Anti-inflammatory effect of glucose-mannose binding lectins isolated from Brazilian beans

Selectins are essential for leukocyte recruitment in inflammation. Because of a lectin domain present in the selectin structure, we investigated the anti-inflammtory activity of six mannose-glucose binding lectins from brazilian beans: Dioclea guianensis-DguiL; D. grandiflora-DgL; Cratylia floribunda-CfL; D. violacea-D.vL; D. virgata-DvirL and Canavalia brasiliensis-ConBr. The lectins were injected intravenously (i.v.) into rats (0.1 and 1.0 mg/kg; 30 min before irritants) and its activities compared to E. coli endotoxin (LPS,30 mug/kg i.v.). Three lectins (DvL, CfL and DguiL), although less intense than LPS, inhibited the neutrophil migration induced by carrageenan (Cg, 300 mug) in a dose-dependent manner (0.1 and 1.0 mg/kg). DvL activity was reversed by 0.1 M alpha-D-methyl-mannoside (alpha-CH3), but not by 0.1 M alpha-D-galactose. The fMLP (44 ng)-induced neutrophil migration was also reduced by these lectins. Endotoxin contamination of lectin samples could be excluded since alpha-CH3 treatment reversed the DvL effect, but did not modify LPS inhibitory activity. Carrageenan (300 mug)-induced paw oedema was also reduced by LPS or lectin treatments. Conversely, none of the tested lectins inhibited dextran (Dex, 300 mug)-induced paw oedema, a classical leukocyte independent model, or zymosan (Zy, 1.0 mg)-induced peritonitis and paw oedema. LPS showed no effect upon Dex-induced paw oedema and barely reduced (25%) the oedematogenic effects of zymosan. As proposed for LPS, the lectin inhibitory activity was better observed on neutrophil-mediated inflammatory reactions. We speculate that the plant lectin antiinflammatory activity is probably due to a competitive blockage of a common leukocyte and/or endothelial selectin carbohydrate ligand.     

Plant‐insect interactions: what can we learn from plant lectins?

Many plant lectins have high anti‐insect potential. Although the effects of most lectins are only moderately influencing development or population growth of the insect, some lectins have strong insecticidal properties. In addition, some studies report a deterrent activity towards feeding and oviposition behavior. Transmission of plant lectins to the next trophic level has been investigated for several tritrophic interactions. Effects of lectins with different sugar specificities can vary substantially with the insect species under investigation and with the experimental setup. Lectin binding in the insect is an essential step in exerting a toxic effect. Attempts have been made to study the interactions of lectins in several insect tissues and to identify lectin‐binding receptors. Ingested lectins generally bind to parts of the insect gut. Furthermore, some lectins such as the Galanthus nivalus agglutinin (GNA) cross the gut epithelium into the hemolymph and other tissues. Recently, several candidate lectin‐binding receptors have been isolated from midgut extracts. To date little is known about the exact mechanism for insecticidal activity of plant lectins. However, insect glycobiology is an emerging research field and the recent technological advances in the analysis of lectin carbohydrate specificities and insect glycobiology will certainly lead to new insights in the interactions between plant lectins and insects, and to a better understanding of the molecular mechanisms involved. © 2010 Wiley Periodicals, Inc.     

The effect of lectins on the attachment and invasion of Enteromyxum scophthalmi (Myxozoa) in turbot (Psetta maxima L.) intestinal epithelium in vitro

The involvement of the lectin/carbohydrate interaction in the invasion of the turbot intestinal epithelium by Enteromyxum scophthalmi was studied in vitro using explants of turbot intestine and pre-treatment of parasite stages with the plant lectins of Canavalia ensiformis (Con A) and Glycine max (SBA). Both lectins inhibited the attachment and invasion of E. scophthalmi stages to the intestinal epithelium, though the inhibitory effect was higher for SBA than for Con A. Such results point to the involvement of N-acetyl-galactosamine (GalNAc) and galactose (Gal) residues and also of mannose/glucose residues in the E. scophthalmi-intestinal epithelium interaction. The inhibitory effect of both lectins on the parasite adhesion and penetration points to the interest of further studies to confirm the presence of putative lectins recognising GalNAc-Gal and mannose/glucose residues in turbot intestine. The obtained results demonstrated also the adequacy of turbot intestinal explants as an in vitro model to study the interaction with E. scophthalmi.     

