Late Embryogenesis Abundant Research Articles

Cross‐disciplinary insights into the mechanisms of plant cold hardiness: From molecules to ecosystems

Cross‐disciplinary insights into the mechanisms of plant cold hardiness: From molecules to ecosystems

Genome-wide identification, characterization, and expression pattern of the late embryogenesis abundant (LEA) gene family in Juglans regia and its wild relatives J. mandshurica

BackgroundLate Embryogenesis Abundant (LEA) proteins are a class of proteins associated with plant stress resistance. Two Juglans species, Juglans regia and J. mandshurica, are both diploid (2n = 32), monoecious perennial economic tree species with high edible, pharmaceutical, and timber value. The identification, characterization, and expression patterns of LEA proteins in J. regia and its wild relative, J. mandshurica, would not only provide the genetic basis of this gene family, but it would also supply clues for further studies of the evolution and regulating mechanisms of LEA proteins in other tree species.ResultsIn this study, we identified 25 and 20 members of the LEA gene family in Juglans regia and its wild relative, Juglans mandshurica, respectively. The results of phylogenetic analysis showed that the LEA members were divided into eight main subgroups. Predictions of their physicochemical properties showed the variable characteristics of LEA proteins, and the subcellular localization analysis indicated that most LEA proteins are localized in the nucleus. Chromosomal localization analysis and gene replication pattern prediction indicated that WGD is the predominant duplication mode of LEA genes. The results of the comparative analysis indicated a high level of collinearity between the two Juglans species. Analysis of cis-acting elements indicated that LEA genes had a relatively wide range of responses to abiotic stresses and phytohormonal processes, particularly in two phytohormones, methyl jasmonate and abscisic acid. Transcriptome profiling and qRT-PCR experiments showed that JrLEAs are commonly expressed in leaves, green husks, and male and female flowers, and most JmLEAs are more highly expressed in male flowers. We also hypothesized that JrLEAs are involved in the process of anthracnose resistance. Anthracnose-resistant varieties of JrLEAs presented relatively high expression levels at later stages.ConclusionIn this study, we provide a theoretical basis for the functional study of LEA genes in J. regia and J. mandshurica. Analysis of cis-acting elements and gene expression indicated that JrLEAs and JmLEAs play important roles in resistance to biotic stresses in these species.

Identification of the BcLEA Gene Family and Functional Analysis of the BcLEA73 Gene in Wucai (Brassica campestris L.).

Late embryogenesis abundant (LEA) proteins are important developmental proteins in the response of plants to abiotic stress. In our previous study, BcLEA73 was differentially expressed under low-temperature stress. Herein, we combined bioinformatics analysis, subcellular localization, expression assays, and stress experiments (including salt, drought, and osmotic stress) to identify and analyze the BcLEA gene family. Gene cloning and functional analysis of BcLEA73 were performed in tobacco and Arabidopsis. Based on the sequence homology and the available conservative motif, 82 BrLEA gene family members were identified and were divided into eight subfamilies in the genome-wide database of Chinese cabbage. The analysis showed that the BrLEA73 gene was located on chromosome A09 and belonged to the LEA_6 subfamily. Quantitative real-time PCR analysis indicated that the BcLEA genes were differentially expressed to varying degrees in the roots, stems, leaves, and petioles of Wucai. The overexpressed BcLEA73 transgenic plants exhibited no significant differences in root length and seed germination rates compared to the wild-type (WT) plants under control conditions. Under salt and osmotic stress treatment, the root length and seed germination rates of the BcLEA73-OE strain were significantly greater than those of WT plants. Under salt stress, the total antioxidant capacity (T-AOC) of the BcLEA73-OE lines increased significantly, and the relative conductivity, (REL), hydrogen peroxide (H2O2) content, and superoxide anion (O2-) production rate decreased significantly. Under drought treatment, the survival rate of the BcLEA73-OE lines was significantly higher than that of WT plants. These results showed that the BcLEA73 gene of Wucai functions in enhancing the tolerance of plants to salt, drought, and osmotic stress. This study provides a theoretical basis to explore the relevant functions of the BcLEA gene family members of Wucai.

