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Pollen of butterfly pea (Clitoria ternatea L.) could germinate in vitro with emergence of 1, 2 and 3 tubes from individual pollen grain on germination medium (modified after Mercado et al. Sci Hortic 57:273–281, 1994). The highest percentage of pollen germination was observed on medium supplemented with 20% sucrose while addition of osmotica such as polyethylene glycol or mannitol did not have positive effect on butterfly pea pollen germination. Starch utilization, through conversion to simpler sugars including sucrose, may be the cause for pollen tube elongation. The optimum pH for maximum germination (42%) was recorded to be 5.7. Addition of various plant growth regulators including brassin-like substance and GA3 singly into medium supplemented with 20% sucrose at pH 5.7 did not result in germination percentage higher than that on the medium supplemented with only 20% sucrose. The germination percentage was increased to 71% and the pollen tube length was the greatest when butterfly pea pollen was cultured on medium supplemented with 20% sucrose, 0.25 mg L− 1 brassin-like substance and 1 mg L− 1 GA3 at pH 5.7. This suggests that brassin-like substance and GA3 may play an important synergistic role on pollen germination of butterfly pea in vitro.

In this study, a simple pH-colorimetric film was developed to detect the spoilage of pasteurized milk. The biosensor was fabricated by incorporating the pH-sensitive anthocyanins (AC) extracted from blue pea (Clitoria ternatea L.) flower into a starch-based edible film. The color responses of both AC aqueous extracts and edible films incorporated with different AC concentrations towards different pH buffers (pH 2–11) were measured using Ultraviolet–visible (UV–Vis) spectrophotometer and colorimeter, respectively. Based on the results, there were significant differences (p

Clitoria ternatea L., known as the butterfly pea plant, is from the Fabaceae family and is commonly used in traditional medicine or as a brain tonic in Asia. The root extracts of C. ternatea have been previously studied for its neuroprotective properties linked to memory enhancement of the mammalian brain, indirectly delaying the occurrence of neurodegenerative diseases. The current study aims to establish in vitro adventitious root cultures of C. ternatea via cotyledon explants and to identify the phytoconstituents of the induced adventitious roots. In this study, the highest percentage of induction (95.24%) was achieved in half-strength MS medium supplemented with 2.50 mg/L NAA (1-naphthaleneacetic acid) after 6 weeks of culture. Results also indicated that the supplementation of 1.50 mg/L NAA induced the highest number of adventitious roots (12.86 ± 2.14), while 2.50 mg/L 4-Cl-IAA (4-chloroindole-3-acetic acid) yielded higher adventitious root length (0.75 ± 0.13 cm) in comparison with the other auxin treatments. Microscopic analysis indicated thicker in vitro root morphology for all NAA treatments in comparison with the roots of in vitro seedling. Ethanolic extract of in vitro-generated adventitious roots showed potential anti-acetylcholinesterase activities (IC50 = 1519.89 ± 0.18 μg/mL) albeit being lower than the control (IC50 = 70.37 ± 0.01 μg/mL), while gas chromatography–mass spectrometry analysis identified the presence of pentacyclic triterpenes, phytosterols and fatty acids in the adventitious root culture that are linked to the mammalian neuroprotective properties of C. ternatea. The current investigation reports on the potential of C. ternatea root cultures being further utilized to produce valuable metabolites linked to the mammalian memory enhancement.

Clitoria ternatea (L.) is a medicinal leguminous plant and is cultivated to cater the need of herbal industries and asthetic purposes. The unavailability of steady molecular marker impedes the genetic improvement of C. ternatea. In the present study, transferability of 98 pairs of Cajanus spp. specific SSR primers were assessed among 14 genotypes of C. ternatea, varied for their flower color, floral architecture and bio-metabolite (taraxerol and delphinidin) content, and out of them 43 had successfully amplified the fragments. Among them, 36 pairs of primers showed 100% transferability, whereas rest seven varied from 42.86 to 92.85% transferability. The transferable 43 pairs of SSR primers generated 196 alleles across the 14 genotypes and the AMOVA analysis showed moderate genetic variation (55.1%) among the genotypes of C. ternatea, which was also reinforced by Nei’s genetic distance and gene identity estimates derived haplotype matrix. Similarly, both the principal coordinate analysis and dendrogram grouped these 14 genotypes of C. ternatea into two major clusters based on SSR allele distribution and frequency, and the clustering pattern is in accordance with petal color but in contrast to floral architecture. MCheza based outlier analysis revealed 16 alleles for balancing selection, which are putatively involved in the maintenance of genetic polymorphism in C. ternatea. Moreover, the estimates of molecular diversity and bio-metabolite content revealed the possible use of these genotypes in future breeding programme of this species.

