ABSTRACT
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.
Subject(s)
Abscisic Acid/pharmacology , Camellia sinensis/growth & development , Monomeric GTP-Binding Proteins/genetics , Monomeric GTP-Binding Proteins/metabolism , Camellia sinensis/drug effects , Camellia sinensis/enzymology , Camellia sinensis/genetics , Cell Membrane/metabolism , Cell Nucleus/metabolism , Droughts , Gene Expression Regulation, Plant/drug effects , Open Reading Frames , Plant Proteins/genetics , Plant Proteins/metabolism , Salt Stress , Signal Transduction/drug effects , Stress, PhysiologicalABSTRACT
Both UV and blue light have been reported to regulate the biosynthesis of flavonoids in tea plants; however, the respective contributions of the corresponding regions of sunlight are unclear. Additionally, different tea cultivars may respond differently to altered light conditions. We investigated the responses of different cultivars ('Longjing 43', 'Zhongming 192', 'Wanghai 1', 'Jingning 1' and 'Zhonghuang 2') to the shade treatments (black and colored nets) regarding the biosynthesis of flavonoids. For all cultivars, flavonol glycosides showed higher sensitivity to light conditions compared with catechins. The levels of total flavonol glycosides in the young shoots of different tea cultivars decreased with the shade percentages of polyethylene nets increasing from 70% to 95%. Myricetin glycosides and quercetin glycosides were more sensitive to light conditions than kaempferol glycosides. The principal component analysis (PCA) result indicated that shade treatment greatly impacted the profiles of flavonoids in different tea samples based on the cultivar characteristics. UV is the crucial region of sunlight enhancing flavonol glycoside biosynthesis in tea shoots, which is also slight impacted by light quality according to the results of the weighted correlation network analysis (WGCNA). This study clarified the contributions of different wavelength regions of sunlight in a field experiment, providing a potential direction for slightly bitter and astringent tea cultivar breeding and instructive guidance for practical field production of premium teas based on light regimes.
Subject(s)
Camellia sinensis/growth & development , Flavonoids/biosynthesis , Glycosides/biosynthesis , Plant Shoots/growth & development , Camellia sinensis/radiation effects , Flavonoids/chemistry , Flavonoids/radiation effects , Glycosides/radiation effects , Kaempferols/chemistry , Plant Shoots/radiation effects , Principal Component Analysis , Sunlight , Ultraviolet RaysABSTRACT
Tea plant (Camellia sinensis (O.) Kuntze) can survive from high levels of aluminum (Al) in strongly acidic soils. However, the mechanism driving its tolerance to Al, the predominant factor limiting plant growth in acid condition, is still not fully understood. Here, two-year-old rooted cuttings of C. sinensis cultivar 'Longjingchangye' were used for Al resistance experiments. We found that the tea plants grew better in the presence of 0.4 mM Al than those grew under lower concentration of Al treatments (0 and 0.1 mM) as well as higher levels treatment (2 and 4 mM), confirming that appropriate Al increased tea plant growth. Hematoxylin staining assay showed that the apical region was the main accumulator in tea plant root. Subsequently, immunolocalization of pectins in the root tip cell wall showed a rise in low-methyl-ester pectin levels and a reduction of high-methyl-ester pectin content with the increasing Al concentration of treatments. Furthermore, we observed the increased expressions of C. sinensis pectin methylesterase (CsPME) genes along with the increasing de-esterified pectin levels during response to Al treatments. Additionally, the levels of organic acids increased steadily after treatment with 0.1, 0.4 or 2 mM Al, while they dropped after treatment with 4 mM Al. The organic acids secretion from root followed a similar trend. Similarly, a gradual increase in malate dehydrogenase (MDH), citrate synthase (CS) and glycolate oxidase (GO) enzyme activities and relevant metabolic genes expression were detected after the treatment of 0.1, 0.4 or 2 mM Al, while a sharp decrease was resulted from treatment with 4 mM Al. These results confirm that both pectin methylesterases and organic acids contribute to Al tolerance in C. sinensis.
