Canavalia Maritima Research Articles

Identifying Calmodulin and Calmodulin-like Protein Members in Canavalia rosea and Exploring Their Potential Roles in Abiotic Stress Tolerance.

Calmodulins (CaMs) and calmodulin-like proteins (CMLs) belong to families of calcium-sensors that act as calcium ion (Ca2+) signal-decoding proteins and regulate downstream target proteins. As a tropical halophyte, Canavalia rosea shows great resistance to multiple abiotic stresses, including high salinity/alkalinity, extreme drought, heat, and intense sunlight. However, investigations of calcium ion signal transduction involved in the stress responses of C. rosea are limited. The CaM and CML gene families have been identified and characterized in many other plant species. Nevertheless, there is limited available information about these genes in C. rosea. In this study, a bioinformatic analysis, including the gene structures, conserved protein domains, phylogenetic relationships, chromosome distribution, and gene synteny, was comprehensively performed to identify and characterize CrCaMs and CrCMLs. A spatio-temporal expression assay in different organs and environmental conditions was then conducted using the RNA sequencing technique. Additionally, several CrCaM and CrCML members were then cloned and functionally characterized using the yeast heterogeneous expression system, and some of them were found to change the tolerance of yeast to heat, salt, alkalinity, and high osmotic stresses. The results of this study provide a foundation for understanding the possible roles of the CrCaM and CrCML genes, especially for halophyte C. rosea's natural ecological adaptability for its native habitats. This study also provides a theoretical basis for further study of the physiological and biochemical functions of plant CaMs and CMLs that are involved in tolerance to multiple abiotic stresses.

Plant-associated bacteria and enzymes support Canavalia rosea growth in coastal hypersaline soils

Canavalia rosea is an extremophilic legume that grows in hypersaline and nutrient-deficient ecosystems. The extremophilic nature of C. rosea may be attributed to its ability to establish symbiotic associations with nutrient mineralizing and plant growth promoting (PGP) bacteria housed in the nodules. This study examined legume-microbe symbiosis and plant nutrition of C. rosea growing in subtropical coastal zone in KwaZulu-Natal province, South Africa. Canavalia rosea adult plants of the same age from Westbrook, Scottburgh and Durban were collected for plant biomass and plant nutrition and root nodules were used for bacterial extraction and identification. Rhizosphere soils sampled from the three localities were used for bacterial extraction and identification, extracellular enzyme assays and soil characteristics (pH, nutrient concentrations, total cation, and exchange acidity). Westbrook, Scottburgh and Durban soils were nutrient-deficient with varying total cations, acid saturation and a pH range of 7.3–7.6. Soil nutrient mineralizing extracellular enzyme activities varied across study sites. The culturable bacterial strains isolated from the sampled soils belonged to the Pseudomonas, Pantoea and Flavobacterium genera. Canavalia rosea root nodules were nodulated by Pseudomonas guariconensis, Pseudomonas fulva, Pseudomonas fluorescens, Pseudomonas chlororaphis and Pseudomonas chlororaphis subsp. aurantiaca. Plants growing in Westbrook soils had a significantly higher total plant biomass compared to Scottburgh and Durban plants. Plant P concentration did not vary significantly between sites while plant N and C concentrations varied significantly. Plant-associated and soil bacteria with phosphorus (P) solubilising, nitrogen (N) cycling, and N fixing functions and associated enzymes seem to facilitate the mobilization of nutrients enabling C. rosea to thrive in hypersaline and low-nutrient environments.

