Lectin Gene Expression Research Articles

Fruit ripening and chitosan coating impacts on the expression profile of Banana Lectin (BanLec) genes

Banana Lectin (BanLec) was found in banana, is capable to identify and attach to carbohydrates without modified them. While the process for the synthesis of lectins remains unidentified, it is understood that certain inducing factors can trigger their expression. This research assessed the gene expression levels of lectins in Cavendish bananas throughout different stages of ripening by contrasting bananas with peels coated in 1.25 % chitosan solution to their uncoated counterparts. This experiment was performed using a transcriptomic approach and conducted in silico. The in silico analysis revealed the induction of 14 distinct lectin genes expression subsequent to the ripening stages in uncoated Cavendish bananas. Along with the ripening process, it was observed that lectins potentially contribute to protein phosphorylation and augment the carbohydrate-binding activity. Conversely, the application of a 1.25 % chitosan coating led to the heightened expression of 17 specific lectin genes. Post-coating, it is hypothesized that modifications to the primary cell wall were effected through the cleavage and recombination of xyloglucan molecules. The qPCR results substantiated the induction of BanLec genes, Ma09_g10350 and Ma11_g22300, by the application of chitosan coating. Findings from this transcriptomic investigation propose that the upregulation of lectin genes induced after chitosan treatment may be implicated in plant defense mechanisms. This study reveals that natural ripening and a 1.25 % chitosan coating influenced lectin gene expression in Cavendish bananas differently. Both ripening and chitosan treatments appear to be viable strategies for enhancing banana lectin expression, which has potential health applications.

Differing Expression and Potential Immunological Role of C-Type Lectin Receptors of Two Different Chicken Breeds against Low Pathogenic H9N2 Avian Influenza Virus.

Diverse immune responses in different chicken lines can result in varying clinical consequences following avian influenza virus (AIV) infection. We compared two widely used layer breeds, Lohmann Brown (LB) and Lohmann White (LW), to examine virus replication and immune responses against H9N2 AIV infection. The transcription profile in the spleen of H9N2-infected chickens was compared using a microarray. Confirmatory real-time RT-PCR was used to measure the expression of C-type lectin, OASL, and MX1 genes. Additionally, to investigate the role of chicken lectin receptors in vitro, two C-type lectin receptors (CLRs) were expressed in DF-1 cells, and the early growth of the H9N2 virus was evaluated. The LB chickens shed a lower amount of virus from the cloaca compared with the LW chickens. Different expression levels of C-type lectin-like genes were observed in the transcription profile, with no significant differences in OASL or MX gene expression. Real-time RT-PCR indicated a sharp decrease in C-type lectin levels in the spleen of H9N2-infected LW chickens. In vitro studies demonstrated that cells overexpressing CLR exhibited lower virus replication, while silencing of homeostatic CLR had no effect on AIV replication. This study demonstrated distinct immune responses to H9N2 avian influenza in LB and LW chickens, particularly with differences in C-type lectin expression, potentially leading to lower virus shedding in LB chickens.

Impact of Exogenous Nitric Oxide Treatment on Vascularization of a Subcutaneous Device for Cell Transplantation

Subcutaneous polymer scaffolds have shown potential for creating an optimal transplantation site in cellular replacement therapy, e.g., when transplanting insulin-producing cells to cure type 1 diabetes. Imperative for these scaffolds is a high degree of vascularization to guarantee long-term functional cellular survival. In this study, the effect of the nitric oxide (NO) donor S-nitroso-N-acetyl-dl-penicillamine (SNAP) on the vascularization degree of a subcutaneous poly(d,l-lactide-co-ε-caprolactone) (PDLLCL) scaffold was investigated. To this end, scaffolds were implanted under the skin of C57BL/6 mice. Each mouse received a control scaffold and a scaffold containing SNAP. At day 7, 14, and 28, the oxygen percentage within the scaffolds was measured and at day 28, the vascularization degree was determined with lectin infusion and gene expression analysis. We measured lower oxygen percentages within the scaffolds containing the NO-donor up to day 14 compared to the control scaffolds, but no differences were found at day 28. Although blood vessels in the scaffolds were well perfused, no differences between the groups were found in the lectin staining and gene expression of vascular markers, such as CD31, CD105, and VEGFa. To conclude, in this biomaterial setting, addition of a NO-donor did not improve the vascularization degree of the subcutaneous scaffold.

