Indian Mustard Seeds Research Articles

Impact Assessment of Lead-Tolerant Rhizobacteria to Improve Soil Health Using Indian Mustard (Brassica juncea) as an Indicator Plant.

Due to ongoing human activities, heavy metals are heavily accumulated in the soil. This leads to an increase in the discharge and the quick spread of heavy metal pollution in human settlements and natural habitats, having a disastrous effect on agricultural products. The current experiment was planned to evaluate the effect of lead-tolerant-plant-growth-promoting rhizobacteria (LTPGPR) on growth, yield, antioxidant activities, physiology, and lead uptake in the root, shoot, and seed of Indian mustard (Brassica juncea) in lead-amended soil. Three pre-isolated well-characterized lead-tolerant rhizobacterial strains-S10, S5, and S2-were used to inoculate seeds of Indian mustard grown at three different levels of lead (300 mg kg-1, 600 mg kg-1, 900 mg kg-1) contaminated soil. The experiment was designed following a completely randomized design (CRD) under factorial arrangements. Lead nitrate was used as a source of lead contamination. At harvesting, data regarding growth, physiology, yield per plant, antioxidant activities, malondialdehyde and proline content, and lead uptake in the root, shoot, and seed of Indian mustard were recorded. Results demonstrated that lead contamination at all levels significantly reduced the plant growth, yield, and physiological processes. Plants inoculated with lead-tolerant rhizobacteria showed a significant improvement in plant growth, yield, antioxidant activities, and physiological attributes and cause a valuable reduction in the malondialdehyde contents of Indian mustard in lead-contaminated soil. Moreover, plants inoculated with lead-tolerant rhizobacteria also showed an increment in lead uptake in the vegetative parts and a significant reduction of lead contents in the seed of Indian mustard.

Nutritional quality evaluation of oil and fatty acid profile in various genotypes/varieties of Indian mustard [Brassica juncea Czern & Coss (L.)

Brassicas are one of the most agronomically eminent oilseeds that are employed as a variety of oilseed, vegetable, and fodder crops. The experiment was conducted with thirty-four genotypes/varieties seeds of Indian mustard [Brassica juncea Czern & Coss (L.)] for oil content, Iodine value, oil stability index and fatty acid composition during 2018-2019. The experiment was laid out in completely randomized design with three replications. The range of variability of contents of oil, palmitate, stearate, oleate, linoleate, linolenate, ecosenate, doecosenate, iodine value and oil stability index varied from 33.52 to 42.15%, 1.53 to 4.98%, 0.16 to 2.71%, 5.06 to 17.78%, 17.88 to 32.15%, 11.82 to 19.85%, 5.44 to 11.89%, 28.82 to 47.66%, 114.43 to 131.71 and 1.08 to 1.99, respectively. The Brassica juncea genotype-KMR-15-6 followed by genotype- KMR-17-6 had the higher oil content, oleic acid content and low value of erucic acid which indicates that seed oil this B. juncea species genotype is possibly suitable for both human consumption and industrial purposes.

Attenuating effect of Indian mustard (Brassica juncea) seed and its nano formulation on arsenic induced-oxidative stress and associated genotoxicity in rat

ObjectiveArsenic (As) has been in the forefront of toxicological research due to its continual worldwide human exposure and its adverse effects including genotoxicity. The study assessed the potential ameliorative effect of Indian mustard seeds (Brassica juncea) and their nanoparticles against arsenic toxicity. MethodsMale Wistar rats were used for the exposure study. Group I was control and group V was exposed only to the nanoparticles. Groups II, III, and IV were exposed to 10 mg/kg sodium arsenite for two weeks. Groups III and IV were treated with mustard seeds and the nanoparticles (500 mg/kg) respectively. ResultsArsenic intoxication caused a marked increase in the levels of As in the brain. Additionally, oxidative damage was exhibited with significantly enhanced levels of serum 8-OHdG and hepatic p53 and pS15. The pictographs and the analyzed parameters of the COMET assay illustrated DNA damage with significantly increased values of tail moment and length. Treatment with mustard seeds and its nanoparticles markedly reversed the adverse effects of As toxicity, which is attributed to the presence of a gamut of constituent phytocompounds in the mustard seeds. ConclusionTaken together the nano formulation was more efficacious and could be introduced as a potential nutraceutical to combat metal toxicity.

