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Clitoria ternatea L. var. angustifolia Hochst. ex Baker f. (Fig. 1) Leguminosae of Tropical Africa: 428 (Baker 1929). SPECIMENS EXAMINED. — India. Gujarat, Morbi District, near Machchu river, Machchu Dam 3, 22°51’40N, 70°49’32E, 92 m a.s.l., 21.IV.2018, PKP 0722 (BSI!, Department of Botany Herbarium, SPT Arts & Science College, Godhra, Gujarat). Ethiopia. Ad radices montium in valle fluvii Tacaze, 18.VIII.1841, Schimper 1736 (G[G00015969 image!], P[P02779519, P02779523]). LECTOTYPE. — Ethiopia. Fluvium Tacaze prope Djeladjeranne, Abyssinia, 1.IX.1841, Schimper 1617 (lecto-, designated here, BM[BM000842757 image]!; isolecto-, P[P02779513, P02779514], TUB[TUB001563]). HABITAT. — Grows near wetlands, edges of crop fields. DISTRIBUTION. — Africa (Angola, Burundi, Chad, Ethiopia, Kenya, Malawi, Mozambique, Sudan, Tanzania, Uganda, Zaire and Zambia) and Asia (India) (Fig. 1). DESCRIPTION A bush-like perennial herb with twining, climbing, or trailing branches up to 5 m long. Stem terete, slender, 1-3 mm thick, weakly striate, much branched near the woody base, infrequently branched above, pubescence strigose.Leaves 5 (7) foliate, rarely with 3 foliate leaf, green to silvery green, polymorphic in shape. Leaflets elongate, narrow, typically 4-15 (25) mm wide, linear, lanceolate, oblong, occasionally narrowly elliptic (leaflet size minute), apex obtuse, base rotund, pubescence on upper surface uncinate, dense to scattered, becoming glabrate, macroscopic trichomes lacking. Petioles 1-4 cm, shorter than the rachis, weakly striate, strigose-pubescent, rachis 2-7 cm, pubescence and striations similar to petiole, internode segments 1-2.5 cm long. Petiolules 1-3 mm long, uncinate-pubescent, densely strigose. Stipules persistent, linear, 4-10 mm long, 0.5-1 mm wide, pubescence uncinate and sparsely strigose. Stipels persistent, 1-3 mm long, acicular, reduced to a nerve, inconspicuous, uncinate-pubescent with sparse appressed trichomes. Inflorescence axillary, solitary, reduced to a short peduncle bearing one flower at its apex; peduncle 0.3-1.5 cm long, sparsely strigose. Pedicel 3-6 mm long, usually borne laterally from peduncle apex, straight or often bending at swollen base to form an obtuse to right angle with peduncle, uncinate-pubescent. Bracts 2, apparently the outer pair, ovate to lanceolate, acuminate to acute, spreading-ascending or weakly concave and appressed to pedicel when pedicel is terminally located, 2-4 (5) mm long, 1 mm wide, pubescence uncinate and more or less strigose. Bracteoles highly variable, broadly ovate to nearly orbicular, 4-11 mm diameter. Flowers small, (2.5) 3-4 cm, typically white, usually papilionaceous. Calyx subpellucid, more or less 10 nerved with a nerve extending to the apex of each lobe, tube 9-14 mm long, 4-5 mm wide near base expanding to 7-9 mm wide at the throat, lobes oblong. Vexillum pubescence uncinate and strigose dorsally, greenish-white medially and white peripherally, 1.5-3.5 cm wide, blade white. Stamens diadelphous, vexillary stamen nearly free, tube nearly straight, 14-18 mm long, incurved slightly last 2-4 mm, filaments free 3-4 mm; anthers lanceolate, 1-1.5 mm long, 0.6-0.9 mm wide. Gynophore 1-2 mm; ovary 5-8 mm long, 1-1.5 mm wide; pubescence appressed, dense, white; style 12-14 mm long, base pubescent like that of ovary; stigma capitate, c. 0.7-0.8 mm in diameter. Legume subsessile, yellowish green to green becoming light brownish to tan, flat, valves linear-oblong to oblanceolate, nearly straight to weakly curved towards apex, margins sometimes wavy, infrequently minutely serrulate, pubescence short, strigose to spreading, and uncinate, typically (5) 6-10 cm long, 8-11 mm wide; stipe enclosed along with legume base within persistent calyx, 1-2 mm; beak 2-6 mm; dehiscence causing valves to twist 1-2 turns. Seeds yellowish brown turning black, smooth, subreniform, compressed, 1.5-2 mm thick, 4-5 mm long, 5-6 mm wide, 6-8 seeds per pod; hilum broad elliptic, 1 × 0.8 mm. NOTES Baker (1929) published Hochstetter’s varietal name, Clitoria ternatea var. angustifolia, and synonymized Vatke’s (1878) name Clitoria zanzibarensis. This variety