A new norneolignan derivative, clitorternalactone (1), has been isolated from the stems of Clitoria ternatea, together with six known compounds, clitorienolactone A (2), myricetin 3-glucoside (3), quercetin 3-glucoside (4), kaempferol 3-glucoside (5), taraxerol (6), and taraxerone (7). The structure of the new compound 1 was determined through spectroscopic and MS analyses. Among the isolates, compound 1 exhibited cytotoxicities with IC50 values of 2.54 ± 0.23, 3.68 ± 0.17, and 4.05 ± 0.43 μM, respectively, against DLD-1, CCRF-CEM, and IMR-32 cell lines. In addition, clitorternalactone (1), clitorienolactone A (2), quercetin 3-glucoside (4), and kaempferol 3-glucoside (5) showed potent inhibition with IC50 values of 3.05 ± 0.12, 5.44 ± 0.46, 10.20 ± 0.69, and 13.57 ± 0.36 μM, respectively, against LPS-induced NO generation.

Clitoria ternatea or commonly known as 'Butterfly pea' has been used traditionally in Ayurvedic medicine in which various parts of the plants are used to treat health issues such as indigestion, constipation, arthritis, skin diseases, liver and intestinal problems. The flowers of C. ternatea are used worldwide as ornamental flowers and traditionally used as a food colorant. This paper reviews the recent advances in the extraction and biological activities of phytochemicals from C. ternatea flowers. The application of maceration or ultrasound assisted extraction greatly increased the yield (16-247% of increase) of phytochemicals from C. ternatea flowers. Various phytochemicals such as kaempferol, quercetin and myricetin glycosides as well as anthocyanins have been isolated from C. ternatea flowers. Clitoria ternatea flower extracts were found to possess antimicrobial, antioxidant, anti-inflammatory, cytotoxic and antidiabetic activities which are beneficial to human health. Clitoria ternatea flower is a promising candidate for functional food applications owing to its wide range of pharmacotherapeutic properties as well as its safety and effectiveness.

The stability of anthocyanins is greatly affected by pH and temperature, and thus their application in food products is limited. This study was aimed to develop an antibacterial food colorant using anthocyanin from Clitoria ternatea flowers by applying microencapsulation technology. Maltodextrin was used as a carrier agent in the spray drying process. The anthocyanin microcapsule was durable to the light exposure up to 21 days, and the best color stability was observed between the temperature range of − 20 °C to 4 °C. However, the color was sustained with approximately 50% of the stability index after 1 day exposure at 80 °C and 180 °C. On disk diffusion assay, the anthocyanin microcapsule manifested a broad-spectrum antibacterial activity against Gram-bacteria. According to minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) assay, anthocyanin microcapsule exhibited bactericidal activity on all test bacteria, with the lowest MIC and MBC observed on Escherichia coli. The application of anthocyanin microcapsule on muffins has shown a significant inhibitory activity on foodborne bacteria. This study revealed the potential of C. ternatea anthocyanin microcapsule as a bio-preservative, which can prolong the shelf life of baked food products.

Part of the native root nodule endophytic microflora referring to members of the genera Proteobacteria and Sphingobacteria were used to test their bioefficacy as seed biopriming. These were quantified for their plant growth promoting (PGP) attributes such as IAA production, P and K-solubilization and ACC deaminase production. Results showed that significantly highest IAA was produced by E. hormaechi RCT10. The highest P-solubilization was observed with S. maltophila RCT31 it was solubilizing all the substrate tri-calcium phosphate, di-calcium phosphate, and zinc phosphate. Significantly highest K-solubilization was observed with S. maltophila RCT31 followed by E. turicensis RCT5. However, the maximum zinc solubilization was reported with S. maltophila RCT31 followed by E. turicensis RCT5. The maximum ACC deaminase was quantified in the bacterium. Results revealed that the E. hormaechi RCT10 utilized seed leechates most effectively while root exudates were maximally utilized by S. maltophila RCT31. The pots experiment proves that S. maltophila RCT31 was the most effective bacterium and it was replication vis-à-vis field experiment. In particular, S. maltophila RCT31 holds strong potential to be possibly used as a bioformulation for the medicinal legume, as an economical and eco-friendly alternative to agrochemicals.

