Elevated Ozone Reduces the Quality of Tea Leaves but May Improve the Resistance of Tea Plants.

Tropospheric ozone (O3) pollution can affect plant nutritional quality and secondary metabolites by altering plant biochemistry and physiology, which may lead to unpredictable effects on crop quality and resistance to pests and diseases. Here, we investigated the effects of O3 (ambient air, Am; ambient air +80 ppb of O3, EO3) on the quality compounds and chemical defenses of a widely cultivated tea variety in China (Camellia sinensis cv. 'Baiye 1 Hao') using open-top chamber (OTC). We found that elevated O3 increased the ratio of total polyphenols to free amino acids while decreasing the value of the catechin quality index, indicating a reduction in leaf quality for green tea. Specifically, elevated O3 reduced concentrations of amino acids and caffeine but shows no impact on the concentrations of total polyphenols in tea leaves. Within individual catechins, elevated O3 increased the concentrations of ester catechins but not non-ester catechins, resulting in a slight increase in total catechins. Moreover, elevated O3 increased the emission of biogenic volatile organic compounds involved in plant defense against herbivores and parasites, including green leaf volatiles, aromatics, and terpenes. Additionally, concentrations of main chemical defenses, represented as condensed tannins and lignin, in tea leaves also increased in response to elevated O3. In conclusion, our results suggest that elevated ground-level O3 may reduce the quality of tea leaves but could potentially enhance the resistance of tea plants to biotic stresses.

Linc01588 deletion inhibits the malignant biological characteristics of hydroquinone-induced leukemic cells via miR-9-5p/SIRT1.

Leukemia caused by environmental chemical pollutants has attracted great attention, the malignant leukemic transformation model of TK6 cells induced by hydroquinone (HQ) has been previously found in our team. However, the type of leukemia corresponding to this malignant transformed cell line model needs further study and interpretation. Furthermore, the molecular mechanism of malignant proliferation of leukemic cells induced by HQ remains unclear. This study is the first to reveal the expression of aberrant genes in leukemic cells of HQ-induced malignant transformation, which may correspond to chronic lymphocytic leukemia (CLL). The expression of Linc01588, a long non-coding RNA (lncRNA), was significantly up-regulated in CLL patients and leukemic cell line model which previously described. After gain-of-function assays and loss-of-function assays, feeble cell viability, severe apoptotic phenotype and the increased secretion of TNF-α were easily observed in malignant leukemic TK6 cells with Linc01588 deletion after HQ intervention. The tumors derived from malignant TK6 cells with Linc01588 deletion inoculated subcutaneously in nude mice were smaller than controls. In CLL and its cell line model, the expression of Linc01588 and miR-9-5p, miR-9-5p and SIRT1 were negative correlation respectively in CLL and cell line model, while the expression of Linc01588 and SIRT1 were positive correlation. The dual-luciferase reporter assay showed that Linc01588 & miR-9-5p, miR-9-5p & SIRT1 could bind directly, respectively. Furthermore, knockdown of miR-9-5p successfully rescued the severe apoptotic phenotype and the increased secretion of TNF-α caused by the Linc01588 deletion, the deletion of Linc01588 in human CLL cell line MEC-2 could also inhibit malignant biological characteristics, and the phenotype caused by the deletion of Linc01588 could also be rescued after overexpression of SIRT1. Moreover, the regulation of SIRT1 expression in HQ19 cells by Linc01588 and miR-9-5 P may be related to the Akt/NF-κB pathway. In brief, Linc01588 deletion inhibits the malignant biological characteristics of HQ-induced leukemic cells via miR-9-5p/SIRT1, and it is a novel and hopeful clue for the clinical targeted therapy of CLL.

Diversity in rhizospheric microbial communities in tea varieties at different locations and tapping potential beneficial microorganisms.