Assessment of the role of ACC deaminase, IAA, and siderophore-producing bacteria isolated from Spratly Islands on the growth of Brassica juncea on the coral sand

Root endophytic and rhizosphere bacteria play a significant role in enhancing plant tolerance and promoting growth by producing 1-aminocyclopropane-1-carboxylate (ACC) deaminase, indole-3-acetic acid (IAA), and siderophore. These bacteria have been isolated and screened from 45 plants’ roots and rhizosphere soil samples of 19 types of plants which are growing on 8 floating islands in Spratley Islands, Viet Nam. In which, 3 root endophytic bacteria strains were selected and identified to be Agrobacterium tumefaciens, Pseudomonas plecoglossicida, and Klebsiella aerogenes from roots of Scaevola taccada, Barringtonia asiatica (L.) Kurz, and Carica papaya, respectively. Correspondingly, 3 rhizosphere bacteria strains were selected and identified to be Bacillus velezensis, Bacillus aryabhattai, and Bacillus velezensis from rhizosphere soils of Canavalia maritima (Aubt) Thouars (C. obtusifolia DC.), Cucumis sativus, Heliotropium foertherianum, respectively. These strains were found could increase the germination rate of Brassica juncea’s seeds and 20-day-old fresh biomass, compared to the control treatment in the same condition of 10‰ saline watering. When these bacterial strains were combined as an inoculant, they indicated the same result on coral sand. Thus, the addition of selected endophytic and rhizosphere bacteria is efficient in improving the coral sand environment to enhance food crops’ development in the Spratly Islands.

Temporal and spatial variation in extrafloral nectar and associated ants in Canavalia rosea (Fabaceae) on coastal dunes along the Gulf of Mexico

ABSTRACT Extrafloral nectar production can vary depending on the structure of the plant in which it is found, as well as environmental conditions. This can in turn influence plant-ant interactions. We evaluated the role of extrafloral nectaries of Canavalia rosea as ant attractants in a coastal zone of the Gulf of Mexico. We compared the productivity of the extrafloral nectaries present on leaves and inflorescences, as well as between day and night, and described the associated ant community. Nectar volume and concentration were higher in inflorescences than in leaves, and the rate of ant visitation was higher in inflorescences during the day. These results suggest congruence with the Optimal Defense Hypothesis. Nectar volume was generally greater during the night in both structures, but sugar concentration did not differ between day and night. We found a turnover in the ant species visiting C. rosea, with S. geminata being the most abundant species. The extrafloral nectar of C. rosea attracts several ant species and probably other arthropods, providing a valuable resource in a stressful system.

Genome-wide functional characterization of Canavalia rosea cysteine-rich trans-membrane module (CrCYSTM) genes to reveal their potential protective roles under extreme abiotic stress

Cysteine-rich transmembrane module (CYSTM) proteins constitute small molecular protein families and have been identified across eukaryotes, including yeast, humans, and several plant species. Plant CYSTMs play vital roles in growth regulation, development, phytohormone signal transduction, pathogen defense, environmental stress response, and even heavy metal binding and detoxification. Canavalia rosea (Sw.) DC is a perennial halophyte with great semi-arid and saline-alkali tolerance. In this study, the CrCYSTM family including 10 members were identified in the C. rosea genome, with the purpose of clarifying the possible roles of CrCYSTMs in C. rosea plants development and stress resistance. The phylogenetic relationships, exon–intron structure, domain structure, chromosomal localization, and putative cis-acting elements in promoter regions were predicted and analyzed. Transcriptome analysis combined with quantitative reverse transcription PCR showed that different CrCYSTM members exhibited varied expression patterns in different tissues and under different abiotic stress challenges. In addition, several CrCYSTMs were cloned and functionally characterized for their roles in abiotic stress tolerance with yeast expression system. Overall, these findings provide a foundation for functionally characterizing plant CYSTMs to unravel their possible roles in the adaptation of C. rosea to tropical coral reefs. Our results also lay the foundation for further research on the roles of plant CYSTM genes in abiotic stress signaling, especially for heavy metal detoxification.

