LiNAC100 contributes to linalool biosynthesis by directly regulating LiLiS in Lilium 'Siberia'.

MAIN CONCLUSION: The transcription factor LiNAC100 has a novel function of regulating floral fragrance by directly regulating linalool synthase gene LiLiS. Lilium 'Siberia', an Oriental hybrid, is renowned as both a cut flower and garden plant, prized for its color and fragrance. The fragrance comprises volatile organic compounds (VOCs), primarily monoterpenes found in the plant. While the primary terpene synthases in Lilium 'Siberia' were identified, the transcriptional regulation of these terpene synthase (TPS) genes remains unclear. Thus, understanding the regulatory mechanisms of monoterpene biosynthesis is crucial for breeding flower fragrance, thereby improving ornamental and commercial values. In this study, we isolated a nuclear-localized LiNAC100 transcription factor from Lilium 'Siberia'. The virus-induced gene silencing (VIGS) of LiNAC100 was found to down-regulate the expression of linalool synthase gene (LiLiS) and significantly inhibit linalool synthesis. Conversely, transient overexpression of LiNAC100 produced opposite effects. Additionally, yeast one-hybrid and dual-luciferase assays confirmed that LiNAC100 directly activates LiLiS expression. Our findings reveal that LiNAC100 plays a key role in monoterpene biosynthesis in Lilium 'Siberia', promoting linalool synthesis through the activation of LiLiS expression. These results offer insights into the molecular mechanisms of terpene biosynthesis in Lilium 'Siberia' and open avenues for biotechnological enhancement of floral scent.

Selection and Validation of Reference Genes for Gene Expression Studies in Euonymus japonicus Based on RNA Sequencing.

Euonymus japonicus is one of the most low-temperature-tolerant evergreen broad-leaved tree species in the world and is widely used in urban greening. However, there are very few molecular biology studies on its low-temperature tolerance mechanism. So far, no researcher has selected and reported on its reference genes. In this study, 21 candidate reference genes (12 traditional housekeeping genes and 9 other genes) were initially selected based on gene expression and coefficient of variation (CV) through RNA-Seq (unpublished data), and qRT-PCR was used to detect the expression levels of candidate reference genes in three different groups of samples (leaves under different temperature stresses, leaves of plants at different growth stages, and different organs). After further evaluating the expression stability of these genes using geNorm, NormFinder, Bestkeeper, and RefFind, the results show that the traditional housekeeping gene eIF5A and the new reference gene RTNLB1 have good stability in the three different groups of samples, so they are reference genes with universality. In addition, we used eIF5A and RTNLB1 as reference genes to calibrate the expression pattern of the target gene EjMAH1, which confirmed this view. This article is the first to select and report on the reference gene of E. japonicus, laying the foundation for its low-temperature tolerance mechanism and other molecular biology research.

Chromosome-scale genome assembly and insights into the metabolome and gene regulation of leaf color transition in an important oak species, Quercus dentata.

Quercus dentata Thunb., a dominant forest tree species in northern China, has significant ecological and ornamental value due to its adaptability and beautiful autumn coloration, with color changes from green to yellow into red resulting from the autumnal shifts in leaf pigmentation. However, the key genes and molecular regulatory mechanisms for leaf color transition remain to be investigated. First, we presented a high-quality chromosome-scale assembly for Q. dentata. This 893.54 Mb sized genome (contig N50 = 4.21 Mb, scaffold N50 = 75.55 Mb; 2n = 24) harbors 31 584 protein-coding genes. Second, our metabolome analyses uncovered pelargonidin-3-O-glucoside, cyanidin-3-O-arabinoside, and cyanidin-3-O-glucoside as the main pigments involved in leaf color transition. Third, gene co-expression further identified the MYB-bHLH-WD40 (MBW) transcription activation complex as central to anthocyanin biosynthesis regulation. Notably, transcription factor (TF) QdNAC (QD08G038820) was highly co-expressed with this MBW complex and may regulate anthocyanin accumulation and chlorophyll degradation during leaf senescence through direct interaction with another TF, QdMYB (QD01G020890), as revealed by our further protein-protein and DNA-protein interaction assays. Our high-quality genome assembly, metabolome, and transcriptome resources further enrich Quercus genomics and will facilitate upcoming exploration of ornamental values and environmental adaptability in this important genus.