Soil microenvironments and plant varieties could largely affect rhizosphere microbial community structure and functions. However, their specific effects on the tea rhizosphere microbial community are yet not clear. Beneficial microorganisms are important groups of microbial communities that hold ecological functionalities by playing critical roles in plant disease resistance, and environmental stress tolerance. Longjing43 and Zhongcha108 are two widely planted tea varieties in China. Although Zhongcha108 shows higher disease resistance than Longjing43, the potential role of beneficial tea rhizosphere microbes in disease resistance is largely unknown. In this study, the structure and function of rhizosphere microbial communities of these two tea varieties were compared by using the Illumina MiSeq sequencing (16S rRNA gene and ITS) technologies. Rhizosphere soil was collected from four independent tea gardens distributed at two locations in Hangzhou and Shengzhou cities in eastern China, Longjing43 and Zhongcha108 are planted at both locations in separate gardens. Significant differences in soil physicochemical properties as demonstrated by ANOVA and PCA, and distinct rhizosphere microbial communities by multiple-biotech analyses (PCoA, LEfSe, Co-occurrence network analyses) between both locations and tea varieties (p < 0.01) were found. Functions of bacteria were annotated by the FAPROTAX database, and a higher abundance of Nitrososphaeraceae relating to soil ecological function was found in rhizosphere soil in Hangzhou. LDA effect size showed that the abundance of arbuscular mycorrhizal fungi (AMF) was higher in Zhongcha108 than that in Longjing43. Field experiments further confirmed that the colonization rate of AMF was higher in Zhongcha108. This finding testified that AMF could be the major beneficial tea rhizosphere microbes that potentially function in enhanced disease resistance. Overall, our results confirmed that locations affected the microbial community greater than that of tea varieties, and fungi might be more sensitive to the change in microenvironments. Furthermore, we found several beneficial microorganisms, which are of great significance in improving the ecological environment of tea gardens and the disease resistance of tea plants. These beneficial microbial communities may also help to further reveal the mechanism of disease resistance in tea and potentially be useful for mitigating climate change-associated challenges to tea gardens in the future.

Genome-Wide Identification and Expression Analysis of Isopentenyl transferase Family Genes during Development and Resistance to Abiotic Stresses in Tea Plant (Camellia sinensis).

The tea plant is an important economic crop and is widely cultivated. Isopentenyl transferase (IPT) is the first and rate-limiting enzyme of cytokinin (CK) signaling, which plays key roles in plant development and abiotic stress. However, the IPT gene family in tea plants has not been systematically investigated until now. The phylogenetic analyses, gene structures, and conserved domains were predicted here. The results showed that a total of 13 CsIPT members were identified from a tea plant genome database and phylogenetically classified into four groups. Furthermore, 10 CsIPT members belonged to plant ADP/ATP-IPT genes, and 3 CsIPTs were tRNA-IPT genes. There is a conserved putative ATP/GTP-binding site (P-loop motif) in all the CsIPT sequences. Based on publicly available transcriptome data as well as through RNA-seq and qRT-PCR analysis, the CsIPT genes which play key roles in the development of different tissues were identified, respectively. Furthermore, CsIPT6.2 may be involved in the response to different light treatments. CsIPT6.4 may play a key role during the dormancy and flush of the lateral buds. CsIPT5.1 may play important regulatory roles during the development of the lateral bud, leaf, and flower. CsIPT5.2 and CsIPT6.2 may both play key roles for increased resistance to cold-stress, whereas CsIPT3.2 may play a key role in improving resistance to high-temperature stress as well as drought-stress and rewatering. This study could provide a reference for further studies of CsIPT family's functions and could contribute to tea molecular breeding.

Expression and functional analysis of CsA-IPT5 splice variants during shoot branching in Camellia sinensis.

Alternative splicing (AS) is a process by which several functional splice variants are generated from the same precursor mRNA. In our recent study, five CsA-IPT5 splice variants with various numbers of ATTTA motifs in the untranslated regions (UTRs) were cloned. Meanwhile, their transient expression, as well as the expression and functional analysis in the two shoot branching processes were studied. Here, we examined how these splice variants regulate the other three important shoot branching processes, including the spring tea development, the distal branching of new shoots, and the shoot branching induced by 2,3,5-triiodobenzoic acid (TIBA) spraying, and thus unraveling the key CsA-IPT5 transcripts which play the most important roles in the shoot branching of tea plants. The results showed that the increased expression of 5' UTR AS3, 3' UTR AS1 and 3' UTR AS2 could contribute to the increased synthesis of tZ/iP-type cytokinins (CKs), thus promoting the spring tea development. Meanwhile, in the TIBA-induced shoot branching or in the distal branching of the new shoots, CsA-IPT5 transcripts regulated the synthesis of CsA-IPT5 protein and CKs through transcriptional regulation of the ratios of its splice variants. Moreover, 3' UTR AS1 and 3' UTR AS2 both play key roles in these two processes. In summary, it is revealed that 3' UTR AS1 and 3' UTR AS2 of CsA-IPT5 might act as the predominant splice variants in shoot branching of the tea plant, and they both can serve as gene resources for tea plant breeding.