Root biomechanical features and wind erosion resistance of three native leguminous psammophytes for coastal dune restoration

Taiwan is a subtropical mountainous island with fragile geological conditions and about 1566 km of coastline. Monsoon usually bring severe coastal erosion and dust storms. Canavalia lineata, Canavalia rosea and Vigna marina, three native pioneer leguminous psammophytes occurring on coastal sand dune areas, play important roles in reducing wind erosion in coastal regions. Nonetheless, its root biomechanical properties and wind erosion resistance are still unexplored. In this study, the root biomechanical properties of these plants were inspected using uprooting and tensile tests. Wind erosion resistance was estimated by wind tunnel tests. Our results showed that the growth charactersitics of C. lineata are superior than those of C. rosea and V. marina. Furthermore, the root pullout force of C. lineata (0.36 ± 0.04 kN) and C. rosea (0.36 ± 0.05 kN) are remarkably higher than that of V. marina (0.22 ± 0.02 kN). Root tensile strength of C. rosea (291.24 ± 11.89 MPa) plants is significantly greater than those of C. lineata (231.33 ± 9.47 MPa) and V. marina (95.78 ± 8.83 MPa) plants. Moreover, wind tunnel tests show that C. lineata has superior wind erosion resistance than C. rosea and V. marina. Altogether, these results clearly demonstrate that the priority species order for coastal dune restoration is: C. lineata > C. rosea > V. marina. These results contribute important strategy for ecological engineering of coastal management in sand dune areas.

Antioxidants and cardioprotective effects of ethyl acetate fraction of Canavalia rosea leaves in myocardial ischemia-reperfusion injury.

Different degrees in the biological activities of Canavalia rosea had been previously reported . In this study, our group assessed the cardioprotective effects of the ethyl acetate fraction (EAcF) of the Canavalia rosea leaves. Firstly, it was confirmed, by in vitro approach, that the EAcF has high antioxidant properties due to the presence of important secondary metabolites, as flavonoids. In order to explore their potential protector against cardiovascular disorders, hearts were previously perfused with EAcF (300 μg.mL-1) and submitted to the global ischemia followed by reperfusion in Langendorff system. The present findings have demonstrated that EAcF restored the left ventricular developed pressure and decreased the arrhythmias severity index. Furthermore, EAcF significantly increased the glutathiones peroxidase activity with decreased malondialdehyde and creatine kinase levels. EAcF was effective upon neither the superoxide dismutase, glutationes reductase nor the catalase activities. In addition, the Western blot analysis revealed that ischemia-reperfusion injury significantly upregulates caspase 3 protein expression, while EAcF abolishes this effect. These results provide evidence that the EAcF reestablishes the cardiac contractility and prevents arrhythmias; it is suggested that EAcF could be used to reduce injury caused by cardiac reperfusion. However more clinical studies should be performed, before applying it in the clinic.

Genome-Wide Identification and Functional Analysis of the GASA Gene Family Responding to Multiple Stressors in Canavalia rosea.

In plants, the Gibberellic Acid-Stimulated Arabidopsis (GASA) gene family is unique and responds to ubiquitous stress and hormones, playing important regulatory roles in the growth and development of plants, as well as in the resistance mechanisms to biotic and abiotic stress. In this study, a total of 23 CrGASAs were characterized in C. rosea using a genome-wide approach, and their phylogenetic relationships, gene structures, conserved motifs, chromosomal locations, gene duplications, and promoter regions were systematically analyzed. Expression profile analysis derived from transcriptome data showed that CrGASAs are expressed at higher levels in the flowers or fruit than in the leaves, vines, and roots. The expression of CrGASAs also showed habitat- and environmental-stress-regulated patterns in C. rosea analyzed by transcriptome and quantitative reverse transcription PCR (qRT-PCR). The heterologous induced expression of some CrGASAs in yeast enhanced the tolerance to H2O2, and some CrGASAs showed elevated heat tolerance and heavy metal (HM) Cd/Cu tolerance. These findings will provide an important foundation to elucidate the biological functions of CrGASA genes, especially their role in the ecological adaptation of specific plant species to tropical islands and reefs in C. rosea.