Related mannose-specific lectins from different species of the family Amaryllidaceae

Bulbs from three species of the plant family Amaryllidaceae ( Narcissus pseudonurcissus L., Leucojum aestivum L. and Leucojum vernum L.) were found to contain mannose-specific lectins. These lectins were serologically identical to a previously reported Amaryllidaceae lectin from Galanthus nivalis L. bulbs, but had a different molecular structure. The lectins described in this paper are dimeric proteins composed of subunits of 13 kDa, which are not held together by disulphide bridges. In hapten-inhibition assays Amaryllidaceae lectins exhibited exclusive specificity towards mannose. Furthermore, they all had a high specific agglutination activity with trypsin-treated rabbit erythrocytes, whereas human red blood cells were not agglutinated.     

Isolation and characterization of a jacalin-related mannose-binding lectin from salt-stressed rice (Oryza sativa) plants

A novel plant lectin was isolated from salt-stressed rice (Oryzasativa L.) plants and partially characterized. The lectin occurs as a natural mixture of two closely related isoforms consisting of two identical non-covalently linked subunits of 15 kDa. Both isoforms are best inhibited by mannose and exhibit potent mitogenic activity towards T-lymphocytes. Biochemical analyses and sequence comparisons further revealed that the rice lectins belong to the subgroup of mannose-binding jacalin-related lectins. In addition, it could be demonstrated that the lectins described here correspond to the protein products of previously described salt-stress-induced genes. Our results not only identify the rice lectin as a stress protein but also highlight the possible importance of protein-carbohydrate interactions in stress responses in plants. Received: 27 July 1999 / Accepted: 11 November 1999     

Nucleocytoplasmic plant lectins

During the last decade it was unambiguously shown that plants synthesize minute amounts of carbohydrate-binding proteins upon exposure to stress situations like drought, high salt, hormone treatment, pathogen attack or insect herbivory. In contrast to the ‘classical’ plant lectins, which are typically found in storage vacuoles or in the extracellular compartment this new class of lectins is located in the cytoplasm and the nucleus. Based on these observations the concept was developed that lectin-mediated protein–carbohydrate interactions in the cytoplasm and the nucleus play an important role in the stress physiology of the plant cell. Hitherto, six families of nucleocytoplasmic lectins have been identified. This review gives an overview of our current knowledge on the occurrence of nucleocytoplasmic plant lectins. The carbohydrate-binding properties of these lectins and potential ligands in the nucleocytoplasmic compartment are discussed in view of the physiological role of the lectins in the plant cell.     

Natal insect experience with Arabidopsis thaliana plant genotypes influences plasticity in oviposition behavior

Previous studies have shown that insect experience with secondary chemicals present in different plant species can induce behavioral changes in female oviposition preferences. However, there is a lack of information on whether insect experience with intraspecific plant variation may influence oviposition behavior. The prediction that experience with plant genotypes would affect the oviposition behavior of two crucifer insect pests was tested using a wild ecotype of Arabidopsis thaliana (L.) Heynh. (Brassicaceae) (Col‐0) and two of its genetically modified lines (35S:ESP and tgg1 tgg2), which differ in their glucosinolate hydrolysis profiles. Choice oviposition assays were performed using both naïve and experienced females of the specialist Plutella xylostella (L.) (Lepidoptera: Plutellidae) and the generalist Trichoplusia ni (Hübner) (Lepidoptera: Noctuidae). In addition to oviposition preferences, the effect of plant genetic lines was assessed on insect traits related to development, growth, survival, and fecundity. Experience with different natal treatments (i.e., artificial diet and plant genotypes) led to changes in oviposition behavior of the diamondback moth P. xylostella; however, this effect was dependent on the specific genetic lines included in dual‐choice oviposition assays. In addition, for both moth species, experience led to female oviposition choices that would have maximized fitness of their offspring. In summary, this article suggests that insect experience with plant genotypes varying in their secondary metabolites can influence subsequent oviposition behavior. This outcome may have implications for plant‐insect coevolution and integrated pest management.     

First report of an arabinose-specific fungal lectin

To date, arabinose-binding lectins have been reported only from the human opportunistic pathogen Pseudomonas aeruginosa, the plant aggressive pathogen Ralstonia solanacearum, and the sponge Pellina semitubulosa. An arabinose-binding lectin with mitogenic activity toward splenocytes and a high specific hemagglutinating activity was isolated in the present study from a wild discomycete mushroom, Peziza sylvestris. The maximal mitogenic activity was induced by a lectin concentration of 8 microM. The lectin was a single-chained protein with a molecular mass of 20 kDa. Its N-terminal sequence showed only slight resemblance to other mushroom lectins. It was adsorbed on both diethylaminoethyl-cellulose and carboxymethyl-cellulose. Unlike previously reported mushroom lectins, the hemagglutinating activity of the lectin was inhibited by arabinose, but not by a large variety of other carbohydrates. The lectin activity was adversely affected in the presence of 0.05 M NaOH or 0.025 M HCl, and when the ambient temperature was elevated above 35 degrees C.     

