Mechanism Underlying the Shading-Induced Chlorophyll Accumulation in Tea Leaves

Besides aroma and taste, the color of dry tea leaves, tea infusion, and infused tea leaves is also an important index for tea quality. Shading can significantly increase the chlorophyll content of tea leaves, leading to enhanced tea leaf coloration. However, the underlying regulatory mechanism remains unclear. In this study, we revealed that the expressions of chlorophyll synthesis genes were significantly induced by shading, specially, the gene encoding protochlorophyllide oxidoreductase (CsPOR). Indoor control experiment showed that decreased light intensity could significantly induce the expression of CsPOR, and thus cause the increase of chlorophyll content. Subsequently, we explored the light signaling pathway transcription factors regulating chlorophyll synthesis, including CsPIFs and CsHY5. Through expression level and subcellular localization analysis, we found that CsPIF3-2, CsPIF7-1, and CsHY5 may be candidate transcriptional regulators. Transcriptional activation experiments proved that CsHY5 inhibits CsPORL-2 transcription. In summary, we concluded that shading might promote the expression of CsPORL-2 by inhibiting the expression of CsHY5, leading to high accumulation of chlorophyll in tea leaves. The results of this study provide insights into the mechanism regulating the improvements to tea plant quality caused by shading.

Herbaria Reveal Herbivory and Pathogen Increases and Shifts in Senescence for Northeastern United States Maples Over 150 Years

Recent studies suggest climate-related delays in the timing of leaf coloration and abscission in maple trees but lack baseline data prior to the late 20th century. To better understand how autumn foliar phenology and late-season damage risks have changed for this genus over the past century, we evaluated 2,972 digitized herbaria specimens of red and sugar maple collected between 1826 and 2016 for the presence of leaves, autumn leaf coloration, and pathogen or herbivore damage. We found that the onset (first appearance) of colored leaves has shifted 0.26 days later each year, leading to a delay of more than a month in autumn phenology since 1880. We find that these shifts are related to precipitation regimes in both the fall and summer seasons and that more severe droughts are associated with higher probabilities of colored leaves. Moreover, we found that the probability of both herbivory and pathogen damage has increased significantly over the study period. In particular, we find a strong association between increasing summer drought conditions and increased probability of herbivory. Furthermore, the presence of foliar damage increased the probability of leaf coloration on herbaria specimens. However, the end-of-season abscission date (last appearance of leaves) was strongly associated with herbivory and climate in a contrary direction: Increasing yearly drought, higher fall temperatures, and the presence of herbivory were associated with earlier abscission. In fact, the last leaf dates for specimens with herbivory were nearly 2 weeks earlier than specimens without herbivore damage. Our study documents significant changes in maple senescence over the last 150 years and suggests that incorporating herbivory into models may improve our ability to predict forest responses to climate shifts.

Biochemical and transcriptomic analyses reveal that critical genes involved in pigment biosynthesis influence leaf color changes in a new sweet osmanthus cultivar ‘Qiannan Guifei’

Background Osmanthus fragrans (Oleaceae) is one of the most important ornamental plant species in China. Many cultivars with different leaf color phenotypes and good ornamental value have recently been developed. For example, a new cultivar ‘Qiannan Guifei’, presents a rich variety of leaf colors, which change from red to yellow-green and ultimately to green as leaves develop, making this cultivar valuable for landscaping. However, the biochemical characteristics and molecular mechanisms underlying leaf color changes of these phenotypes have not been elucidated. It has been hypothesized that the biosynthesis of different pigments in O. fragrans might change during leaf coloration. Here, we analyzed transcriptional changes in genes involved in chlorophyll (Chl), flavonoid, and carotenoid metabolic pathways and identified candidate genes responsible for leaf coloration in the new cultivar ‘Qiannan Guifei’. Methods Leaf samples were collected from ‘Qiannan Guifei’ plants at the red (R), yellow-green (YG) and green (G) leaf stages. We compared the different-colored leaves via leaf pigment concentrations, chloroplast ultrastructure, and transcriptomic data. We further analyzed differentially expressed genes (DEGs) involved in the Chl, flavonoid, and carotenoid metabolic pathways. In addition, we used qRT-PCR to validate expression patterns of the DEGs at the three stages. Results We found that, compared with those at the G stage, chloroplasts at the R and YG stages were less abundant and presented abnormal morphologies. Pigment analyses revealed that the leaves had higher flavonoid and anthocyanin levels at the R stage but lower Chl and carotenoid concentrations. Similarly, Chl and carotenoid concentrations were lower at the YG stage than at the G stage. By using transcriptomic sequencing, we further identified 61 DEGs involved in the three pigment metabolic pathways. Among these DEGs, seven structural genes (OfCHS, OfCHI, OfF3H, OfDFR, OfANS, OfUGT andOf3AT) involved in the flavonoid biosynthesis pathway were expressed at the highest level at the R stage, thereby increasing the biosynthesis of flavonoids, especially anthocyanins. Six putativeOfMYB genes, including three flavonoid-related activators and three repressors, were also highly expressed at the R stage, suggesting that they might coordinately regulate the accumulation of flavonoids, including anthocyanins. Additionally, expressions of the Chl biosynthesis-related genes OfHEMA, OfCHLG and OfCAO and the carotenoid biosynthesis-related genes OfHYB and OfZEP were upregulated from the R stage to the G stage, which increased the accumulation of Chl and carotenoids throughout leaf development. In summary, we screened the candidate genes responsible for the leaf color changes of ‘Qiannan Guifei’, improved current understanding of the regulatory mechanisms underlying leaf coloration and provided potential targets for future leaf color improvement in O. fragrans.