Effect of Ripening Stage on Biochemical Composition of Seeds from three Species of Canavalia spp Consumed as Protein Substitutes in Côte d’Ivoire

Aims: The present study was aimed at investigating the nutritional properties of legume-seeds from Canavalia genus as a function of their ripening stage.
 Methodology: Seeds of three species of canavalia seeds notably Canavalia gladiata, Canavalia ensiformis and Canavalia rosea were obtained from an experimental plot located in Bouaké area’s (7°41′00″ N, 5°01′59″ W) in Côte d’Ivoire. The seeds of these three species were collected at different ripening stages. The seeds were sun dried and ground to obtain the crude flour. Chemical composition and functional properties were investigated using standard methods while amino acid profiles were performed by using HPLC analytical methods.
 Results: Results reveal that the profiles of nutrients in the three Canavalia species varied as a function of ripening stages. The highest fat (3.27 ± 0.01 %), protein (33.20 ± 0.17), fibre (7.51 ± 0.06 %), ash (6.67 ± 0.01 %) and total sugar (25.40 ± 0.05 %) contents were obtained at the last ripening stage. All the legume-seeds investigated are excellent sources of proteins mainly consisting of high amounts of essential amino acids such as lysine (100.23 ± 0.04 mg/g protein), leucine (103.51 ± 0.04 mg/g protein), valine (83.68 ± 0.04 mg/g protein), methionine (27.36 ± 0.04 mg/g protein), tryptophan (74.32 ± 0.04 mg/g protein), cysteine (44.24 ± 0.04 mg/g protein), isoleucine (63.61 ± 0.04 mg/g protein), threonine (40.22 ± 0.04 mg/g protein) and histidine (17.76 ± 0.04 mg/g protein). As regards mineral contents of these seeds, they were higher at penultimate ripening stage and, they partially meet dietary allowance of infants.
 Conclusion: These seeds could be ranked as protein rich food for human and then, used as valuable substitutes for meat or fish.

Functional Characterization of Heat Shock Factor (CrHsf) Families Provide Comprehensive Insight into the Adaptive Mechanisms of Canavalia rosea (Sw.) DC. to Tropical Coral Islands.

Heat shock transcription factors (Hsfs) are key regulators in plant heat stress response, and therefore, they play vital roles in signal transduction pathways in response to environmental stresses, as well as in plant growth and development. Canavalia rosea (Sw.) DC. is an extremophile halophyte with good adaptability to high temperature and salt-drought tolerance, and it can be used as a pioneer species for ecological reconstruction on tropical coral islands. To date, very little is known regarding the functions of Hsfs in the adaptation mechanisms of plant species with specialized habitats, especially in tropical leguminous halophytes. In this study, a genome-wide analysis was performed to identify all the Hsfs in C. rosea based on whole-genome sequencing information. The chromosomal location, protein domain or motif organization, and phylogenetic relationships of 28 CrHsfs were analyzed. Promoter analyses indicated that the expression levels of different CrHsfs were precisely regulated. The expression patterns also revealed clear transcriptional changes among different C. rosea tissues, indicating that the regulation of CrHsf expression varied among organs in a developmental or tissue-specific manner. Furthermore, the expression levels of most CrHsfs in response to environmental conditions or abiotic stresses also implied a possible positive regulatory role of this gene family under abiotic stresses, and suggested roles in adaptation to specialized habitats such as tropical coral islands. In addition, some CrHsfAs were cloned and their possible roles in abiotic stress tolerance were functionally characterized using a yeast expression system. The CrHsfAs significantly enhanced yeast survival under thermal and oxidative stress challenges. Our results contribute to a better understanding of the plant Hsf gene family and provide a basis for further study of CrHsf functions in environmental thermotolerance. Our results also provide valuable information on the evolutionary relationships among CrHsf genes and the functional characteristics of the gene family. These findings are beneficial for further research on the natural ecological adaptability of C. rosea to tropical environments.

Neokomagataea anthophila sp. nov., an osmotolerant acetic acid bacterium isolated in Thailand and emended description of the genus Neokomagataea.