Functional synergy between disordered proteins and sugars in desiccation protection.

Organisms across all biological kingdoms have evolved to survive near complete desiccation. Desiccation tolerance relies on the intracellular accumulation of high levels of specific intrinsically disordered proteins such as late embryogenesis abundant (LEA) proteins as well as non-reducing disaccharides, such as trehalose and sucrose. IDPs have few intramolecular bonds and a large solvent-accessible surface area, making their ensemble more sensitive to the solution environment than their well-folded counterparts. This implies that the ensemble and functions of protective IDPs are likely to be influenced by changes in metabolome induced by desiccation. Here we show that cosolutes enriched during desiccation, augment the protective function of full length IDPs during drying. We found that trehalose, a key molecule enriched in tardigrades, confers a synergistic protective effect when mixed with a model tardigrade IDP, CAHS D, at biological relevant ratios, and this synergy persists in vivo. Interestingly, a non-tardigrade sugar, sucrose, exerts less synergy. Extending these experiments, we found that LEA proteins from an organism synergizes best with the major non-reducing disaccharide (trehalose or sucrose) enriched in that organism during desiccation to promote protection. LEA proteins are made of 11-mer motif repeats; and interestingly, LEA-sugar synergy is not consistent for 11-, 22-, or 44-mer LEA motifs. Our results demonstrate that desiccation tolerant organisms may have evolved to create a specific cosolute environment to precisely tune the function of constituent IDPs. Beyond extending our knowledge of desiccation tolerance, our finding that cosolutes can tune the protective capacity of IDPs provides evidence that changing solution chemistry can modulate IDP function. We further seek to understand the mechanistic basis of IDP-cosolute synergy using biophysical approaches where we speculate that cosolutes modulate the local and global ensemble of IDPs resulting differences in IDP function.

Exogenous gibberellic acid shortening after-ripening process and promoting seed germination in a medicinal plant Panax notoginseng

BackgroundPanax notoginseng (Burk) F.H. Chen is an essential plant in the family of Araliaceae. Its seeds are classified as a type of morphophysiological dormancy (MPD), and are characterized by recalcitrance during the after-ripening process. However, it is not clear about the molecular mechanism on the after-ripening in recalcitrant seeds.ResultsIn this study, exogenous supply of gibberellic acid (GA3) with different concentrations shortened after-ripening process and promoted the germination of P. notoginseng seeds. Among the identified plant hormone metabolites, exogenous GA3 results in an increased level of endogenous hormone GA3 through permeation. A total of 2971 and 9827 differentially expressed genes (DEGs) were identified in response to 50 mg L−1 GA3 (LG) and 500 mg L−1 GA3 (HG) treatment, respectively, and the plant hormone signal and related metabolic pathways regulated by GA3 was significantly enriched. Weighted gene co-expression network analysis (WGCNA) revealed that GA3 treatment enhances GA biosynthesis and accumulation, while inhibiting the gene expression related to ABA signal transduction. This effect was associated with higher expression of crucial seed embryo development and cell wall loosening genes, Leafy Contyledon1 (LEC1), Late Embryogenesis Abundant (LEA), expansins (EXP) and Pectinesterase (PME).ConclusionsExogenous GA3 application promotes germination and shorts the after-ripening process of P. notoginseng seeds by increasing GA3 contents through permeation. Furthermore, the altered ratio of GA and ABA contributes to the development of the embryo, breaks the mechanical constraints of the seed coat and promotes the protrusion of the radicle in recalcitrant P. notoginseng seeds. These findings improve our knowledge of the contribution of GA to regulating the dormancy of MPD seeds during the after-ripening process, and provide new theoretical guidance for the application of recalcitrant seeds in agricultural production and storage.