is characterized by its round bracteoles, small flowers and narrower leaflets. Later botanists included De Wild’s (1925) name Clitoria mearnsii and Micheli’s (1897) name Clitoria tanganicensis as synonyms under the name Clitoria ternatea var. angustifolia. According to Wilczek (1954), this variety is distinguished by its narrow leaflets, rounded leaflet base, smaller white flowers, and smaller bracteoles. Several characteristics are present in these varieties. Leaflet length/width ratio, leaflet pubescence with uncinate hairs on the leaflet surface, leaflet width, flower length, and flower color are all included. Most specimens can be identified by their particular combination of characteristics. There are, however, a number of specimens with intermediate characteristics or with characteristics that are difficult to interpret. According to De Wild. (1925), Clitoria mearnsii is collected from Tanzania and deposited in BR herbarium (BR 0000006252830). As per Micheli’s (1897) Clitoria tanganicensis type material was gathered in Burundi and put away at BR (BR 0000008932587). Clitoria ternatea var. angustifolia (Schimper 1736) verified syntype material was collected in Abyssinia (Ethiopia), and it was deposited at herbaria BM (BM 000842761), G (G 00015967), and K (K 000418286). KEY TO THE INDIAN CLITORIA TERNATEA L. VARIETIES (ADAPTED FROM FANTZ 1977) 1. Flowers double, actinomorphic; stamens all free............................................................... var. pleniflora Fantz — Flowers papilionaceous; stamens diadelphous.............................................................................................. 2 2. Leaflets narrow and elongated; leaflet shape linear, lanceolate, oblong, lanceolate-oblong, or narrow elliptic (leaflet minute size); leaflet pubescence on upper surface moderately dense to scattered, uncinate, macroscopic trichomes absent; flowers small, typically white.......................................................... var. angustifolia Baker f. — Leaflets broad, leaflet shape ovate, ovate-elliptic, obovate, or sometimes oblong-ovate to oblong-elliptic; leaflet pubescence on upper surface moderate to sparse of very short appressed hairs becoming glabrate, uncinate trichomes lacking or sparse............................................................................................................................. 3 3. Flowers large, typically pale blue to purple.............................................................................. var. ternatea L. — Flowers large, typically white............................................................................. var. pilosula (Benth.) Baker f., Published as part of Patel, P. K., 2023, Clitoria ternatea L. var. angustifolia Hochst. ex Baker (Fabaceae): new varietal record to Asia from India, pp. 267-271 in Adansonia (3) (3) 45 (14) on pages 268-270, DOI: 10.5252/adansonia2023v45a14, http://zenodo.org/record/8082956, {"references":["BAKER E. G. 1929. - The Leguminosae of Tropical Africa. Part 2: Suborder Papilionaceae. Erasmus Press, Ostende: 427 - 429.","SCHIMPER G. W. 1736. - In Catalogue des herbiers de Geneve (CHG). Syntype Clitoria ternatea L. var. angustofolia Hochst. ex Baker f., Conservatoire et Jardin botaniques de Geneve. https: // www. ville-ge. ch / musinfo / bd / cjb / chg / index. php? lang","VATKE W. 1878. - Plantas in itinere africano ab J. M. Hildebrandt collectas determinare pergit W. Vatke. Osterreichische botanische Zeitschrift 28: 261 - 264. https: // doi. org / 10.1007 BF 01615120","DE WILD E. 1925. - Sur quelques Legumineuses africaines nouvelles. Journal of Zoology and African Botany, Supplement Botany 13 (8): 8 - 9.","MICHELI M. 1897. - Leguminosaceae, in DURAND TH. & DE WILDEMAN E., Materiaux pour la Flore du Congo. Bulletin de la Societe royale de botanique de Belgique 36: 60 - 61. https: // www. biodiversitylibrary. org / page / 8169094","WILCZEK R. 1954. - Flora of the Congo Belge et Ruanda-Urundi 6. Institut national pour l'etude agronomique du Congo belge: 260 - 269.","FANTZ P. R. 1977. - A Monograph of the Genus Clitoria (Leguminosae: Glycineae). Doctoral Dissertation, University of Florida, Gainesville, FL, 1066 p."]}