Transcriptome analysis identifies key gene LiMYB305 involved in monoterpene biosynthesis in Lilium 'Siberia'.

Lilium is a popular cut flower that is highly favored by consumers due to its snowy white color and strong fragrance, which originates from the release of monoterpenes. However, the underlying molecular mechanism of monoterpene synthesis remains poorly understood. In this study, the content of three main monoterpenes (linalool, ocimene, and myrcene) was examined in Lilium 'Siberia', and RNA sequencing of the 11 stages of flower development was conducted. The biosynthesis of the three monoterpenes increased with flower development, reaching their peak levels at the full flowering stage. Transcriptome data revealed 257,140 unigenes, with an average size of 794 bp, from which 43,934 differentially expressed genes were identified and enriched in the KEGG pathways partly involved in plant hormone signal transduction and monoterpenoid biosynthesis. Furthermore, the essential factor LiMYB305 was identified by WGCNA after the release of the flower fragrance. The transient silencing of LiMYB305 in petals using VIGS technology showed that the mRNA expression levels of LiLiS, LiOcS, and LiMyS were significantly downregulated and that the release of linalool, ocimene, and myrcene had decreased significantly. Y1H, LUC, and EMSA experiments revealed that LiMYB305 directly bound and activated the LiOcS promoter to increase the synthesis of monoterpenes. Taken together, these results provide insight into the molecular mechanism of monoterpene synthesis and provide valuable information to investigate the formation of the flower fragrance in Lilium.

Lilac (Syringa oblata) genome provides insights into its evolution and molecular mechanism of petal color change.

Color change during flower opening is common; however, little is understood on the biochemical and molecular basis related. Lilac (Syringa oblata), a well-known woody ornamental plant with obvious petal color changes, is an ideal model. Here, we presented chromosome-scale genome assembly for lilac, resolved the flavonoids metabolism, and identified key genes and potential regulatory networks related to petal color change. The genome assembly is 1.05 Gb anchored onto 23 chromosomes, with a BUSCO score of 96.6%. Whole-genome duplication (WGD) event shared within Oleaceae was revealed. Metabolome quantification identified delphinidin-3-O-rutinoside (Dp3Ru) and cyanidin-3-O-rutinoside (Cy3Ru) as the major pigments; gene co-expression networks indicated WRKY an essential regulation factor at the early flowering stage, ERF more important in the color transition period (from violet to light nearly white), while the MBW complex participated in the entire process. Our results provide a foundation for functional study and molecular breeding in lilac.

AmMYB24 Regulates Floral Terpenoid Biosynthesis Induced by Blue Light in Snapdragon Flowers.

Floral terpenoid volatiles are impacted by light quality. In snapdragon, blue light can significantly enhance the emissions of ocimene and myrcene and the expression of ocimene synthase (AmOCS) and myrcene synthase (AmMYS). However, the mechanisms underlying the response to blue light are largely unknown. In this study, two transcription factors (TFs), AmMYB24 and AmMYB63 were screened which showed high expression level under blue light. AmMYB24 exhibited synchronous expression with AmOCS. Moreover, AmOCS transcript expression was up-regulated in response to AmMYB24 overexpression. This activation is direct and occurs through binding of AmMYB24 to MYBCORECYCATB1 sites in the AmOCS promoter. In addition, AmMYB24 interacts with the blue light signal key receptor AmCRY1 and the transcriptional activation activity of AmMYB24 was decreased in AmCRY1 silencing flowers. Taken together, our results revealed the regulatory pathway of biosynthesis of ocimene induced by blue light mediated by AmMYB24 and AmCRY1. When snapdragon flowers were exposed to blue light, AmCRY1 was first activated, the light signal is transduced to AmMYB24 through interaction with AmCRY1, and finally AmMYB24 activates AmOCS by binding to its MYBCOREATCYCB1 motif, resulting in abundant ocimene emission.

Highly sensitive determination of trace antimony in water samples by cobalt ion enhanced photochemical vapor generation coupled with atomic fluorescence spectrometry or ICP-MS.