Analyses of leaf morphological changes and expressed genes related to water shortage of Fagus orientalis populations in a drought gradient

Climate change models predict an increase in mean annual temperature, a decrease in precipitation and consequently a scarcity of water resources during the growing season for Iran, including the northern forests of the country. One of the consequences is increased water shortage stress during the summer season. Fagus orientalis Lipsky is one of the most important commercial broad leaf species in the Hyrcanian forests and its potential of phenotypic and genetic adaptation to the climate change is essentially vital. The present research was carried out along two precipitation gradients from very humid to semi-humid conditions on four beech populations. The results of the flexibility of the leaf morphological traits showed that the populations of dryer regions had smaller and more elongated leaves than other populations. Evaluation of the expression of seven candidate genes in response to water shortage showed that the expression of aldehyde dehydrogenase, ascorbate peroxidase, Dehydrin (DHN), early responsive to dehydration, late embryogenesis abundant and Cys/His-Type Zinc-Finger in the eastern population was higher than that in the western population, while a difference in elevation gradient was only observed in the expression of some genes. Therefore, the natural selection pressure in the past evolutionary periods in the westeast gradient seems to be higher than the elevation gradient and has led to greater resistance to drought. Principal component analysis revealed that aldehyde dehydrogenase genes (ALDH), early responsive to dehydration (ERD), late embryogenesis abundant (LEA) and DHN are the most effective genes responding to water shortage stress for Oriental beech in the Hyrcanian forests. Keywords: drought gradient, Fagus orientalis, gene expression, leaf morphology

Overexpression of ZmDHN15 Enhances Cold Tolerance in Yeast and Arabidopsis.

Maize (Zea mays L.) originates from the subtropical region and is a warm-loving crop affected by low-temperature stress. Dehydrin (DHN) protein, a member of the Group 2 LEA (late embryogenesis abundant proteins) family, plays an important role in plant abiotic stress. In this study, five maize DHN genes were screened based on the previous transcriptome sequencing data in our laboratory, and we performed sequence analysis and promoter analysis on these five DHN genes. The results showed that the promoter region has many cis-acting elements related to cold stress. The significantly upregulated ZmDHN15 gene has been further screened by expression pattern analysis. The subcellular localization results show that ZmDHN15 fusion protein is localized in the cytoplasm. To verify the role of ZmDHN15 in cold stress, we overexpressed ZmDHN15 in yeast and Arabidopsis. We found that the expression of ZmDHN15 can significantly improve the cold resistance of yeast. Under cold stress, ZmDHN15-overexpressing Arabidopsis showed lower MDA content, lower relative electrolyte leakage, and less ROS (reactive oxygen species) when compared to wild-type plants, as well as higher seed germination rate, seedling survival rate, and chlorophyll content. Furthermore, analysis of the expression patterns of ROS-associated marker genes and cold-response-related genes indicated that ZmDHN15 genes play an important role in the expression of these genes. In conclusion, the overexpression of the ZmDHN15 gene can effectively improve the tolerance to cold stress in yeast and Arabidopsis. This study is important for maize germplasm innovation and the genetic improvement of crops.

Genome-wide study and functional characterization elucidates the potential association of late embryogenesis abundant (LEA) genes with lotus seed development

Late embryogenesis abundant (LEA) proteins are extremely hydrophilic proteins imperatively associated with plant growth and development, as well as cell protection from abiotic stress. However, the genome-wide characterization of LEA gene family remains limited, especially in aquatic species such as lotus (Nelumbo spp.). Here, 57 putative LEA genes, including 28 NnLEAs and 29 NlLEAs were identified in the N.nucifera and N.lutea genomes, respectively. A total of 27 homologous LEA gene pairs were identified, indicating high degree of sequence homologies between the two Nelumbo species. Secondary structure prediction indicated high prevalence of alpha (α) helix structure among LEA proteins in the LEA_1, LEA_4, and SMP groups. Screening of putative promoter cis-elements revealed that NnLEA genes were involved in diverse biological processes. Most NnLEA genes were predominantly expressed in the late cotyledons and plumules development stages, suggesting their potential vital roles in lotus seed maturation. In addition, genes co-expressed with NnLEAs were involved in ABA signaling, seed maturation, and development processes. Overall, this study provides new insights for the in-depth understanding of the functions of NnLEA proteins in lotus seed development, and could act as a useful reference for the molecular breeding of seeds with prolonged lifespan.