Using FIBexDB for In-Depth Analysis of Flax Lectin Gene Expression in Response to Fusarium oxysporum Infection

Plant proteins with lectin domains play an essential role in plant immunity modulation, but among a plurality of lectins recruited by plants, only a few members have been functionally characterized. For the analysis of flax lectin gene expression, we used FIBexDB, which includes an efficient algorithm for flax gene expression analysis combining gene clustering and coexpression network analysis. We analyzed the lectin gene expression in various flax tissues, including root tips infected with Fusarium oxysporum. Two pools of lectin genes were revealed: downregulated and upregulated during the infection. Lectins with suppressed gene expression are associated with protein biosynthesis (Calreticulin family), cell wall biosynthesis (galactose-binding lectin family) and cytoskeleton functioning (Malectin family). Among the upregulated lectin genes were those encoding lectins from the Hevein, Nictaba, and GNA families. The main participants from each group are discussed. A list of lectin genes, the expression of which can determine the resistance of flax, is proposed, for example, the genes encoding amaranthins. We demonstrate that FIBexDB is an efficient tool both for the visualization of data, and for searching for the general patterns of lectin genes that may play an essential role in normal plant development and defense.

Regulation of mucosal lectins in the oyster Crassostrea virginica in response to food availability and environmental factors

ABSTRACT Lectins are a large and diverse group of sugar-binding proteins involved in nonself recognition and cell-to-cell interactions. Suspension-feeding bivalves, such as the oyster Crassostrea virginica, are capable of using these molecules to bind cell surface carbohydrates of food particles, allowing particle capture and selection. The aim of this project was to assess whether the expression of mucosal lectins in C. virginica is constant or changes with the season, and to determine whether lectin expression is linked to environmental parameters and/or internal biological factors (gametogenesis). A total of 130 oysters were placed in submerged cages at a tidal estuary and monitored for changes in lectin gene expression over a 1-year period. In parallel, environmental parameters prevailing at the field site, including seawater physicochemical characteristics (temperature, salinity and dissolved oxygen), particulate organic matter and chlorophyll contents, were also monitored. Throughout the study, oysters were dissected and the gills were collected and used for the assessment of the expression of three different lectin genes (CvML, CvML3914 and CvML3912). Remaining tissues were processed for histology and the classification of the gonad development stage. Results showed that when food is abundant, such as during the spring bloom, lectin gene expressions are low, and inversely lectin levels increase with lower food levels. These findings suggest that oysters increase lectin expression to enhance the capture and ingestion of scarce food, while during spring, enough food is already being ingested and lectins are not needed. Furthermore, results showed that as the energy demands of oysters increase (gonad maturation), lectin gene expressions also increase to enhance selective ingestion of nutritious food particles. This study, therefore, demonstrates the seasonality of lectin gene expression in C. virginica, and suggests that lectin regulation is related to the reproduction process and abundance of high-quality food.

OsEUL Lectin Gene Expression in Rice: Stress Regulation, Subcellular Localization and Tissue Specificity.