Biosynthesis of silver nanoparticles (CTNP’s) by Clitoria ternatea flower in the aqueous extract was investigated. Synthesized nanoparticles were characterized by using UV–Visible spectroscopy, followed by DLS, Zeta potential, XRD, FTIR, SEM, and AFM. The biocompatibility nature of CTNP’s was determined using erythrocytes model system. Cytotoxicity of CTNP’s against MCF-7 and EAC cells were determined by using MTT and Trypan blue exclusion method and their IC50 was found to be 19.37 µg/mL and 24 µg/mL. Cytotoxic potential of CTNP’s was further confirmed by clonogenic assay. Further in vivo studies using EAC mice model supports the anti-cancer potential of silver nanoparticles. Results found that the CTNP’s effectively control the proliferation rate by inhibiting the ascites secretion and cellular density. Further quantification of VEGF, microvessel density counts and CAM assays show the anti-angiogenic potential of the CTNP’s. The apoptotic inducing activity of CTNP’s was confirmed by DNA fragmentation, fluorescent staining studies. More interestingly, EAC treated mice exhibit significant increase in lifespan (~ 2.25 fold) compared to control EAC mice. Interestingly CTNP’s did not exhibit any secondary complications against normal mice. The present findings give an experimental proof that the CTNP’s could serve as a promising candidate to overcome limitations of existing conventional cancer therapeutics.

Butterfly pea (Clitoria ternatea L.) is a climbing legume grown as a medicinal, ornamental, and forage plant, and used as a natural food colorant. Due to its various medicinal properties in different plant parts, pharmacological potential is evident. However, improvement of the plant for high phytochemical properties has been very limited, hence being pursued. For that improvement effort, butterfly pea germplasm was collected from different parts of Thailand in addition to accessions from 12 other countries acquired from a C. ternatea collection curated by the USDA-Agricultural Research Service National Plant Germplasm System. Characterization and evaluation for phytochemical properties of these germplasm is fundamental for breeding purposes. Floral antioxidant activities (Ferric reducing antioxidant power—FRAP and 2,2-diphenyl-1-picrylhydrazyl—DPPH), total phenolic compound (TPC), total anthocyanin content (TAC), and ascorbic acid content (AAC) were investigated for 126 individuals from 46 accessions of the combined collection. The plants were regenerated from seeds and grown at Kamphaeng Saen, Nakhon Pathom, Thailand, and flowers were harvested in August 2017. Significant differences of TAC and AAC were found among the included accessions. Both variations were correlated with petal color at a moderate level, but not in accordance with geographical locations of the samples. Grouping of the accessions based on antioxidant activities and compounds revealed by cluster and principal component analyses would be useful in the utilization and management of the germplasm for butterfly pea improvement.

Bakery products are a food appreciated by consumers all over the world. There is a great opportunity to incorporate more bioactive compounds to enhance its quality. The objective of this study was to utilize the advantage of CTE in the production of sponge cake. The five different levels of CTE (0, 5, 10, 15 and 20%, w/w) was incorporated into sponge cake. The sponge cakes were evaluated for physicochemical (color, volume, water activity, total phenolic content, and antioxidant properties) and texture characteristics as well as consumer acceptance. Addition of CTE into the sponge cakes increased the polyphenol content and antioxidant activity concomitant with reduced lipid peroxidation. Increasing hardness, adhesiveness, gumminess, and chewiness and decreasing cohesiveness, springiness and resilience of cakes were seen when increasing percentage of CTE in the cake. A significant decrease was observed in the lightness, redness and yellowness in the cake containing CTE. No differences were found in overall acceptability between the control and the cake containing CTE. The findings suggest that CTE could be a potential source for development of sponge cakes with more effective antioxidant properties.

Homogentisate prenyltransferase (HPT) is an important enzyme involved in the α-tocopherol (vitamin E) biosynthetic pathway of all plant taxa. Tocopherol biosynthesis and chlorophyll degradation are related, but more information is needed to explain their relationship. In this study, a candidate gene for HPT from Clitoria ternatea (CtHPT) was isolated and identified via a phylogeny-based approach, and its hypothetical protein sequence was analyzed. Transient expression of CtHPT with Agrobacterium-mediated infiltration into tomato leaves was then performed and observed for the metabolic relationship between the α-tocopherol biosynthesis and chlorophyll degradation by gas chromatography–mass spectrometry. In silico analysis showed that CtHPT contained a chloroplast signal peptide and nine-transmembrane α-helixes. The results showed that, the content of α-tocopherol increased in transient expression of CtHPT, with the increased pool sizes of its biosynthetic intermediates: 2-methyl-6-phythylbenzoquinol and 2,3-dimethyl-5-phythylbenzoquinol, and the increased levels of phytol and various fatty acids. Moreover, the CtHPT transient expression was observed to cause chlorophyll deficiency in the tomato leaves with simultaneous increase of phytol and fatty acids, presumably the degradative products of chlorophyll and chloroplast membranes, respectively. It was concluded that the overexpression of CtHPT may enhance the metabolic flow of the α-tocopherol biosynthetic pathway, causing the degradation of chlorophylls, thereby increasing the supply of the precursor phytol for the α-tocopherol biosynthetic pathway.