Biochemical responses to natural and accelerated ageing on seed quality in Indian mustard (Brassica juncea)

Indian mustard [Brassica juncea (L.) Czern. & Coss.] is an important oilseed crop of many tropical and subtropicalcountries of the world. The present study was carried out at research farm of Department of Seed Scienceand Technology, CCS Haryana Agricultural University, Hisar during winter (rabi) season 2017–18 and 2018–19.The objective of present study was to evaluate the effect of natural and accelerated aging on biochemical constitutes in 3 Indian mustard varieties, viz. RH-30, RH-9304 and RH-0406 with three seed lots (fresh seed, 1-year old seed, 2-year-old seed). Naturally aged seed lots of variety RH-30 was found to meet out Indian Minimum Seed Certification Standard after 1-year under ambient condition. Tetrazolium test indicated that the viability of Indian mustard seeds decreased after increasing the ageing duration. Maximum seed viability recorded in the variety RH-30 and minimum was recorded in variety RH-0406. Activities of all the enzymes, viz. superoxide dismutase, dehydrogenase and peroxidase significantly decreased with natural ageing; RH-30 was found to be a good storer variety. The maximum performance for standard germination and enzymes activity was observed in fresh seed lots. Results showed that among different Indian mustard varieties, RH-30 and RH-9304 were found best responsive with minimal seed deterioration effects, and better germination and seedling establishment potential.

Seed longevity and vigour studies in different types of Indian mustard

An experiment was conducted at ICAR-Indian Agricultural Research Institute, New Delhi (2017-18) to study the possible reason for reduced seed vigour in Indian quality mustard genotypes during storage for the first time. Freshly harvested seeds, one year, two year aged seeds and controlled deteriorated seeds of freshly harvested seeds of conventional and quality Indian mustard were used for the study. Reactive oxygen species (ROS), i.e. hydrogen peroxide (H2O2) and superoxide radical (O2 .-) were estimated along with enzymatic antioxidants such as catalase and peroxidase and redox couples ratio (ASC/DHA and GSH/GSSG) in all the nine genotypes of different age group and correlated with germination percentage, mean germination time and seed vigour indices. The experiment revealed that during storage, the amount of ROS accumulation goes on increasing and both enzymatic and non-enzymatic antioxidants goes on decreasing. In comparison to conventional genotypes, quality genotypes were found to contain significantly lower level of antioxidants and significantly higher level of ROS content during the storage period. Germination percentage as well as seed vigour indices were negatively correlated with ROS and positively correlated with enzymatic and non-enzymatic antioxidants. This experiment revealed that reduced vigour in quality genotypes could be due to faster deterioration during the storage due to increased production of ROS.

Effect of microwave pre-treatment on quality parameters in Indian mustard.

Demand for edible oil from crops like groundnut, mustard and soybean, is increasing and likely to increase further. In this regard, Indian mustard seeds were treated with microwaves (MW), to investigate the possibility of enhancing oil yield and nutritional content. MW pre-treatment was given to the seeds of two Indian mustard varieties i.e. PM21 (V1) and PDZ1 (V2) for 0, 2, 4 and 6min (H0, H2, H4 and H6 min respectively). MW treatment with increasing exposure time showed a linear reduction in the glucosinolate and erucic acid content with concomitant increase in oil yield and moisture loss in both the varieties, as evident from correlation and principal component analysis. Antioxidant capacity has increased with the reduction in phytic acid content (1.82) in V2 at 6min exposure time with respect to untreated control. Free radical scavenging activity was improved with increasing MW treatment in both the varieties. Therefore, from the obtained results, it is advisable to treat mustard seeds with MW before extraction of oil, because it gives a relatively good oil yield, with enhanced nutritional factors. Moreover, microwaving was effective in reducing glucosinolates and erucic acid also.

Deciphering the toxic effects of iprodione, a fungicide and malathion, an insecticide on thiol protease inhibitor isolated from yellow Indian mustard seeds

Pesticides are being used globally to improve agricultural production. They are applied specifically to combat with pathogens that are a major threat for reduced optimum yield of crops. This study was carried out to see the effect of commercially used pesticides on a specific plant protein viz. phytocystatin isolated from yellow mustard seeds (YMP). Phytocystatin is a thiol proteinase inhibitor, which regulates endogenous and exogenous cysteine proteinases and plays a vital physiological role in plants. Different classes of pesticides like fungicide (iprodione) of dicarboximide class and an insecticide (malathion) of class organophosphate are retorted for our study. In the presence of these pesticides, biophysical and biochemical changes were observed in phytocystatin. These changes were evaluated making use of caseinolytic activity assay, UV–vis spectroscopy, fluorescence spectroscopy, FTIR, and circular dichroism. Isothermal titration calorimetry was employed to see interaction pattern of these pesticides with phytocystatin. The results obtained clearly depict that the pesticides bind with the phytocystatin thereby changing its native conformation and reducing its intrinsic property of inhibition on cysteine proteinase as evident by reduced anti-papain inhibition in the presence of pesticides. Furthermore, CD and FTIR spectroscopy results clearly show a decrease in α-helical content upon interaction with malathion and iprodione. Among the two pesticides, iprodione has far more pronounced effect on YMP evident from striking changes in UV, Fluorescence, CD and FTIR spectroscopy. 2,4-dinitrophenylhydrazine spectrophotometric assay was also carried out to check production of ROS, generation of ROS was observed in the presence of these pesticides thus implying that ROS might be responsible for changes in native structure of phytocystatin induced by pesticides.