Streptococcus mutans (S. mutans) and Staphylococcus aureus (S. aureus) pathogenicity that alter biofim, has become one of risk factor in orthodontic treatment. The medicinal plant’s Butterfly pea efficacy as an antibacterial agent should be confirmed in dentistry. The purpose of this study is to investigate the antibacterial activity of Medan butterfly pea corolla extract (BPCE) against S. mutans ATCC®25175™ and S. aureus ATCC®6538™. This is a laboratory experiment with Post Test Only Group Design. The minimum inhibitory concentration of BPCE is 6.25 mg/mL. The best concentration of butterfly pea extract to inhibit biofilm formation (antibiofilm) is 100 mg/mL. There was a significant difference (p < 0.05) for antibiofilm activity assays and determination of intramembrane cellular leakage. Although Medan BPCE was inadequate enough in forming antibiofilm and caused intramembrane leakage of S. mutans and S. aureus, further studies in exploring the potential morphological traits of these herbs related to orthodontic products are quite promising.

Background: While several biologics have been reported from different parts of Clitoria ternatea, a herbaceous climber of the family Fabaceae, specific production of cationic peptides other than cyclotides ( Objective: The study aims to uncover potential bioactivities and characteristics of novel cationic peptides from C. ternatea seeds. Methods: C. ternatea seed cationic peptide purified by simple and cost-effective procedures was analyzed by electrophoresis and mass spectrometry. Antimicrobial efficacy was evaluated against bacterial and fungal pathogens. Antioxidant potential was quantified by in vitro antioxidant assays. Physicochemical characterization and Tandem mass spectrometry were performed. Results: An 8.5 kDa cationic peptide purified from C. ternatea seeds was active against Candida albicans, Staphylococcus aureus, Aeromonas hydrophila and Escherichia coli at a minimum inhibitory concentration in the range of 8-32 μg/ml. This activity was totally uncompromised at pH 5-8 or after 1 h of heat treatment at 70-80ºC, but was sensitive to protease treatment. Concentration-dependent free-radical scavenging activity and ferric-reducing capacity demonstrated the antioxidant potential of the peptide. Tandem MS analysis of trypsin-digested peptide based on shotgun proteomics detected matching peptide sequences with one or two cysteine residues but had low sequence coverage (≤17%) to known sequences in the C. ternatea protein database. Taken together, the distinct characteristics of this novel 8.5 kDa peptide clearly distinguish it from known cyclotides of C. ternatea. Conclusions: Insights have been obtained into the functional characteristics of what appears to be a novel cationic peptide from C. ternatea seeds, exhibiting significant antimicrobial and antioxidant activities.

Cyclotides, a class of macrocyclic plant peptides, characterized by a cyclic backbone and three inter-locking disulfide bonds, may be divided into two major structural subfamilies, Möbius and Bracelet, based on the presence or absence of a specific proline residue. The present study describes the suite of cyclotides obtained from Clitoria ternatea, characterized by LC-MS and MS/MS techniques. Notable variations in product ion distributions were observed in cyclotides belonging to different structural subfamilies based on the number and positions of proline residues. For instance, Cter M which is an abundant Möbius cyclotide in this plant containing three proline residues, displayed distinct b- and y- ion characteristics in the MS/MS spectra compared to Cliotide T1, another commonly identified cyclotide but belonging to the Bracelet subfamily having two proline residues. The distinct fragmentation pattern of prototypical cyclotides of each structural subfamily, determined by Xxx-Pro bond fragmentation, was used to rapidly identify and sequence a novel cyclotide ctr pep 30 from this plant.

Clitoria ternatea L. Climbing herbaceous or small liana, in wooded savannah and grassy savannah; also in fallow and disturbed sites., Published as part of Monteiro, Francisca, Costa, Esperança Da, Kissanga, Raquel, Costa, José Carlos & Catarino, Luís, 2022, An annotated checklist of the vascular flora of Quiçama National Park, Angola, pp. 1-67 in Phytotaxa 557 (1) on page 33, DOI: 10.11646/phytotaxa.557.1.1, http://zenodo.org/record/6985699

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.

Butterfly pea flower have great sensory attraction, but they have not yet been used widely in Vietnam. Extracts of butterfly pea flowers can be used conveniently as a natural blue colorant for food products. In this study, the identification of anthocyanin compounds in butterfly pea flowers was performed by UPLC coupled with a UV and Mass spectrometer instrument. Positive and negative ion electrospray MS/MS chromatograms and spectra of the anthocyanin compounds were determined. By analyzing the chromatograms and spectra for each ion, five anthocyanins were identified in the butterfly pea flower extract, these were delphinidin-3-(6”‐p-coumaroyl)-rutinoside, cyanidin 3-(6”-p-coumaroyl)-rutinoside, delphinidin-3-(p-coumaroyl) glucose in both cis- and trans- isomers, cyanidin-3-(p-coumaroyl-glucoside) and delphinidin-3-pyranoside. Additionally, based on their intensity, it was determined that cyanidin-3-(p-coumaroyl-glucoside) was the most abundant anthocyanin, followed by cyanidin 3-(6”-p-coumaroyl)-rutinoside, delphinidin-3-(p-coumaroyl-glucoside), delphinidin-3-(6”-p-coumaroyl)-rutinoside and delphinidin-3-pyranoside. In this study, cyanidin derivatives were discovered in butterfly pea flower extract, where these compounds had not been detected in previous studies.

Butelase-1 is an efficient ligase from Clitoria ternatea with wide applications in the food and biopharmaceutical fields. This research aimed to achieve high-efficiency expression of butelase-1 and explore its application in food-derived angiotensin I-converting enzyme (ACE) inhibitory peptides. The recombinant butelase-1 zymogen was prepared at a yield of 100 mg/L in Escherichia coli and successfully activated at pH 4.5, resulting in a 6973.8 U/L yield of activated butelase-1 with a specific activity of 348.69 U/mg and a catalytic efficiency of 9956 M-1 s-1. Activated butelase-1 exhibited considerable resistance to Tween-20, Triton X-100, and methanol. The "traceless" cyclization of ACE inhibitory peptides was realized using activated butelase-1, which resulted in higher stability and ACE inhibitory activity than those of the linear peptides. Our work proposed an efficient method for the preparation of butelase-1 and provided a promising model for its application in food fields.

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.