The Euonymus lectin (EUL) family is a unique group of carbohydrate-binding proteins that is omnipresent in plants. Sequences encoding EUL-related lectins have been retrieved from all completely sequenced plant genomes. The rice (Oryza sativa) genome contains 5 functional EUL genes referred to as OsEULS2, OsEULS3, OsEULD1a, OsEULD1b, and OsEULD2. In this study we focused on the tissue specific expression, stress inducibility and subcellular localization of the rice EULs. Even though the EUL domain sequence is highly conserved among the rice EULs (at least 80% sequence similarity) different biotic and abiotic stress treatments yielded unique responses for the different EULs. Transcript levels for OsEULs were differentially affected by drought and salt stress, ABA treatment, pathogen infection or insect infestation. Analysis of promoter activity revealed differential expression and tissue specificity for the 5 OsEUL genes, with most expression observed in the vascular system of roots and shoots, as well as in the root tips and seeds. At cell level, all OsEULs are located in the nucleus whereas OsEULD1b and OsEULD2 also locate to the cytoplasm. This paper contributes to the functional characterization of the EULs and provides insight in the biological importance of this family of proteins for rice.

Changes in Hormonal Status and Expression of Lectin Genes in Wheat Seedlings Treated with Stevioside and Exposed to Low Above-Zero Temperatures

Effect of presowing treatment with stevioside (10–8 M) on activity, content, and expression of the wheat-germ agglutinin (WGA) gene, as well as the content of phytohormones, was studied in 4-day-old seedlings of wheat (Triticum aestivum L.) exposed to low above-zero temperatures. Pretreatment with stevioside considerably increased the protective potential of plants in response to low-temperature stress. The obtained results point to an ABA-dependent effect of stevioside on WGA in spite of the gibberellin-like chemical structure of the examined growth regulator.

Quantification of C-type lectin gene expression during hyperinfection in strongyloidiasis

Quantification of C-type lectin gene expression during hyperinfection in strongyloidiasis

Short Communication: Detection of lectin gene (MLL1 and M35) in mulberry plant (Morus spp.) from Bogor, West Java, Indonesia

Wulandari YRE, Yogiara, Lizar M. 2018. Short Communication: Detection of lectin gene (MLL1 and M35) in mulberry plant (Morus spp.) from Bogor, West Java, Indonesia Biodiversitas 19: 2381-2384. Plant species contains carbohydrate-binding protein known as lectin or agglutinin. Lectin binds to specific carbohydrates, such as monosaccharides or oligosaccharides and initiates agglutination process. Lectin plays an important role as plant defense, so that it can be used to prevent pest attacks. Mulberry leaf lectin 1 (MLL1) from young leaves of Morus alba can be used against phytopathogenic bacteria, Pseudomonas syringae pv. mori. Mannose-binding lectin (M35) was found on stems of M. nigra as protein storage. M35 is also produced on roots of M. alba and induced by mulberry stem cuttings. This research purpose was to isolate and analyze lectin gene expression (MLL1 and M35) in M. alba var. multicaulis, M. cathayana, M. bombycis var. lembang, and M. alba var. kanva-2 from Bogor, West Java, Indonesia. Different plant organ including leaves, stems, and roots were used as source of samples and analyze using Reverse Transcription Polymerase Chain Reaction (RT-PCR). Our results showed that all of MLL1 genes were expressed in young leaves, but not expressed in stems and roots of mulberry plant samples. The M35 gene was expressed in young leaves, stems, and roots of all mulberry plant samples. Reverse Transcription PCR of MLL1 gene exhibited a 350 bp DNA band, while M35 gene exhibited a 99 bp DNA band.

Intestinal Epithelial Wnt Signaling Mediates Acetylcholine-Triggered Host Defense against Infection

Regulated antimicrobial peptide expression in the intestinal epithelium is key to defense against infection and to microbiota homeostasis. Understanding the mechanisms that regulate such expression is necessary for understanding immune homeostasis and inflammatory disease and for developing safe and effective therapies. We used Caenorhabditis elegans in a preclinical approach to discover mechanisms of antimicrobial gene expression control in the intestinal epithelium. We found an unexpected role for the cholinergic nervous system. Infection-induced acetylcholine release from neurons stimulated muscarinic signaling in the epithelium, driving downstream induction of Wnt expression in the same tissue. Wnt induction activated the epithelial canonical Wnt pathway, resulting in the expression of C-type lectin and lysozyme genes that enhanced host defense. Furthermore, the muscarinic and Wnt pathways are linked by conserved transcription factors. These results reveal a tight connection between the nervous system and the intestinal epithelium, with important implications for host defense, immune homeostasis, and cancer.

