The tree peony DREB transcription factor PrDREB2D regulates seed α-linolenic acid accumulation.

α-Linolenic acid (ALA), an essential fatty acid (FA) for human health, serves as the precursor of 2 nutritional benefits, docosahexaenoic acid and eicosapentaenoic acid, and can only be obtained from plant foods. We previously found that phospholipid:diacylglycerol acyltransferase 2 (PrPDAT2) derived from ALA-rich tree peony (Paeonia rockii) can promote seed ALA accumulation. However, the regulatory mechanism underlying its promoting effect on ALA accumulation remains unknown. Here, we revealed a tree peony dehydration-responsive element binding transcription factor, PrDREB2D, as an upstream regulator of PrPDAT2, which is involved in regulating seed ALA accumulation. Our findings demonstrated that PrDREB2D serves as a nucleus-localized transcriptional activator that directly activates PrPDAT2 expression. PrDREB2D altered the FA composition in transient overexpression Nicotiana benthamiana leaves and stable transgenic Arabidopsis (Arabidopsis thaliana) seeds. Repressing PrDREB2D expression in P. rockii resulted in decreased PrPDAT2 expression and ALA accumulation. In addition, PrDREB2D strengthened its regulation of ALA accumulation by recruiting the cofactor ABA-response element binding factor PrABF2b. Collectively, the study findings provide insights into the mechanism of seed ALA accumulation and avenues for enhancing ALA yield via biotechnological manipulation.

The tree peony nuclear factor Y transcription factor PrNF-YC2 promotes seed oil accumulation.

Seed oil not only provides energy for seed postgermination development but also provides essential nutrients and raw materials for human products. However, the transcriptional regulatory mechanism controlling seed oil accumulation remains largely unknown. Tree peony (Paeonia rockii) is an emerging woody oilseed crop in China that is known for its high-quality seed oil. Here, we revealed that a tree peony nuclear factor Y transcription factor, PrNF-YC2, is expressed predominantly in developing seeds and functions as an essential positive regulator of seed oil accumulation. PrNF-YC2 promoted oil accumulation in both transient ectopic overexpression Nicotiana benthamiana leaves and stable transgenic Arabidopsis thaliana seeds, globally upregulating the expression of genes involved in oil accumulation. In contrast, PrNF-YC2-silenced tree peony leaves using a virus-induced gene silencing system showed reduced oil content and expression of oil synthesis-related genes, including four master positive regulators contributing to oil accumulation, namely, LEAFY COTYLEDON1 (LEC1), ABSCISIC ACID INSENSITIVE3 (ABI3), FUSCA3 (FUS3), and WRINKLED1 (WRI1). We demonstrated that PrNF-YC2 directly activates PrLEC1 and PrABI3 alone and indirectly activates PrFUS3 and PrWRI1 by interacting with PrLEC1. Moreover, interaction with PrLEC1 also enhances the activation capacity of PrNF-YC2. The activation of these four master positive regulators by PrNF-YC2 triggered the upregulation of numerous oil synthesis-related genes, thus promoting oil accumulation. These findings provide new insights into the regulatory mechanism of seed oil accumulation and manipulation of PrNF-YC2 may be beneficial for enhancing oil yield in tree peony and other oilseed crops.

Tree peony transcription factor PrWRI1 enhances seed oil accumulation.

BACKGROUND: WRINKLED1 (WRI1) encodes a transcription factor, belonging to the APETALA2 (AP2) family, and plays a key role in regulating plant oil biosynthesis. As a newly woody oil crop, tree peony (Paeonia rockii) was notable for the abundant unsaturated fatty acids in its seed oil. However, the role of WRI1 during the accumulation of P. rockii seeds oil remains largely unknown. RESULTS: In this study, a new member of the WRI1 family was isolated from P. rockii and was named PrWRI1. The ORF of PrWRI1 consisted of 1269 nucleotides, encoding a putative protein of 422 amino acids, and was highly expressed in immature seeds. Subcellular localization analysis in onion inner epidermal cells showed that PrWRI1 was located at the nucleolus. Ectopic overexpression of PrWRI1 could significantly increase the total fatty acid content in Nicotiana benthamiana leaf tissue and even PUFAs in transgenic Arabidopsis thaliana seeds. Furthermore, the transcript levels of most genes related to fatty acids (FA) synthesis and triacylglycerol (TAG) assembly were also up-regulated in transgenic Arabidopsis seeds. CONCLUSIONS: Together, PrWRI1 could push carbon flow to FA biosynthesis and further enhance the TAG amount in seeds with a high proportion of PUFAs.