A method for highly sensitive determination of trace antimony was proposed by using cobalt ion enhanced photochemical vapor generation (PCVG) for sample introduction into atomic fluorescence spectrometry (AFS) or inductively coupled plasma-mass spectrometry (ICP-MS) for elemental detection. During the PCVG process, the sample introduction efficiency of Sb could be significantly improved by addition of 5 mg L-1 Co2+ in the mixed acid medium of 10% (v/v) formic acid and 20% (v/v) acetic acid, with a final 12-fold and 133-fold enhancement of AFS and ICP-MS intensity, respectively. The experimental conditions including enhancement ions, acid medium, UV irradiation, working gas as well as potential interference were investigated in detail. Under the optimal experimental conditions, the limit of detection (LOD) for Sb was 0.05 and 0.001 µg L-1 by using AFS and ICP-MS determination, respectively. The method was successfully used for analysis of real water samples, with satisfactory recoveries of 92-94%.

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We explored the effects of salt stress on the growth of Quercus mongolica and the effects of ectomycorrhizal fungi (ECMF) on the ion balance of Q. mongolica. After inoculating four kinds of ECMFs (Gomphidius visci-dus, Suillus leteus, Suillus grevillea, Boletus edulis) on Q. mongolica seedlings, we treated the annual non-mycorrhizated and mycorrhizated seedlings with NaCl stress (0, 100, 200, 300 mmol·L-1) for 36 days, and then analyzed the mycorrhizal characteristics, growth, leaf injury symptoms, leaf electrolyte permeability, water content, and ion contents in roots, stems, and leaves. The results showed that the four ECMFs could establish a symbiotic system with Q. mongolica, and that root system of mycorrhizal seedlings was stronger than that of non-mycorrhizal seedlings. Under salt stress, the growth of Q. mongolica seedlings was inhibited, with the symptoms of scorched leaves. The damage to leaf plasma membrane and the degree of water loss were aggravated with the increases of salt stress. Under low salt stress (100 mmol·L-1), Q. mongolica preferentially accumulated Na+ in roots and stems. Under medium-high salt stress (200-300 mmol·L-1), roots became the primary organ for accumulating Na+. ECMF regulated ion balance in plant by increasing the Na+ level in roots and reducing the Na+ accumulation in stems and leaves, enhancing the absorption of K+ and Ca2+ to increase the K+/Na+ and Ca2+/Na+. The four ECMFs had different mitigation effects on salt poisoning of Q. mongolica. G. viscidus had the strongest effect, followed by S. leteus, while S. grevillei and B. edulis had relatively little effect.

Event-Triggered Impulsive Control for Nonlinear Stochastic Systems.

We study the stabilization problem for nonlinear stochastic systems via an event-triggered impulsive control (ETIC) scheme, where the impulsive control time sequence is generated by the event-triggered mechanism (ETM). Both continuous ETM and periodic ETM are developed by continuous measuring and periodic sampling, respectively. The continuous ETM with time regularization is proposed to exclude the Zeno behavior. The upper bound of the sampling period is given for the periodic ETM. By means of the continuous ETM and periodic ETM, sufficient conditions are given to guarantee the p th moment uniform stability and the p th moment exponential stability of related systems. Moreover, LMI-based conditions of exponential stability in the mean square are established for linear stochastic systems under ETIC. Finally, two examples are presented to illustrate the proposed ETIC schemes, in which an example of the consensus of linear stochastic multiagent systems is considered.

Transcriptome sequencing reveals terpene biosynthesis pathway genes accounting for volatile terpene of tree peony.