Identification of candidate genes associated with sex differentiation and determination of gender diphasic plant Lilium apertum (Liliaceae)

• This is the first study to identify sex-biased genes in L. apertum, a gender diphasy plant. • Zeatin biosynthesis were upregulated in hermaphroditic flower, in line with high ZT and n levels. • Several signaling pathways related to PCD were up-regulated in male flower. • N deficiency or/and water loss signaling might participate sex change in L. apertum. Gender diphasy is a very rare sexual strategy in flowering plant, in which each individual can reproduce as either male or female phase plants over years according to circumstances. However, the limited genetic information related to the molecular basis of sex determination remains largely unknown. Here, comparative transcriptome analysis on hermaphroditic and male flowers at a critical sex divergence stage was performed to identify sex-biased differentially expressed genes (DEGs) in a gender diphasic lily species Lilium apertum . A total of 2,338 unigenes were identified as DEGs of which the top match specie is oil palm whose gender affected by environmental conditions, such as water deficit. Among seven pathways enriched in KEGG enrichment analysis, many DEGs involved in photosynthesis, protein processing in endoplasmic reticulum, and zeatin biosynthesis were up-regulated in hermaphroditic flower. While many DEGs involved in phenylpropanoid biosynthesis, plant-pathogen interaction, cutin, suberine and wax biosynthesis, and starch and sucrose metabolism were up-regulated in male flower. Moreover, the significant higher level of zeatin (ZT) in hermaphroditic flower and available nitrogen (N) that is kind of necessary synthetic substrate of cytokinin (CK) in hermaphroditic plants may be attributed to, at least in part, sex differentiation of L. apertum . Additionally, the significant sex-biased expression of two important drought-associated proteins, late embryogenesis abundant ( LEA ) proteins and heat-shock proteins ( HSPs ), could be considered as marker to distinguish the hermaphroditic and male flower at earlier developmental stage. We hypothesize that N deficiency or/and water loss signaling induced apoptosis and arrested the pistil development, in turn, affected the sex divergence of L. apertum. Our results demonstrated, for the first time in well-documented gender diphasic plants, that genes related to many pathways might regulate sex changes triggered by environmental conditions.

Cloning and prokaryotic expression of FcLEA-D29 gene of Fritillaria cirrhosa

Fritillaria cirrhosa, the most valuable source of the precious Chinese medicinal material Fritillariae Cirrhosae Bulbus, suffers from various stresses during growth which influence the yield and quality of the medicinal part. This study aims to explore the genes related to stress resistance in this medicinal species. To be specific, based on the transcriptome data of F. cirrhosa, a gene encoding the late embryogenesis abundant(LEA) protein was obtained, which was named as FcLEA-D29. The gene sequence and protein structure were analyzed with bioinformatics methods and qRT-PCR was used to detect the expression of the gene in different tissues and in response to temperature stress. The low-temperature tolerance of FcLEA-D29 was verified by the prokaryotic expression system. The results showed that FcLEA-D29 contained an open reading frame of 777 bp, encoding 258 amino acids. Multiple sequence alignment revealed that FcLEA-D29 protein belonged to LEA-D29 subfamily of LEA3 family. qRT-PCR results showed that FcLEA-D29 was specifically expressed in bulbs and responded to low temperature. The strain with the recombinant plasmid demonstrated better growth status than the control strain in the instance of low temperature stress, suggesting that FcLEA-D29 may play a role in bulb development and low temperature response of F. cirrhosa. This study laid a basis for further research on the role of FcLEA-D29 in the accumulation of secondary metabolites in F. cirrhosa, especially alkaloids, under low temperature and provided evidence for the low-temperature adaptation of F. cirrhosa.