Currently, silver nanoparticles (AgNPs) have been providing unique approach to the treatment of infectious diseases. Therefore, herein we demonstrate AgNPs synthesis by conventional bioreduction approach using quercetin compound from Clitoria ternatea L. methanolic petal extract. Bioreduction of silver nitrate (AgNO3) with the quercetin extract, was incubated for 240 min and the mixture were characterized by UV–Vis spectroscopy, FTIR, XRD, DLS, SEM, EDS and Zeta potential analysis. The results were confirmed the presence of silver ion. Furthermore, the biosynthesized AgNPs possess significant antimicrobial activity against multidrug resistant bacteria such as, Staphylococcus aureus and Shigella sp. with maximum concentration (100 µl) showed strong inhibitory action. In addition, the hybrid AgNPs showed potential Quorum Sensing Inhibition activity with different concentration (25–100 µg/ml) against the AHL-mediated violacein production in C. violaceum (14–86%) and biofilm formation (19–92%), extracellular polymeric substances (16–72%), protease (17–93%) productions in S. aureus and. It was again confirmed by spectrophotometry and microscopy. Further, the synthesized hybrid AgNPs showed less hemolysis activity 4.58%, compared with American Society for Testing and Materials range

Recently, in the field of natural product drug discovery, there has been increasing interest in effective extraction and isolation of bioactive phytomolecules from plants for use as important starting materials or chemical intermediates for new drug development. In this investigation, various solvent extracts of butterfly pea (Clitoria ternatea Linn.) were evaluated in terms of phenolic content and antioxidant activity. Moreover, various parameters influencing optimal flavonoid extraction were studied based on determination of the quercetin and kaempferol yields by an improved high-performance liquid chromatography (HPLC) method. The results indicated that the total phenolics and 2,2′-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) scavenging activity were maximum for the 80 % aqueous methanol extract, while the 1,1-diphenyl-2-picrylhydrazyl (DPPH) reducing power and flavonoid content were maximum in the methanol extract. Different solvent extracts showed significantly (p

Saccharomyces cerevisiae was able to degrade a highly toxic textile dye malachite green (MG) at 100 mg/L concentration. Although 99% decolourization was observed, a tremendous metabolic and oxidative stress was exerted on the cells. Ethanolic extracts of Terminalia chebula, Clitoria ternatea and Boerhaavia diffusa at a concentration of 1 mg/mL were independently supplied to S. cerevisiae cells to counter the stress. T. chebula, C. ternatea and B. diffusa extracts reduced the activities of glutathione peroxidase (67, 8 and 71%), superoxide dismutase (2, 7 and 16%) and catalase (16, 52 and 57%), respectively. Inductions in the activities of laccase (66, 82 and 50%), lignin peroxidase (35, 75 and 10%), NADH-DCIP reductase (43, 52 and 91%) and MG reductase (66, 126 and 117%) were observed respectively. Presence of dye (MG) extended the lag phase of the growth cycle of S. cerevisiae up to 36 h, which was observed to be restored to normal (4 h) after phytoextract supplementation. Scanning electron microscope imaging revealed the restored cell morphology upon exposure to plant extracts. The accumulation of reactive oxygen species (ROS) was observed to be 355% greater in cells exposed to MG, which was significantly reduced after phytoextracts augmentation when compared to control cells. Phytoextracts proved to be beneficial in increasing the viability of S. cerevisiae cells and reduced the intracellular ROS and nuclear damage. Inclusion of plant extracts during decolourization proved to be beneficial and protected the cells so that 20 treatment cycles could be run achieving significant removal of MG.

Ayurveda is a renowned traditional medicine practiced in India from ancient times and Clitoria ternatea is one such prospective medicinal herb incorporated as an essential constituent in a brain tonic called as medhya rasayan for treating neurological disorders. This work emphasises the significance of the plant as a brain drug there by upholding Indian medicine. The phytochemicals from the root extract were extricated using gas chromatography–mass spectrometry assay and molecular docking against the protein Monoamine oxidase was performed with four potential compounds along with four reference compounds of the plant. This persuades the prospect of C. ternatea as a remedy for neurodegenerative diseases and depression. The in silico assay enumerates that a major compound (Z)-9,17-octadecadienal obtained from the chromatogram with a elevated retention time of 32.99 furnished a minimum binding affinity energy value of −6.5 kcal/mol against monoamine oxidase (MAO-A). The interactions with the amino acid residues ALA 68, TYR 60 and TYR 69 were analogous to the reference compound kaempferol-3-monoglucoside with a least score of −13.90/−12.95 kcal/mol against the isoforms (MAO) A and B. This study fortifies the phytocompounds of C. ternatea as MAO-inhibitors and to acquire a pharmaceutical approach in rejuvenating Ayurvedic medicine. Graphical Abstract

