Chenopodium murale L. is an invasive weed species significantly interfering with wheat crop. However, the complete nature of its allelopathic influence on crops is not yet fully understood. In the present study, the focus is made on establishing the relation between plant morphophysiological changes and oxidative stress, induced by allelopathic extract. Phytotoxic medium of C. murale hairy root clone R5 reduced the germination rate (24% less than control value) of wheat cv. Nataša seeds, as well as seedling growth, diminishing shoot and root length significantly, decreased total chlorophyll content, and induced abnormal root gravitropism. The R5 treatment caused cellular structural abnormalities, reflecting on the root and leaf cell shape and organization. These abnormalities mostly included the increased number of mitochondria and reorganization of the vacuolar compartment, changes in nucleus shape, and chloroplast organization and distribution. The most significant structural changes were observed in cell wall in the form of amoeboid protrusions and folds leading to its irregular shape. These structural alterations were accompanied by an oxidative stress in tissues of treated wheat seedlings, reflected as increased level of H 2 O 2 and other ROS molecules, an increase of radical scavenging capacity and total phenolic content. Accordingly, the retardation of wheat seedling growth by C. murale allelochemicals may represent a consequence of complex activity involving both cell structure alteration and physiological processes.
Since the genetic transformation of the apple is strongly genotype-dependent and generally inefficient, the evaluation of factors affecting shoot regeneration are crucial for the establishment of a successful transformation process. In this report, we evaluated the effects of the ?-lactam antibiotics meropenem and timentin on in vitro regeneration via de novo shoot organogenesis from leaf explants of apple cv. Golden Delicious, as well as on the growth of the Agrobacterium tumefaciens strain EHA 105, and compared them with the commonly used ?-lactam cefotaxime. Also, we report for the first time the effect of hygromycin B as a selective agent in the domesticated apple, as regards shoot regeneration and shoot multiplication efficiency. We observed that cefotaxime and timentin at concentrations higher than 100 mg L-1 were sufficient to prevent Agrobacterium growth during a two-week period, while meropenem exhibited an inhibitory effect on bacterial growth at all tested concentrations (25-150 mg L-1). Cefotaxime at a concentration of 300 mg L-1 increased the number of regenerated shoots per explant (9.39) in comparison with the control (7.67). In contrast to cefotaxime, meropenem and timentin caused a decrease in shoot regeneration efficiency, but larger and more developed shoots were obtained on meropenem (25-125 mg L-1) after the same period of cultivation. Hygromycin B at a concentration of 5 mg L-1 or higher completely inhibited shoot regeneration and induced explant tissue necrosis. Therefore, the selection procedure with a final concentration of 4 mg L-1 throughout organogenesis and 10 mg L-1 for further shoot growth and multiplication is recommended for an efficient transformation process in apple cv. Golden Delicious.
The aim of the present study was to investigate the effect of sucrose on shoot regeneration potential in Hypericum perforatum L. roots obtained by Agrobacterium rhizogenes transformation. The morphological evaluation of transgenic roots grown on media supplemented with sucrose (0.5, 1, 2, 4, 6 and 8 %) indicated that both genotype and sucrose concentration significantly affected root elongation and branching, as well as shoot regeneration. For two of five analyzed clones, lower sucrose concentrations (up to 2 %) led to intensive shoot regeneration, while the other three clones intensified shoot development only at elevated sucrose concentrations (4 %). For all clones, concentrations above 4 % had a deleterious effect on both root and shoot development. Genetic characterization of regenerated shoots revealed that all tested clones were diploid with an average of 0.670 ± 0.002 pg of DNA per nucleus, with no significant differences between transgenic and non-transformed plants and, according to PCR, with integrated A. rhizogenes rolA, -B, -C and -D genes. Real-time RT-PCR confirmed the expression of rolA, -B and -C, while expression of the rolD gene was not detected. Differences were detected in the absolute amounts of transcripts between analyzed clones, with the highest levels of expression for all three analyzed rol genes in a clone previously defined as having high root differentiation and less effective shoot regeneration potential. The observed variations in morphogenesis potential could be attributed to different levels of expression of integrated rolA, -B and -C genes; while sucrose additionally pointed out these trends.