A novel Gram-stain-negative, rod-shaped, non-motile, aerobic bacterium isolated from a sea bean flower [Canavalia rosea (Sw.) DC.] collected in Surat Thani Province, Thailand, and designated as AH18T was characterized on the basis of polyphasic taxonomy. The phylogenetic analysis of 16S rRNA gene revealed that strain AH18T represented a member of the genus Neokomagataea. In the 16S rRNA gene sequence analysis, the strain's closest phylogenetic neighbour was Neokomagataea thailandica TBRC 376T. The draft genome size of strain AH18T was 2613495 bp, and its DNA G+C content was 52.0 mol%. The strain showed 90.3 and 76.3% pairwise-determined whole-genome average nucleotide identity and 39.8 and 19.6% digital DNA-DNA hybridization values with N. thailandica TBRC 376T and N. tanensis TBRC 7768T, respectively. The 16S rRNA gene sequences and phylogenomic analysis revealed that the strain clustered with the members of the genus Neokomagataea but was located in a distinct branch closely related to N. thailandica TBRC 376T. The predominant cellular fatty acids of the strain were summed feature 8 (C18:1 ω6c and/or C18:1 ω7c), C16:0 and C18:1 2OH (>5%). The major respiratory ubiquinone was Q-10. In addition, strain AH18T was substantiated by differences in several physiological characteristics and by MALDI-TOF profiling. On the basis of the results obtained from phenotypic, chemotaxonomic, phylogenetic and genomic analyses, the strain clearly represented a novel species within the genus Neokomagataea, for which the name Neokomagataea anthophila sp. nov. (AH18T=TBRC 2177T=NBRC 115156T) is proposed. An emended description of the genus Neokomagataea is also given.

Comprehensive Analysis of the Hsp20 Gene Family in Canavalia rosea Indicates Its Roles in the Response to Multiple Abiotic Stresses and Adaptation to Tropical Coral Islands

Heat shock protein 20 (Hsp20) is a major family of heat shock proteins that mainly function as molecular chaperones and are markedly accumulated in cells when organisms are subjected to environmental stress, particularly heat. Canavalia rosea is an extremophile halophyte with good adaptability to environmental high temperature and is widely distributed in coastal areas or islands in tropical and subtropical regions. In this study, we identified a total of 41 CrHsp20 genes in the C. rosea genome. The gene structures, phylogenetic relationships, chromosome locations, and conserved motifs of each CrHsp20 or encoding protein were analyzed. The promoters of CrHsp20s contained a series of predicted cis-acting elements, which indicates that the expression of different CrHsp20 members is regulated precisely. The expression patterns of the CrHsp20 family were analyzed by RNA sequencing both at the tissue-specific level and under different abiotic stresses, and were further validated by quantitative reverse transcription PCR. The integrated expression profiles of the CrHsp20s indicated that most CrHsp20 genes were greatly upregulated (up to dozens to thousands of times) after 2 h of heat stress. However, some of the heat-upregulated CrHsp20 genes showed completely different expression patterns in response to salt, alkaline, or high osmotic stresses, which indicates their potential specific function in mediating the response of C. rosea to abiotic stresses. In addition, some of CrHsp20s were cloned and functionally characterized for their roles in abiotic stress tolerance in yeast. Taken together, these findings provide a foundation for functionally characterizing Hsp20s to unravel their possible roles in the adaptation of this species to tropical coral reefs. Our results also contribute to the understanding of the complexity of the response of CrHsp20 genes to other abiotic stresses and may help in future studies evaluating the functional characteristics of CrHsp20s for crop genetic improvement.

Lectins ConA and ConM extracted from Canavalia ensiformis (L.) DC and Canavalia rosea (Sw.) DC inhibit planktonic Candida albicans and Candida tropicalis.