Comprehensive Metabolite Profile Uncovers the Bioactive Components, Antioxidant and Antibacterial Activities in Wild Tree Peony Leaves.

Tree peonies (Paeonia Section Moutan)-including nine wild species, which belong to subsections Vaginatae and Delavayanae-are economically important plants with ornamental, nutritional, and medicinal applications. In this study, for the first time, we determined the bioactive components and antioxidant activities and antibacterial activities of the newly grown leaves of nine wild tree peony species (WTPS). A total of 276 bioactive components were identified through non-targeted metabolomics; more than 80% of the 276 metabolites identified are terpenoids and flavonoids. A total of 42 differential metabolites were quantitatively determined. The main differential metabolites were Paeoniflorin, Luteoloside, Hyperin, Apigenin-7-glucoside, Rhoifolin, and Cantharidin. Such a high terpenoid and flavonoid content of the leaf extracts renders them as species with strong antibacterial capacities, and most of the bacteria tested showed greater sensitivity derived from the members of subsection Vaginatae than those of subsection Delavayanae. All WTPS have significant antioxidant activity; this activity is attributed to high levels of the total phenolic content (TPC) and total flavonoid content (TFC), of which, among the nine WTPS, P. lutea has the strongest antioxidant capacity. Our results provided a theoretical basis for the in-deep application of tree peony leaves for food, medical, and pharmaceutical industries.

Investigation of Volatile Components and Assessment of Antioxidant Potential in Seven Lamiaceae Plant Hydrosols.

Aromatic plants of the family Lamiaceae, especially of the genus Thymus, have promising antioxidant applications in pharmacology, medicine, food, cosmetology, and aromatherapy. Hydrosols (HDs) were extracted by hydrodistillation from seven species of Lamiaceae, including Thymus vulgaris, Thymus mongolicus, Mentha × piperita, Melissa officinalis, Rosmarinus officinali, Salvia elegans, and Leonurus artemisia. In total, 369 volatile components were determined and analyzed by gas chromatography-mass spectrometry (GC-MS). Among them, alcohols (2.86-28.48%), ethers (2.46-10.69%), and phenols (0.11-21.78%) constituted a large proportion, mainly linalool (0.28-19.27%), eucalyptol (0.16-6.97%), thymol (0-19.54%), and carvacrol (0-26.82%). Multivariate statistical analyses were performed and 27 differential metabolites were screened. Three different methods (ABTS+•, DPPH•, and FRAP) were used to determine the in vitro antioxidant activity of seven HDs. Thymus vulgaris hydrosols (Tv HDs) and Thymus mongolicus hydrosols (Tm HDs) had the strongest antioxidant activity and their stronger antioxidant capacity was related to their high levels of phenolic constituents, mainly thymol. The antioxidant activity of the other five Lamiaceae HDs was associated with their high alcohol (mainly linalool and eucalyptol) content, and the alcohol constituents may synergistically affect their antioxidant capacity. Therefore, the present study suggests that Lamiaceae plants can be utilized as antioxidant products or antioxidants in different industrial sectors including pharmaceuticals, food, cosmetics, and agrochemicals.

Integrative lipidomics profile uncovers the mechanisms underlying high-level α-linolenic acid accumulation in Paeonia rockii seeds.