Imbibition behaviour and germination response in conventional and quality of Indian mustard (Brassica juncea) seeds

Indian mustard [Brassica juncea (L.) Czern and Coss] seeds are well known for their multipurpose uses, such as oilseeds, spices etc. Being extensively cultivated in northern and western part of the country it constitutes major share in country's oilseeds production. Developing cultivars having low erucic acid and glucosinolates (single zero and double zero) is a major objective of worldwide Brassica breeding. Such cultivars are also known as Quality Indian mustard. Seed; being vital input in agriculture, its quality determines the performance of a given genotype and efficiency of all other inputs too. Color variation among the seed coat is known to affect the rate of water uptake and thereby the vigor of seeds and these color variation is mainly because of pigments such as melanin and phenolics compounds. No such studies are reported in Indian mustard, though there is variability with respect to seed coat colour in both conventional and quality type of Indian mustard seeds. Thus the present investigation was carried out with a view to evaluate the imbibitional behaviour in black and yellow seeds of Indian mustard. The material had eight varieties of Indian mustard of which four genotypes each were of conventional and quality types. The seed of these genotypesvaried for seed coat color, i.e. four black seeded and four yellow seeded genotypes. Thus, the material has two genotypes each with black and yellow seed coat from conventional and quality groups. We studied the rate of imbibition in black and yellow seeds and correlated them with the melanin and phenol content. We found that the black seeds were having higher amounts of melanin and phenol content and thus imbibing slowly than yellow seeds. On the other hand, faster imbibition in yellow seeds leads to imbibitional injury which is known to reduce the vigor in Brassica. Use of controlled deterioration revealed the proneness of yellow seeds to ageing and thus reduction in vigor of such seeds. Few yellow seeded cultivars showed resistance to such deterioration and thus could be used in improvement programme in Brassica.

Phospholipid Hydrolysis and Transphosphatidylation by Phospholipase D from Indian Mustard Seeds in Two‐Phase Systems

AbstractPhospholipase D (PLD), isolated from Indian mustard seeds and purified to homogeneity, has recently been identified as typical α‐type PLD with a high activity toward phosphatidylcholine (PC) in an aqueous mixed micellar system (Khatoon et al. 2014, Phytochemistry 117, 65–75). In light of biocatalytical application, we have now studied the enzyme in aqueous‐organic two‐phase systems and compared the results with the properties of the enzyme in aqueous mixed micellar systems. n‐Hexane containing 5–10 mol% of 1‐ or 2‐octanol proved to be optimal as an organic solvent in the two‐phase system, whereas anionic detergents such as sodium dodecyl sulfate or sodium deoxycholate were favorable components in the mixed micellar system. The optimum pH value (5.5–5.6) and the optimum Ca2+ ion concentration (70 mM) were independent of the reaction system. The head group selectivity in terms of activity toward different phospholipids (PC > phosphatidylethanolamine > phosphatidylglycerol) was similar in both types of reaction systems. Also, the KM values toward PC were on the same order of magnitude. In contrast, the Vmax value in the two‐phase system was 20‐fold lower than in the mixed micellar system. The enzyme was able to catalyze the substitution of choline in PC by ethanolamine, glycerol, and ethylene glycol with high efficiency. l‐Serine was exchanged for choline with low activity. Myo‐inositol was not an alcohol acceptor, but promoted the hydrolysis of PC.