Metabolome and Transcriptome Analyses Reveal Different Flavonoid Biosynthesis and Chlorophyll Metabolism Profiles between Red Leaf and Green Leaf of Eucommia ulmoides

Flavonoids are natural antioxidants in plants that affect the color of plant tissues. Flavonoids can be divided into eight subgroups, including flavonols, anthocyanins, and proanthocyanidins. The mechanisms of flavonoid synthesis in model plants have been widely studied. However, there are a limited number of reports on the synthesis of flavonoids in the red leaf varieties of woody plants. In this study, we combined morphological observation, pigment content determination, metabolomics, and transcriptomics to investigate the metabolites and gene regulation present in the red and green leaves of Eucommia ulmoides Oliv. The results showed that the red leaves have a lower chlorophyll content and a higher anthocyanin content. Metabonomic analysis identified that the relative content of most flavonoids is up-regulated in red leaves based on UPLC-ESI-MS/MS, which included the apigenin class, quercetin class, kaempferol class, and procyanidins. Transcriptome data suggested that the differentially up-regulated genes are enriched in flavonoid and anthocyanin synthesis pathways, ABC transport, and GST pathways. The integrative analysis of the transcriptome and metabolome showed that the up-regulation of flavonoid metabolism and a high expression of chlorophyll degradation with the down-regulation of chlorophyll biosynthesis genes are detected in E. ulmoides red leaves compared with green leaves. In addition, the co-expression networks implied that cyanidin 3-5-O-diglucoside, EuDR5, EuPAL2, EuDFR11, Eu3MaT1, and EuF3′H are likely associated with the red leaf coloration of E. ulmoides. In summary, this research provided a reference for studying the mechanism of red leaf coloration in woody plants and the use of E. ulmoides red leaves as feedstock for bioactive products.

Simultaneous changes in anthocyanin, chlorophyll, and carotenoid contents produce green variegation in pink–leaved ornamental kale

Background Anthocyanin, chlorophyll, and carotenoid pigments are widely distributed in plants, producing various colors. Ornamental kale (Brassica oleracea var. acephala DC) which has colorful inner leaves is an ideal plant to explore how these three pigments contribute to leaf color. The molecular mechanisms of the coloration in ornamental kale could provide reference for exploring the mechanisms of pigmentation in other plants. Results In this study, we sequenced the transcriptome and determined the pigment contents of an unusual cultivar of ornamental kale with three different types of leaf coloration: pink (C3), light pink (C2), and variegated pink–green (C1). A total of 23,965 differentially expressed genes were detected in pairwise comparisons among the three types of leaves. The results indicate that Bo9g058630 coding dihydroflavonol 4–reductase (DFR) and Bo3g019080 coding shikimate O–hydroxycinnamoyltransferase (HCT) acted in anthocyanin biosynthesis in pink leaves. Bo1g053420 coding pheophorbidase (PPD) and Bo3g012430 coding 15–cis–phytoene synthase (crtB) were identified as candidate genes for chlorophyll metabolism and carotenoid biosynthesis, respectively. The transcription factors TT8, MYBL2, GATA21, GLK2, and RR1 might participate in triggering the leaf color change in ornamental kale. Anthocyanin content was highest in C3 and lowest in C1. Chlorophyll and carotenoid contents were lowest in C2 and highest in C1. Conclusions Based on these findings, we suspected that the decrease in anthocyanin biosynthesis and the increase in chlorophyll and carotenoid biosynthesis might be the reason for the leaf changing from pink to variegate pink–green in this unusual cultivar. Our research provides insight into the molecular mechanisms of leaf coloration in ornamental kale, contributing to a theoretical foundation for breeding new varieties.