MAIN CONCLUSION: Transcriptomic and volatile component analyses showed that high expression levels of genes from the terpenoid backbone biosynthesis pathway and the monoterpene metabolic pathway can strengthen the floral fragrance of tree peony. Floral fragrance is a crucial ornamental trait whose improvement is one of the primary objectives of tree peony breeding. So far, exploration of the floral fragrance of tree peony has focused on the identification of its volatile components, but the molecular mechanisms responsible for their formation remain unclear. Here, we identified 128 volatile components from the petals of tree peony and found that they consisted primarily of terpenes, alcohols, and esters. Based on the distribution pattern of these major fragrance components, 24 tree peony cultivars were classified into 4 types: grassy scent (ocimene), woody scent (longifolene), lily of the valley scent (linalool), and fruity scent (2-ethyl hexanol). We used RNA-seq to explore the mechanistic basis of terpenoid metabolism in tree peony petals with various scents. The expression levels of AACT, HMGR, PMK, DXS, DXR, HDS, HDR, and GGPS, which encode key enzymes of terpenoid backbone biosynthesis, were upregulated in 'Huangguan' (strong fragrance) compared to 'Fengdan' (faint fragrance). Moreover, the transcript abundance of LIS and MYS, two monoterpene synthase genes, was also enhanced in petals of 'Huangguan' compared to those of 'Fengdan'. Together, these results demonstrate that differences in the expression of genes from the monoterpene synthesis and terpenoid backbone pathways are associated with differences in the fragrance of tree peony. This research provides crucial genetic resources for fragrance improvement and also lays a foundation for further clarification of the mechanisms that underlie tree peony fragrance.

Ectomycorrhizal Inoculation Enhances the Salt Tolerance of Quercus mongolica Seedlings.

Salt stress harms the growth and development of plants, and the degree of soil salinization in North China is becoming increasingly severe. Ectomycorrhiza (ECM) is a symbiotic system formed by fungi and plants that can improve the growth and salt tolerance of plants. No studies to date have examined the salt tolerance of Quercus mongolica, a typical ectomycorrhizal tree species of temperate forests in the northern hemisphere. Here, we inoculated Q. mongolica with two ectomycorrhizal fungi (Gomphidius viscidus; Suillus luteus) under NaCl stress to characterize the effects of ECM. The results showed that the symbiotic relationship of Q. mongolica with G. viscidus was more stable than that with S. luteus. The cross-sectional area of roots increased after inoculation with the two types of ectomycorrhizal fungi. Compared with the control group, plant height, soluble sugar content, and soluble protein content of leaves were 1.62, 2.41, and 2.04 times higher in the G. viscidus group, respectively. Chlorophyll (Chl) content, stomatal conductance (Gs), and intracellular CO2 concentration (Ci) were significantly higher in Q. mongolica inoculated with ectomycorrhizal fungi than in the control, but differences in the net photosynthetic rate (Pn), transpiration rate (Tr), and photosystem II maximum photochemical efficiency (Fv/Fm) were lower. The relative conductivity of Q. mongolica inoculated with the two ectomycorrhizal fungi was consistently lower than that of non-mycorrhizal seedlings, with the effect of G. viscidus more pronounced than that of S. luteus. The malondialdehyde (MDA) content showed a similar pattern. Peroxidase (POD) and catylase (CAT) levels in mycorrhizal seedlings were generally higher than those of non-mycorrhizal seedlings under normal conditions, and were significantly higher than those of non-mycorrhizal seedlings on the 36th and 48th day after salt treatment, respectively. Overall, the results indicated that the salt tolerance of Q. mongolica seedlings was improved by ectomycorrhizal inoculation.

Development of EST-SSR markers and association mapping with floral traits in Syringa oblata.

BACKGROUND: Lilac (Syringa oblata) is an important woody plant with high ornamental value. However, very limited genetic marker resources are currently available, and little is known about the genetic architecture of important ornamental traits for S. oblata, which is hindering its genetic studies. Therefore, it is of great significance to develop effective molecular markers and understand the genetic architecture of complex floral traits for the genetic research of S. oblata. RESULTS: In this study, a total of 10,988 SSRs were obtained from 9864 unigene sequences with an average of one SSR per 8.13 kb, of which di-nucleotide repeats were the dominant type (32.86%, 3611). A set of 2042 primer pairs were validated, out of which 932 (45.7%) exhibited successful amplifications, and 248 (12.1%) were polymorphic in eight S. oblata individuals. In addition, 30 polymorphic EST-SSR markers were further used to assess the genetic diversity and the population structure of 192 cultivated S. oblata individuals. Two hundred thirty-four alleles were detected, and the PIC values ranged from 0.23 to 0.88 with an average of 0.51, indicating a high level of genetic diversity within this cultivated population. The analysis of population structure showed two major subgroups in the association population. Finally, 20 significant associations were identified involving 17 markers with nine floral traits using the mixed linear model. Moreover, marker SO104, SO695 and SO790 had significant relationship with more than one trait. CONCLUSION: The results showed newly developed markers were valuable resource and provided powerful tools for genetic breeding of lilac. Beyond that, our study could serve an efficient foundation for further facilitate genetic improvement of floral traits for lilac.