Carbohydrate binding properties of banana (Musa acuminata) lectin I. Novel recognition of internal alpha1,3-linked glucosyl residues.

Examination of lectins of banana (Musa acuminata) and the closely related plantain (Musa spp.) by the techniques of quantitative precipitation, hapten inhibition of precipitation, and isothermal titration calorimetry showed that they are mannose/glucose binding proteins with a preference for the alpha-anomeric form of these sugars. Both generate precipitin curves with branched chain alpha-mannans (yeast mannans) and alpha-glucans (glycogens, dextrans, and starches), but not with linear alpha-glucans containing only alpha1,4- and alpha1,6-glucosidic bonds (isolichenan and pullulan). The novel observation was made that banana and plantain lectins recognize internal alpha1,3-linked glucosyl residues, which occur in the linear polysaccharides elsinan and nigeran. Concanavalin A and lectins from pea and lentil, also mannose/glucose binding lectins, did not precipitate with any of these linear alpha-glucans. This is, the authors believe, the first report of the recognition of internal alpha1,3-glucosidic bonds by a plant lectin. It is possible that these lectins are present in the pulp of their respective fruit, complexed with starch.     

Novel effect of plant lectins on the inhibition of Streptococcus mutans biofilm formation on saliva-coated surface

Aims:  Dental caries is caused by the disturbance in oral homeostasis, marked by a notable increase in the population of Streptococcus mutans . Lectins are a group of plant proteins that are capable of recognizing the glycoconjugates present on the bacterial surface. The aim of this study was to evaluate the effect of seven plant lectins on the growth and initial adhesion of S. mutans .
Methods and Results:  Lectins of different carbohydrate specificities were isolated from plant sources by conventional methods of protein purification. The effect on growth of S. mutans was evaluated following CLSI guidelines. None of the lectins used in this study inhibited the bacterial growth and multiplication. The adherence and biofilm formation of bacteria to saliva-coated polystyrene plates was tested in the presence of plant lectins. All the plant lectins tested, inhibited both the adherence and biofilm in a concentration dependent manner. Confocal microscopy and scanning electron microscopy were employed to assess the biofilm formation in the presence of plant lectin (glucose/mannose-specific) at sub-minimal inhibitory concentrations. These evaluations revealed that lectins inhibited the clumping and attachment of S. mutans .
Conclusions:  Lectins tested here inhibited initial biofilm formation by S. mutans. Glucose/Mannose-specific lectin altered the adhesion arrangement of the bacteria on the saliva-coated surfaces.
Significance and Impact of the Study:  The plant lectins used in this study may offer a novel strategy to reduce development of dental caries by inhibiting the initial adhesion and subsequent biofilm formation of S. mutans.     

Plant lectins as defense proteins against phytophagous insects

One of the most important direct defense responses in plants against the attack by phytophagous insects is the production of insecticidal peptides or proteins. One particular class of entomotoxic proteins present in many plant species is the group of carbohydrate-binding proteins or lectins. During the last decade a lot of progress was made in the study of a few lectins that are expressed in response to herbivory by phytophagous insects and the insecticidal properties of plant lectins in general. This review gives an overview of lectins with high potential for the use in pest control strategies based on their activity towards pest insects. In addition, potential target sites for lectins inside the insect and the mode of action are discussed. In addition, the effect of plant lectins on non-target organisms such as beneficial insects as well as on human/animal consumers is discussed. It can be concluded that some insecticidal lectins are useful tools that can contribute to the development of integrated pest management strategies with minimal effect(s) on non-target organisms.     

The biology and preliminary host range of Megacopta cribraria (Heteroptera: Plataspidae) and its impact on kudzu growth

