Seeds Of Momordica Charantia Research Articles

Standardization and Phytochemical Investigation of Calendula Officinalis, Cassia Tora and Momordica Charantia Seed Extract

Purpose : Standardization the process of developing and implementing technical standards and to maximize compatibility, safety, repeatability and quality of the drugs is one of the important measures in view of the various practical problems encountered from time to time especially in the field of herbal drugs and Ayurveda.Scientific data pertaining to the standardization of the herbal drugs could be of immense value to substantiate efficacy, safety or toxicity of an herb. Hence present study was intended to standardize Calendula officinalis flowers, Cassia tora and Momordicacharantia seed extracts, since these are widely used in large number of herbal, ayurvedic and homeopathic formulations. Methodology : Systematic investigations were carried out to assess various quality parameters as per WHO guidelines such as morphological, microscopical and proximate analysis including ash values, moisture contents and extractive values and TLC for Calendula officinalis flowers, Cassia tora and Momordicacharantia seed extracts. Results : Results indicated the authenticity of the herbal drugs used in the study. Organoleptic and proximate analyseswere well within the acceptable limits. TLC system was developed for identification of the extracts. Quantitative estimation of total saponins, phenols and flavonoid contents were determined. Herbs tested indicated absence of heavy metals and arsenic. Qualitative phyto chemical analysis of extracts obtained using different solvents revealed presence of glycosides, saponins, triterpenes, phenols, tannins and carbohydrates in different extracts. Conclusion : Study was successful in establishing quality standards for the flowers of Calendula officinalis, seed of Momordicacharantia and Cassia tora. These preliminary studies may offer great help in initial procurement and assessment of quality of the crude drugs when these are being used as raw materials for preparations of herbal formulations.

Optimization of spray drying process for formulation of solid dispersion containing polypeptide-k powder through quality by design approach

Polypeptide-k is an antidiabetic phytochemical isolated from dried and ripened seeds of Momordica charantia. The peptide has not been able to find much clinical use despite its established therapeutic effect. This is mainly attributed to its poor aqueous solubility. Present study presents the formulation of solid dispersion containing polypeptide-k to enhance its aqueous solubility. The study was carried out by employing the optimization approach using spray drying technique. Variables were evaluated using Box Behnken design. These include trehalose to drug ratio, tween-80 to drug ratio, inlet air temperature and feed flow rate. Responses measured were moisture content, solubility, product yield and angle of repose. Data analysis by ANOVA test indicated that ratio of trehalose to PPK significantly affected product flow properties and yield, whereas ratio of tween-80 to PPK was found to significantly affect moisture content, solubility and flow properties of the resultant formulation. Inlet air temperature significantly influenced moisture content while feed flow significantly affected moisture content and product yield. The optimized batch of formulation exhibited higher solubility in water as well as various aqueous buffers as compared to pure polypeptide-k. Through PXRD and SEM, it was inferred that enhanced solubility was due to reduced particle size and increased surface area of polypeptide-k present in the formulation.

Momordica charantia Seed Lectin: Toxicity, Bacterial Agglutination and Antitumor Properties

In last three decades, several studies were carried out on the D-galactose-specific lectin of Momordica charantia seeds (MCL). In the present study, in vitro growth inhibition (8-23 %) at different concentrations (6-24 μg/ml) of MCL was observed against Ehrlich ascites carcinoma (EAC) cells by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. MCL also showed 28, 45, and 75 % growth inhibitions against EAC cells when administered 1.2, 2.0, and 2.8 mg/kg/day (i.p.), respectively for five consequent days in vivo in mice. After lectin treatment, the level of red blood cell and hemoglobin was increased significantly with the decrease of white blood cell and maintained the normal level when compared with EAC-bearing control and normal mice without EAC cells. Although MCL caused cell cycle arrest at G0/G1 phase of EAC cells, any irregular shape or apoptotic morphological alterations in the lectin-treated EAC cells was not observed by an optical and fluorescence microscope. Lectin showed toxicity against brine shrimp nauplii with an LC50 value of 49.7 μg/ml. Four out of seven pathogenic bacteria were agglutinated by MCL in the absence of inhibitory sugar D-lactose/D-galactose. In conclusion, MCL showed strong cytotoxic effect and therefore can be used as a potent anticancer chemotherapeutic agent.