Functional analysis of a late embryogenesis abundant protein ZmNHL1 in maize under drought stress

Maize is an important feed and industrial cereal crop and is crucial for global food security. The development of drought-tolerant genotypes is a major aim of breeding programs to fight water scarcity and maintain sustainable maize production. Late embryogenesis abundant (LEA) proteins are a family of proteins related to osmotic regulation that widely exist in organisms. Here, we implemented a previously generated maize transcriptomic dataset to identify a drought-responsive gene designated ZmNHL1. Bioinformatics analysis of ZmNHL1 showed that the protein encoded by ZmNHL1 belongs to the LEA-2 protein family. Tissue specific expression analysis showed that ZmNHL1 is relatively abundant in stems and leaves, highly expressed in tassels and only slightly expressed in roots, pollens and ears. Moreover, the activity of SOD and POD of plants from three 35S::ZmNHL1 transgenic lines under either the induced drought stress conditions (by 20% PEG6000) or the natural water deficit treatment (by water withholding) were higher than that of the WT plants, while the electrolyte leakage of the 35S::ZmNHL1 transgenic plants was lower than that of the WT plants under both drought treatments. Our data further revealed that ZmNHL1 promotes maize tolerance to drought stress in 35S::ZmNHL1 transgenic plants by improving ROS scavenging and maintaining the cell membrane permeability. Overall, our data revealed that ZmNHL1 promotes maize tolerance to drought stress and contributes to provide elite germplasm resources for maize drought tolerance breeding programs.

Comparative Seeds Storage Transcriptome Analysis of Astronium fraxinifolium Schott, a Threatened Tree Species from Brazil.

Astronium fraxinifolium Schott (Anacardiaceae), also known as a 'gonçalo-alves', is a tree of the American tropics, with distribution in Mexico, part of Central America, Argentina, Bolivia, Brazil and Paraguay. In Brazil it is an endangered species that occurs in the Cerrado, Caatinga and in the Amazon biomes. In support of ex situ conservation, this work aimed to study two accessions with different longevity (p50) of A. fraxinifolium collected from two different geographic regions, and to evaluate the transcriptome during aging of the seeds in order to identify genes related to seed longevity. Artificial ageing was performed at a constant temperature of 45 °C and 60% relative humidity. RNA was extracted from 100 embryonic axes exposed to control and aging conditions for 21 days. The transcriptome analysis revealed differentially expressed genes such as Late Embryogenesis Abundant (LEA) genes, genes involved in the photosystem, glycine rich protein (GRP) genes, and several transcription factors associated with embryo development and ubiquitin-conjugating enzymes. Thus, these results contribute to understanding which genes play a role in seed ageing, and may serve as a basis for future functional characterization of the seed aging process in A. fraxinifolium.

Drought Stress Priming Improved the Drought Tolerance of Soybean.

The capability of a plant to protect itself from stress-related damages is termed "adaptability" and the phenomenon of showing better performance in subsequent stress is termed "stress memory". While drought is one of the most serious disasters to result from climate change, the current understanding of drought stress priming in soybean is still inadequate for effective crop improvement. To fill this gap, in this study, the drought memory response was evaluated in cultivated soybean (Glycine max). To determine if a priming stress prior to a drought stress would be beneficial to the survival of soybean, plants were divided into three treatment groups: the unprimed group receiving one cycle of stress (1S), the primed group receiving two cycles of stress (2S), and the unstressed control group not subjected to any stress (US). When compared with the unprimed plants, priming led to a reduction of drought stress index (DSI) by 3, resulting in more than 14% increase in surviving leaves, more than 13% increase in leaf water content, slight increase in shoot water content and a slower rate of loss of water from the detached leaves. Primed plants had less than 60% the transpiration rate and stomatal conductance compared to the unprimed plants, accompanied by a slight drop in photosynthesis rate, and about a 30% increase in water usage efficiency (WUE). Priming also increased the root-to-shoot ratio, potentially improving water uptake. Selected genes encoding late embryogenesis abundant (LEA) proteins and MYB, NAC and PP2C domain-containing transcription factors were shown to be highly induced in primed plants compared to the unprimed group. In conclusion, priming significantly improved the drought stress response in soybean during recurrent drought, partially through the maintenance of water status and stronger expression of stress related genes. In sum, we have identified key physiological parameters for soybean which may be used as indicators for future genetic study to identify the genetic element controlling the drought stress priming.