The bean plataspid, Megacopta cribraria (F.), recently was discovered in the United States feeding on kudzu, Pueraria montana Lour. (Merr.) variety lobata (Willd.), an economically important invasive vine. We studied its biology on kudzu and its impact on kudzu growth. We also tested its ability to use other common forest legumes for oviposition and development. Flight intercept traps operated from 17 May 2010 to 31 May 2011 in a kudzu field near Athens, GA showed three peaks of adult flight activity suggesting there are two generations per year on kudzu. Vine samples examined for eggs from April 2010 to April 2011 and June to October 2011 showed two periods of oviposition activity in 2010, which coincided with the peaks in adult activity. In 2011, the second period of oviposition began on or before 24 June and then egg abundance declined gradually thereafter until late August when we recovered <2 eggs/0.5 m of vine. Samples of the five nymphal instars and adults on vines did not show similar trends in abundance. Adults did not lay eggs on the various legume species tested in 2010 in a no-choice test possibly because the cages were too small. In the 2011 field host range experiments conducted in a kudzu field by using 12 legume species, M. cribraria preferentially oviposited on kudzu over soybean, Glycine max Merrill., but they still laid 320 eggs per plant on soybean. Lespedeza hirta (L.) Hornem. and Lespedeza cuneata (Dum. Cours.) G. Don had 122.2 and 108.4 eggs per plant, respectively. Kudzu and soybean were the only species M. cribraria completed development on. Plots protected from M. cribraria feeding by biweekly insecticide applications had 32.8% more kudzu biomass than unprotected plots. Our results show that M. cribraria has a significant impact on kudzu growth and could help suppress this pest weed.     

Plant wounding affects the oviposition of Hellula undalis(F.). The impact of leaf chemistry

Crucifer specialists, such as the tropical pest Hellula undalis (F.), developed a close association to their hosts and depend on characteristic secondary plant compounds, the glucosinolates (GS), in many cases. Following tissue damage, GS undergo hydrolysis catalyzed by the enzyme myrosinase to produce a complex array of products which include isothiocyantes, cyanides, and thiocyanates depending on reaction condition and the chemical nature of the side chain. We found that mechanical plant damage to the tropical crop plant Brassica campestris L. ssp. chinensis (pak-choi) leads to increased GS content, especially of indolyl GS and affected the oviposition behaviour of H. undalis negatively. GS were detected in methanolic leaf surface extracts from intact B chinensis and Sinapis alba (L.) plants, but no hydrolysis products of GS were found in leaf extracts with methylene chloride. Furthermore, the oviposition stimulant activity of GS extracts for H. undalis was greatly reduced after hydrolysis of GS by myrosinase. These results suggest the importance of GS for inducing oviposition, thus excluding hydrolysis products as active principle. Specific produced slightly volatile hydrolysis products of GS (3-indolylmethyl-cyanide and 5-vinyloxazolidine-2-thione) reduced oviposition, when applied to the host-plant. Leaf chemistry, including volatile and non-volatile compounds, may determine oviposition in H. undalis indicating a suitable host-plant.     

A novel mitogenic and antiproliferative lectin from a wild cobra lily, Arisaema flavum

A novel lectin having specificity towards a complex glycoprotein asialofetuin was purified from tubers of Arisaema flavum (Schott.) by affinity chromatography on asialofetuin-linked amino-activated silica beads. A. flavum gave a single peak on HPLC size exclusion and a single band on non-denatured PAGE at pH 4.5. The molecular mass of the lectin, as determined by gel filtration chromatography, was 56 kDa. In SDS-PAGE, pH 8.3, the lectin migrated as a single band of 13.5 kDa, under reducing and non-reducing conditions, indicating the homotetrameric nature. A. flavum lectin (AFL) readily agglutinated rabbit, rat, sheep, goat, and guinea pig erythrocytes but not human ABO blood group erythrocytes even after neuraminidase treatment. This lectin is stable up to 55 degrees C and does not require metal ions for its hemagglutination activity. AFL was completely devoid of sulphur containing amino acids and was rich in aspartic acid and glycine. In Oucterlony's double immunodiffusion, the antisera raised against A. flavum lectin showed distinct lines of identity with those of other araceous lectins. AFL showed potent mitogenic activity towards BALB/c splenocytes and human lymphocytes in comparison to Con A, a well-known plant mitogen. AFL also showed significant in vitro antiproliferative activity towards J774 and P388D1 murine cancer cell lines.     