Tree peony (Paeonia rockii) is an excellent woody oilseed crop, known for its high α-linolenic acid (ALA, ~45%) content, which is of great value for human health. However, the mechanisms underlying this high-level ALA accumulation in tree peony seeds are poorly understood. In this study, we evaluated the dynamic changes in the lipidomic profile of P. rockii seeds during development. A total of 760 lipid molecules were identified in P. rockii seeds; triacylglycerol (TAG) lipid molecules showed the highest abundance and diversity, both increasing during seed development. Particularly, ALA was the predominant fatty acid at the TAG sn-3 position. We further characterized two diacylglycerol acyltransferase (DGAT) genes and three phospholipid:diacylglycerol acyltransferase (PDAT) genes involved in the transfer of fatty acids to the TAG sn-3 position. Gene expression and subcellular localization analyses suggested that PrDGATs and PrPDATs may function as endoplasmic reticulum-localized proteins in seed TAG biosynthesis. In vitro functional complementation analysis showed different substrate specificities, with PrPDAT2 having a specific preference for ALA. Multiple biological assays demonstrated that PrDGAT1, PrDGAT2, PrPDAT1-2, and PrPDAT2 promote oil synthesis. Specifically, PrPDAT2 leads to preferential ALA in the oil. Our findings provide novel functional evidence of the roles of PrDGAT1 and PrPDAT2, which are potential targets for increasing the ALA yield in tree peony and other oilseed crops.

Intra- and interspecific ecophysiological responses to waterlogging stress in two contrasting waterlogging-tolerant arbor species.

At present, establishing planted forests, typically composed of not more than two tree species, to avoid forest losses has received increasing attention. In addition, investigating the impact of environmental stress such as waterlogging on different planting patterns is essential for improving wetland ecosystem resilience. Knowledge about the impact of waterlogging on planted forests is crucial for developing strategies to mitigate its adverse effects. Here, we conducted experimentally a simulated pure and mixed planting system composed of two contrasting WL-tolerant species (Cleistocalyx operculatus and Syzygium cumini) to determine their ecophysiological responses based on the type of interaction. Results showed that the aboveground growth performance of S. cumini was better than that of C. operculatus under well-watered conditions regardless of the planting model, which is contrary to the belowground accumulation that was significantly improved in C. operculatus. Intra- and interspecific interactions in different planting models facilitated the growth performance of C. operculatus while provoking a significant competition in S. cumini under waterlogging. Such phenomenon was explained through the remarkable ability of C. operculatus to naturally increase its root network under stress on non-stress conditions compared with S. cumini. In this study, two main factors are proposed to play key roles in the remarkable performance of C. operculatus compared with S. cumini following the planting model under waterlogging. The high level of nitrogen and phosphor absorption through C. operculatus primary roots and the significant starch biosynthesis constituted the key element that characterized the facilitation or competition within the intra- or interspecific interactions shown in C. operculatus compared with S. cumini. Furthermore, the intraspecific competition is more pronounced in S. cumini than in C. operculatus when grown in a pure planting pattern, particularly when subjected to waterlogging. However, when the two species are planted together, this competition is alleviated, resulting in enhanced waterlogging tolerance.

Corrigendum: A Tree Peony Trihelix Transcription Factor PrASIL1 Represses Seed Oil Accumulation.

[This corrects the article DOI: 10.3389/fpls.2021.796181.].

A Tree Peony Trihelix Transcription Factor PrASIL1 Represses Seed Oil Accumulation.