Study describes protocols for in vitro propagation of Gentiana utriculosa L. via axillary shoot multiplication and indirect somatic embryogenesis. Shoot cultures were established from seedling epicotyl explants cultured on MS medium supplemented with 0.25 mg L−1 BA and 0.1 mg L-1 IAA. Medium containing 2% sucrose and 0.2 mg L−1 BA improved multiple shoot production, providing 2.3 shoots per explant. The highest rooting (29.6%) was obtained on medium with 1/2 MS mineral salts and 0.5 mg L−1 NAA. Somatic embryogenesis was induced using different explants, including immature seeds as well as leaves and roots from shoot cultures. Following auxin treatment with either 1.0 mg L−1 2,4-D (immature seeds and leaves) or 0.1 mg L−1 NAA (roots), explants produced embryogenic calli which upon transfer to plant growth regulator-free medium allowed embryo conversion into plantlets. The best embryogenic response (82%) was obtained in calli derived from leaves cultured with their abaxial surface in contact with medium, whereas the highest embryo conversion rate (68%) was recorded for calli induced on immature seed explants. Histological analysis in all explant types revealed development of proembryogenic cell complexes at callus periphery, giving rise to somatic embryos. The presence of embryos at various stages of development indicated asynchronous somatic embryogenesis in G. utriculosa. Derooted embryo-derived plantlets placed on medium with 0.2 mg L−1 BA multiplied further as shoot cultures.
Unlike garlic and onion, the regeneration of chive (Allium schoenoprasum L.), cultivated both for culinary and ornamental purposes, has not been intensively studied. The effects of the eight cytokinins and the plant basal section thickness on regeneration efficiency and subsequent plant growth were studied. Representatives of all cytokinin structural groups: isoprenoide side chain (trans-zeatin) and aromatic side chain (benziladenine, kinetin, meta-topolin) adenine derivatives, and phenylurea derivatives (thidiazuron and N-(2-chloro-4-pyridyl)-N′-phenylurea) at 0, 1, 5 or 10 μM were used. Histological analysis revealed adventitious buds formation from the leaf sheaths’ bases and the basal plate. The highest regeneration frequency (100 %) and the mean bud number per explant (20.0) were achieved with 10 μM thidiazuron (TDZ), and 5 mm-thick basal sections were the most responsive explants. Inferior shoot and root growth characteristics of plants regenerated by this treatment was avoided by exclusion or replacement of 10 μM TDZ with 5 μM kinetin (Kin) after a 4-week bud induction period, without consequences on the regeneration efficiency. In addition, a positive correlation between peroxidase, catalase and superoxide dismutase activity and the regeneration capacity was observed. All antioxidative enzymes activity changed much faster with 10 μM TDZ than with 1 μM Kin, which provoked the weakest regeneration response. Moreover, a unique peroxidase isoform was observed only in TDZ-treated explants after 3rd day of treatment. This work is useful for genetic engineering and virus-free plant production advancement, and for the knowledge expansion regarding the role of antioxidative enzymes in plant organogenesis.
The effects of Chenopodium murale root exudates, applied as phytotoxic medias (PMs), were tested on Arabidopsis thaliana and Triticum aestivum. The effects of PMs, where wild-type roots (K), hairy roots derived from roots (R clones) or from cotyledons (C clones) were cultured, were different. K medium suppressed Arabidopsis germination, while other PMs reduced root and leaf elongation and the number of rosette leaves. R media were more phytotoxic than C media. Treatment of Arabidopsis with R8 down-regulated expression of core cell cycle genes: cyclin-dependent kinase (CDK) A1;1, four B-class CDKs, and cyclins CYCA3;1, CYCB2;4, CYCD4;2 and CYCH1 in root and shoot tips. Only CYCD2;1 transcript was elevated in treated shoots, but down-regulated in roots. Wheat Ta-CDC2 and Ta-CYCD2 genes showed the same expression profiles as their Arabidopsis counterparts, CDKA1;1 and CYCD2;1. PMs also caused increase of antioxidative enzyme activities in both plants. Exposure of Arabidopsis to PMs induced one catalase isoform, but repressed another, resulting in no net change of catalase activity. Wheat seedlings treated with PMs had catalase activity significantly elevated in all treatments, particularly in shoots. In both plants, PMs induced the activity of different peroxidase isozymes and total peroxidase activity. Both plants responded to phytotoxic treatments by induction of CuZn-superoxide dismutase. Thus, the phytotoxicity of C. murale root exudates is, at least partially, based on down-regulation of the cell cycle regulators and on generation of oxidative stress in the affected plants. We propose that C. murale root exudates should be considered as means of biological weed control.