Ectopic Overexpression of Lectin in Transgenic Brassica juncea Plants Exhibit Resistance to Fungal Phytopathogen and Showed Alleviation to Salt and Drought Stress

Lectins are ubiquitous in nature and one of the most important secondary metabolites in plants. They are heterogeneous class of proteins that bind specifically and reversibly to carbohydrates and are used as a defence tool against various abiotic and biotic stresses. In this study, we describe generation of transgenic Brassica juncea plants over-expressing chickpea lectin in conferring fungal resistance. In addition, the transgenic plants also showed resistance against salinity (NaCl) and mannitol induced drought stress. The stable integration and transcript expression of the transgene was confirmed by southern blot and RT-PCR analysis in the transgenic plants. The transgenic lines were evaluated for resistance against Alternaria brassicae as assessed by the number and size of lesions and time taken for leaf necrosis, the Alternaria infection was delayed and restricted in the transgenic plants with the protection varying from 36-60% in different transgenic lines compared to the Untransformed Control (UC) plants. In addition, the transgenic Brassica juncea plants also showed tolerance against high salinity and drought stress. The Expression of lectin gene imparts greater water retention capacity, higher proline content and more chlorophyll retention with less membrane damage and reduction in electrolyte leakage in the transgenic plants compared to UC. Together, these findings suggest the potential use of chickpea lectin gene to engineer the crop plants for broad spectrum biotic and abiotic stress tolerance to improve crop productivity.

Functional assay using lectin gene targeting technologies (over-expression).

Function of lectin depends on its amino acid sequence of carbohydrate-recognition domain (CRD), conformation, and extracellular/intracellular localization. Altering lectin gene expression by over-expression or knockdown is a powerful tool for analyzing its cellular function. Here, we describe a method of lectin gene over-expression, taking a C-type lectin, mannan-binding protein (MBP), as an example. Carbohydrate-binding ability of MBP, its subcellular localization, and functional co-localization with ligand glycoprotein are assayed comparing with an inactive mutant MBP.

Prokaryotic expression of C-type lectin gene ofLitopenaeus vannameiand immunologic activity evaluation of recombinant protein

The studies on innate immune responses of Litopenaeus vannamei C-type lectin,were conducted by recombinant C-type lectin protein through expression in Escherichia coli.We used cell suspension culture to study the effect of C-type lectin on the cellular immune response of Litopenaeus vannamei.The experiment was divided into two groups:control group and recombinant protein(1.0 mg/mL).After 0,3,6,9,12,24 hours,we harvested the solutions and then assessed parameters respectively.The results showed that SDS-PAGE revealed the molecular weight of LvLec protein was 39 ku,which was consistent with the predicted size.Western-blotting revealed it could bind anti-His antibody,which proved that C-type lectin recombinant protein in Escherichia coli succeeded in expression.For the hemagglutination assay,massive agglutination could be observed after Escherichia coli was incubated with LvLec for 1 h.At the same time,LvLec protein had a significant effect on haemocyte count,cell viability and phagocytic activity of white shrimp Litopenaeus vannamei(P0.05).The extracelluar phenoloxidase activity of the LvLec protein group showed peak changes during 3-12 h,and at 9 h the extracelluar phenoloxidase activity was highest,then returned to the control level(P0.05).Therefore,crustacean C-type lectin not only had the function of hemagglutination,but also activated prophenoloxidase activating system,and probably played an important role in immune response.