Transcriptomic analysis reveals the mechanism of low/high temperature resistance in an outstanding diet alga Nannochloropsis oceanica

Low/high temperature stress is one of the key abiotic factors restricting the implementation of microalgal large-scale production. Metabolism, involved in abiotic stress resistances, have been reported in plants, but the related research in microalgae is far behind. This study analyzed the transcriptional changes in Nannochloropsis oceanica under low and high temperature stresses by RNA-seq technology. Results indicated that the total differentially expressed gene (DEG) amount under high temperature stress was much more than low temperature stress. GO enrichment analysis suggested that DEGs under low temperature were mainly concentrated in protein modification and phosphorus metabolic process, while transmembrane transport, lipid localization and transport, and cellular homeostasis were the most enriched terms under high temperature. Moreover, “membrane” was the most enriched term in both low and high temperature groups. The gene expressions of 37 stress resistance-related enzymes/proteins (172 genes) were analyzed concretely in N. oceanica for the first time. Results revealed that ornithine decarboxylase (ODC) was significantly down-regulated, while choline dehydrogenase (CHDH), late embryogenesis abundant (LEA), dehydrin (DHN), chitinase (CHI), calmodulin (CaM), and lipid transfer protein (LTP) were extremely up-regulated under low temperature. Under high temperature, more genes with drastic changes were identified. Superoxide dismutase (SOD), ascorbate peroxidase (APX), glutathione synthetase (GSS), and glyoxalase II (Gly II) exhibited down-regulated significantly, while glutathione S-transferase (GST), CHDH, L-asparaginase (L-ASP), LEA, DHN, early response to dehydration protein (ERD), heat shock protein (HSP), and CaM were dramatically up-regulated. No significant changes were detected in glutathione peroxidase (GPX), tocopherol cyclase (TC), gamma-glutamylcysteine synthetase (γ-GCS), pyrroline‐5–carboxylate reductase (P5CR), asparagine synthetase (ASN ), succinate-semialdehyde dehydrogenase (SSADH), pyruvate decarboxylase (PDC), and S-adenosylmethionine synthetase (SAMS) under low or high temperature. Furthermore, 166 transcription factor (TF) genes related to plant stress resistance were characterized and categorized into 20 TF families. MYB was the most abundant and active one, revealing that the MYB family played a critical role of temperature resistance in N. oceanica. These new findings could benefit the understanding of the temperature resistance mechanism in Nannochloropsis. The significantly changed genes might be ideal candidates for breeding elite Nannochloropsis strains with stronger resistance to temperature stress using genetic engineering methods.

Genomic Analysis of LEA Genes in Carica papaya and Insight into Lineage-Specific Family Evolution in Brassicales.

Late embryogenesis abundant (LEA) proteins comprise a diverse superfamily involved in plant development and stress responses. This study presents a first genome-wide analysis of LEA genes in papaya (Carica papaya L., Caricaceae), an economically important tree fruit crop widely cultivated in the tropics and subtropics. A total of 28 members were identified from the papaya genome, which belong to eight families with defined Pfam domains, i.e., LEA_1 (3), LEA_2 (4), LEA_3 (5), LEA_4 (5), LEA_5 (2), LEA_6 (2), DHN (4), and SMP (3). The family numbers are comparable to those present in Ricinus communis (Euphorbiaceae, 28) and Moringa oleifera (Moringaceae, 29), but relatively less than that found in Moringa oleifera (Cleomaceae, 39) and Arabidopsis thaliana (Brassicaceae, 51), implying lineage-specific evolution in Brassicales. Indeed, best-reciprocal-hit-based sequence comparison and synteny analysis revealed the presence of 29 orthogroups, and significant gene expansion in Tarenaya and Arabidopsis was mainly contributed by whole-genome duplications that occurred sometime after their split with the papaya. Though a role of transposed duplication was also observed, tandem duplication was shown to be a key contributor in gene expansion of most species examined. Further comparative analyses of exon-intron structures and protein motifs supported fast evolution of this special superfamily, especially in Arabidopsis. Transcriptional profiling revealed diverse expression patterns of CpLEA genes over various tissues and different stages of developmental fruit. Moreover, the transcript level of most genes appeared to be significantly regulated by drought, cold, and salt stresses, corresponding to the presence of cis-acting elements associated with stress response in their promoter regions. These findings not only improve our knowledge on lineage-specific family evolution in Brassicales, but also provide valuable information for further functional analysis of LEA genes in papaya.

