Biomolecular condensates in plant cells: mediating and integrating environmental signals and development.

In response to the spatiotemporal coordination of various biochemical reactions and membrane-encapsulated organelles, plants appear to provide another effective mechanism for cellular organization by phase separation that allows the internal compartmentalization of cells to form a variety of membrane-less organelles. Most of the research on phase separation has centralized in various non-plant systems, such as yeast and animal systems. Recent studies have shown a remarkable correlation between the formation of condensates in plant systems and the formation of condensates in these systems. Moreover, the last decade has made new advances in phase separation research in the context of plant biology. Here, we provide an overview of the physicochemical forces and molecular factors that drive liquid-liquid phase separation in plant cells and the biochemical characterization of condensates. We then explore new developments in phase separation research specific to plants, discussing examples of condensates found in green plants and detailing their role in plant growth and development. We propose that phase separation may be a conserved organizational mechanism in plant evolution to help plants respond rapidly and effectively to various environmental stresses as sessile organisms.

The property and function of proteins undergoing liquid-liquid phase separation in plants.

A wide variety of membrane-less organelles in cells play an essential role in regulating gene expression, RNA processing, plant growth and development, and helping organisms cope with changing external environments. In biology, liquid-liquid phase separation (LLPS) usually refers to a reversible process in which one or more specific molecular components are spontaneously separated from the bulk environment, producing two distinct liquid phases: concentrated and dilute. LLPS may be a powerful cellular compartmentalisation mechanism whereby biocondensates formed via LLPS when biomolecules exceed critical or saturating concentrations in the environment where they are found will be generated. It has been widely used to explain the formation of membrane-less organelles in organisms. LLPS studies in the context of plant physiology are now widespread, but most of the research is still focused on non-plant systems; the study of phase separation in plants needs to be more thorough. Proteins and nucleic acids are the main components involved in LLPS. This review summarises the specific features and properties of biomolecules undergoing LLPS in plants. We describe in detail these biomolecules' structural characteristics, the mechanism of formation of condensates, and the functions of these condensates. Finally, We summarised the phase separation mechanisms in plant growth, development, and stress adaptation.

Temporal control of the Aux/IAA genes BnIAA32 and BnIAA34 mediates Brassica napus dual shade responses.

Precise responses to changes in light quality are crucial for plant growth and development. For example, hypocotyls of shade-avoiding plants typically elongate under shade conditions. Although this typical shade-avoidance response (TSR) has been studied in Arabidopsis (Arabidopsis thaliana), the molecular mechanisms underlying shade tolerance are poorly understood. Here we report that B. napus (Brassica napus) seedlings exhibit dual shade responses. In addition to the TSR, B. napus seedlings also display an atypical shade response (ASR), with shorter hypocotyls upon perception of early-shade cues. Genome-wide selective sweep analysis indicated that ASR is associated with light and auxin signaling. Moreover, genetic studies demonstrated that phytochrome A (BnphyA) promotes ASR, whereas BnphyB inhibits it. During ASR, YUCCA8 expression is activated by early-shade cues, leading to increased auxin biosynthesis. This inhibits hypocotyl elongation, as young B. napus seedlings are highly sensitive to auxin. Notably, two non-canonical AUXIN/INDOLE-3-ACETIC ACID (Aux/IAA) repressor genes, BnIAA32 and BnIAA34, are expressed during this early stage. BnIAA32 and BnIAA34 inhibit hypocotyl elongation under shade conditions, and mutations in BnIAA32 and BnIAA34 suppress ASR. Collectively, our study demonstrates that the temporal expression of BnIAA32 and BnIAA34 determines the behavior of B. napus seedlings following shade-induced auxin biosynthesis.

The Potential Relevance of PnDREBs to Panax notoginseng Nitrogen Sensitiveness.