Chenopodium murale L. wild-type roots (WR) and hairy roots (HR) exudates have been previously shown to exert allelopathic activity against test plant species lettuce, wheat and Arabidopsis. To further investigate their allelopathic nature, a comparative morpho-histological characterization and total phenolic content (TPC) evaluation were performed. Furthermore, the phytotoxic activity of WR and HR clone R5 against three weed species and their antioxidant responses were also assessed. Except for the higher degree of branching and root hair incidence in HR clones, both WR and HR showed similar anatomical features, typical for higher plants. No significant difference in total phenolic content between WR and HRs, nor their exudates was found. Root exudates of WR and R5, applied as growth medium (GM) wherein these C. murale roots were cultured, displayed selective phytotoxic activity depending on the target weed species, ranging from suppression of germination to inhibition of seedling growth, that were linked with alterations in antioxidant enzyme activities. R5 greatly inhibited germination and seedling growth of Capsella bursa-pastoris, while WR inhibited those of Vicia angustifolia and Amaranthus retroflexus. Such phytotoxic properties (of WR and R5) qualify them as a promising natural resource in the management of weeds. Differences in allelopathic activity between HR and WR should be searched for in their allelochemical profile and the content of each allelopathic substance. The lack of anatomical abnormalities in hairy roots renders them an efficient tool for functional-genomic studies of C. murale root genes.
Goosefoot (Chenopodium murale L.) is a cosmopolitan weed species whose root produces substances with allelopathic effects to various plant species, including agricultural ones. To investigate the allelopathic effect of C. murale on vegetable plant collard greens (Brassica oleracea L. var. acephala), the sterilized seeds of collard greens were treated with liquid nutrient medium in which C. murale hairy root clone R5 was grown for 4 weeks and which contained its exudates. Results indicated that final germination rate was not affected by R5 while the growth and development of collard greens seedlings were significantly inhibited. The roots were more affected than shoots. Comparing to the control, seedlings treated with R5 had reduced number of roots per seedling (27%), root length (33%) and root fresh weight (59%). Further, R5 led to the seedlings bending, loss of chlorophyll in the leaves, root necrosis and finally seedlings decay, indicating the ultimate lethal effect of C. murale allelochemicals. The seedling growth inhibition was accompanied with alteration in antioxidant enzymes activities illustrated by increased peroxidase (POD), superoxide dismutase (SOD) and catalase (CAT) activity in shoots and emphasized CAT and decreased SOD activity in roots. The results could be contributed to the dissemination of knowledge about allelopathic influence of C. murale on cultivated plants and can be helpful in determining the mode of action against this weed species to protect collard greens plants.
The examinations of conditions for establishing a variety independent Agrobacterium-mediated transformation procedure for wheat are preferable since many of cultivars and breeding lines remain recalcitrant for biotechnological manipulation, mainly due to low efficiency of plant regeneration in vitro, which is highly genotype specific. This paper describes and discusses an improved protocol for enhanced and low-genotype dependent Agrobacterium-mediated transformation using a super-binary vector LBA4404/pTOK233 carrying reporter gus-intron gene and hygromycin (hpt) and kanamicyn (nptII) selectable marker genes. The protocol was optimized on highly responsive common wheat cv. Vesna. Transient expression monitored by the gus-intron on explants after 3, 6 and 25 days of co-cultivation, followed by GUS expression and hygromycin resistance in whole plants indicated the protocol including a co-cultivation of freshly isolated immature embryos in the presence of ascorbic acid, and acetosyringone added only in the bacteria-containing infection medium combined with a delayed and stepwise increasing hygromycin B selection procedure significantly enhanced the transformation efficiency in cv. Vesna that exceed 7% of treated explants from previously 0.41%. Explant pre-cultivation did not additionally improve transformation efficiency. The optimized protocol was successful in evoking satisfactory transformation efficiencies from 3.6% to 10.8% in 5 less-responsive wheat genotypes. All 57 T0 hygromycin-resistant and GUS-positive lines were phenotypically normal and fertile. Therefore, the conditions employed in this study may serve as a base to facilitate the transformation in other, particularly recalcitrant wheat cultivars.