This research evaluated the changes on populations of culturable N-fixing free bacteria (NFFB) and P-solubilizing bacteria (PSB), as well as on the root nodulation by native rhizobia, the root colonization and spore number of arbuscular mycorrhizal fungi (AMF), in the rhizosphere of Clitoria ternatea and Brachiaria brizantha grown in mesocosms contaminated with crude oil (0, 3000, 6000, 9000, and 12000 mg kg -1 ), for 240 days. After 24 h of soil contamination, the highest populations of NFFB and PSB (5.5 and 4.9 Log UFC, respectively) were found in control, and the lowest populations were obtained at 12000 mg kg -1 (5.1 and 4.2 Log UFC, respectively). In contrast, at 60 and 240 days, the control showed lower populations of NFFB and PSB (5.4 and 4.8 Log UFC, respectively) than contaminated treatments. The highest number or root nodules in C. ternatea was quantified in control at 60 and 240 days (25 and 27 nodules, respectively) in comparison to those observed at the treatment with 12000 mg kg -1 (7 and 1 nodule, respectively). At 60 days, AMF colonization in both plant species, and the number of spores significantly decreased as the crude oil concentration increased; however, at 240 days, the highest number of AMF spores was recorded at treatments with 6000 and 12000 mg kg -1 . The dry weight of both plant species significantly decreased as crude oil concentrations increased. Although C. ternatea was more susceptible to the toxic effects of crude oil, this plant species showed greater content of total chlorophyll than B. brizantha .

A 2-year field experiment (2015/2016) was conducted on clitoria planted on May 20 as early (PE) and June 20 as late (PL) under 100% = I100%, 80% = I80% and 60% = I60% of allowable soil moisture dep...

Background Methylglyoxal (MG) is a highly reactive dicarbonyl precursor for the formation of advanced glycation end products (AGEs) associated with age-related diseases, including diabetes and its complications. Clitoria ternatea L. flower has been reported to possess antioxidant and antiglycating properties. Evidence indicates that the extract of Clitoria ternatea L. flower inhibits fructose-induced protein glycation and oxidative damage to bovine serum albumin (BSA). However, there is no evidence to support the inhibitory effect of CTE against MG-mediated protein glycation and oxidative damage to protein and DNA. Therefore, the aim of the present study was to investigate whether C. ternatea flower extract (CTE) prevents MG-induced protein glycation and oxidative DNA damage. Methods The formation of fluorescent AGEs in BSA was evaluated using spectrofluorometer. The protein carbonyl and thiol group content were used for detecting protein oxidation. DNA strand breakage in a glycation model comprising of MG, lysine and Cu2+ or a free radical generator 2,2′-azobis(2-methylpropionamidine) dihydrochloride (AAPH) systems was investigated using gel electrophoresis. Generation of superoxide anions and hydroxyl radicals in the MG/lysine system was assessed by the cytochrome c reduction assay and thiobarbituric acid reactive substances assay, respectively. High performance liquid chromatography (HPLC) was used to measure the MG-trapping ability. Results In the BSA/MG system, CTE (0.25–1 mg/mL) significantly inhibited the formation of fluorescent AGEs and protein oxidation by reducing protein carbonyl content as well as preventing the protein thiol depletion. The concentration of CTE at 0.125–1 mg/mL prevented oxidative DNA cleavage in MG/lysine and AAPH systems associated with the inhibition of superoxide anion and hydroxyl radical formation. It also directly trapped MG in a concentration-dependent manner, ranging from 15 to 43%. Conclusions The study findings suggest that the direct carbonyl trapping ability and the free radical scavenging activity of CTE are the underlying mechanisms responsible for the prevention of protein glycation and oxidative DNA damage.

Clitoria ternatia L. (CT) has been reported to have anti-inflammatory and antioxidant effects. This study investigated the effect of CT aqueous flower extract on blood pressure and renal alterations in Nω-nitro- l -arginine methyl ester hydrochloride ( l -NAME)-induced hypertensive rats. Male Sprague Dawley rats received l -NAME in drinking water and were treated with CT flower extract or lisinopril. CT aqueous flower extract and lisinopril alleviated l -NAME-induced hypertension (p l -NAME-induced hypertensive rats and attenuated by CT flower extract or lisinopril co-treatment (p l -NAME were alleviated by CT flower extract or lisinopril co-treatment (p l -NAME-induced hypertensive rats (p l -NAME-induced renal injury and dysfunction in rats. The possible mechanism may be related to the suppression of Ang II-mediated Nox4 expression and the oxidative stress cascade in rats.

Biodegradable chitosan-poly(vinyl alcohol) films containing natural anthocyanin-rich extracts were prepared using solvent casting method and employed as intelligent indicators for monitoring beverages freshness. The surface and cross-sectional scanning electron micrograph indicated a compact structure for the intelligent films, whereas the atomic force micrograph indicated a 16.22 and 20.31 nm increase in surface roughness for Clitoria ternatea and Carissa carandas extract incorporated films, respectively. Moreover, the test films demonstrated enhanced radical scavenging efficacy. The extracts and anthocyanin incorporated films presented excellent colorimetric changes at pH 2 to 8. In addition, the C. ternatea test films showed changes in color for juice stored at 25 °C after 72 h. Photo-degradability results indicated stability of test films stored in dark at 4 °C and 25 °C, whereas leaching study indicated the release of ≤2.0% anthocyanin after 24 h. The cytocompatibilty assay showed that the test and control films were biocompatible with a viability of >80% on HaCat cells. The results demonstrated that the incorporation of anthocyanins-rich extracts into chitosan-poly(vinyl alcohol) did not significantly interfere with the films properties (p > 0.05). The natural anthocyanin incorporated films demonstrated good pH sensing property that could be further explored for monitoring of beverages freshness.

