CsCIPK11-Regulated Metalloprotease CsFtsH5 Mediates the Cold Response of Tea Plants.

Photosystem II repair in chloroplasts is a critical process involved in maintaining a plant's photosynthetic activity under cold stress. FtsH (filamentation temperature-sensitive H) is an essential metalloprotease that is required for chloroplast photosystem II repair. However, the role of FtsH in tea plants and its regulatory mechanism under cold stress remains elusive. In this study, we cloned a FtsH homolog gene in tea plants, named CsFtsH5, and found that CsFtsH5 was located in the chloroplast and cytomembrane. RT-qPCR showed that the expression of CsFtsH5 was increased with leaf maturity and was significantly induced by light and cold stress. Transient knockdown CsFtsH5 expression in tea leaves using antisense oligonucleotides resulted in hypersensitivity to cold stress, along with higher relative electrolyte leakage and lower Fv/Fm values. To investigate the molecular mechanism underlying CsFtsH5 involvement in the cold stress, we focused on the calcineurin B-like-interacting protein kinase 11 (CsCIPK11), which had a tissue expression pattern similar to that of CsFtsH5 and was also upregulated by light and cold stress. Yeast two-hybrid and dual luciferase (Luc) complementation assays revealed that CsFtsH5 interacted with CsCIPK11. Furthermore, the Dual-Luc assay showed that CsCIPK11-CsFtsH5 interaction might enhance CsFtsH5 stability. Altogether, our study demonstrates that CsFtsH5 is associated with CsCIPK11 and plays a positive role in maintaining the photosynthetic activity of tea plants in response to low temperatures.

The New Insight into the Effects of Different Fixing Technology on Flavor and Bioactivities of Orange Dark Tea.

Peach leaf orange dark tea (ODT) is a fruity tea made by removing the pulp from peach leaf orange and placing dry Qingzhuan tea into the husk, followed by fixing them together and drying. Since the quality of traditional outdoor sunlight fixing (SL) is affected by weather instability, this study explored the feasibility of two new fixing methods, including hot air fixing (HA) and steam fixing (ST). Results showed that fixing method had a great impact on ODT shape, aroma, and taste. Compared with SL and ST, HA endowed ODT with higher fruit aroma, mellow taste, better coordination, and higher sensory evaluation score. Physical-chemical composition analysis showed that SL-fixed orange peel was higher than HA- or ST-fixed peel in the content of polyphenols, flavonoids, soluble protein, hesperidin and limonin, while HA has a higher content of volatile substances and contains more alcohols, aldehydes and ketones, and acid and esters than ST and SL. Activity analysis showed that HA was superior to ST or SL in comprehensive antioxidant activity and inhibitory activity against α-glucosidase. Comprehensive results demonstrated that HA has better performance in improving ODT quality and can replace the traditional SL method in production.

Genome-wide identification of PME gene family and expression of candidate genes associated with aluminum tolerance in tea plant (Camellia sinensis).

BACKGROUND: The major aluminum (Al) detoxication mechanism of tea plant (Camellia sinensis), as an Al hyperaccumulator plant, is the fixation of almost 70% of Al in the cell walls. Pectin is the primary constituent of cell walls, a degree of methylation of pectin polysaccharides regulated by the pectin methylesterase (PME) genes can greatly affect the Al binding capacity. The knowledge on PME gene family in tea plant is still poor. RESULTS: We identified 66 (CsPME1-CsPME66) PME genes from C. sinensis genome. We studied their protein characterization, conserved motifs, gene structure, systematic evolution and gene expression under Al treatments, to establish a basis for in-depth research on the function of PMEs in tea plant. Gene structures analysis revealed that the majority of PME genes had 2-4 exons. Phylogenetic results pointed out that the PME genes from the same species displayed comparatively high sequence consistency and genetic similarity. Selective pressure investigation suggested that the Ka/Ks value for homologous genes of PME family was less than one. The expression of CsPMEs under three Al concentration treatments was tissue specific, eight PME genes in leaves and 15 in roots displayed a trend similar to of the Al contents and PME activities under Al concentration treatments, indicating that the degree of pectin de-esterification regulated by PME was crucial for Al tolerance of tea plant. CONCLUSIONS: Sixty-six CsPME genes were identified for the first time in tea plant. The genome-wide identification, classification, evolutionary and transcription analyses of the PME gene family provided a new direction for further research on the function of PME gene in Al tolerance of tea plant.