The dehydration response element-binding (DREB) transcription factor is a subfamily of AP2/ERF. It actively responds to various abiotic stresses in plants. As one of the representative plants, Panax notoginseng is sensitive to Nitrogen (N). Here, bioinformatics analysis, the identification, chromosomal location, phylogeny, structure, cis-acting elements, and collinearity of PnDREBs were analyzed. In addition, the expression levels of PnDREBs were analyzed by quantitative reverse transcription PCR. In this study, 54 PnDREBs were identified and defined as PnDREB1 to PnDREB54. They were divided into 6 subfamilies (A1-A6). And 44 PnDREBs were irregularly distributed on 10 of 12 chromosomes. Each group showed specific motifs and exon-intron structures. By predicting cis-acting elements, the PnDREBs may participate in biotic stress, abiotic stress, and hormone induction. Collinear analysis showed that fragment duplication events were beneficial to the amplification and evolution of PnDREB members. The expression of PnDREBs showed obvious tissue specificity in its roots, flowers, and leaves. In addition, under the action of ammonium nitrogen and nitrate nitrogen at the 15 mM level, the level of PnDREB genes expression in roots varied to different degrees. In this study, we identified and characterized PnDREBs for the first time, and analyzed that PnDREBs may be related to the response of P. Notoginseng to N sensitiveness. The results of this study lay a foundation for further research on the function of PnDREBs in P. Notoginseng.

The DREB transcription factor, a biomacromolecule, responds to abiotic stress by regulating the expression of stress-related genes.

Abiotic stress is a crucial factor that affects plant survival and growth and even leads to plant death in severe cases. Transcription factors can enhance the ability of plants to fight against various stresses by controlling the expression of downstream genes. The dehydration response element binding protein (DREB) is the most extensive subfamily of AP2/ERF transcription factors involved in abiotic stress. However, insufficient research on the signal network of DREB transcription factors has limited plant growth and reproduction. Furthermore, field planting of DREB transcription factors and their roles under multiple stress also require extensive research. Previous reports on DREB transcription factors have focused on the regulation of DREB expression and its roles in plant abiotic stress. In recent years, there has been new progress in DREB transcription factors. Here, the structure and classification, evolution and regulation, role in abiotic stress, and application in crops of DREB transcription factors were reviewed. And this paper highlighted the evolution of DREB1/CBF, as well as the regulation of DREB transcription factors under the participation of plant hormone signals and the roles of subgroups in abiotic stress. In the future, it will lay a solid foundation for further study of DREB transcription factors and pave the way for the cultivation of resistant plants.

Genetic relationship of Pleione based on the chloroplast genome.

Pleione (Orchidaceae) is a very famous horticultural plant with a high international reputation for its unique flower shape and abundant color. The small difference in morphological characteristics among Pleione species caused by weak reproductive isolation and easy hybridization makes the taxonomic status of individual species very confusing. Chloroplast (cp) genome information is of great significance for the study of plant phylogeny and taxonomy. In this study, the cp genomes of Pleione were sequenced and the complete cp structure and sequence characteristics of 19 species were compared and analyzed. The cp genome of Pleione species exhibited a conserved tetrad structure and each species encoded 135 protein-coding genes, 38 tRNA and 8 RNA genes. The cp genome sizes of 19 Pleione were 157964-159269 bp and the length of LSC, SSC and IR were 85953-87172 bp, 18499-18712 bp, 26459-26756 bp, respectively. Palindromic and forward repeats accounted for a high proportion and the SSRs were mainly mononucleotide repeats in Pleione. Analysis of chloroplast sequence differences indicated that the differences in coding regions were smaller than those in non-coding regions, and the variation in LSC and SSC regions was greater than that in IR regions. Phylogenetic analysis showed that all Pleione species inferred from the cp genome were clustered together and received high support. However, the genetic relationship of Pleione plants is different from the current update system of this genus. Therefore, the demarcation of Pleione interspecific relationships still needs further investigation due to the lack of sufficient evidence. The cp genome serves as valuable information for the identification of Pleione species and the study of phylogenetic relationships.

Gnawing pressure led to the expansion of JAZ genes in angiosperms.