The Orange Spiny Whitefly (OSW) Aleurocanthus spiniferus (Hemiptera: Aleyrodidae) represents a new serious threat to Citrus spp., grapevine and ornamental plants in the whole Mediterranean area. Such threat urgently calls for the development of a sustainable control strategy, including insecticides compatible with biological control, and applicable also in organic citrus farming that represent an essential part of Mediterranean agricultural economy. Therefore, we evaluated the toxicity and the effects on host searching, oviposition, and probing and feeding behavior exerted on OSW by organic insecticides supposed to have limited side effects on environment and ecosystem services, i.e. sweet orange essential oil (EO), extract of Clitoria ternatea (CT), mineral oil, pyrethrin and azadirachtin. Despite none of the compounds caused a significant mortality of any of the OSW instars, we observed interesting effects on whitefly behavior: (i) EO and pyrethrin showed a relevant repellent effect, with impairment of both adults landing and oviposition on treated plants; (ii) CT and pyrethrin strongly affected probing behavior. Here, in the light of our findings, we discuss possible OSW sustainable control strategies and further research perspectives.

Clitoria ternatea a perennial climber of the Fabaceae family, is well known for its agricultural and medical applications. It is also currently the only known member of the Fabaceae family that produces abundant amounts of the ultra-stable macrocyclic peptides, cyclotides, across all tissues. Cyclotides are a class of gene-encoded, disulphide-rich, macrocyclic peptides (26–37 residues) acting as defensive metabolites in several plant species. Previous transcriptomic studies have demonstrated the genetic origin of cyclotides from the Fabaceae plant family to be embedded in the albumin-1 genes, unlike its counterparts in other plant families. However, the complete mechanism of its biosynthesis and the repertoire of enzymes involved in cyclotide folding and processing remains to be understood. In this study, using RNA-Seq data and de novo transcriptome assembly of Clitoria ternatea, we have identified 71 precursor genes of cyclotides. Out of 71 unique cyclotide precursor genes obtained, 51 sequences display unique cyclotide domains, of which 26 are novel cyclotide sequences, arising from four individual tissues. MALDI-TOF mass spectrometry analysis of fractions from different tissue extracts, coupled with precursor protein sequences obtained from transcriptomic data, established the cyclotide diversity in this plant species. Special focus in this study has also been on identifying possible enzymes responsible for proper folding and processing of cyclotides in the cell. Transcriptomic mining for oxidative folding enzymes such as protein-disulphide isomerases (PDI), ER oxidoreductin-1 (ERO1) and peptidylprolyl cis-trans isomerases (PPIases)/cyclophilins, and their levels of expression are also reported. In particular, it was observed that the CtPDI genes formed plant-specific clusters among PDI genes as compared to those from other plant species. Collectively, this work provides insights into the biogenesis of the medicinally important cyclotides and establishes the expression of certain key enzymes participating in peptide biosynthesis. Also, several novel cyclotide sequences are reported and precursor sequences are analysed in detail. In the absence of a published reference genome, a comprehensive transcriptomics approach was adopted to provide an overview of diverse properties and constituents of C. ternatea.

Clitoria ternatea is a vigorous, herbaceous perennial legume that belongs to the Fabaceae family. All parts of the plant are used in the preparations of Ayurvedic drugs. It is an astringent, an aphrodisiac, a rejuvenator, and a brain tonic. It also has anti-inflammatory, analgesic, and antipyretic properties. Baidyanath Shankapushpi, which contains extracts of herbs such as C. ternatea, Bacopa monnieri, Withania somnifera and Asparagus racemosus, is clinically administered for memory improvement, blood purification and to improve digestion. However, its neuroprotective effect has not been reported so far. In the present study, the neuroprotective effect of C. ternatea root (CTR) extract on hippocampal CA3 neurons was investigated. Three-month-old albino mice were divided into four groups. Group I was the normal control, group II was the saline control, group III was the stress group, and group IV was the stress + CTR-treated group. Group-III mice were stressed in a wire mesh restrainer for 6 hours/day for 6 weeks. Grou-IV mice were also stressed like group III, but received CTR extract orally throughout the stress period. After 6 weeks, their brain was removed, and their hippocampi were dissected and processed for Golgi staining. The hippocampal neurons were traced using a camera lucida focused at 400x magnification. The Sholl concentric circle method was used to quantify the dendrites. The results showed a decrease in the number of dendritic branching points and of dendritic intersections in the stressed group. On the other hand, there was an increase in the number of dendritic branching points and of dendritic intersections of hippocampal CA3 neurons in group IV, which was subjected to restraint stress and was treated with the CTR extract. The results showed that the oral administration of CTR significantly increased the dendritic branching points and the dendritic intersections of hippocampal CA3 neurons.

Aim: An anthocyanin complex (AC), combined Zea mays and Clitoria ternatea extracts, was evaluated for topical oral wound healing in rats and a clinical trial in orthodontic patients. Methods/results: AC enhanced anthocyanin permeation in vitro. In rats, 10% w/w of AC in a mucoadhesive gel (AG) reduced erythema and sizes of oral wounds after topical applications at higher extent than its placebo gel. Acute orthodontic wounds in 68 volunteers were randomly assigned to topically receive either AG or placebo gel and double-blind assessed. Wound size reduction and wound closure enhancement were obvious in AG-treated group on day 3 (p