Random amplified polymorphic DNA (RAPD) and inter simple sequence repeats (ISSR) markers were used to compare genetic diversity of 17 accessions of Clitoria ternatea (Fabaceae) populations, collected from nine different states of India. The populations included three different flower colours viz., blue, white and white with blue tinge. Twenty-three RAPD primers and eighteen ISSR primers amplified a total of 137 and 105 reproducible DNA fragments, respectively with fragment sizes ranged from 150 to 3,000 bp. ISSR showed higher polymorphism (29.52 %) in comparison to RAPD (27.73 %). In RAPD analysis, maximum polymorphic information content value (0.66) was observed in primer OPC 10 but in ISSR study, it was (0.55) in primer UBC 889. Jaccard’s coefficient of similarity showed that pair-wise genetic similarity coefficients ranged between 81 and 97 % in RAPD analysis, whereas 80 and 98 % in ISSR analysis, which were in close range to each other. Similarly, clustering pattern showed same trend in both the markers analysis which also revealed that all the accessions were grouped according to their geographical locations rather than on the basis of different flower colours. Mental Z test indicated highly significant co-relation between geographical distance and genetic distance among the populations (r = 0.8321, P

The life cycle of Tetranychus macfarlanei Baker and Pritchard was studied on two different medicinal plants, Clitoria ternatea L. and Justicia adhatoda L. Nees, in BOD at 32.5 °C and 75 % RH during April 2007 to May 2007. Observations towards duration of different stages like egg, larva, protonymph, deutonymph, adult, total life cycle, preoviposition, oviposition, postoviposition periods, longevity of female and male, and fecundity, sex ratio were recorded. Total developmental period of T. macfarlanei from egg to adult was 6.4 ± 0.37 (Mean ± SE) and 10.6 ± 0.56 days on C. ternatea and J. adhatoda, respectively. On C. ternatea, the fecundity in case of fertilized and unfertilized female were 91.6 ± 11.61 and 80 ± 21.64 eggs, respectively and longevity of fertilized and unfertilized female was 16.4 ± 1.44 and 8.6 ± 2.32 days, respectively. The corresponding figure on J. adhatoda for fecundity in case of fertilized and unfertilized female were 39 ± 2.85 and 19.8 ± 3.90 eggs, respectively and for longevity was 16 ± 0.37 and 11 ± 086 days, respectively. Among the two hosts, C. ternatea appears to be more preferred to J. adhatoda because life cycle was completed in shorter time and fecundity and female longevity were for longer duration.

Independent transformed root somaclones (rhizoclones) of butterfly pea (Clitoria ternatea L.) were established using explant co-cultivation with Agrobacterium rhizogenes. Rhizoclones capable of sustained growth were maintained under low illumination in auxin-free agar-solidified MS medium through subcultures at periodic intervals. Integration of T L -DNA rolB gene in the transformed rhizoclone genome was verified by Southern blot hybridization, and the transcript expression of T R -DNA ags and man2 genes was ascertained by reverse transcription polymerase chain reaction analysis. The major compound isolated and purified from the transformed root extracts was identified as the pentacyclic triterpenoid compound taraxerol using IR, 1H-NMR, and 13C-NMR spectroscopy. The taraxerol yield in cultured hairy roots, as quantified by HPTLC analysis, was up to 4-fold on dry weight basis compared to that in natural roots. Scanning of bands from cultured transformed roots and natural roots gave super-imposable spectra with standard taraxerol, suggesting a remarkable homology in composition. To date, this is the first report claiming production of the cancer therapeutic phytochemical taraxerol in genetically transformed root cultures as a viable alternative to in vivo roots of naturally occurring plant species.

An examination of Australian herbarium specimens of powdery mildew on Clitoria revealed two species. Oidium clitoriae is reported for the first time in Australia causing powdery mildew on Clitoria sp. in northern Queensland. Golovinomyces cf. orontii is reported for the first time on the genus Clitoria, from Clitoria ternatea in Queensland and the Northern Territory.