A novel mannose-binding tuber lectin from Typhonium divaricatum (L.) Decne (family Araceae) with antiviral activity against HSV-II and anti-proliferative effect on human cancer cell lines

A novel mannose-binding tuber lectin with in vitro antiproliferative activity towards human cancer cell lines and antiviral activity against HSV-II was isolated from fresh tubers of a traditional Chinese medicinal herb, Typhonium divaricatum (L.) Decne by a combined procedure involving extraction, ammonium sulfate precipitation, ion exchange chromatography on DEAE-SEPHAROSE, CM-SEPHAROSE and gel-filtration on sephacryl S-200. The apparent molecular mass of the purified Typhonium divaricatum lectin (TDL) was 48 kDa. TDL exhibits hemagglutinating activity toward rabbit erythrocytes at 0.95 microg/ml, and its activity could be strongly inhibited by mannan, ovomucoid, asialofetuin and thyroglobulin. TDL showed antiproliferative activity towards some well established human cancer cell lines, e.g. Pro-01 (56.7 +/- 6.8), Bre-04 (41.5 +/- 4.8), and Lu-04 (11.4 +/- 0.3). The anti-HSV-II activity of TDL was elucidated by testing its HSV-II infection inhibitory activity in Vero cells with TC(50) and EC(50) of 5.176 mg/ml and 3.054 microg/ml respectively. The full-length cDNA sequence of TDL was 1145 bp and contained an 813-bp open reading frame (ORF) encoding a 271 amino acid precursor of 29-kDa. Homology analysis showed that TDL had high homology with many other mannose-binding lectins. Secondary and three-dimensional structures analyses showed that TDL is heterotetramer and similar with lectins from mannose-binding lectin superfamily, especially those from family Araceae.     

Feeding and oviposition preference of Frankliniella occidentalis for crops and weeds in Kenyan French bean fields

Western flower thrips (WFT), Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), is an important pest of French beans in Kenya. However, information on the feeding activity and oviposition preference of WFT on crop and weed hosts associated with French beans in Kenya and other parts of the world is lacking. To determine the feeding and oviposition preference of WFT for crop and weed plants commonly encountered in French bean fields in Kenya, no‐choice and choice experiments were conducted using four important crop and weed plants. Among the crop plants tested, highest feeding and oviposition activity of WFT was recorded on courgette/zucchini (Cucurbita pepo L.) and French beans (Phaseolus vulgaris L.). Spinach beet (Beta vulgaris L.) and sweet pepper (Capsicum annuum L.) were of relatively minor importance for feeding and oviposition. Among the weeds tested, gallant soldier (Galinsoga parviflora Cav.) was the most preferred host plant for feeding and oviposition compared with Chinese lantern (Nicandra physaloides L.), wild crucifer (Erucastrum arabicum Fisch. & C.A. Mey.) and pigweed (Amaranthus hybridus L.). Phaseolus vulgaris was the most preferred host for feeding and oviposition in the presence of G. parviflora, E. arabicum and A. hybridus. A positive correlation between the number of feeding punctures and the number of eggs oviposited by WFT on crop and weed plants was observed. The results of this study show that P. vulgaris, C. pepo and G. parviflora are both relatively good feeding and oviposition hosts of WFT. Cucurbita pepo and G. parviflora may serve as potential sources of WFT outbreaks within French bean fields.     

Antagonistic effects of floral scent in an insect–plant interaction

In southwestern USA, the jimsonweed Datura wrightii and the nocturnal moth Manduca sexta form a pollinator–plant and herbivore–plant association. Because the floral scent is probably important in mediating this interaction, we investigated the floral volatiles that might attract M. sexta for feeding and oviposition. We found that flower volatiles increase oviposition and include small amounts of both enantiomers of linalool, a common component of the scent of hawkmoth-pollinated flowers. Because (+)-linalool is processed in a female-specific glomerulus in the primary olfactory centre of M. sexta, we hypothesized that the enantiomers of linalool differentially modulate feeding and oviposition. Using a synthetic mixture that mimics the D. wrightii floral scent, we found that the presence of linalool was not necessary to evoke feeding and that mixtures containing (+)- and/or (−)-linalool were equally effective in mediating this behaviour. By contrast, females oviposited more on plants emitting (+)-linalool (alone or in mixtures) over control plants, while plants emitting (−)-linalool (alone or in mixtures) were less preferred than control plants. Together with our previous investigations, these results show that linalool has differential effects in feeding and oviposition through two neural pathways: one that is sexually isomorphic and non-enantioselective, and another that is female-specific and enantioselective.     

Oviposition behaviour of four ant parasitoids (Hymenoptera, Braconidae, Euphorinae, Neoneurini and Ichneumonidae, Hybrizontinae), with the description of three new European species

The oviposition behaviour of four ant parasitoids was observed and filmed for the first time. The movies are available from YouTube (search for Elasmosoma, Hybrizon, Kollasmosoma and Neoneurus). Two of the observed species (Neoneurus vesculussp. n. and Kollasmosoma sentumsp. n.) are new to science. A third species (Neoneurus recticalcarsp. n.) is described from Slovakia and Norway. Keys to the Palaearctic species of the genera Neoneurus and Kollasmosoma are added.