Ethnopharmacological relevance Clitoria ternatea Linn. (C. ternatea) is a traditionally used herb in arthritis, and its anti-arthritic activity has been attributed to polyphenols (e.g. quercetins) from its flower petal. Aim of the study The present study was designed to investigate whether C. ternatea or quercetin-3s-D-glucoside (QG) support the antibody mediated TNFα-receptor 1 (TNFR1) neutralization to ameliorate arthritis in mice. Materials and methods Development of collagen-induced arthritis (CIA) in male Swiss mice (20–22 g, 3–4 weeks of age) was followed by estimation of synovial polymorphonuclear cell (PMN) accumulation (in terms of myeloperoxidase activity), synovial and systemic release of cytokines, chemokines and C-reactive protein (CRP) by enzyme-linked immunosorbent assay (ELISA), biochemical estimation of synovial free radical generation and antioxidant status, as well as immunoblot assessment of synovial TNFR1, toll-like receptor 2(TLR2), cyclooxygenase-2(COX-2) and inducible nitric oxide synthase (iNOS) expression; and zymographic analysis of synovial matrix-metalloprotease-2 (MMP-2) activity. Results CIA was induced from day 2 post-secondary immunizations as evidenced from arthritic scores and joint swelling in parallel to increased inflammatory and oxidative stress parameters in synovial joints. Long term supplementation with extract from Clitoria ternatea flower petals CTE (50 mg/kg) and QG (2.5 mg/kg) upto 24 days post booster immunization augmented anti-arthritic potential of TNFR1 neutralization with anti-TNFR1 antibody (10 μg per mice) in terms of reduced MPO activity, decrease in release of pro-inflammatory cytokines, chemokines, reactive oxygen species (ROS)/ reactive nitrogen species (RNS) production in parallel to significant (p Conclusion CTE and QG possess potential anti-arthritic activity which targets synovial MMP-2 in arthritic joints and TNFR1 targeting followed by CTE or QG treatment might become a combinatorial approach in future therapeutic research in treatment of arthritis.

INTRODUCTION Perinatal maternal separation stress (PMSS) induces brain lipid peroxidation and reduction in endogenous antioxidants. The present study was designed to assess the brain oxidative stress (MDA) and protein thiol levels through various stages of aging in PMSS rat pups supplemented with choline with docosahexaenoic acid (DHA) or Clitoria ternatea (Linn) aqueous root extract (CTR). MATERIAL AND METHODS Study groups, control, PMSS, PMSS + choline with DHA, PMSS + CTR (n = 6/group) were included in the study. Pups of PMSS groups were separated from their mothers for a period of 6 h/day for 30 days. PMSS + supplemented groups were treated as appropriate during the same period. Rats were sacrificed on day 30, 60, 90, 210 and 360. Brains were processed for MDA and protein thiol levels. RESULTS Brain MDA levels were significantly increased in PMSS rats at day 30, 60 (p < 0.001), 90 (p < 0.01) and attenuated in PMSS pups supplemented with choline with DHA and CTR at day 30, 60 (p < 0.01), 90 (p < 0.01, p < 0.05) and 360 (p < 0.001) when compared to the same in age-matched controls and PMSS rats, respectively. Alternatively, brain protein thiol levels in PMSS rats were reduced in all age groups when compared to the same in age-matched controls. A significant increase in brain thiol levels was observed in supplemented groups at day 60 (p < 0.01) and 210 (p < 0.01, p < 0.05) when compared to the same in age-matched PMSS rats. CONCLUSIONS PMSS causes enhanced brain lipid peroxidation (MDA levels) and reduces endogenous antioxidants. Supplementation of choline and DHA or CTR during PMSS in rats persistently attenuates brain oxidative stress through aging.

Anthocyanins from flowers of the butterfly pea (Clitoria ternatea L.) are promising edible blue food colorants. Food processing often faces extreme pHs and temperatures, which greatly affects the color and nutritional values of anthocyanins. This study explored the color, spectra, storage stability, and antioxidant properties of C. ternatea anthocyanin extract (CTAE) at different pHs. The color and absorption spectra of CTAEs at a pH of 0.5–13 were shown, with their underlying structures analyzed. Then, the storage stability of CTAEs were explored under a combination of pHs and temperatures. The stability of CTAE declines with the increase in temperature, and it can be stored stably for months at 4 °C. CTAEs also bear much resistance to acidic and alkaline conditions but exhibit higher thermal stability at pH 7 (blue) than at pH 0.5 (magenta) or pH 10 (blue-green), which is a great advantage in food making. Antioxidant abilities for flower extracts from the butterfly pea were high at pH 4–7, as assessed by DPPH free radical scavenging assays, and decreased sharply when the pH value exceeded 7. The above results provide a theoretical basis for the application of butterfly pea flowers and imply their great prospect in the food industry.

A spectroscopic study was conducted to evaluate the colour degradation mechanism of anthocyanin-rich extract from butterfly pea petal. The extract was diluted in four different solvent systems, which were buffer solution pH 7 (AQ7) and the mixture of organic solvent with buffer solution pH 7 (4:1 v/v). The organic cosolvent involved were methanol (ME7), ethanol (ET7) and acetone (AC7). The samples were stored in containers with 0% and 50% headspace, and their colour intensity, total anthocyanin and hypsochromic shift were evaluated periodically. The rank of colour and anthocyanin degradation from the biggest was AQ7 > ME7 > ET7 > AC7. The longest hypsochromic shift was AQ7 > ME7 > ET7, while in AC7 the shift was absent. There was evidence that the volume of package headspace provoked colour stability. The colour degradation in AC7 was proposed to occur through hydrophobic interaction unfolding, and in AQ7 was through the deacylation, while in ME7 and ET7 was due to both mechanisms.