The energy dispersive X-ray fluorescence set-up incorporating a molybdenum secondary exciter was used for quantitative determination of major and minor elements in genetically transformed root somaclones (rhizoclones) of butterfly pea (Clitoria ternatea L.) which had been established via explant co-cultivation with Agrobacterium rhizogenes. The multi-elemental composition of these transformed rhizoclones was compared with that of the naturally grown in vivo donor plant. Trace elements namely Cr, Mn, Fe, Co, Ni, Cu, Zn, Se, Rb, Sr and Pb in addition to two macro-elements K and Ca were identified and quantified in root tissues of both sources. The elemental content of transformed root cultures was found to be at par with that of the natural roots of in vivo grown plants of the same species. These findings are implicated on the context of utilization of such Agrobacterium-mediated genetically transformed root cultures as a viable alternative to natural roots, the former being a fast-proliferating renewable resource of medicinally useful minerals essential for designing of effective drugs, besides providing an ex situ means for plant conservation.

An efficient plant regeneration protocol has been developed from embryogenic callus derived from cotyledonary explants of Clitoria ternatea Linn., an important medicinal climber species. Optimum embryogenic callus (75 %) was induced on Murashige and Skoog (MS) medium supplemented with 2 mg/l 2, 4-dichlorophenoxyacetic acid (2, 4-D). On subculturing the callus on MS medium supplemented with 2 mg/l 6-benzyladenine (BA) and 0.5 mg/l α-naphthalene acetic acid (NAA), 61 % of cultures responded with a mean number of 22 somatic embryos per gram callus at different stages of development after 45 days of culture. The addition of higher concentrations of sucrose and abscisic acid (ABA) significantly increased the embryogenic response. MS medium supplemented with 2 mg/l BA, 0.2 mg/l NAA and 4 % of sucrose resulted in 76 % of cultures responding with a mean number of 28 embryos per one gram callus. The highest embryogenic response, frequency of 83 % and mean number of 37 embryos per gram callus, was observed when the MS medium was supplemented with 2 mg/l BA, 0.2 mg/l NAA, and 3 mg/l ABA. Synthetic seeds were produced by encapsulating embryos in calcium alginate gel. The gel contained MS medium with 3 % of sucrose, 1.0 mg/l BA and 0.2 mg/l NAA. The synthetic seeds germinated on MS medium supplemented with BA (1.0-4.0 mg/l) alone or in combination with NAA (0.1–0.7 mg/l) or indole-3-butyric acid (IBA; 0.1–0.7 mg/l). The highest synthetic seed germination (92 %) was observed on MS medium supplemented with 2 mg/l BA and 0.5 mg/l NAA. The synthetic seeds were stored at 4 °C and lab conditions (25 ± 2 °C) up to 5 months. The synthetic seeds kept at 4 °C showed 86 % viability even after 5 months of storage. Both somatic embryos and synthetic seeds germinated and were transferred to soil successfully.

Glyphosate tolerance by Clitoria ternatea, Neonotonia wightii and Amaranthus hybridus was studied in whole plants from Mexico. Experiments in a controlled growth chamber showed both legumes to be highly tolerant of glyphosate, with and ED50 values of 600.18 g ae ha–1 for C. ternatea and 362.94 g ae ha–1 for N. wightii. On the other hand, A. hybridus was highly susceptible to the herbicide (ED50 = 42.22 g ae ha–1). Shikimate accumulation peaked 96 h after treatment in the tolerant plants and the susceptible weed under 500 g ae ha–1 glyphosate. The shikimic acid content of whole leaves was 4.0 and 5.0 times higher in the susceptible weed than in N. wightii and C. ternatea, respectively. 14C-glyphosate absorption and translocation tests showed A. hybridus to absorb 30% more herbicide than the legumes 24 h after glyphosate foliar application. 14C-glyphosate translocation as measured by quantified autoradiography revealed increased translocation of the herbicide to untreated leaves and roots in A. hybridus relative to the two legumes. The cuticular surface of A. hybridus exhibited very low wax coverage relative to the epicuticular surface of N. wightii and, especially, C. ternatea. No significant degradation of glyphosate to aminomethylphosphonic acid and glyoxylate metabolites was detected among the tolerant leguminous plants or the susceptible weed population. These results indicate that the high glyphosate tolerance of Clitoria ternatea and Neonotonia wightii is mainly a result of poor penetration and translocation of the herbicide to apical growing points in their plants.