Lectins participate in the defense against microorganisms and in signaling the damage caused by pathogens to the cell surface and/or intracellular in plants. This study aims to analyze the antifungal potential of lectins extracted from seeds of Canavalia ensiformis (L.) DC and Canavalia rosea (Sw.) DC, against Candida albicans and Candida tropicalis. The antimicrobial tests were performed by microdilution against Candida spp. The test to verify the combined lectin/fluconazole effect was performed using subinhibitory concentrations of lectins and with antifungal ranging from 0.5 to 512 µg/mL. The ability to inhibit the morphological transition of Candida spp. was evaluated by microcultivation in a moist chamber. The results of the minimum inhibitory concentration revealed no antifungal activity against the tested strains. However, lectins modified the action of fluconazole, reducing the IC50 of the drug against C. albicans. Lectins were also able to discretely modulate the morphological transition of the tested strains.Supplementary InformationThe online version contains supplementary material available at 10.1007/s00203-022-02959-x.

Relationship between the distribution of vegetation and the environment in the coastal embryo dunes of Jalisco, México.

BackgroundThe poorly developed soils of the embryo dunes imply little capacity for plant support, however, the adaptation mechanisms of plants respond sensitively to environmental variations, even when these variations are small, which results in a set of specialized habitats and flora that are rarely shared with other terrestrial ecosystems. The coastal dunes of the Mexican Pacific remain vaguely studied, this is why this research explored the relationship between environmental properties and the presence of plant species in the embryo dunes of the coast of Jalisco, Mexico.MethodsTwenty-nine sites were sampled, one or two sites per embryo dune, with a random stratified design. Geomorphological and vegetation data were collected at site. Laboratory determinations included soil color, particle size, organic matter, pH, electrical conductivity, magnetite content, and moisture retention. Statistical analysis included correlation analysis to identify relationships between environmental variables; principal component analysis (PCA) and cluster analysis to group dune sites by environmental properties; canonical correspondence analysis (CCA) to determine a possible significant relationship between the presence of plant species and environmental variables; cluster analysis to group dune sites by presence/absence of plant species and correlate both clusters to validate the relationship between them, the salient aspects of this relationship were described and the spatial distribution of the groups was mapped.ResultsEleven plant species were identified, six of them exclusive to the embryo dunes and the rest ubiquitous. The incipient development of these soils is reflected in a low content of organic matter, silt, clay, and moisture retention, with scattered data on granulometry, electrical conductivity, organic matter, and magnetite. Some significant correlations were found between some environmental properties, and the CCA showed a significant relationship between the presence of plant species and environmental variables (p-value of the Monte Carlo test = 0.026). The cluster analysis of dune sites according to environmental variables and the cluster analysis by presence/absence of plant species produces the formation of five groups of sites with significant environmental differences and five groups of sites with significant floristic differences. A significant connection (r = 0.471, p = 0.01) between the two clustering schemes also evidences the meaningful relationship between the presence of plant species and the environmental characteristics of the embryo dunes of Jalisco, Mexico. Differences in habitat preferences were observed among plant species exclusive to the embryo dunes; thus, Abronia maritima, Uniola pittieri, and Pectis arenaria showed a preference for embryo dunes with poor edaphic conditions, in contrast to Okenia hypogaea, Canavalia rosea, and Scaevola plumieri, which were mostly found in embryo dunes with higher fertility.

Genome-Wide Identification and Functional Analysis of the Gasa Gene Family Responding to Multiple Stressors in Canavalia Rosea

Genome-Wide Identification and Functional Analysis of the Gasa Gene Family Responding to Multiple Stressors in Canavalia Rosea

Distributional Data and New Records of Seed Beetles (Coleoptera: Chrysomelidae: Bruchinae) from Mexico Deposited in the National Museum of Natural History (USNM)