Identification of aroma-active compounds responsible for the floral and sweet odors of Congou black teas using gas chromatography-mass spectrometry/olfactometry, odor activity value, and chemometrics.

BACKGROUND: Floral and sweet odors are two typical characteristic aromas of Congou black tea, but their aroma-active compounds are still unclear. Characterizing the key aroma-active compounds can provide a theoretical foundation for the practical aroma quality evaluation of Congou black tea and directional processing technology of high-quality black tea with floral or sweet odors. Gas chromatography-olfactometry (GC-O) combined with odor activity value (OAV) is often used to screen key aroma-active substances, but the interaction between aroma components and their impact on the overall sensory quality is ignored. Therefore, in this study, OAV combined with variable importance in projection (VIP) and Spearman correlation analysis (SCA) were used to characterize the aroma-active components of Congou black teas with floral and sweet odors. RESULTS: Eighty-five volatiles were identified in these samples using gas chromatography-mass spectrometry (GC-MS). Twenty-three compounds were identified as potential markers for the floral and sweet odors of Congou black teas from orthogonal partial least squares discriminant analysis (OPLS-DA). Eighteen compounds were selected as candidate aroma compounds based on GC-O analysis and OAV calculations. In addition, 26 compounds were screened as crucial aroma compounds based on SCA. Finally, 19 compounds were evaluated as key aroma compounds by the comprehensive evaluation of VIP, OAV, and SCA. Terpenoids are the main active compounds that contribute to the floral odor of Congou black tea, whereas aldehydes are the key compounds for the sweet odor. CONCLUSION: The proposed method can effectively screen the aroma-active compounds and can be used for comprehensive quality control of products. © 2022 Society of Chemical Industry.

Biochemical characterization of specific Alanine Decarboxylase (AlaDC) and its ancestral enzyme Serine Decarboxylase (SDC) in tea plants (Camellia sinensis).

BACKGROUND: Alanine decarboxylase (AlaDC), specifically present in tea plants, is crucial for theanine biosynthesis. Serine decarboxylase (SDC), found in many plants, is a protein most closely related to AlaDC. To investigate whether the new gene AlaDC originate from gene SDC and to determine the biochemical properties of the two proteins from Camellia sinensis, the sequences of CsAlaDC and CsSDC were analyzed and the two proteins were over-expressed, purified, and characterized. RESULTS: The results showed that exon-intron structures of AlaDC and SDC were quite similar and the protein sequences, encoded by the two genes, shared a high similarity of 85.1%, revealing that new gene AlaDC originated from SDC by gene duplication. CsAlaDC and CsSDC catalyzed the decarboxylation of alanine and serine, respectively. CsAlaDC and CsSDC exhibited the optimal activities at 45 °C (pH 8.0) and 40 °C (pH 7.0), respectively. CsAlaDC was stable under 30 °C (pH 7.0) and CsSDC was stable under 40 °C (pH 6.0-8.0). The activities of the two enzymes were greatly enhanced by the presence of pyridoxal-5'-phosphate. The specific activity of CsSDC (30,488 IU/mg) was 8.8-fold higher than that of CsAlaDC (3467 IU/mg). CONCLUSIONS: Comparing to CsAlaDC, its ancestral enzyme CsSDC exhibited a higher specific activity and a better thermal and pH stability, indicating that CsSDC acquired the optimized function after a longer evolutionary period. The biochemical properties of CsAlaDC might offer reference for theanine industrial production.

Inhibition of the facilitative sugar transporters (GLUTs) by tea extracts and catechins.