Screening of natural products for mosquito larvicidal activity against three major mosquito vectors Aedes aegypti, Culex quinquefasciatus, and Anopheles stephensi resulted in the identification of three potential plant extracts viz., Saraca indica/asoca, Nyctanthes arbor-tristis, and Clitoria ternatea for mosquito larval control. In the case of S. indica/asoca, the petroleum ether extract of the leaves and the chloroform extract of the bark were effective against the larvae of C. quinquefasciatus with respective LC(50) values 228.9 and 291.5 ppm. The LC(50) values of chloroform extract of N. arbor-tristis leaves were 303.2, 518.2, and 420.2 ppm against A. aegypti, A. stephensi, and C. quinquefasciatus, respectively. The methanol and chloroform extracts of flowers of N. arbor-tristis showed larvicidal activity against larvae of A. stephensi with the respective LC(50) values of 244.4 and 747.7 ppm. Among the methanol extracts of C. ternatea leaves, roots, flowers, and seeds, the seed extract was effective against the larvae of all the three species with LC(50) values 65.2, 154.5, and 54.4 ppm, respectively, for A. stephensi, A. aegypti, and C. quinquefasciatus. Among the three plant species studied for mosquito larvicidal activity, C. ternatea was showing the most promising mosquito larvicidal activity. The phytochemical analysis of the promising methanolic extract of the seed extract was positive for carbohydrates, saponins, terpenoids, tannins, and proteins. In conclusion, bioassay-guided fractionation of effective extracts may result in identification of a useful molecule for the control of mosquito vectors.

Butterfly pea, Clitoria ternatea is used in Africa as a companion crop and in the United States as an ornamental. The USDA, ARS, PGRCU curates 28 butterfly pea accessions. Butterfly pea accessions were transplanted from about 30-day-old seedlings to the field in Griffin, GA around 01 June 1999, 2003, 2006-2007 or directly sown in 2001. At 50% maturity, 19 accessions were characterized for morphology, phenology, and evaluated for regeneration. High quality plants regenerated from all accessions produced 6 to more than 3,400 total seeds. Butterfly pea can be successfully grown and regenerated in Griffin, GA. Coefficients of variation and principal component analysis revealed considerable variability among accessions for morphological and reproductive traits. Butterfly pea has potential to be used as a nutraceutical, pharmaceutical, or food. The flavonoid quercetin has been shown to reduce upper respiratory infections in humans while delphinidin and malvidin identified in butterfly pea flowers may inhibit various forms of cancer.

A UDP-glucose: anthocyanin 3′,5′-O-glucosyltransferase (UA3′5′GT) (EC 2.4.1.-) was purified from the petals of Clitoria ternatea L. (Phaseoleae), which accumulate polyacylated anthocyanins named ternatins. In the biosynthesis of ternatins, delphinidin 3-O-(6″-O-malonyl)-β-glucoside (1) is first converted to delphinidin 3-O-(6″-O-malonyl)-β-glucoside-3′-O-β-glucoside (2). Then 2 is converted to ternatin C5 (3), which is delphinidin 3-O-(6″-O-malonyl)-β-glucoside-3′,5′-di-O-β-glucoside. UA3′5′GT is responsible for these two steps by transferring two glucosyl groups in a stepwise manner. Its substrate specificity revealed the regioselectivity to the anthocyanin′s 3′- or 5′-OH groups. Its kinetic properties showed comparable k cat values for 1 and 2, suggesting the subequality of these anthocyanins as substrates. However, the apparent K m value for 1 (3.89 × 10−5 M), which is lower than that for 2 (1.38 × 10−4 M), renders the k cat/K m value for 1 smaller, making 1 catalytically more efficient than 2. Although the apparent K m value for UDP-glucose (6.18 × 10−3 M) with saturated 2 is larger than that for UDP-glucose (1.49 × 10−3 M) with saturated 1, the k cat values are almost the same, suggesting the UDP-glucose binding inhibition by 2 as a product. UA3′5′GT turns the product 2 into a substrate possibly by reversing the B-ring of 2 along the C2-C1′ single bond axis so that the 5′-OH group of 2 can point toward the catalytic center.

A lectin present in seeds of Clitoria ternatea agglutinated trypsin-treated human B erythrocytes. The sugar specificity assay indicated that lectin belongs to Gal/Gal NAc-specific group. Hence the lectin, designated C. ternatea agglutinin (CTA), was purified by the combination of acetic acid precipitation, salt fractionation and affinity chromatography. HPLC gel filtration, SDS-polyacrylamide gel electrophoresis and mass spectrometry indicated that the native lectin is composed of two identical subunits of molecular weight 34.7 kDa associated by non covalent bonds. The N-terminal sequence of CTA shared homology with Glycine max and Pisum sativum. Complete sequence was also found to be homologous to S-64 protein of Glycine max, suggesting that CTA probably exhibits both hemagglutination and probably sugar uptake activity. The carbohydrate binding specificity of the lectin was investigated by quantitative turbidity measurements, and percent inhibition assays. Based on these assays, we conclude that CTA binds beta-D: -galactosides, and also may has an extended specificity towards non-reducing terminal Neu5Acalpha2,6Gal.