Examination of 1,805 specimens of seed beetles (Coleoptera: Chrysomelidae: Bruchinae) from Mexico in the National Museum of Natural History (USNM), Washington, DC, resulted in discovery of unpublished data from 11 genera and 63 species. New country records for Mexico include Amblycerus lineolatus (Motschulsky, 1874) and Amblycerus luctuosus (Sharp, 1885). Seven plant species are reported as new host records: Prosopis articulata S. Watson and Senna mollissima var. glabrata (Benth.) H.S. Irwin & Barneby (both Leguminosae) for Amblycerus obscurus (Sharp, 1885), Rhynchosia pyramidalis (Lam.) Urb. (Leguminosae) for Zabrotes subfasciatus (Boheman, 1833), Abutilon palmeriA. Gray (Malvaceae) for Abutiloneus flavicornis (Sharp, 1885), Prosopis chilensis (Molina) Stuntz for Algarobius johnsoni Kingsolver, 1986, Prosopis articulata S. Watson for Algarobius prosopis (LeConte, 1858), and Bauhinia macranthera Hemsl. (Leguminosae) for Gibbobruchus divaricatae Whitehead and Kingsolver, 1975. Canavalia maritima Thouars (Leguminosae) is confirmed as a host for Caryedes longicollis (Fahraeus, 1839). Twenty-four new state records are reported for more localized distributions within Mexico. Updated information from specimens previously published is included.

Genome-Wide Identification, Primary Functional Characterization of the NHX Gene Family in Canavalia rosea, and Their Possible Roles for Adaptation to Tropical Coral Reefs.

Canavalia rosea, distributed in the coastal areas of tropical and subtropical regions, is an extremophile halophyte with good adaptability to high salinity/alkaline and drought tolerance. Plant sodium/hydrogen (Na+/H+) exchanger (NHX) genes encode membrane transporters involved in sodium ion (Na+), potassium ion (K+), and lithium ion (Li+) transport and pH homeostasis, thereby playing key roles in salinity tolerance. However, the NHX family has not been reported in this leguminous halophyte. In the present study, a genome-wide comprehensive analysis was conducted and finally eight CrNHXs were identified in C. rosea genome. Based on the bioinformatics analysis about the chromosomal location, protein domain, motif organization, and phylogenetic relationships of CrNHXs and their coding proteins, as well as the comparison with plant NHXs from other species, the CrNHXs were grouped into three major subfamilies (Vac-, Endo-, and PM-NHX). Promoter analyses of cis-regulatory elements indicated that the expression of different CrNHXs was affected by a series of stress challenges. Six CrNHXs showed high expression levels in five tested tissues of C. rosea in different levels, while CrNHX1 and CrNHX3 were expressed at extremely low levels, indicating that CrNHXs might be involved in regulating the development of C. rosea plant. The expression analysis based on RNA-seq showed that the transcripts of most CrNHXs were obviously decreased in mature leaves of C. rosea plant growing on tropical coral reefs, which suggested their involvement in this species’ adaptation to reefs and specialized islands habitats. Furthermore, in the single-factor stress treatments mimicking the extreme environments of tropical coral reefs, the RNA-seq data also implied CrNHXs holding possible gene-specific regulatory roles in the environmental adaptation. The qRT-PCR based expression profiling exhibited that CrNHXs responded to different stresses to varying degrees, which further confirmed the specificity of CrNHXs’ in responding to abiotic stresses. Moreover, the yeast functional complementation test proved that some CrNHXs could partially restore the salt tolerance of the salt-sensitive yeast mutant AXT3. This study provides comprehensive bio-information and primary functional identification of NHXs in C. rosea, which could help improve the salt/alkaline tolerance of genetically modified plants for further studies. This research also contributes to our understanding of the possible molecular mechanism whereby NHXs maintain the ion balance in the natural ecological adaptability of C. rosea to tropical coral islands and reefs.