A long-standing problem in evolutionary biology is why some populations differentiate into many species while the majority do not. Angiosperms is an excellent group for investigating this problem because their diversity is unevenly distributed in space and phylogeny. Plant hormone participates in growth, development and defense. However, jasmonic acid (JA) was the only hormone response to bites. We first searched jasmonate ZIM-domain (JAZ), AUXIN/INDOLE ACETIC ACID (IAA / aux), PYR/PYL/RCAR (PYL), DELLA, and SUPPRESSOR OF MAX2 1-like (SMAX) in 272 plant species. We found the gene number change trends were consistent with origination rates and species numbers of angiosperms. So, 26 representative species were selected as an example for further analysis. The results showed JAZ had experienced two lineage-specific gene expansion events in angiosperms, which coincided with increases in mammalian body size and dental diversity. The proliferation of large herbivores as a results of mammalian prosperity after dinosaur extinction may be related to angiosperm evolution and bursting. The proliferation of large herbivores as the result of mammalian prosperity after the extinction of the dinosaurs was related to angiosperm evolution and bursting. Overall, our study uncovered a previously unknown co-evolution mechanism in terrestrial plants exposed to extreme environmental conditions.

Genome-wide identification and expression analysis of MYB gene family under nitrogen stress in Panax notoginseng.

The myeloblastosis (MYB) gene family, involved in regulating many important physiological and biochemical processes, is one of the largest transcript factor superfamilies in plants. Since the identification of genome sequencing of Panax notoginseng has been completed, there was little known about the whole genome of its specific MYB gene family and the response to abiotic stresses, in consideration of the excessive application of nitrogen fertilizers in P. notoginseng. In this study, 123 PnMYB genes (MYB genes of P. notoginseng) have been identified and divided into 3 subfamilies by the phylogenetic analysis. These PnMYB genes were unevenly located on 12 chromosomes. Meanwhile, the gene structure and protein conserved domain were established by MEME Suite. The analysis of collinear relationships reflected that there were 121 homologous genes between P. notoginseng and Arabidopsis and 30 between P. notoginseng and rice. Moreover, cis-acting elements of PnMYB gene promoters were predicted which indicated that PnMYBs are involved in biotic, abiotic stress, and hormone induction. The expressions of PnMYB transcription factors in its roots, flowers, and leaves were detected by qRT-PCR and they had tissue-specific expressions and related to the growth of different tissues. Under nitrogen stress, MYB transcription factors had great feedback. Ten R2R3-MYB subfamily genes were significantly induced and indicated the possible function of protecting P. notoginseng from excess nitrogen. With further knowledge on identification of PnMYB gene related to tissue selectivity and abiotic stresses, this study laid the foundation for the functional development of PnMYB gene family and improved the cultivation of P. notoginseng.

Multi-algorithm cooperation research of WRKY genes under nitrogen stress in Panax notoginseng.

WRKY transcription factors play an important role in the immune system and the innate defense response of plants. WRKY transcription factors have great feedback on nitrogen stress. In this study, bioinformatics was used to detect the WRKYs of Panax notoginseng (PnWRKYs). The response of PnWRKYs under nitrogen stress was also well studied. PnWRKYs were distributed on 11 chromosomes. According to PnWRKY and Arabidopsis thaliana WRKY (AtWRKY) domains, these PnWRKY proteins were divided into three groups by phylogenetic analysis. MEME analysis showed that almost every member contained motif 1 and motif 2. PlantCARE online predicted the cis-acting elements of the promoter. PnWRKY gene family members obtained 22 pairs of repeat fragments by collinearity analysis. The expression levels of PnWRKYs in different parts (roots, flowers, and leafs) were analyzed by the gene expression pattern. They reflected tissue-specific expressions. The qRT-PCR experiments were used to detect 74 PnWRKYs under nitrogen stress. The results showed that the expression levels of 8 PnWRKYs were significantly induced. The PnWRKY gene family may be involved in biotic/abiotic stresses and hormone induction. This study will not only lay the foundation to explore the functions of PnWRKYs but also provide candidate genes for the future improvement of P. notoginseng.

Characterization of the complete chloroplast genome sequence of Elaeagnus henryi Warb. ex Diels (Elaeagnaceae).