Effects of elevated ozone on the uptake and allocation of macronutrients in poplar saplings above- and belowground.

Ground-level ozone (O3) is a secondary air pollutant and affects the roots and soil processes of trees. Therefore, O3 can affect the uptake and allocation of nutrients in trees, which merits further clarification. A fumigation experiment with five O3 levels was conducted in 15 open top chambers for two poplar clones, and the concentrations of six macronutrients (N, P, K, S, Ca, Mg) in different organs and leaf positions were determined. Under all O3 levels, the concentration of mobile nutrients (N and P) was higher in upper leaves than in lower leaves, while the non-mobile nutrients (Ca and S) concentration was the opposite. Relative to charcoal filtered ambient air (CF), high O3 treatment (NF60) significantly increased the concentration of mobile nutrients K and Mg in upper leaves by 38 % and 33 %, in lower leaves by 142 % and 65 %, respectively, which suggested the effect of O3 on their concentrations was greater at the lower leaf position than at the upper leaf position. Elevated O3 significantly increased the macronutrient concentrations in most organs. The effects of O3 on nutrient concentrations were attributed using graphical vector analysis, suggested that the increase of nutrient concentration in the shoots was attributed to excessive nutrient stocks, while their increase in root was attributed to the "concentration" effect. Compared to CF, NF60 also reduced the root-to-shoot ratio of N, P, S, K, Ca and Mg stocks by 34 %, 39 %, 37 %, 64 %, 46 % and 42 %, respectively, indicating the allocation of increased nutrients to shoots in response to O3 stress. Changes in the allocation pattern of nutrients in different leaf positions and organs of poplar were primarily in response to O3 stress since these nutrients play important roles in some physiological processes. These results will help improve the plantation nutrient utilization by optimizing fertilizer management regimes under O3 pollution.

Control of Newly-Designed Wearable Robotic Hand Exoskeleton Based on Surface Electromyographic Signals.

The human hand plays a role in a variety of daily activities. This intricate instrument is vulnerable to trauma or neuromuscular disorders. Wearable robotic exoskeletons are an advanced technology with the potential to remarkably promote the recovery of hand function. However, the still face persistent challenges in mechanical and functional integration, with real-time control of the multiactuators in accordance with the motion intentions of the user being a particular sticking point. In this study, we demonstrated a newly-designed wearable robotic hand exoskeleton with multijoints, more degrees of freedom (DOFs), and a larger range of motion (ROM). The exoskeleton hand comprises six linear actuators (two for the thumb and the other four for the fingers) and can realize both independent movements of each digit and coordinative movement involving multiple fingers for grasp and pinch. The kinematic parameters of the hand exoskeleton were analyzed by a motion capture system. The exoskeleton showed higher ROM of the proximal interphalangeal and distal interphalangeal joints compared with the other exoskeletons. Five classifiers including support vector machine (SVM), K-near neighbor (KNN), decision tree (DT), multilayer perceptron (MLP), and multichannel convolutional neural networks (multichannel CNN) were compared for the offline classification. The SVM and KNN had a higher accuracy than the others, reaching up to 99%. For the online classification, three out of the five subjects showed an accuracy of about 80%, and one subject showed an accuracy over 90%. These results suggest that the new wearable exoskeleton could facilitate hand rehabilitation for a larger ROM and higher dexterity and could be controlled according to the motion intention of the subjects.

Biogenic volatile organic compound emissions from leaves and fruits of apple and peach trees during fruit development.