Tea polyphenolics have been suggested to possess blood glucose lowering properties by inhibiting sugar transporters in the small intestine and improving insulin sensitivity. In this report, we studied the effects of teas and tea catechins on the small intestinal sugar transporters, SGLT1 and GLUTs (GLUT1, 2 and 5). Green tea extract (GT), oolong tea extract (OT), and black tea extract (BT) inhibited glucose uptake into the intestinal Caco-2 cells with GT being the most potent inhibitor (IC50 : 0.077 mg/mL), followed by OT (IC50 : 0.136 mg/mL) and BT (IC50 : 0.56 mg/mL). GT and OT inhibition of glucose uptake was partial non-competitive, with an inhibitor constant (Ki ) = 0.0317 and 0.0571 mg/mL, respectively, whereas BT was pure non-competitive, Ki  = 0.36 mg/mL. Oocytes injected to express small intestinal GLUTs were inhibited by teas, but SGLT1 was not. Furthermore, catechins present in teas were the predominant inhibitor of glucose uptake into Caco-2 cells, and gallated catechins the most potent: CG > ECG > EGCG ≥ GCG when compared to the non-gallated catechins (C, EC, GC, and EGC). In Caco-2 cells, individual tea catechins reduced the SGLT1 gene, but not protein expression levels. In contrast, GLUT2 gene and protein expression levels were reduced after 2 hours exposure to catechins but increased after 24 hours. These in vitro studies suggest teas containing catechins may be useful dietary supplements capable of blunting postprandial glycaemia in humans, including those with or at risk to Type 2 diabetes mellitus.

Controllable Self-Assembly of SERS Hotspots in Liquid Environment.

Controllable synthesis of novel metal nanoparticles and effective capture of hotspots are of great significance for SERS (surface-enhanced Raman spectroscopy) detection. Therefore, in this paper, a green controllable synthesis method of gold nanoparticle was achieved via epigallocatechin gallate reduction. Different morphologies of gold nanoparticles were synthesized just by changing the solution pH values, and the growth kinetics of AuNPs (gold nanoparticles) were systematically studied. The synthetic AuNPs were put in a droplet to study dynamic variations of self-assembly SERS hotspots from the liquid sol state to the solid dry state. The addition of halogen ions in the droplet can controllably regulate the self-assembly three-dimensional hotspot model of gold nanoparticles in the evaporation process of a droplet, during which the most enhancement effect can be easily captured. The dynamically changing images of nanoparticles in the process were graphically described based on the internal interaction forces of a droplet. Two stronger areas in the changes of SERS intensity were achieved with a high concentration of halogen ions, while only one maximum intensity area was obtained with a low concentration of halogen ions added. This method can effectively avoid complex and unpredictable microenvironments of SERS substrates in the liquid drop, further improving the reproducibility of SERS detection as well as broadening it to biological applications.

Rapid field trace detection of pesticide residue in food based on surface-enhanced Raman spectroscopy.

Surface-enhanced Raman spectroscopy is an alternative detection tool for monitoring food security. However, there is still a lack of a conclusion of SERS detection with respect to pesticides and real sample analysis, and the summary of intelligent algorithms in SERS is also a blank. In this review, a comprehensive report of pesticides detection using SERS technology is given. The SERS detection characteristics of different types of pesticides and the influence of substrate on inspection are discussed and compared by the typical ways of classification. The key points, including the progress in real sample analysis and Raman data processing methods with intelligent algorithm, are highlighted. Lastly, major challenges and future research trends of SERS analysis of pesticide residue are also addressed. SERS has been proven to be a powerful technique for rapid test of residue pesticides in complex food matrices, but there still is a tremendous development space for future research.

Ectopic Overexpression of Histone H3K4 Methyltransferase CsSDG36 from Tea Plant Decreases Hyperosmotic Stress Tolerance in Arabidopsis thaliana.

Histone methylation plays an important regulatory role in the drought response of many plants, but its regulatory mechanism in the drought response of the tea plant remains poorly understood. Here, drought stress was shown to induce lower relative water content and significantly downregulate the methylations of histone H3K4 in the tea plant. Based on our previous analysis of the SET Domain Group (SDG) gene family, the full-length coding sequence (CDS) of CsSDG36 was cloned from the tea cultivar 'Fuding Dabaicha'. Bioinformatics analysis showed that the open reading frame (ORF) of the CsSDG36 gene was 3138 bp, encoding 1045 amino acids and containing the conserved structural domains of PWWP, PHD, SET and PostSET. The CsSDG36 protein showed a close relationship to AtATX4 of the TRX subfamily, with a molecular weight of 118,249.89 Da, and a theoretical isoelectric point of 8.87, belonging to a hydrophilic protein without a transmembrane domain, probably located on the nucleus. The expression of CsSDG36 was not detected in the wild type, while it was clearly detected in the over-expression lines of Arabidopsis. Compared with the wild type, the over-expression lines exhibited lower hyperosmotic resistance by accelerating plant water loss, increasing reactive oxygen species (ROS) pressure, and increasing leaf stomatal density. RNA-seq analysis suggested that the CsSDG36 overexpression caused the differential expression of genes related to chromatin assembly, microtubule assembly, and leaf stomatal development pathways. qRT-PCR analysis revealed the significant down-regulation of stomatal development-related genes (BASL, SBT1.2(SDD1), EPF2, TCX3, CHAL, TMM, SPCH, ERL1, and EPFL9) in the overexpression lines. This study provides a novel sight on the function of histone methyltransferase CsSDG36 under drought stress.