The aim of this study was to evaluate the effect of association of forage cactus meal (CM) and cunha hay (CH) on carcass traits, meat quality, and fatty acid (FA) profile of goats. Twenty-four goats were finished on diets composed with the association of CM and CH (83% CH + 17% CM, 67% CH + 33% CM, or 50% CH + 50% CM), corresponding to 60% of the total diet; the remainder was composed of 10% elephant grass and 30% concentrate. The control treatment consisted of 70% elephant grass and 30% concentrate. Carcass yield and commercial cut weight were higher in the animals fed CH and CM at any concentration (P

Plant asparaginyl endopeptidases (AEPs) are expressed as inactive zymogens that perform seed storage protein maturation upon cleavage dependent auto-activation in the low pH environment of storage vacuoles. AEPs have attracted attention for their macrocyclization reactions and have been classified as cleavage or ligation specialists. However, we have recently shown that the ability of AEPs to produce either cyclic or acyclic products can be altered by mutations to the active site region, and that several AEPs are capable of macrocyclization given favorable pH conditions. One AEP extracted from Clitoria ternatea seeds (butelase 1) is classified as a ligase rather than a protease, presenting an opportunity to test for loss of cleavage activity. Here, making recombinant butelase 1 and rescuing an Arabidopsis thaliana mutant lacking AEP, we show butelase 1 retains cleavage functions in vitro and in vivo. The in vivo rescue was incomplete, consistent with some trade-off for butelase 1 specialization toward macrocyclization. Its crystal structure showed an active site with only subtle differences from cleaving AEPs, suggesting the many differences in its peptide binding region are the source of its efficient macrocyclization. All considered, it seems either butelase 1 has not fully specialized or a requirement for auto-catalytic cleavage is an evolutionary constraint upon macrocyclizing AEPs.

The aim of the present study was to evaluate the phenolic composition of crude lyophilized extracts (CLE) and partially purified (PPE) extracts of C. ternatea blue petals as well as the anthocyanin stability against pH, temperature and light in the presence and absence of fructooligosaccharides. Twelve compounds were tentatively identified by UHPLC-Q-TOF-MS/MS in CLE and PPE extracts. In direct/reverse spectrophotometric titration, anthocyanins showed colour changes between pH 2.25 to 10.20, and colour reversibility, maintaining antioxidant activity against the DPPH radical. The aqueous extracts at pH 3.6 and 5.4 exhibited thermal stability with the presence and absence of fructooligosaccharides with activation energy higher than 99 kJ/mol. The addition of fructooligosaccharides in the extracts at pH 5.4 exposed to light provided a protective effect against anthocyanin photodegradation. The data show the technological potential of aqueous extract of C. ternatea blue petals as a natural colourant in a functional beverage model system.

The aim of this work was to investigate the effect of sucrose on the stability of Clitoria ternate extract against thermal and pH degradations. Lyophilised extract of the flower (1 mg/ml) was added into a series of sucrose solutions with concentrations ranging from 0.1% to 20% at pH7. The thermal stability of the extract in the solutions at 60oC was monitored using a UV-VIS spectrophotometer over 24 days. High temperature (60oC) accelerated degradation of the anthocyanin-rich extract but the presence of sucrose appeared to have slowed down the degradation process. However, sucrose asserted no protective effect against pH even at a concentration of 20%. It was thought that sucrose enhanced the thermal stability of anthocyanins by reducing water activity, partially preventing nucleophilic attack at the pyrylium ring of anthocyanins by water molecules. The present work provides some useful information for evaluating the potential of C. ternatea extract on food applications.

Background Clitoria ternatea (CT) is an herbal plant that has been used as a memory booster in folk medicine. CT root extract has been proven to restore chronic cerebral hypoperfusion (CCH)-induced memory deficits in a rat model, but the underlying mechanisms and the toxicity profile following repeated exposure have yet to be explored. The aim of the study To investigate the effects of the chronic (28 days) oral administration of CT root extract on CCH-induced cognitive impairment, neuronal damage and cholinergic deficit, and its toxicity profile in the CCH rat model. Materials and methods The permanent bilateral occlusion of common carotid arteries (PBOCCA) surgery method was employed to develop a CCH model in male Sprague Dawley (SD) rats. Then, these rats were given oral administration of CT root extract at doses of 100, 200, and 300 mg/kg, respectively for 28 days and subjected to behavioural tests. At the end of the experiment, the brain was harvested for histological analysis and cholinesterase activities. Then, blood samples were collected and organs such as liver, kidney, lung, heart, and spleen were procured for toxicity assessment. Results Chronic treatment of CT root extract at doses of 200 and 300 mg/kg, restored memory impairments induced by CCH. CT root extract was also found to diminish CCH-induced neuronal damage in the CA1 region of the hippocampus. High dose (300 mg/kg) of the CT root extract was significantly inhibited the increased acetylcholinesterase (AChE) activity in the frontal cortex and hippocampus of the PBOCCA rats. In toxicity study, repeated doses of CT root extract were found to be safe in PBOCCA rats after 28 days of treatment. Conclusions Our findings provided scientific evidence supporting the therapeutic potential of CT root extract in the treatment of vascular dementia (VaD)-related cholinergic abnormalities and subsequent cognitive decline.