Nitric oxide modulating ion balance in Hylotelephium erythrostictum roots subjected to NaCl stress based on the analysis of transcriptome, fluorescence, and ion fluxes.

Soil salinization is one of the main stress factors that affect both growth and development of plants. Hylotelephium erythrostictum exhibits strong resistance to salt, but the underlying genetic mechanisms remain unclear. In this study, hydroponically cultured seedlings of H. erythrostictum were exposed to 200 mM NaCl. RNA-Seq was used to determine root transcriptomes at 0, 5, and 10 days, and potential candidate genes with differential expression were analyzed. Transcriptome sequencing generated 89.413 Gb of raw data, which were assembled into 111,341 unigenes, 82,081 of which were annotated. Differentially expressed genes associated to Na+ and K+ transport, Ca2+ channel, calcium binding protein, and nitric oxide (NO) biosynthesis had high expression levels in response to salt stress. An increased fluorescence intensity of NO indicated that it played an important role in the regulation of the cytosolic K+/Na+ balance in response to salt stress. Exogenous NO donor and NO biosynthesis inhibitors significantly increased and decreased the Na+ efflux, respectively, thus causing the opposite effect for K+ efflux. Moreover, under salt stress, exogenous NO donors and NO biosynthesis inhibitors enhanced and reduced Ca2+ influx, respectively. Combined with Ca2+ reagent regulation of Na+ and K+ fluxes, this study identifies how NaCl-induced NO may function as a signaling messenger that modulates the K+/Na+ balance in the cytoplasm via the Ca2+ signaling pathway. This enhances the salt resistance in H. erythrostictum roots.

Evaluation of the environmental and plant growth effectiveness of a new substrate consisting of municipal sludge and fly ash.

In order to seek a safe, sustainable, and low-cost method for reuse of municipal sewage sludge, four species of native plants, i.e., Forsythia suspensa, Sophora japonica, Cotinus coggygria, and Ailanthus altissima were planted in flowerpots containing 4 growth substrates consisting of raw sludge and fly ash at volume/volume ratios of 20:80, 40:60, 60:40, and 80:20, respectively. The results showed that the physiochemical characteristics of the sewage sludge and fly ash were complementary. The sludge supplied the nutrients and the fly ash maintained air permeability in the mixed substrate. The mixed substrates containing 40-60% sewage sludge that belonged to sand clay loam were suitable for the seedling growth of the four species. After the end of the growing season, the electrical conductivity, pH, and contents of organic matter, nitrogen, potassium, and heavy metals in the four growth substrates decreased significantly. Moreover, most of the heavy metals were removed from the substrates by seedling root system. A. altissima grew best, and heavy metal enrichments of F. suspense and C. coggygria were stronger than other two species. The results indicate that the new substrates containing 40-60% sludge exhibiting good physiochemical properties, are environmentally friendly, and suitable for landscape planting.

Feasible and effective reuse of municipal sludge for vegetation restoration: physiochemical characteristics and microbial diversity.

The large volume of municipal sludge causes environmental problems in cities worldwide. In this study, municipal sludge, mixed with construction waste residue, was used as substrate to plant Ailanthus altissima. The growth of A. altissima, the substrate characteristics, and substrate microbial diversity were measured to investigate potential recycling and reusing pathways of municipal sludge. The obtained results showed that compared to garden soil, the mixed substrate was weakly alkaline, and had higher nutrient contents, which is beneficial for A. altissima, and results in better growth in mixed substrate. Although the contents of the main heavy metals in the mixed substrate were significantly higher than in garden soil, the values met the criterion of Class II soil in the Environmental Quality Standard for the Soils of China (GB15618-1995). Different substrates showed a variety of microbial diversities. Proteobacteria was the top microbial phylum in all samples, and higher relative abundances were found in samples containing municipal sludge. After growing A. altissima, the relative abundances of Acidobacteria and Gemmatimonadetes increased in the mixed substrate. Therefore, addition of construction waste residue and growth of A. altissima caused a difference. The microbial communities in the mixed substrate with A. altissima are both plant friendly and environmentally friendly. These results suggest this mixed substrate as a potentially feasible and effective pathway for the reuse and recycling of municipal sludge for vegetation restoration.