Subject(s)
Aluminum/pharmacology , Camellia sinensis/metabolism , Meristem/metabolism , Pectins/metabolism , Plant Proteins/metabolism , Esterification/drug effectsABSTRACT
BACKGROUND: Nitric oxide (NO) functions as a critical signaling molecule in the low-temperature stress responses in plants, including polarized pollen tube growth in Camellia sinensis. Despite this, the potential mechanisms underlying the participation of NO in pollen tube responses to low temperature remain unclear. Here, we investigate alterations to gene expression in C. sinensis pollen tubes exposed to low-temperature stress and NO using RNA-Seq technology, in order to find the potential candidate genes related to the regulation of pollen tube elongation by NO under low-temperature stress. RESULTS: Three libraries were generated from C. sinensis cv. 'Longjingchangye' pollen tubes cultured at 25 °C (CsPT-CK) and 4 °C (CsPT-LT) or with 25 µM DEA NONOate (CsPT-NO). The number of unigenes found for the three biological replications were 39,726, 40,440 and 41,626 for CsPT-CK; 36,993, 39,070 and 39,439 for CsPT-LT; and 39,514, 38,298 and 39,061 for CsPT-NO. A total of 36,097 unique assembled and annotated sequences from C. sinensis pollen tube reads were found in a BLAST search of the following databases: NCBI non-redundant nucleotide, Swiss-prot protein, Kyoto Encyclopedia of Genes and Genomes, Cluster of Orthologous Groups of proteins, and Gene Ontology. The absolute values of log2Ratio > 1 and probability > 0.7 were used as the thresholds for significantly differential gene expression, and 766, 497 and 929 differentially expressed genes (DEGs) were found from the comparison analyses of the CK-VS-LT, CK-VS-NO and LT-VS-NO libraries, respectively. Genes related to metabolism and signaling pathways of plant hormones, transcription factors (TFs), vesicle polarized trafficking, cell wall biosynthesis, the ubiquitination machinery of the ubiquitin system and species-specific secondary metabolite pathways were mainly observed in the CK-VS-LT and CK-VS-NO libraries. CONCLUSION: Differentially expressed unigenes related to the inhibition of C. sinensis pollen tube growth under low temperature and NO are identified in this study. The transcriptomic gene expression profiles present a valuable genomic tool to improve studying the molecular mechanisms underlying low-temperature tolerance in pollen tube.
Subject(s)
Camellia sinensis/genetics , Camellia sinensis/metabolism , Cold Temperature , Nitric Oxide/metabolism , Pollen Tube/genetics , Pollen Tube/metabolism , Transcriptome , Cluster Analysis , Computational Biology/methods , Gene Expression Profiling , Gene Expression Regulation, Plant , Gene Ontology , High-Throughput Nucleotide Sequencing , Molecular Sequence Annotation , Pollen Tube/growth & developmentABSTRACT
KEY MESSAGE: Overexpression of CsHis in tobacco promoted chromatin condensation, but did not affect the phenotype. It also conferred tolerance to low-temperature, high-salinity, ABA, drought and oxidative stress in transgenic tobacco. H1 histone, as a major structural protein of higher-order chromatin, is associated with stress responses in plants. Here, we describe the functions of the Camellia sinensis H1 Histone gene (CsHis) to illustrate its roles in plant responses to stresses. Subcellular localization and prokaryotic expression assays showed that the CsHis protein is localized in the nucleus, and its molecular size is approximately 22.5 kD. The expression levels of CsHis in C. sinensis leaves under various conditions were investigated by qRT-PCR, and the results indicated that CsHis was strongly induced by various abiotic stresses such as low-temperature, high-salinity, ABA, drought and oxidative stress. Overexpression of CsHis in tobacco (Nicotiana tabacum) promoted chromatin condensation, while there were almost no changes in the growth and development of transgenic tobacco plants. Phylogenetic analysis showed that CsHis belongs to the H1C and H1D variants of H1 histones, which are stress-induced variants and not the key variants required for growth and development. Stress tolerance analysis indicated that the transgenic tobacco plants exhibited higher tolerance than the WT plants upon exposure to various abiotic stresses; the transgenic plants displayed reduced wilting and senescence and exhibited greater net photosynthetic rate (Pn), stomatal conductance (Gs) and maximal photochemical efficiency (Fv/Fm) values. All the above results suggest that CsHis is a stress-induced gene and that its overexpression improves the tolerance to various abiotic stresses in the transgenic tobacco plants, possibly through the maintenance of photosynthetic efficiency.