(Z)-3-Hexen-1-ol accumulation enhances hyperosmotic stress tolerance in Camellia sinensis.

Volatile components in fresh leaves are involved in the regulation of many stress responses, such as insect damage, fungal infection and high temperature. However, the potential function of volatile components in hyperosmotic response is largely unknown. Here, we found that 7-day hyperosmotic treatment specifically led to the accumulation of (Z)-3-hexen-1-ol, (E)-2-hexenal and methyl salicylate. Transcriptome and qRT-PCR analyses suggested the activation of linolenic acid degradation and methyl salicylate processes. Importantly, exogenous (Z)-3-hexen-1-ol pretreatment dramatically enhanced the hyperosmotic stress tolerance of tea plants and decreased stomatal conductance, whereas (E)-2-hexenal and methyl salicylate pretreatments did not exhibit such a function. qRT-PCR analysis revealed that exogenous ABA induced the expressions of related enzyme genes, and (Z)-3-hexen-1-ol could up-regulate the expressions of many DREB and RD genes. Moreover, exogenous (Z)-3-hexen-1-ol tremendously induced the expressions of specific LOX and ADH genes within 24 h. Taken together, hyperosmotic stress induced (Z)-3-hexen-1-ol accumulation in tea plant via the activation of most LOX, HPL and ADH genes, while (Z)-3-hexen-1-ol could dramatically enhance the hyperosmotic stress tolerance via the decrease of stomatal conductance and MDA, accumulation of ABA and proline, activation of DREB and RD gene expressions, and probably positive feedback regulation of LOXs and ADHs. KEY MESSAGE: Hyperosmotic stress induced (Z)-3-hexen-1-ol accumulation in Camellia sinensis via the up-regulation of most LOX, HPL and ADH genes, while (Z)-3-hexen-1-ol could dramatically enhance the hyperosmotic stress tolerance via the decrease of stomatal conductance, accumulation of proline, activation of DREB and RD gene expressions, and probably positive feedback regulation of LOXs and ADHs.

Genome-wide characterization of tea plant (Camellia sinensis) Hsf transcription factor family and role of CsHsfA2 in heat tolerance.

BACKGROUND: Heat stress factors (Hsfs) play vital roles in signal transduction pathways operating in responses to environmental stresses. However, Hsf gene family has not been thoroughly explored in tea plant (Camellia sinensis L.). RESULTS: In this study, we identified 25 CsHsf genes in C. sinensis that were separated by phylogenetic analysis into three sub-families (i.e., A, B, and C). Gene structures, conserved domains and motifs analyses indicated that the CsHsf members in each class were relatively conserved. Various cis-acting elements involved in plant growth regulation, hormone responses, stress responses, and light responses were located in the promoter regions of CsHsfs. Furthermore, degradome sequencing analysis revealed that 7 CsHsfs could be targeted by 9 miRNAs. The expression pattern of each CsHsf gene was significantly different in eight tissues. Many CsHsfs were differentially regulated by drought, salt, and heat stresses, as well as exogenous abscisic acid (ABA) and Ca2+. In addition, CsHsfA2 was located in the nucleus. Heterologous expression of CsHsfA2 improved thermotolerance in transgenic yeast, suggesting its potential role in the regulation of heat stress response. CONCLUSIONS: A comprehensive genome-wide analysis of Hsf in C. sinensis present the global identification and functional prediction of CsHsfs. Most of them were implicated in a complex gene regulatory network controlling various abiotic stress responses and signal transduction pathways in tea plants. Additionally, heterologous expression of CsHsfA2 increased thermotolerance of transgenic yeast. These findings provide new insights into the functional divergence of CsHsfs and a basis for further research on CsHsfs functions.