Alpha-momorcharin (α-MMC) exerts effective anti-human breast tumor activities but has a narrow therapeutic window in vivo

Alpha-momorcharin (α-MMC), a ribosome inactivating protein (RIP) extracted from the seeds of Momordica charantia, exerts anti-tumor, antiviral, and anti-fungal activities. However, α-MMC has an obvious toxicity that limits its clinical application. We examined the effect of α-MMC on the inhibition of human breast cancer and assessed its general toxicity to find the therapeutic window in vivo for its potential clinical use. It was purified using column chromatography, and then injected into the xenograft nude mouse model induced by MDA-MB-231 and MCF-7. The anti-tumor efficacy was evaluated with T/C%. Next, the α-MMC was injected at a series of doses to Balb/C mice to assess its general toxicity. The MTT assay, the apoptosis test, and the cell cycle inhibition of α-MMC in human breast cancer cells were performed. In the xenografted tumors induced by MDA-MB-231 and MCF-7, α-MMC exerted an obvious inhibition effects on tumor growth at the dosage of 1.2mg/kg and 0.8mg/kg. For in vivo toxicity experiments of α-MMC in Balb/C mice, the minimal toxic dose of α-MMC was 1.2mg/kg. Alpha-MMC induced apoptosis by increasing caspase3 activities, and the cell cycle was arrested at the G0/G1 or G2/M phases. The measurements of IC50 were 15.07μg/mL, 33.66μg/mL, 42.94μg/mL for MDA-MB-231, MCF-7 and MDA-MB-453 respectively. Alpha-MMC exhibits anti-tumor effects in human breast cancer in vivo and in vitro. It inhibits breast cancer cells through the inhibition of tumor growth and induction of cell apoptosis. However, due to its obvious toxicity, α-MMC has a relatively narrow therapeutic window in vivo.

Conjugated linolenic acids (CLnA, super CLA)--natural sources and biological activity

Polyunsaturated fatty acids (PUFA) have a wide range of biological activity. Among them conjugated fatty acids are of great interest. Conjugated linoleic acids (CLA), which exert a multidirectional health-benefiting influence, and conjugated linolenic acids (CLnA, super CLA) are examples of this group of fatty acids. CLnA are a group of positional and geometric isomers of octadecatrienoic acid (C18:3), which possess double bonds at positions 9, 11, 13 or 8, 10, 12 of their chain. Some vegetable oils are rich sources of CLnA, e.g. bitter melon oil (from Momordica charantia seeds) and pomegranate oil (from Punica granatum seeds). The aim of this paper was to present information concerning natural sources and health-promoting activities of conjugated linolenic acids. The presented data reveal that conjugated linolenic acids may be very useful in prevention and treatment of many diseases, especially diabetes, arteriosclerosis , obesity and cancers (mammary, prostate and colon cancer). Among many potential mechanisms of their action, the fact that some CLnA are converted by oxidoreductases into CLA is very important. It seems to be very reasonable to conduct research concerning the possibility of CLnA use in prevention of many diseases.

Two new cucurbitane triterpenoids from the seeds of Momordica charantia

Two new cucurbitane triterpenoids 1 and 2 were isolated, together with six known compounds, from the seeds of Momordica charantia L. The structures of new compounds were determined to be 3-O-{[β-d-galactopyranosyl(1 → 6)]-O-β-d-galactopyranosyl}-23(R), 24(R), 25-trihydroxycucur-bit-5-ene (1), 3-O-[β-d-galactopyranosyl]-25-O-β-d-galactopyranosyl-7(R), 22(S), 23(R), 24(R), 25-pentahydroxycucurbit-5-ene (2), respectively. Their structures were elucidated by the combination of mass spectrometry, one- and two-dimensional NMR experiments and chemical reactions.