This study assessed the shoot regeneration capacity of root-tips isolated from single seed-derived individual plants, obtained from a wild, open-pollinated micropopulation of shallot (Allium ascalonicum). Considerable variation was observed in the regeneration capacities of individual lines, ranging from 0.93 to 100 %, and a mean bud number per root explant between 0.09 and 20.67. One line was found to be superior, and was chosen for protocol optimization, focusing on the 2,4-D/BA ratio, duration of the CI phase and light conditions. The application of the optimized protocol to other lines, selected for their variable regeneration capacities, enhanced the process of regeneration and shortened the time required to obtain healthy plantlets, even in inferior lines. However, highly responsive lines remained superior, indicating that differences at the individual level must not be overlooked. The conditions employed in this study may serve as a base to facilitate the application of molecular breeding methods in shallot.
A simple and efficient protocol for direct somatic embryogenesis and plant regeneration of kohlrabi (Brassica oleracea var. gongylodes) was developed. Somatic embryos were induced from immature zygotic embryos at different developmental stages cultured on Murashige and Skoog medium supplemented with 0, 0.5, 1.0, or 1.5 mg/l 2,4-dichlorophenoxyacetic acid. Zygotic embryos at the early cotyledonary stage, which were cultured for 4 wk on plant growth regulator-free (PGR-free) medium, displayed the highest percentage of somatic embryogenesis (80.7%). Embryogenic tissue could be subcultured on the same medium for over 1 yr. Embryogenic lines derived from early cotyledonary stage zygotic embryos displayed the highest intensity of secondary embryogenesis (highest mean number of new somatic embryos per responsive somatic embryo explant). Histological analyses confirmed the direct origin of the secondary somatic embryos. Prolonged culturing of embryogenic tissue on PGR-free medium led to somatic embryo development into plantlets that were successfully acclimated in the greenhouse with a survival rate of 72.5%. Flow cytometry analysis showed no ploidy variation in 96.7% of the acclimated plants.
An efficient plant propagation system through somatic embryogenesis by using immature zygotic embryos was established in Savoy cabbage (Brassica oleracea var. sabauda L.). The effects of zygotic embryo age, presence of 1 mg/L 2,4-dichlorophenoxy acetic acid (2,4-D) (B5-D) in the culture medium, and the pH (5.0 and 5.8) of the B5 plant growth regulator (PGR)-free (B5-0) induction media were investigated. It appeared that the developmental stage of the immature zygotic embryos used as explants was the most important factor for somatic embryogenesis. Highest frequency of somatic embryogenesis (86.67%) was achieved directly on zygotic embryos at cotyledonary stage (1.8 mm long) of development that cultured on B5-0 medium. Optimal pH of the medium was 5.0. Mean number of primary somatic embryos per explant on B5-0 pH 5.0 medium was 7.2. The inclusion of 2,4-D into the induction medium significantly reduced both frequency of somatic embryogenesis (to 53.34 %) and mean number of embryos per explant (to 2.62) in cotyledonary zygotic embryos. After transferring to PGR-free Murashige and Skoog (MS-0) medium 69.69% of the primary somatic embryos of the Savoy cabbage produced secondary somatic embryos (SSEs) Effective recurrent somatic embryogenesis may be an appropriate practical solution for clonal propagation and genetic modifications of Savoy cabbage.
Resistance to the non-selective herbicide dl-phosphinothricin (PPT) was introduced into commercial Lotus corniculatus cv. Bokor by co-cultivation of cotyledons with Agrobacterium tumefaciensAGL1 harbouring the binary vector pDM805 which contains the bialaphos resistance gene (bar) from Streptomyces hygroscopicus encoding phosphinothricin acetyltransferase (PAT) and the uidA gene encoding -glucuronidase. The half-cotyledon explants were precultured on regeneration Murashige and Skoog's (MS) medium supplemented with 6-benzyladenine (BA) and 1-naphthaleneacetic acid (NAA) at 0.5mgL(-1) each, 3days prior to infection. Upon co-cultivation, the explants were cultured on PPT-free regeneration medium for 10days, and then subcultured on regeneration/selection media with increasing PPT concentrations (5-7mgL(-1)) for about 18weeks. Out of 480 initially co-cultivated explants, 272 regenerated shoots survived the entire PPT selection procedure. Resistant shoots were grown further, multiplied by tillering that was additionally promoted by PPT and rooted on hormone-free MS medium containing 5mgL(-1) PPT. Established shoot cultures, continuously maintained on the same medium, have preserved PPT resistance up to now (more than 2years). Transformed plants assessed in vitro and in a greenhouse were tolerant to the herbicide PPT at 300mgL(-1) equivalent to more than twofold the recommended field dosage for weed eradication. Applied PPT treatment did not affect the activities of glutamine synthetase (GS; EC 6.3.1.2) and NADH-dependent glutamate dehydrogenase (NADH-GDH; EC 1.4.1.2) in transformed plants. However, PPT did increase the mobility of glutamine synthetase isoforms GS1 and GS2 as well as the inhibition of an additional high mobility GS (hmGS) activity. In untransformed plants, PPT treatment reduced total GS activity by 4.4-fold while contrary the activity of NADH-GDH was increased by ninefold. All transformed herbicide-resistant plants were phenotypically normal and exhibited genomic stability, as were the untransformed plants analysed by flow cytometry. Under greenhouse conditions, they grew to maturity, flowered and set seeds. Stable integration and expression of the bar gene in T0 and T1 plants were confirmed by Southern and Western blot analysis, while integration of the reporter uidA gene did not occur. The bar gene was inherited in a Mendelian fashion by the progeny, as detected by PPT resistance. The production of PPT-resistant plants may have significant practical applications in weed control in fields of L. corniculatus.