Biogenic volatile organic compounds (BVOCs) are widely involved in a variety of atmospheric chemical processes due to their high reactivity and species diversity. To date, however, research on BVOCs in agroecosystems, particularly fruit trees, remains scarce despite their large cultivation area and economic interest. BVOC emissions from different organs (leaf or fruit) of apple and peach trees were investigated throughout the stages of fruit development (FS, fruit swelling; FC, fruit coloration; FM, fruit maturity; and FP, fruit postharvest) using a proton-transfer-reaction mass spectrometer. Results indicated that methanol was the most abundant compound emitted by the leaf (apple tree leaf 492.5 ± 47.9 ng/(g·hr), peach tree leaf 938.8 ±  154.5 ng/(g·hr)), followed by acetic acid and green leaf volatiles. Beside the above three compounds, acetaldehyde had an important contribution to the emissions from the fruit. Overall, the total BVOCs (sum of eight compounds studied in this paper) emitted by both leaf and fruit gradually decreased along the fruit development, although the effect was significant only for the leaf. The leaf (2020.8 ±  258.8 ng/(g·hr)) was a stronger BVOC emitter than the fruit (146.0 ± 45.7 ng/(g·hr)) (P = 0.006), and there were no significant differences in total BVOC emission rates between apple and peach trees. These findings contribute to our understanding on BVOC emissions from different plant organs and provide important insights into the variation of BVOC emissions across different fruit developmental stages.

Root Secondary Metabolites in Populus tremuloides: Effects of Simulated Climate Warming, Defoliation, and Genotype.

Climate warming can influence interactions between plants and associated organisms by altering levels of plant secondary metabolites. In contrast to studies of elevated temperature on aboveground phytochemistry, the consequences of warming on root chemistry have received little attention. Herein, we investigated the effects of elevated temperature, defoliation, and genotype on root biomass and phenolic compounds in trembling aspen (Populus tremuloides). We grew saplings of three aspen genotypes under ambient or elevated temperatures (+4-6 °C), and defoliated (by 75%) half of the trees in each treatment. After 4 months, we harvested roots and determined their condensed tannin and salicinoid (phenolic glycoside) concentrations. Defoliation reduced root biomass, with a slightly larger impact under elevated, relative to ambient, temperature. Elevated temperature decreased condensed tannin concentrations by 21-43% across the various treatment combinations. Warming alone did not alter salicinoid concentrations but eliminated a small negative impact of defoliation on those compounds. Graphical vector analysis suggests that effects of warming and defoliation on condensed tannins and salicinoids were predominantly due to reduced biosynthesis of these metabolites in roots, rather than to changes in root biomass. In general, genotypes did not differ in their responses to temperature or temperature by defoliation interactions. Collectively, our results suggest that future climate warming will alter root phytochemistry, and that effects will vary among different classes of secondary metabolites and be influenced by concurrent ecological interactions such as herbivory. Temperature- and herbivory-mediated changes in root chemistry have the potential to influence belowground trophic interactions and soil nutrient dynamics.

Nonlinear responses of foliar phenylpropanoids to increasing O3 exposure: Ecological implications in a Populus model system.

Plant phenolic compounds (phenylpropanoids) act as defense chemicals against herbivores and can mediate ecosystem processes. Tropospheric ozone (O3) pollution alters concentrations of plant phenolics; however, little is known about how these phytochemicals respond to different levels of O3 exposure. Here, we investigated the effects of five different O3 exposure levels on foliar concentrations of phenylpropanoids (53 compounds in total) and antioxidative capacity in hybrid Populus (Populus euramericana cv. '74/76') saplings grown in the presence of high or low soil nitrogen (N) load. Increasing O3 exposure initially increased and then decreased total concentrations of phenolic compounds, revealing a biphasic exposure-response profile (hormetic zone: 1.1-36.3 ppm h AOT40). This biphasic response pattern was driven by changes in a subset of phenylpropanoids with high antioxidative capacity (e.g. condensed tannins) but not in phenolics with low antioxidative capacity (e.g. salicinoids). The O3 exposure-response relationships of some phenylpropanoids (e.g. flavonoids and chlorogenic acids) varied in response to soil N, with hormesis occurring in high N soil but not in low N soil. Collectively, our findings indicated that plant phenolic compounds exhibit nonlinear responses to increasing O3 exposure, and that the responses vary in relation to phenolic compound class, antioxidative capacity, and soil nitrogen conditions. Our findings further suggest that the impact of O3 on ecological processes mediated by phenolics will be concentration-dependent, highlighting the complexity of the ecological effects of ground-level O3 pollution.