Preferential cytotoxicity of the type I ribosome inactivating protein alpha-momorcharin on human nasopharyngeal carcinoma cells under normoxia and hypoxia

All primary nasopharyngeal carcinoma (NPC) tumors contain hypoxic regions which are implicated in decreased local control and increased distant metastases, as well as resistance to chemotherapy in advanced NPC patients. One of the promising therapeutic approaches for NPC is to use drugs that can target hypoxic factors in tumors. In the present investigation, the type I ribosome inactivating protein α-momorcharin (α-MMC), isolated from seeds of the bitter gourd Momordica charantia, reduced cell viability and inhibited clonogenic formation of human NPC CNE2 and HONE1 cells under normoxia and cobalt chloride-induced hypoxia. By comparison, α-MMC exhibited only slight cytotoxicity on human nasopharyngeal epithelial NP69 cells under normoxia. Interestingly, α-MMC suppressed the expression levels of hypoxia-inducible factor 1-alpha (HIF1α) and vascular endothelial growth factor (VEGF) in hypoxic NPC, as well as the growth of human umbilical vein endothelial cells. Further study disclosed that α-MMC targeted endoplasmic reticulum and down-regulated unfolded protein response (UPR) in NPC cells. Moreover, α-MMC induced apoptosis in NPC cells in a dose- and time-dependent manner. It initiated mitochondrial- and death receptor-mediated apoptotic signaling in CNE2 cells, but there was hardly any effect on HONE1 cells. In addition, α-MMC brought about G0/G1 phase cell cycle arrest in CNE2 cells and S phase arrest in HONE1 cells. Collectively, α-MMC preferentially exhibited inhibitory effect on normoxic and hypoxic NPC cells partly by blocking survival signaling (e.g. HIF1α, VEGF and UPR), and triggering apoptotic pathways mediated by mitochondria or death receptor. These observations indicate the potential utility of α-MMC for prophylaxis and therapy of NPC.

Two New Bidesmoside Triterpenoid Saponins from the Seeds of Momordica charantia L.

Two new bidesmoside triterpenoid saponins which were identifed as 28-O-β-d-xylopyranosyl(1→3)-β-d-xylopyranosyl(1→4)-α-l-rhamnopyranosyl(1→2)-[α-l-rhamno-pyranosyl(1→3)]-β-d-fucopyranosyl gypsogenin 3-O-β-d-glucopyranosyl (1→2)-β-d-glucopyranosiduronic acid (C1) and 28-O-β-d-xylopyranosyl(1→4)-α-l-rhamnopyranosyl (1→2)-[α-l-rhamnopyranosyl(1→3)]-β-d-fucopyranosyl gypsogenin 3-O-β-d-gluco-pyranosyl(1→2)-β-d-glucopyranosiduronic acid (C2) were isolated together with two known compounds from the seeds of Momordica charantia L. Their structures were elucidated by the combination of mass spectrometry (MS), one and two-dimensional NMR experiments and chemical reactions.

Phytochemical and medicinal investigations ofMomordica charantia

Crude ethanolic extracts of Momordica charantia fruit and seeds were analyzed for antibacterial, antifungal and phytotoxic properties. Antibacterial activity was carried out using well diffusion method. Ethanolic fruits and seeds extract of Momordica charantia were found effective against all bacterial species studied. Momordica charantia fruit and seed extracts showed moderate antifungal activity against Alternaria alternata, Fusarium oxysporum and Trichoderma harizanum. From the results obtained it is clear that the extracts of Momordica charantia could further be exploited for isolation of active ingredients which could be utilized in medicinal formulations to develop cheaper natural drugs against bacterial and fungal infections. Crude ethanolic extracts of Momordica charantia also showed moderate phytotoxic activity at a concentration of 1000 μg/L, encouraging their utilization in production of eco-friendly herbicides.

Extract of Momordica charantia (Karela) Seeds as Corrosion Inhibitor for P110SS Steel in Co2 Saturated 3.5% NaCl Solution

Extract of Momordica charantia (Karela) Seeds as Corrosion Inhibitor for P110SS Steel in Co2 Saturated 3.5% NaCl Solution

Didecanoate compound: Isolated from Momordica charantia Linn. seeds from Nigeria

The seeds of Momordica charantia were extracted with hexane, ethyl acetate and 95% ethanol successively by percolation and concentrated at 37°C. Fractions 19-21 and 247 gave two compounds. However, only one has been elucidated to be 4a-phorbol-12, 13-didecanoate using the state- of –art- tools of spectrometry. The phytochemical screening of seeds of M. charantia showed the presence of flavonoids, glycosides, sterols, fat and oil in hexane, ethyl acetate and ethanol extracts. Anthraquinone is only present in hexane and ethyl acetate while alkaloid is in hexane and ethanol. And also phyto-medicine should be integrated into the health system of Nigeria and developed in such a way to bring harmony between the traditional and modern system of health care with minimum threat to each other. Key words: Momordica charantia, Cucurbitaceae, gas chromatography, thin layer chromatography, phytochemicals.