Elaeagnus henryi Warb. ex Diels belongs to the Elaeagnaceae. Here, we reveal the complete chloroplast genome of Elaeagnus henryi. The complete chloroplast genome is 152,244 bp in length and contains a large single-copy (LSC) region of 82,235 bp, a small single-copy (SSC) region of 18,279 bp and a pair of inverted repeats (IRs) of 25,865 bp. There are 126 genes, including 81 protein-coding, 37 transfer RNA (tRNA), and eight ribosomal RNA (rRNA) genes. The total GC content of the chloroplast genome sequence is 37.1%. The maximum-likelihood phylogenetic analysis indicated that E. henryi was sister to Elaeagnus pungens (MW145133). The result may be because the species are advanced and developed from the same ancestor.

Physiological and Transcriptional Analysis Reveals the Response Mechanism of Camellia vietnamensis Huang to Drought Stress.

Drought stress is considered the main obstacle restricting Camellia vietnamensis Huang (C. vietnamensis) yield. Hainan is the southernmost distribution region of C. vietnamensis in China and experiences a drought period annually. To study the drought-stress-response mechanism of C. vietnamensis, we treated seedlings of drought-tolerant (HD1) and drought-sensitive (WH1) cultivars with PEG-6000 (PEG) to simulate drought stress and compared the physiology and transcriptome of their leaves at 0 d, 3 d and 6 d posttreatment. Under drought stress, the growth of C. vietnamensis was inhibited, the relative water content (RWC) of leaves decreased and the contents of malondialdehyde (MDA), antioxidant enzyme activities, osmotic regulatory substances and secondary metabolites increased. Compared with those of WH1, the leaf RWC, osmotic-regulation substance content (proline, soluble protein and soluble sugar) and antioxidant enzyme activity (superoxide dismutase, peroxidase and catalase) of HD1 were significantly increased, while the relative electrical conductivity and MDA content were significantly decreased. Compared with WH1, 2812, 2070 and 919, differentially expressed genes (DEGs) were detected in HD1 0 d, 3 d and 6 d posttreatment, respectively, and the number of DEGs increased with increasing treatment time. The detected DEGs are involved in the drought stress response of C. vietnamensis mainly through plant-hormone signal transduction and lignin and flavonoid biosynthesis pathways. Drought stress significantly activated the expression of several lignin and flavonoid biosynthesis genes in HD1. Moreover, total flavonoid and total polyphenol contents in HD1 were significantly increased, suggesting that the accumulation of flavonoids may be a key factor in the drought stress response of C. vietnamensis. Additionally, 191 DEGs were associated with coding transcription factors (TFs). This study provides insight into the molecular mechanism of the drought stress response of C. vietnamensis and provides a theoretical basis for the development and cultivation of new drought-resistant cultivars.

Integrative Metabolome and Transcriptome Analysis Reveals the Regulatory Network of Flavonoid Biosynthesis in Response to MeJA in Camelliavietnamensis Huang.

Flavonoids are secondary metabolites widely found in plants, which perform various biological activities, such as antiinflammation, antioxidation, antitumor, and so on. Camellia vietnamensis Huang, a species of oil-tea Camellia tree, is an important woody oil crop species widely planted on Hainan Island, which provides health benefits with its high antioxidant activity and abundant flavonoid content. However, very little is known about the overall molecular mechanism of flavonoid biosynthesis in C. vietnamensis Huang. In this study, methyl jasmonate (MeJA) is used as an inducer to change the content of secondary metabolites in C. vietnamensis. Then, the potential mechanisms of flavonoid biosynthesis in C. vietnamensis leaves in response to MeJA were analyzed by metabolomics and transcriptomics (RNA sequencing). The results showed that metabolome analysis detected 104 flavonoids and 74 fatty acyls which showed different expression patterns (increased or decreased expression). It was discovered by KEGG analysis that three differentially accumulated metabolites (cinnamaldehyde, kaempferol and quercitrin) were annotated in the phenylpropanoid biosynthesis (ko00940), flavonoid biosynthesis (ko00941), and flavone and flavonol biosynthesis (ko00944) pathways. In the transcriptome analysis, 35 different genes involved in the synthesis of flavonoids were identified by MapMan analysis. The key genes (PAL, 4CL, CCR, CHI, CHS, C4H, FLS) that might be involved in the formation of flavonoid were highly expressed after 2 h of MeJA treatment. This study provides new insights and data supporting the molecular mechanism underlying the metabolism and synthesis of flavonoids in C. vietnamensis under MeJA treatment.