This study describes a reproducible protocol for rapid mass propagation of a multipurpose legume, Clitoria ternatea L., using cotyledonary node explants derived from axenic seedlings. Multiple shoots were induced in Murashige and Skoog (MS) medium supplemented with N6-benzyladenine (BA), zeatin riboside, or thidiazuron. N6-Benzyladenine at 1.0 mg l−1 (4.44 μM) was most effective for shoot proliferation. Multiple shoots were also induced in nodal segments of in vitro-raised shoots grown on MS medium containing 1.0 mg l−1 (4.44 μM) of BA. Rooting was best induced in shoots grown on half-strength MS medium with 0.25 mg l−1 (1.42 μM) of indole-3-butyric acid. Plants were acclimatized in vermicompost and established in soil where they flowered and formed mature seeds.

An efficient protocol was established for in vitro shoot multiplication from nodal explants of Clitoria ternatea on semisolid Murashige and Skoog (MS) basal medium supplemented with 8.9μM 6-benzylaminopurine (BA). Inclusion of 1-naphthaleneacetic acid (NAA) in the culture medium along with BA promoted higher rates of shoot multiplication than BA alone. The rate of shoot multiplication was maximum (5.21) after 4 wk of culture on MS basal medium supplemented with 8.9μM BA and 1.34μM NAA. The elongated shoots rooted within 7–8d in half-strength MS basal salts supplemented with 1.34μM NAA and 2% (w/v) sucrose. About 85% of the rooted plantlets were acclimatized and transferred to the greenhouse.

A tobamovirus was isolated from feral plants of the African forage legume Clitoria ternatea collected in 1995 from two out of nineteen sites in Northern Australia. This virus, which we call Clitoria yellow mottle virus, is most closely related to, although distant from, sunnhemp mosaic virus in all regions of its genome.

Inadequate supply of fodder is a serious constraint to the potentially-promising small-holder-dairy production system in coastal Kenya. Alley farming could be an approach to addressing this problem. A study of forage production based on Napier grass and leucaena in an alley cropping system was conducted on an infertile sandy soil in lowland coastal Kenya. The effects of leucaena hedgerows,Clitoria ternatea (L.) intercropping, addition of slurry (110 t ha−1 yr−1) and two harvesting managements (severe and lenient) on the yield of Napier grass fodder, were assessed. The study was initiated in 1989 and three years results are reported.

This study demonstrates variability in amino acid composition among accessions of several Philippine indigenous legumes. Moreover, two accessions of D. lablab were identified to have high level of methionine (greater than 2%). Methionine was identified as the first limiting amino acid with leucine, and threonine as the second limiting amino acids for most legumes studies. The IVPDs of the legumes under study ranged from greater than 70 to 79%. Raw mature seeds had relatively low RNVs of 11 to 68% which increased to 68 to 94% and 51 to 89% after boiling and roasting, respectively.

Effects of P and S on growth and regrowth were assessed in a greenhouse trial in which Clitoria was grown through three cuttings on a deficient soil with a factorial combination of 4 levels of P and 3 levels of S. Phosphorus deficiency caused stunting, flowering delay, and a rusty appearance of leaflets. Sulfur deficiency caused stunting and chlorosis. With successive cuttings, P deficiency became less severe and S deficiency more severe. Nodulation observed at the end of the trial, was influenced more by S than by P. Maximal nodulation was achieved with the highest S level, at any of the highest three P levels. Root and stubble dry weight, and food reserves evaluated by etiolated regrowth and by soluble sugar in roots and stubble, responded positively to both P and S, and correlated with each other and with shoot growth preceding the final cutting. For maximal etiolated regrowth, the S requirement was higher than the P requirement.

THE plant Clitoria marina Linn. belongs to the Papilionaceae sub-group of Leguminosae, and is distinguished from Clitoria ternatea Linn. by its light-blue to almost white flowers. The leaves of the plant are used in medicine in India. A systematic chemical examination of the leaves has therefore been undertaken, and isolation of a δ-lactone compound from the alcoholic extract of the leaves has been achieved.

ZermahleneClitoria ternatea-Wurzeln oder deren Extrakt in 95% Alkohol zeigten bei oraler Verabreichung in nicht-toxischer Dosis keine nennenswerte diuretische oder natriuretische Wirkung beim Hund. Intravenose Gaben des Extraktes fuhrten zu massiger Steigerung der Ausscheidung von Natrium und Kalium im Harn, zeigten aber gleichzeitig auch Anzeichen von Nierenschadigung.