Interaction of ciprofloxacin with the activated sludge of the sewage treatment plant.

Interaction of ciprofloxacin with the activated sludge of the sewage treatment plant is of importance for the ciprofloxacin migration and risk control. More than 96.0% ciprofloxacin was removed through the sludge adsorption. The sludge surface charge varied little with ciprofloxacin since most ciprofloxacin was dissociated into the neutral one. No obvious shift was observed for the soluble carbohydrate concentration and composition with the addition of ciprofloxacin, indicating the weak interaction between the carbohydrates and ciprofloxacin. The introduction of ciprofloxacin resulted in a reduction of the soluble protein concentration, a marked increase of the extracellular protein fluorescence intensities, and a dramatic emergence of new extracellular proteins. The alteration of the proteins highlights the strong interaction between the extracellular proteins and ciprofloxacin, and the consequent integration of certain soluble proteins and original unextractable inner layer extracellular proteins into the extractable extracellular proteins. Different types of interactions are suggested to dominate between the extracellular proteins and the differently dissociated ciprofloxacin.

Antioxidant effect of aromatic volatiles emitted by Lavandula dentata, Mentha spicata, and M. piperita on mouse subjected to low oxygen condition.

This study aims to investigate the antioxidant effect of aromatic volatiles of three common aromatic plants, Lavandula dentata, Mentha spicata, and M. piperita. In this study, kunming mice subjected to low oxygen condition were treated with the volatiles emitted from these aromatic plants through inhalation administration. Then the blood cell counts, and the activities and gene expressions of antioxidant enzymes in different tissues were tested. The results showed that low oxygen increased the counts of red blood cells, white blood cells, and blood platelets of mice, and aromatic volatiles decreased their counts. Exposure to aromatic volatiles resulted in decreases in the malonaldehyde contents, and increases in the activities and gene expressions of superoxide dismutase, glutathione peroxidase, and catalase in different tissues under low oxygen. In addition, as the main component of aromatic volatiles, eucalyptol was the potential source that imparted positive antioxidant effect.

Molecular characterization and functional analysis of chalcone synthase from Syringa oblata Lindl. in the flavonoid biosynthetic pathway.

The flower color of Syringa oblata Lindl., which is often modulated by the flavonoid content, varies and is an important ornamental feature. Chalcone synthase (CHS) catalyzes the first key step in the flavonoid biosynthetic pathway. However, little is known about the role of S. oblata CHS (SoCHS) in flavonoid biosynthesis in this species. Here, we isolate and analyze the cDNA (SoCHS1) that encodes CHS in S. oblata. We also sought to analyzed the molecular characteristics and function of flavonoid metabolism by SoCHS1. We successfully isolated the CHS-encoding genomic DNA (gDNA) in S. oblata (SoCHS1), and the gene structural analysis indicated it had no intron. The opening reading frame (ORF) sequence of SoCHS1 was 1170bp long and encoded a 389-amino acid polypeptide. Multiple sequence alignment revealed that both the conserved CHS active site residues and CHS signature sequence were in the deduced amino acid sequence of SoCHS1. Crystallographic analysis revealed that the protein structure of SoCHS1 is highly similar to that of FnCHS1 in Freesia hybrida. The quantitative real-time polymerase chain reaction (PCR) performed to detect the SoCHS1 transcript expression levels in flowers, and other tissues revealed the expression was significantly correlated with anthocyanin accumulation during flower development. The ectopic expression results of Nicotiana tabacum showed that SoCHS1 overexpression in transgenic tobacco changed the flower color from pale pink to pink. In conclusion, these results suggest that SoCHS1 plays an essential role in flavonoid biosynthesis in S. oblata, and could be used to modify flavonoid components in other plant species.