Phytochemicals, anti-microbial and free radical scavenging activities of Momordica charantia Linn (Palisota Reichb) seeds

Extractives from the seeds of Momordica charantia Linn were screened for phytochemical, antioxidant and anti-microbial activities against Escherichia coli, Candida albicans, Staphylococcus aureus, Staphylococcus epidedermis, Klebsiella pneumonia and Proteus mirabilis. The phytochemical screening of Momordica charantia seeds showed the presence of flavonoids, glycosides, sterols, fat and oil in hexane, ethyl acetate and ethanol extracts. At concentration of 1 mg/ml the radical scavenging activities of the plant extract was comparable to that of vitamin C used as control but at lower concentrations, the scavenging activity decreases. Therefore, it is a concentration dependent. The crude extract was found active against E. coli at 325 μg/ml, C. albicans at 162.5 μg/ml, S. aureus at 325 μg/ml, S. epidedermis at 500 μg/ml and K. pneumonia at 1500 μg/ml. These results suggest that the active principles in the extract possess broad spectrum antimicrobial activity, thus the plant could be used as a crude herbal drug. Key words: Momordica charantia, diphenyl-1-picrylhyrazyl radical (DPPH), inhibition, Cucurbitaceae, phytochemicals.

Enhanced resistance to blast fungus in rice (Oryza sativa L.) by expressing the ribosome-inactivating protein alpha-momorcharin

Rice blast caused by Magnaporthe grisea is one of the three major diseases that seriously affect the rice production. Alpha-momorcharin (α-MC), a ribosome-inactivating protein (RIP) isolated from Momordica charantia seeds, has antifungal effects in vitro. In this study, the α-MC gene was constitutively expressed under the control of the 2×35S promoter in transgenic rice (Oryza sativa L.) using an Agrobacterium tumefaciens-mediated method. The nine transgenic plants were obtained and confirmed by PCR and RT-PCR, and the four (B2, B4, B7 and B9) of them whose copy numbers were 1, 2, 3 and 3, respectively, were shown to express the α-MC protein by Western blot. The molecular weight of α-MC in transgenic plants was approximately 38kDa larger than the purified α-MC protein (28kDa) in vitro. When the confirmed T1 generations were inoculated with a suspension of M. grisea spores for ten days, the lesions on leaves of transgenic plants were much lesser than those found on wild type (WT). According to the criteria of International Rice Research Institute standard, the mean values for morbidity and disease index numbers were 29.8% and 14.9%, respectively, which were lower than for WT. It is unclear whether RIPs could impact plant fitness and however our results suggest that the α-MC protein is an effective antifungal protein preventing rice blast in transgenic rice.

Novel Inhibitor Cystine Knot Peptides from Momordica charantia

Two new peptides, MCh-1 and MCh-2, along with three known trypsin inhibitors (MCTI-I, MCTI-II and MCTI-III), were isolated from the seeds of the tropical vine Momordica charantia. The sequences of the peptides were determined using mass spectrometry and NMR spectroscopy. Using a strategy involving partial reduction and stepwise alkylation of the peptides, followed by enzymatic digestion and tandem mass spectrometry sequencing, the disulfide connectivity of MCh-1 was elucidated to be CysI-CysIV, CysII-CysV and CysIII-CysVI. The three-dimensional structures of MCh-1 and MCh-2 were determined using NMR spectroscopy and found to contain the inhibitor cystine knot (ICK) motif. The sequences of the novel peptides differ significantly from peptides previously isolated from this plant. Therefore, this study expands the known peptide diversity in M. charantia and the range of sequences that can be accommodated by the ICK motif. Furthermore, we show that a stable two-disulfide intermediate is involved in the oxidative folding of MCh-1. This disulfide intermediate is structurally homologous to the proposed ancestral fold of ICK peptides, and provides a possible pathway for the evolution of this structural motif, which is highly prevalent in nature.