Herbicide phosphinothricin (PPT) inhibits glutamine synthetase (GS), a key enzyme in nitrogen assimilation, thus causing ammonia accumulation, glutamine depletion and eventually plant death. However, the growth response of Lotus corniculatus L. plants immersed in solutions with a broad range of PPT concentrations is biphasic, with pronounced stimulating effect on biomass production at concentrations ≤ 50 μM and growth inhibition at higher concentrations. The growth stimulation at low PPT concentrations is a result of activation of chloroplastic isoform GS2, while the growth suppression is caused by inhibition of both cytosolic GS1 and GS2 at higher PPT concentrations. Since the results are obtained in cell-free system (e.g. protein extracts), to which the principles of homeostasis are not applicable, this PPT effect is an unambiguous example of direct stimulation hormesis. A detailed molecular mechanism of concentration-dependent interaction of both PPT and a related GS inhibitor, methionine sulfoximine, with GS holoenzymes is proposed. The mechanism is in concurrence with all experimental and literature data.
A simple and rapid protocol was established for repetitive somatic embryogenesis and subsequent plant regeneration in two important Brassica oleracea varieties, cabbage and cauliflower. Direct regeneration of somatic embryos (SEs) was achieved from immature zygotic embryos cultured on B5 plant growth regulator (PGR)-free (B5-0) induction medium and on B5 medium supplemented with 1 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D) (B5-D). Zygotic embryos of both cabbage and cauliflower at the cotyledonary (C) stage (1.8 mm long) incubated on B5-0 medium displayed the highest embryo-forming capacities (EFCs) of 11.84 and 11.95, respectively. Secondary somatic embryos (SSEs) appeared on the cabbage and cauliflower’s primary embryos at a high frequency (83.3 and 87.5 %, respectively), and this process continued in a repetitive way on PGR-free Murashige and Skoog (MS-0) medium. The embryogenic potential of the cultures with a gradual diminution was maintained for 10 months (ten cycles). A total of 20 % of the mature SSEs from cabbage and 55 % from cauliflower spontaneously regenerated plantlets on MS-0 medium. The addition of 1 mg l−1 6-benzyladenine (BA) or 6-furfurylaminopurine (Kin) in the regeneration medium significantly improved somatic embryo conversion into plantlets by up to 56 % in cabbage and 79 % in cauliflower. Regenerated plants acclimated successfully to ex vitro conditions and displayed morphological and reproductive characteristics similar to seed-derived plants. Effective recurrent somatic embryogenesis may be an appropriate practical solution for clonal propagation and genetic modifications of cabbage and cauliflower.
As has been previously shown, the genotype at the individual plant level has a strong effect on the regeneration capacity of spinach, and this effect quite often masks the impact of other factors, including the light environment. Therefore, apical sections of the lateral roots of the same seedling (line) were exposed in parallel to both long day (LD, 16-h photoperiod) and short day (SD, 8-h photoperiod) conditions to assess the effect of photoperiod on somatic embryo (SE) regeneration. The root sections in most lines responded with significantly higher frequencies of regeneration and mean SE numbers with LD compared to roots of the same origin with SD. while 7 of 40 lines were only able to regenerate when exposed to LD. Moreover, in all lines cultured with SD, regeneration was delayed by approximately 4 weeks. Obtained SEs were further grown and multiplied, providing material for the evaluation of the effect of light intensity on regeneration capacity. Maximal regeneration capacity was recorded at 100 mu mol m(-2) s(-1) in all lines tested. The present research confirmed the previously observed strong effect of genotype on regeneration capacity and undoubtedly showed a significantly better response in explants exposed to LD than to SD. These results could contribute to better understanding the process of SE induction in spinach and further improve the efficiency of this process. (c) 2012 Elsevier B.V. All rights reserved.