Sex Differences in Carbohydrate Metabolism Are Linked to Gene Expression in Caenorhabditis elegans

The male and the hermaphrodite forms of the nematode Caenorhabditis elegans (C. elegans) differ markedly in anatomy, nervous system and behavior at adulthood. Using the male mutants fog-2, him-5, and him-8, we compared body proportions and composition, and aspects of carbohydrate metabolism and gene expression between the C. elegans sexes in three adult stages. In all experiments, both sexes were grown on the same plate and separated using flow cytometry. The fat to fat-free mass ratio and the body volume-adjusted fat mass is similar between the sexes, although the body size is more than 50% smaller in adult males than in age-matched hermaphrodites. The volume-adjusted total RNA content is approximately 2-fold lower in males. Biochemical and NMR-based analyses reveal higher trehalose levels and much lower glucose levels in males than in hermaphrodites. The resulting trehalose-to-glucose ratio is 5.4-fold higher in males. These sex differences are reflected in gene expression data because the genes encoding key enzymes of the glycolysis and trehalose synthesis pathways are more highly expressed in males than in hermaphrodites. Notably, expression of the phosphofructokinase gene (C50F4.2) is 29-fold higher in males. Comparative analysis of gene expression data identifies 285 male-specific and 160 hermaphrodite-specific genes. These include transcription factor and C-type lectin-encoding genes. More than 35% of all C-type lectin genes are more highly expressed in males. The expression of many C-type lectin genes differs by a factor of >100 between the sexes. In conclusion, we found sex differences in carbohydrate metabolism that are linked to gene expression and identified certain lectin genes that are differentially expressed by the C. elegans sexes.

Expression of putative immune response genes during early ontogeny in the coral Acropora millepora.

BackgroundCorals, like many other marine invertebrates, lack a mature allorecognition system in early life history stages. Indeed, in early ontogeny, when corals acquire and establish associations with various surface microbiota and dinoflagellate endosymbionts, they do not efficiently distinguish between closely and distantly related individuals from the same population. However, very little is known about the molecular components that underpin allorecognition and immunity responses or how they change through early ontogeny in corals.Methodology/Principal FindingsPatterns in the expression of four putative immune response genes (apextrin, complement C3, and two CELIII type lectin genes) were examined in juvenile colonies of Acropora millepora throughout a six-month post-settlement period using quantitative real-time PCR (qPCR). Expression of a CELIII type lectin gene peaked in the fourth month for most of the coral juveniles sampled and was significantly higher at this time than at any other sampling time during the six months following settlement. The timing of this increase in expression levels of putative immune response genes may be linked to allorecognition maturation which occurs around this time in A.millepora. Alternatively, the increase may represent a response to immune challenges, such as would be involved in the recognition of symbionts (such as Symbiodinium spp. or bacteria) during winnowing processes as symbioses are fine-tuned.Conclusions/SignificanceOur data, although preliminary, are consistent with the hypothesis that lectins may play an important role in the maturation of allorecognition responses in corals. The co-expression of lectins with apextrin during development of coral juveniles also raises the possibility that these proteins, which are components of innate immunity in other invertebrates, may influence the innate immune systems of corals through a common pathway or system. However, further studies investigating the expression of these genes in alloimmune-challenged corals are needed to further clarify emerging evidence of a complex innate immunity system in corals.

Bioengineering of symbiotic systems: Creation of new associative symbiosis with the use of lectins on the example of tobacco and oil seed rape

"Barbate roots" in tobacco and colza transgenic on lectin gene were obtained with the use of a wild strain of Agrobacterium rhizogenes 15834 transformed with pCAMBIA1305.1 plasmid containing the full-size lectin gene (psl) from the Pisum sativum. Influence of expression oflectin gene on colonization oftransgenic roots with symbiont of pea (Rhizobium leguminosarum) was investigated. The number of adhered bacteria onto the roots transformed with lectin gene was 14-fold and 37-fold higher in comparison with the control; this confirms the interaction of R. leguminosarum with pea lectin at the surface of the transformed roots of tobacco and colza. The developed experimental approach, based on the simulation of recognition processes and early symbiotic interactions with lectins of pea plants, may, in perspective, be used for obtaining stable associations of economically valuable, nonsymbiotrophic plant species with rhizobia.