In many higher plants, seed oil accumulation is governed by complex multilevel regulatory networks including transcriptional regulation, which primarily affects fatty acid biosynthesis. Tree peony (Paeonia rockii), a perennial deciduous shrub endemic to China is notable for its seed oil that is abundant in unsaturated fatty acids. We discovered that a tree peony trihelix transcription factor, PrASIL1, localized in the nucleus, is expressed predominantly in developing seeds during maturation. Ectopic overexpression of PrASIL1 in Nicotiana benthamiana leaf tissue and Arabidopsis thaliana seeds significantly reduced total fatty acids and altered the fatty acid composition. These changes were in turn associated with the decreased expression of multitudinous genes involved in plastidial fatty acid synthesis and oil accumulation. Thus, we inferred that PrASIL1 is a critical transcription factor that represses oil accumulation by down-regulating numerous key genes during seed oil biosynthesis. In contrary, up-regulation of oil biosynthesis genes and a significant increase in total lipids and several major fatty acids were observed in PrASIL1-silenced tree peony leaves. Together, these results provide insights into the role of trihelix transcription factor PrASIL1 in controlling seed oil accumulation. PrASIL1 can be targeted potentially for oil enhancement in tree peony and other crops through gene manipulation.

Virus-induced gene silencing in the perennial woody Paeonia ostii.

Tree peony is a perennial deciduous shrub with great ornamental and medicinal value. A limitation of its current functional genomic research is the lack of effective molecular genetic tools. Here, the first application of a Tobacco rattle virus (TRV)-based virus-induced gene silencing (VIGS) in the tree peony species Paeonia ostii is presented. Two different approaches, leaf syringe-infiltration and seedling vacuum-infiltration, were utilized for Agrobacterium-mediated inoculation. The vacuum-infiltration was shown to result in a more complete Agrobacterium penetration than syringe-infiltration, and thereby determined as an appropriate inoculation method. The silencing of reporter gene PoPDS encoding phytoene desaturase was achieved in TRV-PoPDS-infected triennial tree peony plantlets, with a typical photobleaching phenotype shown in uppermost newly-sprouted leaves. The endogenous PoPDS transcripts were remarkably down-regulated in VIGS photobleached leaves. Moreover, the green fluorescent protein (GFP) fluorescence was detected in leaves and roots of plants inoculated with TRV-GFP, suggesting the capability of TRV to silence genes in various tissues. Taken together, the data demonstrated that the TRV-based VIGS technique could be adapted for high-throughput functional characterization of genes in tree peony.

[Emission of Particulate Matter, Organic and Elemental Carbon from Burning of Fallen Leaves].

The two kinds of burning conditions, i. e., flaming and smoldering, were selected to investigate the particulate matter (PM), organic carbon (OC), and elemental carbon ( EC) from burning of ten kinds of fallen leaves. In the experiment, the emission smoke was sampled from the fallen leaves burning, in which the OC and EC loadings were measured by the Thermal Optical Carbon Analyzer. The results showed that the emission factors of PM, OC, and EC were 7.9-31.9, 0.9-9.7, and 3.6-13.9 g x kg(-1), with the average values of 19.7, 5.2, and 6.8 g x kg(-1), respectively, under the flaming condition. The emission factors of PM, OC, and EC were 61.3-128.9, 31.7-60.4, and 1.9-6.0 g x kg(-1), with the average values of 91.0, 43.0, and 4.0 g x kg(-1), respectively, under the smoldering condition. The OC/EC ratio ranged from 0.21 to 1.82 and from 8.16 to 16.84 under the flaming and smoldering condition, respectively. The OC/PM and EC/PM ratios ranged from 0.11 to 0.41 and from 0.18 to 0.56, respectively under the flaming condition. The OC/PM and EC/PM ratios, however, ranged from 0.43 to 0.53 and from 0.03 to 0.06, respectively, under the smoldering condition. The OC emission factor was well correlated with the PM emission factor in the two burning conditions. Those results indicated that rather different emission factors occurred in all kind of components in different burning emission. In addition, the OC emission factor was higher under the smoldering condition than that under the flaming condition. However, the EC emission factor was higher under flaming condition, compared with that under smoldering condition. Analysis of the PM, OC, and EC emission factor and their ratios was beneficial for building the emission list from the biomass burning and the sources apportionment.

[Organic Carbon and Elemental Carbon in Forest Biomass Burning Smoke].