Interactive effects of ozone exposure and nitrogen addition on the rhizosphere bacterial community of poplar saplings.

It is widely documented that elevated ground-level ozone (O3) has negative effects on tree physiological characteristics, and in return, affects forest ecosystem function. However, the effect may be modified by soil nitrogen (N) availability. Numerous studies have focused on the aboveground part of trees under elevated O3 alone or in combination with soil N; however, little is known about the response of soil bacterial communities. Here, we investigated the effects of O3 (charcoal-filtered air, CF, versus ambient air +40 ppb of O3, E-O3), N addition (0 kg ha-1 yr-1, N0, versus 200 kg ha-1 yr-1, N200), and their combination on rhizosphere soil bacterial communities of hybrid poplar, using an MiSeq targeted amplicon sequencing of the bacterial 16S rRNA gene. E-O3 significantly decreased bacterial abundance, and N200 significantly decreased the α-diversity. The negative impacts of N200 on α-diversity were alleviated by E-O3. Nitrogen and E-O3-N200 combination altered bacterial community composition, with a significant increase in the relative abundance of Proteobacteria and Bacteroidetes and a decrease in the abundance of Firmicutes. From an ecological network analysis, E-O3, alone and in combination with N200, complicated the co-occurrence network of bacterial communities by inducing a microbial survival strategy, shifting the hub species from RB41 to Bacillus and Blastococcus. Conversely, N200 led to simplification and decentralization of the co-occurrence network. These findings demonstrate that the rhizosphere bacterial communities exhibit divergent responses to E-O3 and N200, suggesting the need to consider the stability of the belowground ecosystem to optimize plantation management in response to environmental changes.

Surface-based morphometry study of the brain in benign childhood epilepsy with centrotemporal spikes.

BACKGROUND: The study aimed to explore cortical morphology in benign childhood epilepsy with centrotemporal spikes (BECTS) and the relationship between cortical characteristics and age of onset and intelligence quotient (IQ). METHODS: Cortical morphometry with surface-based morphometry (SBM) was used to compare changes in cortical thickness, gyrification, sulcal depth, and fractal dimension of the cerebral cortex between 25 BECTS patients and 20 healthy controls (HCs) with two-sample t-tests [P<0.05, family-wise error (FWE) corrected]. Relationships between abnormal cortical morphological changes and age of onset and IQ, which included verbal intelligence quotient (VIQ), performance intelligence quotient (PIQ), and full-scale intelligence quotient (FIQ) were investigated with Spearman correlation analysis (P<0.05, uncorrected). RESULTS: The BECTS patients showed extensive cortical thinning predominantly in bilateral frontal, temporal regions, and limbic system. Cortical gyrification increased in the left hemisphere and partial right hemisphere, and the decreased cortical gyrification was only in the left hemisphere. The increased sulcal depth was the left fusiform gyrus. There are no statistically significant differences in the fractal dimension. Correlation analysis revealed the negative correlation between age of onset and cortical thickness in the right precentral gyrus. It also revealed the negative correlation between the age of onset and cortical gyrification in the left inferior parietal gyrus. Also, there was negative correlation between VIQ and cortical gyrification in the left supramarginal gyrus of BECTS patients. CONCLUSIONS: This study reveals aberrant cortical thickness, cortical gyrification, and sulcal depth of BECTS in areas related to cognitive functions including language, attention and memory, and the correlation between some brain regions and VIQ and age of onset, providing a potential marker of early neurodevelopmental disturbance and cognitive dysfunction in BECTS.

Ozone affects plant, insect, and soil microbial communities: A threat to terrestrial ecosystems and biodiversity.