The present study was conducted to evaluate the influence of ultrasound on bioactive compounds and biological activities of blue butterfly pea flower (Clitoria ternatea L.). For this purpose, optimized conditions (temperature 50 °C, time 150 min, solid to liquid ratio 1 g:15 ml, 70% amplitude and 240 W, 20 kHz frequency, 3 s on and 3 s off) of ultrasound (US) and conventional extraction (AGE: Agitation, water bath for 150 min, 50 °C at 150 rmp) were used. The results showed significant (p 0.05) effect of US and AGE over TFlaCs and PACs were observed. Moreover, the results of Fourier-transform infrared spectroscopy (FTIR) showed little changes in spectrum and US does not affect the functional group of bioactive compounds structure. Additionally, extracts (500–2000 µg/ml) protect pBR322 plasmid DNA damage induced by (1 mM H2O2 and 1 mM FeSO4), plasma oxidation (induced by 250 µM CuCl2) and inhibit erythrocyte hemolysis (induced by 200 mM AAPH, 34.6 to 66.73%). Sonication can be applied successfully for the extraction of bioactive compounds from plant materials with high biological activities.

The binding of an extract from the flowers of Clitoria ternatea L. to the digestive enzyme α-amylase was investigated. This extract is a mixture of flavonoids, including anthocyanins, and has been previously shown to inhibit the activity this enzyme. This has implications for modulating starch digestion. In order to investigate the kinetics, we made use of time-resolved fluorescence to simultaneously monitor two different emission bands emanating from the extract. This measurement was enabled by the use of a “photon streaming” approach and changes in fluorescence lifetime and intensity were used to follow the interaction. A longer wavelength band (655 nm) was ascribed to anthocyanins in the mixture and these were observed to bind at a rate an order of magnitude slower than other flavonoids present in the extract, monitored at a shorter wavelength (485 nm). Changes in the fluorescence emission of the extract upon binding were further assessed by the use of decay associated spectra.

Air pollutants especially polyaromatic hydrocarbons pose countless threats to the environment. This issue demands for an effective phytoremediation technology. In this study we report the beneficial interactions of Clitoria ternatea and its plant growth promoting endophytic bacteria Bacillus cereus ERBP by inoculating it for the remediation of 5 ppm airborne ethylbenzene (EB). The percentage efficiency for ethylbenzene removal among B. cereus ERBP inoculated and non-inoculated sterile and natural C. ternatea has also been determined. The inoculation of B. cereus ERBP has significantly increased EB removal efficiency of both sterile and natural C. ternatea. The inoculated natural C. ternatea seedlings showed 100% removal efficiency within 84 h for the aforementioned pollutant compared with the sterile inoculated C. ternatea seedlings (108 h). The degradation of EB by C. ternatea seedlings with and without B. cereus ERBP was assessed by measuring the intermediates of EB including 1-phenylethanol, acetophenon, benzaldehyde and benzoic acid. In addition, cytochrome P450s monooxygenase (CYP83D1) and dehydrogenases (LOC100783159) involved in the oxidation of hydrocarbons are well reported for their bio catalytic activities under xenobiotic stress conditions. Hence, the co-effect of the native endophyte B. cereus ERBP inoculation and EB exposure on the expression level of CYP83D1 and dehydrogenase were also determined. The targeted genes CYP83D1and dehydrogenases have shown an increased expression level under the 5 ppm of EB exposure enabling C. ternatea to withstand and remediate the pollutant.

Background Clitoria ternatea L., a natural food-colorant containing anthocyanin, demonstrated antioxidant and antihyperglycemic activity. The aim of this study was to determine the effects of Clitoria ternatea flower extract (CTE) on postprandial plasma glycemia response and antioxidant status in healthy men. Methods In a randomized, crossover study, 15 healthy men (ages 22.53 ± 0.30 years; with body mass index of 21.57 ± 0.54 kg/m2) consumed five beverages: (1) 50 g sucrose in 400 mL water; (2) 1 g CTE in 400 mL of water; (3) 2 g CTE in 400 mL of water; (4) 50 g sucrose and 1 g CTE in 400 mL of water; and (5) 50 g sucrose and 2 g CTE in 400 mL of water. Incremental postprandial plasma glucose, insulin, uric acid, antioxidant capacities and lipid peroxidation were measured during 3 h of administration. Results After 30 min ingestion, the postprandial plasma glucose and insulin levels were suppressed when consuming sucrose plus 1 g and 2 g CTE. In addition, consumption of CTE alone did not alter plasma glucose and insulin concentration in the fasting state. The significant increase in plasma antioxidant capacity (ferric reducing ability of plasma (FRAP), oxygen radical absorbance capacity (ORAC), trolox equivalent antioxidant capacity (TEAC), and protein thiol) and the decrease in malondialdehyde (MDA) level were observed in the subjects who received 1 g and 2 g CTE. Furthermore, consumption of CTE protected sucrose-induced reduction in ORAC and TEAC and increase in plasma MDA. Conclusions These findings suggest that an acute ingestion of CTE increases plasma antioxidant capacity without hypoglycemia in the fasting state. It also improves postprandial glucose, insulin and antioxidant status when consumed with sucrose. Trial registration Thai Clinical Trials Registry: TCTR20170609003 . Registered 09 September 2017. ‘retrospectively registered’.