Transcriptomic analysis reveals mechanism of light-sensitive albinism in tea plant Camellia sinensis 'Huangjinju'.

BACKGROUND: Camellia sinensis 'Huangjinju' is an albino tea variety developed recently in China. Young leaves of 'Huangjinju' demonstrate bright yellow when cultivated under natural sunlight, but regreens under reduced light intensity. To elucidate the physiological and molecular mechanisms of this light-sensitive albinism, we compared leaf pigmentation, metabolites, cellular ultrastructure and transcriptome between plants cultured under natural sunlight and shade. RESULTS: Shading treatment doubled the chlorophyll concentration and regreened albino leaves; carotenoid also increased by 30%. Electron microscopy analyses showed that chloroplast not only increased in number but also in size with a complete set of components. In addition, regreened leaves also had a significantly higher concentration of polyphenols and catechins than albino leaves. At transcriptomic level, a total of 507 genes were differentially expressed in response to light condition changes. The most enriched pathways include light harvest protein complex, response to stimuli, oxidation-reduction process, generation of precursor metabolites and energy response. CONCLUSION: The integrated strategy in this study allows a mechanistic understanding of leaf albinism in light-sensitive tea plants and suggested the regulation of gene networks involved in pigmentation and protein processing. Results from this study provide valuable information to this area and can benefit the domestication and artificial breeding to develop new albino tea varieties.

Identification of MTP gene family in tea plant (Camellia sinensis L.) and characterization of CsMTP8.2 in manganese toxicity.

Cation diffusion facilitators (CDFs) play central roles in metal homeostasis and tolerance in plants, but the specific functions of Camellia sinensis CDF-encoding genes and the underlying mechanisms remain unknown. Previously, transcriptome sequencing results in our lab indicated that the expression of CsMTP8.2 in tea plant shoots was down-regulated exposed to excessive amount of Mn2+ conditions. To elucidate the possible mechanisms involved, we systematically identified 13 C. sinensis CsMTP genes from three subfamilies and characterized their phylogeny, structures, and the features of the encoded proteins. The transcription of CsMTP genes was differentially regulated in C. sinensis shoots and roots in responses to high concentrations of Mn, Zn, Fe, and Al. Differences in the cis-acting regulatory elements in the CsMTP8.1 and CsMTP8.2 promoters suggested the expression of these two genes may be differentially regulated. Transient expression analysis indicated that CsMTP8.2 was localized to the plasma membrane in tobacco and onion epidermal cells. Moreover, when heterologously expressed in yeast, CsMTP8.2 conferred tolerance to Ni and Mn but not to Zn. Additionally, heterologous expression of CsMTP8.2 in Arabidopsis thaliana revealed that CsMTP8.2 positively regulated the response to manganese toxicity by decreasing the accumulation of Mn in plants. However, there was no difference in the accumulation of other metals, including Cu, Fe, and Zn. These results suggest that CsMTP8.2 is a Mn-specific transporter that contributes to the efflux of excess Mn2+ from plant cells.

Genome-Wide Identification of CsATGs in Tea Plant and the Involvement of CsATG8e in Nitrogen Utilization.

Nitrogen (N) is a macroelement with an indispensable role in the growth and development of plants, and tea plant (Camellia sinensis) is an evergreen perennial woody species with young shoots for harvest. During senescence or upon N stress, autophagy has been shown to be induced in leaves, involving a variety of autophagy-related genes (ATGs), which have not been characterized in tea plant yet. In this study, a genome-wide survey in tea plant genome identified a total of 80 Camellia Sinensis autophagy-related genes, CsATGs. The expression of CsATG8s in the tea plant showed an obvious increase from S1 (stage 1) to S4 (stage 4), especially for CsATG8e. The expression levels of AtATGs (Arabidopsis thaliana) and genes involved in N transport and assimilation were greatly improved in CsATG8e-overexpressed Arabidopsis. Compared with wild type, the overexpression plants showed earlier bolting, an increase in amino N content, as well as a decrease in biomass and the levels of N, phosphorus and potassium. However, the N level was found significantly higher in APER (aerial part excluding rosette) in the overexpression plants relative to wild type. All these results demonstrated a convincing function of CsATG8e in N remobilization and plant development, indicating CsATG8e as a potential gene for modifying plant nutrient utilization.