Effects of Different Processing Methods Based on Different Drying Conditions on the Active Ingredients of Salvia miltiorrhiza Bunge.

Compared to the traditional processing method, fresh processing can significantly enhance the preservation of biologically active ingredients and reduce processing time. This study evaluated the influences of fresh and traditional processing based on different drying conditions (sun drying, oven drying and shade drying) on the active ingredients in the roots and rhizomes of S. miltiorrhiza. High-performance liquid chromatography (HPLC) was utilized to determine the contents of six active ingredients in the roots and rhizomes of S. miltiorrhiza. The data were analyzed by fingerprint similarity evaluation, hierarchical cluster analysis (HCA) and principal component analysis (PCA). The results suggest that compared to the traditional processing method, the fresh processing method may significantly increase the preservation of biologically active ingredients. Furthermore, the findings demonstrated that among the three drying methods under fresh processing conditions, the shade-drying (21.02-26.38%) method is most beneficial for retaining the active ingredients in the roots and rhizomes of S. miltiorrhiza. Moreover, the fingerprint analysis identified 17 common peaks, and the similarity of fingerprints among samples processed by different methods ranged from 0.989 to 1.000. Collectively, these results suggest novel processing methods that may improve the yield of active ingredients for S. miltiorrhiza and may be implemented for industrial production.

Complete chloroplast genome of Holboellia grandiflora Réaubourg (Lardizabalaceae), a plant with an edible fruit.

Holboellia grandiflora Réaubourg (Lardizabalaceae) is an evergreen twining perennial woody vine. To our knowledge, this is the first report on the complete chloroplast genome sequence of H. grandiflora. The complete chloroplast genome sequence was 157,811 bp in length and contained a large single-copy region of 86,554 bp and a small single-copy region of 18,975 bp. A pair of inverted repeats of 26,141 bp were included. It contained 130 genes, comprising 37 transfer RNA genes, and eight ribosomal RNA genes, as well as 85 coding sequences (CDSs). The GC content of the complete chloroplast genome sequence was 38.7%. The phylogenetic tree showed a close relationship among the three species of Holboellia (H. grandiflora, H. angustifolia, and H. latifolia). These findings provide a reference for phylogenetic relationships and assessment of the genetic structure of the Lardizabalaceae family.

The whole chloroplast genome of Neomartinella yungshunensis (Brassicaceae), an unusual wild plant.

Neomartinella yungshunensis (W. T. Wang) Al-Shehbaz 2000 is a kind of perennial herb usually distributed in Yongshun County, Xiangxi Tujia Miao Autonomous Prefecture, Hunan Province. It was the first time to report the complete chloroplast genome sequence of N. yungshunensis. The complete chloroplast genome was 152,597 bp in size, including a large single-copy (LSC) region of 83,145 bp, a small single copy region (SSC) of 17,400 bp, and a pair of reverse repeats (IR) of 26,026 bp. It contained 133 genes in the chloroplast genome, including 87 protein-coding genes (PCGs), 37 transfer RNA (tRNA) genes, and 8 ribosomal RNA (rRNA) genes. The GC content of the chloroplast genome was 36.4%. The phylogenetic analysis showed that N. yungshunensis is closely related to Eutrema integrifolium (NC_049636).

Quality Evaluation of the Oil of Camellia spp.

The oil of Camellia spp. has become a well-known high-quality edible oil because of its rich nutrition. It is of great significance to breed fine varieties of Camellia spp. for the sustainable growth of the Camellia spp. industry. This study mainly evaluated the quality and antioxidant capacity of the camellia seed from several sources. The fatty acid composition and main active components of 40 kinds of C. oleifera, C. vietnamensis, C. osmantha, and C. gigantocarpa seeds, and so on, from different regions, were tested using GC-MS and HPLC. The quality of different Camellia spp. germplasm resources was comprehensively evaluated using multiple indices. The unsaturated fatty acid content and the antioxidant capacity of C. vietnamensis from Hainan were higher than those of C. oleifera Abel. In addition, there were a few differences in the fatty acid compositions of Camellia spp. oil from different species. Correlation analysis confirmed that rutin, total saponin, total flavonoids, squalene, and vitamin E were strongly correlated to the antioxidant capacity of Camellia spp. In the comprehensive evaluation, the best quality and strongest antioxidant activity were found for Chengmai Dafeng (C. vietnamensis). These methods in the study were applied for the first time for the quality evaluation of the Camellia spp. species. This study provided new insights into the quality evaluation of the Camellia spp. species, thus facilitating further development of variety breeding along with quality evaluation.