Alpha-momorcharin: A Ribosome-Inactivating Protein from Momordica charantia, Possessing DNA Cleavage Properties

Ribosome-inactivating proteins (RIPs) function to inhibit protein synthesis through the removal of specific adenine residues from eukaryotic ribosomal RNA and rending the 60S subunit unable to bind elongation factor 2. They have received much attention in biological and biomedical research due to their unique activities toward tumor cells, as well as the important roles in plant defense. Alpha-momorcharin (α-MC), a member of the type I family of RIPs, is rich in the seeds of Momordica charantia L. Previous studies demonstrated that α-MC is an effective antifungal and antibacterial protein. In this study, a detailed analysis of the DNase-like activity of α-MC was conducted. Results showed that the DNase-like activity toward plasmid DNA was time-dependent, temperature-related, and pH-stable. Moreover, a requirement for divalent metal ions in the catalytic domain of α-MC was confirmed. Additionally, Tyr(93) was found to be a critical residue for the DNase-like activity, while Tyr(134), Glu(183), Arg(186), and Trp(215) were activity-related residues. This study on the chemico-physical properties and mechanism of action of α-MC will improve its utilization in scientific research, as well as its potential industrial uses. These results may also assist in the characterization and elucidation of the DNase-like enzymatic properties of other RIPs.

Momordica charantia and Its Novel Polypeptide Regulate Glucose Homeostasis in Mice via Binding to Insulin Receptor

Momordica charantia (MC) has been used as an alternative therapy for diabetes mellitus. This study analyzed and elucidated therapeutic targets contributing to the hypoglycemic effect of aqueous extract of MC seeds (MCSE) by transcriptomic analysis. Protein ingredients aimed at the hypoglycemic target were further identified by proteomic, docking, and receptor-binding assays. The data showed that MSCE (1 g/kg) significantly lowered the blood glucose level in normal and diabetic mice. Moreover, MCSE primarily regulated the insulin signaling pathway in muscles and adipose tissues, suggesting that MCSE might target insulin receptor (IR), stimulate the IR-downstream pathway, and subsequently display hypoglycemic activity in mice. It was further revealed that inhibitor against trypsin (TI) of MC directly docked into IR and activated the kinase activity of IR in a dose-dependent manner. In conclusion, the findings suggested that MCSE regulated glucose metabolism mainly via the insulin signaling pathway. Moreover, TI was newly identified as a novel IR-binding protein of MC that triggered the insulin signaling pathway via binding to IR.

Alpha-momorcharin, a RIP produced by bitter melon, enhances defense response in tobacco plants against diverse plant viruses and shows antifungal activity in vitro

Alpha-momorcharin (α-MMC) is type-1 ribosome inactivating proteins (RIPs) with molecular weight of 29kDa and has lots of biological activity. Our recent study indicated that the α-MMC purified from seeds of Momordica charantia exhibited distinct antiviral and antifungal activity. Tobacco plants pre-treated with 0.5mg/mL α-MMC 3days before inoculation with various viruses showed less-severe symptom and less reactive oxygen species (ROS) accumulation compared to that inoculated with viruses only. Quantitative real-time PCR analysis revealed that the replication levels of viruses were lower in the plants treated with the α-MMC than control plants at 15days post inoculation. Moreover, the coat protein expression of viruses was almost completely inhibited in plants which were treated with the α-MMC compared with control plants. Furthermore, the SA-responsive defense-related genes including non-expressor of pathogenesis-related genes 1 (NPR1), PR1, PR2 were up-regulated and activities of some antioxidant enzymes including superoxide dismutase (SOD), catalase (CAT), peroxidase (POD) were increased after the α-MMC treatment. In addition, the α-MMC (500μg/mL) revealed remarkable antifungal effect against phytopathogenic fungi, in the growth inhibition range 50.35-67.21%, along with their MIC values ranging from 100 to 500μg/mL. The α-MMC had also a strong detrimental effect on spore germination of all the tested plant pathogens along with concentration as well as time-dependent kinetic inhibition of Sclerotinia sclerotiorum. The α-MMC showed a remarkable antiviral and antifungal effect and hence could possibly be exploited in crop protection for controlling certain important plant diseases.