Stress-dependent cell stiffening by tardigrade tolerance proteins that reversibly form a filamentous network and gel.

Tardigrades are able to tolerate almost complete dehydration by entering a reversible ametabolic state called anhydrobiosis and resume their animation upon rehydration. Dehydrated tardigrades are exceptionally stable and withstand various physical extremes. Although trehalose and late embryogenesis abundant (LEA) proteins have been extensively studied as potent protectants against dehydration in other anhydrobiotic organisms, tardigrades produce high amounts of tardigrade-unique protective proteins. Cytoplasmic-abundant heat-soluble (CAHS) proteins are uniquely invented in the lineage of eutardigrades, a major class of the phylum Tardigrada and are essential for their anhydrobiotic survival. However, the precise mechanisms of their action in this protective role are not fully understood. In the present study, we first postulated the presence of tolerance proteins that form protective condensates via phase separation in a stress-dependent manner and searched for tardigrade proteins that reversibly form condensates upon dehydration-like stress. Through a comprehensive search using a desolvating agent, trifluoroethanol (TFE), we identified 336 proteins, collectively dubbed "TFE-Dependent ReversiblY condensing Proteins (T-DRYPs)." Unexpectedly, we rediscovered CAHS proteins as highly enriched in T-DRYPs, 3 of which were major components of T-DRYPs. We revealed that these CAHS proteins reversibly polymerize into many cytoskeleton-like filaments depending on hyperosmotic stress in cultured cells and undergo reversible gel-transition in vitro. Furthermore, CAHS proteins increased cell stiffness in a hyperosmotic stress-dependent manner and counteract the cell shrinkage caused by osmotic pressure, and even improved the survival against hyperosmotic stress. The conserved putative helical C-terminal region is necessary and sufficient for filament formation by CAHS proteins, and mutations disrupting the secondary structure of this region impaired both the filament formation and the gel transition. On the basis of these results, we propose that CAHS proteins are novel cytoskeleton-like proteins that form filamentous networks and undergo gel-transition in a stress-dependent manner to provide on-demand physical stabilization of cell integrity against deformative forces during dehydration and could contribute to the exceptional physical stability in a dehydrated state.

Classification of LEA proteins using Support Vector Machine Algorithm

The tolerance of plants to environmental stress is aided by numerous proteins, one such protein is a LateEmbryogenesis Abundant (LEA) Protein. Thus, it is very essential to identify and classify LEA proteins.For this purpose, machine learning would be greatly useful. In this study, an attempt was made to developSVM classifiers for the classification of LEA proteins based on their physico-chemical properties. The physico -chemicalproperties were analyzed from protein sequences and then the dataset is split into training andtest data. The model is developed and evaluated using cross-validation techniques like 5-fold. The resultsshow that the accuracy of developed SVM classifiers ranges from 60 to 97 per cent.

Proteomics Evidence of a Systemic Response to Desiccation in the Resurrection Plant Haberlea rhodopensis.