Multi-algorithm cooperation comprehensive research of bZIP genes under nitrogen stress in Panax notoginseng.

Basic leucine zipper (bZIP) transcription factors play an irreplaceable position in the regulation of plant secondary metabolism, growth and development, and resistance to abiotic stress. Panax notoginseng is a traditional medicinal plant in China, but the systematic identification and the resistance of Panax notoginseng bZIP (PnbZIP) family under nitrogen stress have not been reported before, considering the excessive application of N fertilizers. In this study, we conducted a genome-wide identification of the PnbZIP family and analyzed its phylogeny, tissue selectivity, and abiotic resistence. 74 PnbZIPs were distributed on 12 chromosomes and 8 were not successfully located. Through phylogenetic analysis of Arabidopsis and Panax notoginseng, we divided them into 14 subgroups. In the same subgroup, bZIPs had similiar intron/exon structure and conserved motifs. In the analysis of chromosome structure, two PnbZIP genes were duplicated in tandem on chromosome 3. Intraspecific collinearity analysis showed that 28 PnbZIPs participated in segmental replication. Each PnbZIP promoter contained at least one stress response element or stress-related hormone response element. RNA-seq and qRT-PCR methods were used to analyze the expression patterns of the PnbZIP gene in different tissues (roots, flowers, and leaves) and under different nitrogen stresses. The results showed that the PnbZIP gene had the highest expression level in flowers and reflected tissue-specific expressions. Meanwhile, under the stress of ammonium nitrogen fertilizer and nitrate nitrogen fertilizer, PnbZIPs in roots were differently expressed. 10 PnbZIP stress-responsive genes were screened for significant expression, among which PnbZIP46 was significantly up-regulated, which could be a candidate gene for resistance to Nitrogen stress. This study laid the foundation for functional identification of PnbZIPs and improved the cultivation of Panax notoginseng.

Characterization of the complete chloroplast genome sequence of Cardamine lyrata Bunge(Brassicaceae).

Cardamine lyrata Bunge 1833 grows near paddy fields, streams and shallow water. Its young stems and leaves can be eaten. It can also be used as medicine and has the effect of clearing away heat and dampness. The complete chloroplast genome sequence of the C. lyrata was determined and assembled. The complete genome is 155,170 bp in length, including a large single-copy region (LSC) of 84,270 bp, a small single-copy region (SSC) of 17,918 bp and two copies of inverted repeat (IR) regions of 26,491 bp. The overall GC content of C. lyrata is 36.2%. The genome of C. lyrata contains 131 genes, including 85 protein-coding genes (PCGs), 37 tRNAs, and 8 rRNAs. Phylogenetic analysis suggested that the ten species in Cardamine were clustered together into a single branch within the Brassicaceae family and C. amariformis is at the base of the tree and C. lyrata and C. fallax are sister groups of the inner clade.

Characterization of the complete chloroplast genome sequence of Maesa hupehensis Rehd (primulaceae).

Maesa hupehensis Rehd 1916 is mainly planted in Hubei and Sichuan. In this study, we assembled and characterized the complete chloroplast genome of M. hupehensis as resources for the future study. The chloroplast genome was 157,005 bp in length, with 37.3% GC content, composing of one large single copy (87,628 bp) and one small single copy (18,111 bp), separated by two inverted repeats (25,633 bp). A total of 130 genes were predicted, including 8 rRNAs, 37 tRNAs, and 84 protein-coding genes. Phylogenetic analysis showed that M. hupehensis was closely related to Maesa montana.