Ten kinds of trees were selected for preparing dry and wet stick samples. Concentrations of organic carbon (OC), elemental carbon (EC) in particular matter produced by sticks samples in the flaming and smoldering were analyzed through the Thermal Optical Carbon Analyzer (Model 2001A). The results showed that mean values of OC (EF(OC)), EC (EF(EC)), PM (EF(PM)) emission factors were 6.8, 2.1, 16.5 g x kg(-1) in the dry stick flaming smoke, 57.5, 11.1, 130.9 g x kg(-1) in the dry stick smoldering smoke, 13.6, 3.3, 30.5 g x kg(-1) in the wet stick flaming smoke, 57.6, 9.6, 125.6 g x kg(-1) in the wet stick smoldering smoke. Compared to the flaming condition, EF(OC), EF(EC), EF(PM), were much higher in the smoldering condition. In the flaming condition, EF(OC), EF(EC), EF(PM) had positive correlations with the moisture content. The mean values of OC/PM, EC/PM, TC/PM (TC = OC + EC) were 45%, 10%, 55%, and the mass fractions of OC was much higher in smoldering condition than those in flaming condition, but the mass fractions of EC was lower in the smoldering condition. Compared to dry sticks, the smoke of wet sticks combustion had higher mass fractions of OC and lower mass fractions of EC. The mean value of OC/EC was 3.3 (2.5-5.2) in the dry stick flaming smoke, and was 5.2 (4.3-6.3) in the dry stick smoldering smoke, in the wet stick flaming smoke was 4.1 (3.1-5.3), and was 6.2 (4.2-8.4) in the wet stick smoldering smoke. Compared to the flaming condition, the mean value of OC/EC was higher in the smoldering condition, and the mean value of OC/EC was much higher in high moisture content stick combustion smoke. The correlation coefficient between OC and EC was 0.985 in dry stick combustions, and was 0.915 in wet stick combustions. So, based on the flaming and smoldering condition, the correlation between OC and EC was significant in different moisture contents of sticks.

[Composition of organic carbon/elemental carbon and water-soluble ions in rice straw burning].

Six types of rice straw were selected in China in this paper, the homemade biomass combustion devices were used to simulate the outdoor burning. The concentrations of organic carbon (OC), elemental carbon (C) and water-soluble ions in particular matter produced by the flaming and smoldering were analyzed using Thermal Optical Carbon Analyzer (Model 2001A) and Ion Chromatography(ISC 2000/ISC 3000). The results showed that the mean value of OC (EFoc) and EC (EFEC) emission factors were (6.37 +/- 1.86) g x kg(-1) and (1.07 +/- 0.30) g x kg(-1) under the flaming conditions, respectively, while under the smoldering conditions the two mean values were (37.63 +/- 6.26) g x kg(-1) and (4.98 x 1.42) g x kg(-1). PM, OC and EC emitted from the same kind of rice straw had similar change trends. The average values of OC/EC under flaming and smoldering were 5.96 and 7.80, and the value of OC/PM was almost unchanged along with the combustion state. Nevertheless, the values of EC/PM under flaming and smoldering were 0.06-0.08 and 0.08-0.11, respectively. The trend of combustion state could be determined using the ratio of EC/PM and the RZ of emitted OC and EC through those two types of combustion reached 0. 97, which was significantly correlated at the 0. 01 level. Among the anions, Cl- showed the highest concentration, the results indicated that the average value of of Cl- emission factor was (0.246 +/- 0.150) g x kg(-1) under flaming, while it was (0.301 +/- 0.274) g x kg(-1) under smoldering. However, A big difference between flaming and smoldering was found in the average value of K+ emission factor, where (0.118 +/- 0.051) g x kg(-1) of the former was significantly higher than the latter (0.053 +/- 0.031) g x kg(-1). When it came to Na, the result of smoldering was significantly higher than that of flaming. The correlation between water-soluble ions in flaming was more significant than smoldering. Rice straw burning could be distinguished from fossil fuels and some other biomass burning by the OC/EC ratio, and it could also be distinguished from sort of trees burning by the ratio of K+ /Na+, Cl-/Na+ in water-soluble ions.