Dehydroascorbic Acid Affects the Stability of Catechins by Forming Conjunctions.

Although tea catechins in green tea and green tea beverages must be stable to deliver good sensory quality and healthy benefits, they are always unstable during processing and storage. Ascorbic acid (AA) is often used to protect catechins in green tea beverages, and AA is easily oxidized to form dehydroascorbic acid (DHAA). However, the function of DHAA on the stability of catechins is not clear. The objective of this study was to determine the effects of DHAA on the stability of catechins and clarify the mechanism of effects by conducting a series of experiments that incubate DHAA with epigallocatechin gallate (EGCG) or catechins. Results showed that DHAA had a dual function on EGCG stability, protecting its stability by inhibiting hydrolysis and promoting EGCG consumption by forming ascorbyl adducts. DHAA also reacted with (-)-epicatechin (EC), (-)-epicatechin gallate (ECG), and (-)-epigallocatechin (EGC) to form ascorbyl adducts, which destabilized them. After 9 h of reaction with DHAA, the depletion rates of EGCG, ECG, EC, and EGC were 30.08%, 22.78%, 21.45%, and 13.55%, respectively. The ability of DHAA to promote catechins depletion went from high to low: EGCG, ECG, EGC, and EC. The results are important for the processing and storage of tea and tea beverages, as well as the general exploration of synergistic functions of AA and catechins.

Identification of Aroma Composition and Key Odorants Contributing to Aroma Characteristics of White Teas.

The identification of aroma composition and key odorants contributing to aroma characteristics of white tea is urgently needed, owing to white tea's charming flavors and significant health benefits. In this study, a total of 238 volatile components were identified in the three subtypes of white teas using headspace solid-phase microextraction (HS-SPME) combined with comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry (GC × GC-TOFMS). The multivariate statistical analysis demonstrated that the contents of 103 volatile compounds showed extremely significant differences, of which 44 compounds presented higher contents in Baihaoyinzhen and Baimudan, while the other 59 compounds exhibited higher contents in Shoumei. The sensory evaluation experiment carried out by gas chromatography-olfactometry/mass spectrometry (GC-O/MS) revealed 44 aroma-active compounds, of which 25 compounds were identified, including 9 alcohols, 6 aldehydes, 5 ketones, and 5 other compounds. These odorants mostly presented green, fresh, floral, fruity, or sweet odors. Multivariate analyses of chemical characterization and sensory evaluation results showed that high proportions of alcohols and aldehydes form the basis of green and fresh aroma characteristic of white teas, and phenylethyl alcohol, γ-Nonalactone, trans-ß-ionone, trans-linalool oxide (furanoid), α-ionone, and cis-3-hexenyl butyrate were considered as the key odorants accounting for the different aroma characteristics of the three subtypes of white tea. The results will contribute to in-depth understand chemical and sensory markers associated with different subtypes of white tea, and provide a solid foundation for tea aroma quality control and improvement.

Facile Reduction Method Synthesis of Defective MoO2- x Nanospheres Used for SERS Detection with High Chemical Enhancement.

Recently, more and more attention has been given to a semiconductor oxide-based surface-enhanced Raman spectroscopy substrate for its great stability and biocompatibility. However, its poor SERS sensitivity limits the applications of semiconductor oxide SERS substrates. In this paper, we provide a facile reduction method to modulate oxygen vacancy concentrations in oxide SERS substrates. Using MoO2 as an example, the resonance coupling as well as charge transfer between the semiconductor oxide SERS substrate and the target molecules were promoted for the reason of artificial oxygen vacancy embodied in the Raman signals being improved. By using the TEM, SEM, and XPS measurements, we confirmed that we successfully prepared defective MoO2- x with a polycrystalline surface. MoO2- x modulated oxygen vacancy treated with 6 wt % Li shows a very high detection sensitivity of 10-8 M (4.79 ug/L) for R6G, and the intensity of the Raman signal was highly enhanced. Because of the existence of defective energy levels, resonance coupling, as well as charge transfer between semiconductor and molecules, was obviously promoted. More importantly, the method of modulating oxygen vacancy can be widely used in semiconductor oxide materials for its chemical enhancement capacity can be promoted by artificial oxygen vacancy.