Hygromycin (hyg) at low doses (0.5–1.0 mg l−1) promoted somatic embryogenesis from apical sections of spinach lateral roots. The highest promoting effect on both the frequency of regeneration and the mean number of somatic embryos (SE) per explant was achieved at 0.5 mg l−1 hyg. With increasing the concentration of hyg to 1 mg l−1, the regeneration frequency decreased, while the mean SE number remained significantly higher than in control (hyg-free medium). Complete inhibition of SE regeneration started at 7.5 mg l−1 hyg. Moreover, hyg efficiently promoted the process of secondary somatic embryogenesis. Compared to control, a 2.75-fold increase in the secondary somatic embryo (SSE) mean number was obtained at 0.5 mg l−1 hyg, and the increment was still discernible at 1.0 and 2.5 mg l−1 hyg. Both primary SE and SSE explants became completely necrotic at 12.5 mg l−1 hyg. Since attempts with direct selection at 20 mg l−1 hyg proved unsuccessful, the results obtained in this study suggest that a stepwise selection procedure is suitable, starting with selection at 0.5 mg l−1 hyg, to exploit the promoting effect of low hyg doses on SE regeneration from transformed cells, then gradually increasing the hyg concentration to 20 mg l−1 for final selection. Complete SE and SSE explant mortality at hyg above 12.5 mg l−1 guarantees a low possibility of escape during the selection process. This study will be useful for increasing the efficiency of transgenic plant regeneration following genetic transformation in spinach.
An efficient in vitro shoot regeneration method from leaf explants of apple cultivars Golden Delicious and Melrose by optimization of regeneration medium, explant type and orientation, dark pre-treatment, and gelling agent is presented. Murashige and Skoog’s regeneration medium containing 22 μM thidiazuron (TDZ) and 1.5 μM indole-3-butyric acid (IBA) (M2 medium) was superior for regeneration as well as for subsequent shoot multiplication in both cultivars, providing regeneration frequency of 95% or higher in the best combination with other factors. Pre-incubation in the dark proved to be an essential factor for regeneration. The use of agar as a gelling agent provides satisfactory regeneration frequency compared with media gelled with Phytagel TM . Leaf explants of cv. Melrose with adaxial surface in contact with M2 medium and those of cv. Golden Delicious orientated contrary regenerated the highest mean number of shoots per explant. Under optimal conditions, a maximal index of shoot-forming capacity of 11.44 and 6.30 for ‘Melrose’ and ‘Golden Delicious’, respectively, was achieved. Regenerated shoots were successfully rooted and acclimated ex vitro.
This study was aimed at obtaining insight into the diversity of sialic acids in cancer-and non-cancer-related CA125 antigen, tumour marker of serous ovarian cancer. Starting from available data suggesting the possible relevance of sialic acids for discriminating CA125 antigens of different origin, we have employed a new experimental approach based on the use of human sialic acid-binding Ig-like lectins, Siglecs, as tools for the investigation of sialylation.Siglec-2, belonging to the group of evolutionarily conserved Siglecs, and Siglec-3, -6, -7, -9 and -10, which are CD33-like Siglecs, were probed in solid-phase binding assays with cancer-related CA125 antigens from pleural fluid of patients with ovarian carcinoma (pfCA125), the OVCAR-3 ovarian carcinoma cell line (clCA125) and a non-cancer-related CA125 antigen, i.e. pregnancy-associated pCA125 antigen.All Siglecs used showed detectable binding to pCA125 antigen. Siglec-3, Siglec-7 and Siglec-2 exhibited moderately stronger binding to pCA125 antigen than the others. In contrast to this, Siglec-2 and Siglec-3 preferentially recognized pfCA125 with greater total binding than for pCA125, whereas Siglec-9 and Siglec-10 were highly selective for clCA125.Siglecs promise to be powerful tools for discriminating CA125 of different origin and could propagate further research on other molecular markers of biomedical and diagnostic importance.