Global warming and drought stress are expected to have a negative impact on agricultural productivity. Desiccation-tolerant species, which are able to tolerate the almost complete desiccation of their vegetative tissues, are appropriate models to study extreme drought tolerance and identify novel approaches to improve the resistance of crops to drought stress. In the present study, to better understand what makes resurrection plants extremely tolerant to drought, we performed transmission electron microscopy and integrative large-scale proteomics, including organellar and phosphorylation proteomics, and combined these investigations with previously published transcriptomic and metabolomics data from the resurrection plant Haberlea rhodopensis. The results revealed new evidence about organelle and cell preservation, posttranscriptional and posttranslational regulation, photosynthesis, primary metabolism, autophagy, and cell death in response to desiccation in H. rhodopensis. Different protective intrinsically disordered proteins, such as late embryogenesis abundant (LEA) proteins, thaumatin-like proteins (TLPs), and heat shock proteins (HSPs), were detected. We also found a constitutively abundant dehydrin in H. rhodopensis whose phosphorylation levels increased under stress in the chloroplast fraction. This integrative multi-omics analysis revealed a systemic response to desiccation in H. rhodopensis and certain targets for further genomic and evolutionary studies on DT mechanisms and genetic engineering towards the improvement of drought tolerance in crops.

Evaluation of a bacterial group 1 LEA protein as an enzyme protectant from stress-induced inactivation.

Late embryogenesis abundant (LEA) proteins are hydrophilic proteins that lack a well-ordered tertiary structure and accumulate to high levels in response to water deficit, in organisms such as plants, fungi, and bacteria. The mechanisms proposed to protect cellular structures and enzymes are water replacement, ion sequestering, and membrane stabilization. The activity of some proteins has a limited shelf-life due to instability that can be caused by their structure or the presence of a stress condition that limits their activity; several LEA proteins have been shown to behave as cryoprotectants in vitro. Here, we report a group1 LEA from Azotobacter vinelandii AvLEA1, capable of conferring protection to lactate dehydrogenase, catechol dioxygenase, and Baylase peroxidase against freeze-thaw treatments, desiccation, and oxidative damage, making AvLEA a promising biological stabilizer reagent. This is the first evidence of protection provided by this LEA on enzymes with biotechnological potential, such as dioxygenase and peroxidase under in vitro stress conditions. Our results suggest that AvLEA could act as a molecular chaperone, or a "molecular shield," preventing either dissociation or antiaggregation, or as a radical scavenger, thus preventing damage to these target enzymes during induced stress. KEY POINTS: • This work expands the basic knowledge of the less-known bacterial LEA proteins and their in vitro protection potential. • AvLEA is a bacterial protein that confers in vitro protection to three enzymes with different characteristics and oligomeric arrangement. • The use of AvLEA as a stabilizer agent could be further explored using dioxygenase and peroxidase in bioremediation treatments. AvLEA1 protects against freeze-thaw treatments, desiccation, and oxidative damage on three different enzymes with biotechnological potential.

Transcriptome Analysis of Moso Bamboo (Phyllostachys edulis) Reveals Candidate Genes Involved in Response to Dehydration and Cold Stresses.

Bamboo (Bambusoideae) belongs to the grass family (Poaceae) and has been utilized as one of the most important nontimber forest resources in the world. Moso bamboo (Phyllostachys edulis) is a large woody bamboo with high ecological and economic values. Global climate change brings potential challenges to the normal growth of moso bamboo, and hence its production. Despite the release of moso bamboo genome sequence, the knowledge on genome-wide responses to abiotic stress is still limited. In this study, we generated a transcriptome data set with respect to dehydration and cold responses of moso bamboo using RNA-seq technology. The differentially expressed genes (DEGs) under treatments of dehydration and cold stresses were identified. By combining comprehensive gene ontology (GO) analysis, time-series analysis, and co-expression analysis, candidate genes involved in dehydration and cold responses were identified, which encode abscisic acid (ABA)/water deficit stress (WDS)-induced protein, late embryogenesis abundant (LEA) protein, 9-cis-epoxycarotenoid dioxygenase (NCED), anti-oxidation enzymes, transcription factors, etc. Additionally, we used PeLEA14, a dehydration-induced gene encoding an “atypical” LEA protein, as an example to validate the function of the identified stress-related gene in tolerance to abiotic stresses, such as drought and salt. In this study, we provided a valuable genomic resource for future excavation of key genes involved in abiotic stress responses and genetic improvement of moso bamboo to meet the requirement for environmental resilience and sustainable production.