Leaf structural traits of two Restinga plant species with different resistance patterns to iron toxicity

Iron mining activities are a source of particulate iron, which contaminates soil and plants of Restinga biome (Brazil). To investigate the possible effects of iron toxicity on Ipomoea pes-caprae (L.) R. Br. and Canavalia rosea (Sw.) DC. leaves, plants were submitted to different exposure times (12, 36, 108, and 228 hours) and iron concentrations (0.5 or 150 mg·L−1 Fe2+ as FeSO4·7H2O). After 108 hours of iron excess treatment, C. rosea leaves were chlorotic, whereas I. pes-caprae leaves presented venal chlorosis and bronzing after 228 hours of iron excess treatment. The anatomical alterations in I. pes-caprae were more intense and appeared after 36 hours, which was earlier than in C. rosea. The leaf epidermal cells of I. pes-caprae presented alterations in organization, size, and shape, and for both species the epicuticular wax was altered and wax rupture occurred close to the stomata. The positive staining for the presence of iron in leaf tissues matches with damaged areas in I. pes-caprae leaves, indicating direct iron toxicity. Ipomoea pes-caprae presented the most severe symptoms, whereas C. rosea presented the most resistance. The results suggest that, over time, the emission of particulate matter may negatively impact the ecological succession and biodiversity of Restinga.

Roles of Canavalia rosea metallothioneins in metal tolerance and extreme environmental adaptation to tropical coral reefs

Canavalia rosea (Sw.) DC is a perennial twining herb distributed in the semi-arid and saline–alkali areas of coastal regions and has evolved halotolerance. In this study, we present the first comprehensive survey of the metallothionein (MT) gene family in C. rosea. MT proteins belong to a family of low-molecular-weight polypeptides with a high content of cysteine residues, which have an affinity to bind with heavy metal ions. MTs also play important roles in stress responses as reactive oxygen species (ROS) scavengers. A total of six CrMTs were identified in the C. rosea genome and classified into four subgroups by phylogenetic analysis. An analysis of the cis-acting elements revealed that a series of hormone-, stress-, and development-related cis-acting elements were present in the promoter regions of CrMTs. The expression of CrMTs also showed habitat- and environmental stress-regulated patterns in C. rosea. CrMT overexpression in yeast enhanced tolerance to heavy metals and ROS, as well as high osmotic and alkalinity stress, which is consistent with their predicted roles as metal-chelating proteins and ROS scavengers. Our results indicate that the CrMT genes might contribute to the detoxification of plants to metals and provide marked tolerance against abiotic stress. The expression patterns of CrMTs in C. rosea also indicate that CrMTs play important roles in this species’ response to extreme environments on tropical islands and reefs, probably by improving the thermotolerance of C. rosea plants.

Genome-Wide Identification, Structure Characterization, Expression Pattern Profiling, and Substrate Specificity of the Metal Tolerance Protein Family in Canavalia rosea (Sw.) DC.

Plant metal tolerance proteins (MTPs) play key roles in heavy metal absorption and homeostasis in plants. By using genome-wide and phylogenetic approaches, the origin and diversification of MTPs from Canavalia rosea (Sw.) DC. was explored. Canavalia rosea (bay bean) is an extremophile halophyte with strong adaptability to seawater and drought and thereby shows specific metal tolerance with a potential phytoremediation ability. However, MTP genes in leguminous plants remain poorly understood. In our study, a total of 12 MTP genes were identified in C. rosea. Multiple sequence alignments showed that all CrMTP proteins possessed the conserved transmembrane domains (TM1 to TM6) and could be classified into three subfamilies: Zn-CDFs (five members), Fe/Zn-CDFs (five members), and Mn-CDFs (two members). Promoter cis-acting element analyses revealed that a distinct number and composition of heavy metal regulated elements and other stress-responsive elements existed in different promoter regions of CrMTPs. Analysis of transcriptome data revealed organ-specific expression of CrMTP genes and the involvement of this family in heavy metal stress responses and adaptation of C. rosea to extreme coral reef environments. Furthermore, the metal-specific activity of several functionally unknown CrMTPs was investigated in yeast. These results will contribute to uncovering the potential functions and molecular mechanisms of heavy metal absorption, translocation, and accumulation in C. rosea plants.