Standardized flux seasonality metrics: a companion dataset for FLUXNET annual product

Phenological events are integrative and sensitive indicators of ecosystem processes that respond to climate, water and nutrient availability, disturbance, and environmental change. The seasonality of ecosystem processes, including biogeochemical fluxes, can similarly be decomposed to identify key transition points and phase durations, which can be determined with high accuracy, and are specific to the processes of interest. As the seasonality of different processes differ, it can be argued that the interannual trends and responses to environmental forcings can be better described through the fluxes' own temporal characteristics than through correlation to traditional phenological events like bud break or leaf coloration. Here we present a global dataset of seasonality or phenological metrics calculated for gross primary productivity (GPP), ecosystem respiration (RE), latent heat (LE), and sensible heat (H) calculated for the FLUXNET2015 Dataset of about 200 sites and 1500 site years of data. The database includes metrics (i) on an absolute flux scale for comparisons with flux magnitudes and (ii) on a normalized scale for comparisons of change rates across different fluxes. Flux seasonality was characterized by fitting a single-pass double-logistic model to daily flux integrals, and the derivatives of the fitted time series were used to extract the phenological metrics marking key turning points, season lengths, and rates of change. Seasonal transition points could be determined with a 90 % confidence interval of 6–11 d for GPP, 8–14 d for RE, 10–15 d for LE, and 15–23 d for H. The phenology metrics derived from different partitioning methods diverged, at times significantly. This Flux Seasonality Metrics Database (FSMD) can be accessed at the US Department of Energy's (DOE) Environmental Systems Science Data Infrastructure for a Virtual Ecosystem (ESS-DIVE, https://doi.org/10.15485/1602532; Yang and Noormets, 2020). We hope that it will facilitate new lines of research, including (1) validating and benchmarking ecosystem process models, (2) parameterizing satellite remote sensing phenology and PhenoCam products, (3) optimizing phenological models, and (4) generally expanding the toolset for interpreting ecosystems responses to changing climate.

Metabolomic and transcriptomic analyses of mutant yellow leaves provide insights into pigment synthesis and metabolism in Ginkgo biloba

Background Ginkgo (Ginkgo biloba L.) is an excellent landscape species. Its yellow-green leaf mutants are ideal materials for research on pigment synthesis, but the regulatory mechanism of leaf coloration in these ginkgo mutants remains unclear. Results We compared the metabolomes and transcriptomes of green and mutant yellow leaves of ginkgo over the same period in this study. The results showed that the chlorophyll content of normal green leaves was significantly higher than that of mutant yellow leaves of ginkgo. We obtained 931.52M clean reads from different color leaves of ginkgo. A total of 283 substances in the metabolic profiles were finally detected, including 50 significantly differentially expressed metabolites (DEMs). We identified these DEMs and 1361 differentially expressed genes (DEGs), with 37, 4, 3 and 13 DEGs involved in the photosynthesis, chlorophyll, carotenoid, and flavonoid biosynthesis pathways, respectively. Moreover, integrative analysis of the metabolomes and transcriptomes revealed that the flavonoid pathway contained the upregulated DEM (−)-epicatechin. Fourteen DEGs from the photosynthesis pathway were positively or negatively correlated with the DEMs. Conclusions Our findings suggest a complex metabolic network in mutant yellow leaves. This study will provide a basis for studies of leaf color variation and regulation.

Metabolic profiling and gene expression analysis provides insights into flavonoid and anthocyanin metabolism in poplar

Poplar, a woody perennial model, is a common and widespread tree genus. We cultivated two red leaf poplar varieties from bud mutation of Populus sp. Linn. ‘2025’ (also known as Zhonglin 2025, L2025 for shot): Populus deltoides varieties with bright red leaves (LHY) and completely red leaves (QHY). After measuring total contents of flavonoid, anthocyanin, chlorophyll, and carotenoid metabolites, a liquid chromatography-electrospray ionization-tandem mass spectrometry system was used for the relative quantification of widely targeted metabolites in leaves of three poplar varieties. A total of 210 flavonoid metabolites (89 flavones, 40 flavonols, 25 flavanones, 18 anthocyanins, 16 isoflavones, 7 dihydroflavonols, 7 chalcones, 5 proanthocyanidins, and 3 other flavonoid metabolites) were identified. Compared with L2025, 48 and 8 flavonoids were more and less abundant, respectively, in LHY, whereas 51 and 9 flavonoids were more and less abundant in QHY, respectively. On the basis of a comprehensive analysis of the metabolic network, gene expression levels were analyzed by deep-sequencing to screen for potential reference genes for the red leaves. Most phenylpropanoid biosynthesis pathway involved genes were differentially expressed among the examined varieties. Gene expression analysis also revealed several potential anthocyanin biosynthesis regulators including three MYB genes. The study results provide new insights into poplar flavonoid metabolites and represent the theoretical basis for future studies on leaf coloration in this model tree species.