Elevated tropospheric ozone concentrations induce adverse effects in plants. We reviewed how ozone affects (i) the composition and diversity of plant communities by affecting key physiological traits; (ii) foliar chemistry and the emission of volatiles, thereby affecting plant-plant competition, plant-insect interactions, and the composition of insect communities; and (iii) plant-soil-microbe interactions and the composition of soil communities by disrupting plant litterfall and altering root exudation, soil enzymatic activities, decomposition, and nutrient cycling. The community composition of soil microbes is consequently changed, and alpha diversity is often reduced. The effects depend on the environment and vary across space and time. We suggest that Atlantic islands in the Northern Hemisphere, the Mediterranean Basin, equatorial Africa, Ethiopia, the Indian coastline, the Himalayan region, southern Asia, and Japan have high endemic richness at high ozone risk by 2100.

Water stress rather than N addition mitigates impacts of elevated O3 on foliar chemical profiles in poplar saplings.

Tropospheric ozone (O3) pollution can alter tree chemical profiles, and in turn, affect forest ecosystem function. However, the magnitude of these effects may be modified by variations in soil water and nutrient availability, which makes it difficult to predict the impacts of O3 in reality. Here we assessed the effects of elevated O3 alone, and in combination with soil water deficit and N addition, on the phytochemical composition of hybrid poplar (Populus deltoides cv. '55/56' × P. deltoides cv. 'Imperial'). Potted trees were grown in open-top chambers (OTCs) under either charcoal-filtered air or elevated O3 (non-filtered air +40 ppb of O3), and trees within each OTC were grown with four combinations of water (well-watered or water deficit) and nitrogen (with or without N addition) levels. We found that elevated O3 alone stimulated the accumulation of foliar nitrogen, soluble sugar, and lignin while inhibiting the accumulation of starch, but had limited impacts on condensed tannins and salicinoids in poplar saplings. Graphical vector analysis revealed that these changes in concentrations of nitrogen, starch and lignin were due largely to altered metabolic processes, while increased soluble sugar concentration related mainly to decreased leaf biomass in most cases. The effects of O3 on poplar foliar chemical profiles depended on soil water, but not soil N, availability. Specifically, O3-mediated changes in carbohydrates and lignin were mitigated by decreased soil water content. Taken together, these results suggested that nitrogen acquisition, carbohydrates mobilization and lignification play a role in poplar tolerance to O3. Moreover, the impacts of elevated O3 on phytochemistry of poplar leaves can be context-dependent, with potential consequences for ecosystem processes under future global change scenarios. Our results highlight the needs to consider multi-factors environments to optimize the management of plantations under changing environments.

Late growing season carbon subsidy in native gymnosperms in a northern temperate forest.

Evergreen tree species that maintain positive carbon balance during the late growing season may subsidize extra carbon in a mixed forest. To test this concept of 'carbon subsidy', leaf gas exchange characteristics and related leaf traits were measured for three gymnosperm evergreen species (Chamaecyparis thyoides, Tsuga canadensis and Pinus strobus) native to the oak-hickory deciduous forest in northeast USA from March (early Spring) to October (late Autumn) in a single year. All three species were photosynthetically active in Autumn. During the Summer-Autumn transition, photosynthetic capacity (Amax) of T. canadensis and P. strobus increased (T-test, P < 0.001) and was maintained in C. thyoides (T-test, P = 0.49), while dark respiration at 20 °C (Rn) and its thermal sensitivity were generally unchanged for all species (one-way ANOVA, P > 0.05). In Autumn, reductions in mitochondrial respiration rate in the daylight (RL) and the ratio of RL to Rn (RL/Rn) were observed in P. strobus (46.3% and 44.0% compared to Summer, respectively). Collectively, these physiological adjustments resulted in higher ratios of photosynthesis to respiration (A/Rnand A/RL) in Autumn for all species. Across season, photosynthetic biochemistry and respiratory variables were not correlated with prevailing growth temperature. Physiological adjustments allowed all three gymnosperm species to maintain positive carbon balance into late Autumn, suggesting that gymnosperm evergreens may benefit from Autumn warming trends relative to deciduous trees that have already lost their leaves.