In Vitro Anti-diabetic Activities and Chemical Analysis of Polypeptide-k and Oil Isolated from Seeds of Momordica charantia (Bitter Gourd)

The amino acid and fatty acid composition of polypeptide k and oil isolated from the seeds of Momordica charantia was analysed. The analysis revealed polypeptide k contained 9 out of 11 essential amino acids, among a total of 18 types of amino acids. Glutamic acid, aspartic acid, arginine and glycine were the most abundant (17.08%, 9.71%, 9.50% and 8.90% of total amino acids, respectively). Fatty acid analysis showed unusually high amounts of C18-0 (stearic acid, 62.31% of total fatty acid). C18-1 (oleic acid) and C18-2 (linoleic acid) were the other major fatty acid detected (12.53% and 10.40%, respectively). The oil was devoid of the short fatty acids (C4-0 to C8-0). Polypeptide k and oil were also subjected to in vitro α-glucosidase and α-amylase inhibition assays. Both polypeptide k and seed oil showed potent inhibition of α-glucosidase enzyme (79.18% and 53.55% inhibition, respectively). α-Amylase was inhibited by 35.58% and 38.02%, respectively. Collectively, the in vitro assay strongly suggests that both polypeptide k and seed oil from Momordica charantia are potent potential hypoglycemic agents.

Preparation of an antitumor and antivirus agent: chemical modification of α-MMC and MAP30 from Momordica Charantia L. with covalent conjugation of polyethyelene glycol

Alpha-momorcharin (α-MMC) and momordica anti-HIV protein (MAP30) derived from Momordica charantia L. have been confirmed to possess antitumor and antivirus activities due to their RNA-N-glycosidase activity. However, strong immunogenicity and short plasma half-life limit their clinical application. To solve this problem, the two proteins were modified with (mPEG)(2)-Lys-NHS (20 kDa). In this article, a novel purification strategy for the two main type I ribosome-inactivating proteins (RIPs), α-MMC and MAP30, was successfully developed for laboratory-scale preparation. Using this dramatic method, 200 mg of α-MMC and about 120 mg of MAP30 was obtained in only one purification process from 200 g of Momordica charantia seeds. The homogeneity and some other properties of the two proteins were assessed by gradient SDS-PAGE, electrospray ionization quadruple mass spectrometry, and N-terminal sequence analysis as well as Western blot. Two polyethylene glycol (PEG)ylated proteins were synthesized and purified. Homogeneous mono-, di-, or tri-PEGylated proteins were characterized by matrix-assisted laser desorption ionization-time of flight mass spectrometry. The analysis of antitumor and antivirus activities indicated that the serial PEGylated RIPs preserved moderate activities on JAR choriocarcinoma cells and herpes simplex virus-1. Furthermore, both PEGylated proteins showed about 60%-70% antitumor and antivirus activities, and at the same time decreased 50%-70% immunogenicity when compared with their unmodified counterparts. α-MMC and MAP30 obtained from this novel purification strategy can meet the requirement of a large amount of samples for research. Their chemical modification can solve the problem of strong immunogenicity and meanwhile preserve moderate activities. All these findings suggest the potential application of PEGylated α-MMC and PEGylated MAP30 as antitumor and antivirus agents. According to these results, PEGylated RIPs can be constructed with nanomaterials to be a targeting drug that can further decrease immunogenicity and side effects. Through nanotechnology we can make them low-release drugs, which can further prolong their half-life period in the human body.

Gene cloning and expression of a novel hypoglycaemic peptide from Momordica charantia

Momordica charantia (MC) is used in many Asian countries as a traditional functional food and medicine. Polypeptide-P, a 166 amino acid (AA) polypeptide isolated from MC seeds, has been reported to show hypoglycaemic effects in patients with type I or type II diabetes. The AA sequence of this peptide has been determined, but its gene sequence has yet to be published. In this study a gene-cloning strategy was employed to obtain the polypeptide-P gene sequence using degenerate reverse transcription polymer chain reaction and genome-walking methods. A complete 498 bp sequence encoding the polypeptide-P protein was cloned from MC seeds. Subsequent assays of the bioactivity of the expressed recombinant protein revealed that it had significant hypoglycaemic activity in alloxan-induced diabetic mice. This result suggests that recombinant polypeptide-P has hypoglycaemic effects. This is the first report of cloning and expression of the MC polypeptide-P gene. The cloned gene could be helpful for exploring the mechanisms of polypeptide-P gene expression and regulation in MC. Furthermore, this gene could be used as a potential tool both for screening MC varieties with high hypoglycaemically active substance content and for breeding new varieties of MC with high economic value, which could in turn be beneficial to farmers.