Detection of systemic pesticide residues in tea products at trace level based on SERS and verified by GC-MS.

Surface-enhanced Raman spectroscopy (SERS) has the potential to detect pesticide residues in agricultural products. However, some systemic pesticides, such as chlorpyrifos, can enter the plant tissue, and not just stay on the surface. Consequently, many SERS studies halted at practical application because of its complexity. In this work, SERS technology was used to detect chlorpyrifos residues in tea products at the semiquantitative level. A simple pretreatment method effectively avoided interference of other fluorescent substances, and all major peaks could be distinguished on the basis of a novel substrate. A principal component analysis algorithm was applied to form a regression model, and a nanogram detection limit was obtained. Furthermore, chlorpyrifos residues in the same tea products were also measured by gas chromatography-mass spectrometry, and the results show a small range of errors. From the comparative study of the two detection methods, the results suggest the great promise of SERS technology for rapid inspection of agricultural products.

SERS based determination of vanillin and its methyl and ethyl derivatives using flower-like silver nanoparticles on a silicon wafer.

A surface enhanced Raman scattering (SERS) method is described for the determination of vanillin, methyl vanillin and ethyl vanillin at trace levels. Flower-like silver nanoparticles on a silicon wafer are used as the SERS substrate, and the analytes can be specifically and non-destructively recognized by their specific Raman bands. The molecules can be recognized rapidly by identifying the characteristic bands. The SERS spectra of vanillin (C8H8O3) were used as mid-contrast, and specific bands of methyl vanillin and ethyl vanillin (C9H10O3) were acquired at 775 cm-1, 1350 cm-1 and 1282 cm-1, 1382 cm-1, respectively. In addition, by using an improved principal component analysis (PCA) algorithm, the organic molecule can be quantitatively determined. Dissolved in water, vanillin, methyl vanillin and ethyl vanillin still can be detected at a concentration of 10-8 M, at which their characteristic Raman peaks are still visible. The method was successfully applied to the determination of vanillin in milk powder products. Graphical abstract Vanillin can be identified at trace levels by laser irradiation of milk and by using flower-like silver nanoparticles as the surface enhanced Raman scattering (SERS) substrate. Vanillin and its methyl and ethyl derivatives can be quantitatively analyzed by the principal component analysis (PCA) algorithm.

Isolation and Characterization of CsWRKY7, a Subgroup IId WRKY Transcription Factor from Camellia sinensis, Linked to Development in Arabidopsis.

WRKY transcription factors (TFs) containing one or two WRKY domains are a class of plant TFs that respond to diverse abiotic stresses and are associated with developmental processes. However, little has been known about the function of WRKY gene in tea plant. In this study, a subgroup IId WRKY gene CsWRKY7 was isolated from Camellia sinensis, which displayed amino acid sequence homology with Arabidopsis AtWRKY7 and AtWRKY15. Subcellular localization prediction indicated that CsWRKY7 localized to nucleus. Cis-acting elements detected in the promotor region of CsWRKY7 are mainly involved in plant response to environmental stress and growth. Consistently, expression analysis showed that CsWRKY7 transcripts responded to NaCl, mannitol, PEG, and diverse hormones treatments. Additionally, CsWRKY7 exhibited a higher accumulation both in old leaves and roots compared to bud. Seed germination and root growth assay indicated that overexpressed CsWRKY7 in transgenic Arabidopsis was not sensitive to NaCl, mannitol, PEG, and low concentration of ABA treatments. CsWRKY7 overexpressing Arabidopsis showed a late-flowering phenotype under normal conditions compared to wild type. Furthermore, gene expression analysis showed that the transcription levels of the flowering time integrator gene FLOWERING LOCUS T (FT) and the floral meristem identity genes APETALA1 (AP1) and LEAFY (LFY) were lower in WRKY7-OE than in the WT. Taken together, these findings indicate that CsWRKY7 TF may participate in plant growth. This study provides a potential strategy to breed late-blooming tea cultivar.