked. CONCLUSION: In conclusion, CsXTH activities were regulated by Al or F via controlling the expressions of CsXTH genes and the content of xyloglucan oligosaccharide in C. sinensis roots was affected by Al or F, which might finally influence the elongation of roots and the growth of plants.

MicroRNA Omics Analysis of Camellia sinesis Pollen Tubes in Response to Low-Temperature and Nitric Oxide.

Nitric oxide (NO) as a momentous signal molecule participates in plant reproductive development and responds to various abiotic stresses. Here, the inhibitory effects of the NO-dominated signal network on the pollen tube growth of Camellia sinensis under low temperature (LT) were studied by microRNA (miRNA) omics analysis. The results showed that 77 and 71 differentially expressed miRNAs (DEMs) were induced by LT and NO treatment, respectively. Gene ontology (GO) analysis showed that DEM target genes related to microtubules and actin were enriched uniquely under LT treatment, while DEM target genes related to redox process were enriched uniquely under NO treatment. In addition, the target genes of miRNA co-regulated by LT and NO are only located on the cell membrane and cell wall, and most of them are enriched in metal ion binding and/or transport and cell wall organization. Furthermore, DEM and its target genes related to metal ion binding/transport, redox process, actin, cell wall organization and carbohydrate metabolism were identified and quantified by functional analysis and qRT-PCR. In conclusion, miRNA omics analysis provides a complex signal network regulated by NO-mediated miRNA, which changes cell structure and component distribution by adjusting Ca2+ gradient, thus affecting the polar growth of the C. sinensis pollen tube tip under LT.

Genome-wide identification and expression analysis of the CLC superfamily genes in tea plants (Camellia sinensis).

The voltage-gated chloride channel (CLC) superfamily is one of the most important anion channels that is widely distributed in bacteria and plants. CLC is involved in transporting various anions such as chloride (Cl-) and fluoride (F-) in and out of cells. Although Camellia sinensis is a hyper-accumulated F plant, there is no studies on the CLC gene superfamily in the tea plant. Here, 8 CLC genes were identified from C. sinensis and they were named CsCLC1-8. The structure of CsCLC genes and the proteins were not conserved; the number of exons varied from 3 to 24, and the number of transmembrane domains contained 2 to 10. Furthermore, phylogenetic analysis revealed that CsCLC4-8 in subclass I contained the typical conserved domains GxGIPE (I), GKxGPxxH (II) and PxxGxLF (III), and CsCLC1-3 in subclass II did not contain any of the three conserved residues. We measured the expression levels of CsCLCs in roots, stems and leaves to assess the responses to different concentrations of Cl- and F-. The result indicated that CsCLCs participated in subfunctionalization in response to Cl- and F-, and CsCLC1-3 was more sensitive to F- treatments than CsCLC4-8, CsCLC6 and CsCLC7 may participate in absorption and long-distance transport of Cl-.

CsFEX, a Fluoride Export Protein Gene from Camellia sinensis, Alleviates Fluoride Toxicity in Transgenic Escherichia coli and Arabidopsis thaliana.

A fluoride export gene ( CsFEX) was newly found and isolated from Camellia sinensis, and its functions in detoxifying F were investigated in transgenic Escherichia coli and Arabidopsis thaliana. CsFEX contains two crcB domains, which is the typical structure in plants. The expression of CsFEX in C. sinensis is tissue-specific and related to maturity of leaves, and its expression is significantly induced by F treatments in different tissues of C. sinensis, particularly in leaves. Additionally, the growth of C. sinensis, E. coli, and A. thaliana can all be inhibited by F treatment. However, the growth of CsFEX-overexpression E. coli was increased with lower F content under F treatment compared to the control. Similarly, the germination and growth of CsFEX-overexpression A. thaliana were enhanced with lower F content under F treatment compared to the wild type. CsFEX relieves F toxicity in the transgenic E. coli and A. thaliana by alleviating F accumulation.