Proteomic analysis of Potentilla fruticosa L. leaves by iTRAQ reveals responses to heat stress.

High temperature is an important environmental factor that affects plant growth and crop yield. Potentilla fruticosa L. has a developed root system and characteristics of resistance to several stresses (e.g., high temperature, cold, drought) that are shared by native shrubs in the north and west of China. To investigate thermotolerance mechanisms in P. fruticosa, 3-year-old plants were subjected to a high temperature of 42°C for 1, 2, and 3 days respectively before analysis. Then, we studied changes in cell ultrastructure using electron microscopy and investigated physiological changes in the leaves of P. fruticosa. Additionally, we used isobaric tags for relative and absolute quantification (iTRAQ) coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS) to study proteomic changes in P. fruticosa leaves after 3 d of 42°C heat stress. we found that the cell membrane and structure of chloroplasts, especially the thylakoids in P. fruticosa leaves, was destroyed by a high temperature stress, which might affect the photosynthesis in this species. We identified 35 up-regulated and 23 down-regulated proteins after the heat treatment. Gene Ontology (GO) analysis indicated that these 58 differentially abundant proteins were involved mainly in protein synthesis, protein folding and degradation, abiotic stress defense, photosynthesis, RNA process, signal transduction, and other functions. The 58 proteins fell into different categories based on their subcellular localization mainly in the chloroplast envelope, cytoplasm, nucleus, cytosol, chloroplast, mitochondrion and cell membrane. Five proteins were selected for analysis at the mRNA level; this analysis showed that gene transcription levels were not completely consistent with protein abundance. These results provide valuable information for Potentilla thermotolerance breeding.

Transcriptome Sequencing Analysis Reveals a Difference in Monoterpene Biosynthesis between Scented Lilium 'Siberia' and Unscented Lilium 'Novano'.

Lilium is a world famous fragrant bulb flower with high ornamental and economic values, and significant differences in fragrance are found among different Lilium genotypes. In order to explore the mechanism underlying the different fragrances, the floral scents of Lilium 'Sibeia', with a strong fragrance, and Lilium 'Novano', with a very faint fragrance, were collected in vivo using a dynamic headspace technique. These scents were identified using automated thermal desorption-gas chromatography/mass spectrometry (ATD-GC/MS) at different flowering stages. We used RNA-Seq technique to determine the petal transcriptome at the full-bloom stage and analyzed differentially expressed genes (DEGs) to investigate the molecular mechanism of floral scent biosynthesis. The results showed that a significantly higher amount of Lilium 'Siberia' floral scent was released compared with Lilium 'Novano'. Moreover, monoterpenes played a dominant role in the floral scent of Lilium 'Siberia'; therefore, it is believed that the different emissions of monoterpenes mainly contributed to the difference in the floral scent between the two Lilium genotypes. Transcriptome sequencing analysis indicated that ~29.24 Gb of raw data were generated and assembled into 124,233 unigenes, of which 35,749 unigenes were annotated. Through a comparison of gene expression between these two Lilium genotypes, 6,496 DEGs were identified. The genes in the terpenoid backbone biosynthesis pathway showed significantly different expression levels. The gene expressions of 1-deoxy-D-xylulose 5-phosphate synthase (DXS), 1-deoxy-D-xylulose-5-phosphate reductoisomerase (DXR), 4-hydroxy-3-methylbut-2-enyl diphosphate synthase (HDS), 4-hydroxy-3-methylbut-2-enyl diphosphate reductase (HDR), isopentenyl diphosphate isomerase (IDI), and geranyl diphosphate synthase (GPS/GGPS), were upregulated in Lilium 'Siberia' compared to Lilium 'Novano', and two monoterpene synthase genes, ocimene synthase gene (OCS) and myrcene synthase gene (MYS), were also expressed at higher levels in the tepals of Lilium 'Siberia', which was consistent with the monoterpene release amounts. We demonstrated that the high activation levels of the pathways contributed to monoterpene biosynthesis in Lilium 'Siberia' resulting in high accumulations and emissions of monoterpenes, which led to the difference in fragrance between these two Lilium genotypes.