Characterization of a mannose-binding lectin from channel catfish (Ictalurus punctatus)

Mannose-binding lectin (MBL) is an important component of innate immunity capable of activating the lectin pathway of the complement system. A MBL gene was isolated from channel catfish (Ictalurus punctatus). The deduced protein contains a canonical collagen-like domain, a carbohydrate recognition domain (CRD), and a neck region similar to mammalian mannose-binding lectin. The catfish mannose-binding lectin CRD contains the EPN motif shown previously to mediate mannose specificity. The catfish mannose-binding lectin showed 30–43% identity with MBL protein sequences of rainbow trout, zebrafish, common carp, and goldfish, and 33–35% identity with sequences of mammalian species. In this study, while liver was the predominant source of mannose-binding lectin gene expression in healthy tissues, mannose-binding lectin expression in spleen rose sharply following challenge with a Gram-negative bacterium.

SUMO fusion facilitates expression and purification of garlic leaf lectin but modifies some of its properties

Over expression of lectin genes in E. coli often gives inclusion bodies that are solubilised to characterize lectins. We made N-terminal fusion of the Allium sativum leaf agglutinin (ASAL) with SUMO (small ubiquitin related modifier) peptide. The SUMO peptide allowed expression of the recombinant lectin in E. coli, predominantly in soluble form. The soluble fusion protein could be purified by immobilized metal affinity column (IMAC), followed by size exclusion chromatography. The SUMO protease failed to cleave the SUMO peptide from ASAL. This may be due to steric hindrance caused by the homodimer structure of the chimeric ASAL. Some properties like dimerization, haemagglutination and insecticidal properties of the recombinant SUMO–ASAL fusion protein were comparable to the plant derived native lectin. However, glycan array analysis revealed that the carbohydrate binding specificity of the recombinant SUMO–ASAL was altered. Further, the fusion protein was not toxic to E. coli (native ASAL exhibited toxicity). The recombinant lectin was more thermo-labile as compared to the native lectin. Three important findings of this study are: (1) sugar specificity of ASAL can be altered by amino-terminal fusion; (2) anti- E. coli activity of ASAL can be eliminated by N-terminal SUMO fusion and (3) SUMO–ASAL may be a preferred candidate insecticidal protein for the development of transgenic plants.

Экспрессия генов лектина и дефенсина у сортов пшеницы Мироновская 808 и Roazon при инфицировании Pseudocercosporella herpotrichoides

Экспрессия генов лектина и дефенсина у сортов пшеницы Мироновская 808 и Roazon при инфицировании Pseudocercosporella herpotrichoides

Symbiotic reactions of sea-buckthorn roots transformed with the pea lectin gene

Sea-buckthorn (Hyppopha L.) transgenic roots transformed with the lectin gene were obtained using the wild-type strain of Agrobacterium rhizogenes 15834 preliminary transformed with the plasmid pCAMBIA 1305.1, which contained the full-size pea lectin gene. Effects of lectin gene expression on symbiotic responses of sea-buckthorn to inoculation with rhizobia (Rhizobium leguminosarum, pea symbiont) and actinomycetes of genus Frankia (sea-buckthorn symbiont) were studied. In sea-buckthorn seedlings, whose transgenic roots were inoculated with both microsymbionts simultaneously, atypical nodule-like structures were found along with typical actinorhizal nodules. Random amplified polymorphic DNA (RAPD) analysis of bacteria, isolated from these structures, revealed the presence of R. leguminosarum rhizobia and the absence of Frankia actinomycetes.