Biomass Quality of Brassica Oilseed Crops in Response to Sulfur Fertilization

Rapeseed [Brassica napus (L.)], white mustard [Sinapis alba (L.)], and Indian mustard [Brassica juncea (L.) Czern. et Coss] belong to the family Brassicaceae. These crop species synthesize substantial amounts of fat, protein, and glucosinolates (GLS). Brassica species can differ in their concentrations of fat, protein, and GLS in response to S fertilization. Plant material for chemical analyses was harvested from a field experiment conducted in northeastern Poland in 2005 to 2008. The examined species of oilseed crops were fertilized or not fertilized with S applied to soil at optimal rates for each species based on S uptake and forecast seed yield. On average, S fertilization increased seed yield of oilseed crops by 8%, but it did not induce significant differences in the concentrations of fat and protein in seeds. Sulfur fertilization increased total GLS concentrations by 0.22 and 0.27 µmol g−1 dry matter (DM) in the root residues of white mustard and Indian mustard, respectively, and by 0.52 µmol g−1 DM in winter rapeseed. Total GLS concentration in winter rapeseed straw decreased by 0.18 µmol g−1 DM with S application. In white mustard straw, S fertilization increased total GLS concentration by 0.27 µmol g−1 DM. Sulfur fertilization increased total GLS concentrations in the seeds of spring rapeseed, white mustard and Indian mustard by 11.43, 41.75 and 32.05 µmol g−1 DM, respectively. Sulfur fertilization has a significant effect on the seed yield and biomass quality of oilseed crops of the family Brassicaceae.

The effect of sulphur fertilization on macronutrient concentrations in the post-harvest biomass of mustard

The aim of this study was to determine the effect of sulphur (S) fertilization on macronutrient concentrations in the roots, straw and oil cake of white mustard and Indian mustard. The experiment was conducted in northeastern Poland (2006-2008). The highest content of N, P, K, Mg and S was noted in the oil cake of both mustard species. Sulphur fertilization increased S concentrations and had no effect on Mg concentrations in the roots of white mustard and Indian mustard. Sulphur fertilizers applied to soil significantly increased the content of N, K and Ca in the roots of Indian mustard, whereas they had no significant effect on the content of N, P and Ca, and decreased K concentrations in the roots of white mustard. Sulphur fertilization led to a significant decrease in N content, and an increase in the content of K and Ca in the straw of both mustard species. Sulphur fertilization significantly decreased P concentrations and increased S concentrations in the oil cake of both mustard species. In response to S fertilization, the content of N and K decreased in the oil cake of Indian mustard, but did not change significantly in the oil cake of white mustard. Therefore, S fertilization significantly differentiated the fertilizing value of post-harvest residues and the feeding value of white mustard and Indian mustard seeds.

Purification, sequencing and characterization of phospholipase D from Indian mustard seeds

Phospholipase D (PLD; E.C. 3.1.4.4) is widespread in plants where it fulfills diverse functions in growth and in the response to stresses. The enzyme occurs in multiple forms that differ in their biochemical properties. In the present paper PLD from medicinally relevant Indian mustard seeds was purified by Ca2+-mediated hydrophobic interaction and anion exchange chromatography to electrophoretic homogeneity. Based on mass-spectrometric sequence analysis of tryptic protein fragments, oligonucleotide primers for cloning genomic DNA fragments that encoded the enzyme were designed and used to derive the complete amino acid sequence of this PLD. The sequence data, as well as the molecular properties (molecular mass of 92.0kDa, pI 5.39, maximum activity at pH 5.5–6.0 and Ca2+ ion concentrations ⩾60mM), allowed the assignment of this enzyme to the class of α-type PLDs. The apparent kinetic parameters Vmax and Km, determined for the hydrolysis of phosphatidylcholine (PC) in an aqueous mixed-micellar system were 356±15μmolmin−1mg−1 and 1.84±0.17mM, respectively. Phosphate analogs such as NaAlF4 and Na3VO4 displayed strong inhibition of the enzyme. Phosphatidylinositol 4,5-bisphosphate had a strong activating effect at 2–10mM CaCl2. PLD was inactivated at temperatures >45°C. The enzyme exhibited the highest activity toward PC followed by phosphatidylethanolamine and phosphatidylglycerol. PCs with short-chain fatty acids were better substrates than PCs with long fatty acid chains. Lyso-PC was not accepted as substrate.

Phytoextract of Indian mustard seeds acts by suppressing the generation of ROS against acetaminophen-induced hepatotoxicity in HepG2 cells

Context: Indian mustard [Brassica juncea (L.) Czern. & Coss. (Brassicaceae)] is reported to possess diverse pharmacological properties. However, limited information is available concerning its hepatoprotective activity and mechanism of action.Objective: To study the protective mechanism of mustard seed extract against acetaminophen (APAP) toxicity in a hepatocellular carcinoma (HepG2) cell line.Materials and methods: Hepatotoxicity models were established using APAP (2.5–22.5 mM) based on the cytotoxicity profile. An antioxidant-rich fraction from mustard seeds was extracted and evaluated for its hepatoprotective potential. The mechanism of action was elucidated using various in vitro antioxidant assays, the detection of intracellular generation of reactive oxygen species (ROS), and cell cycle analysis. The phytoconstituents isolated via HPLC-DAD were also evaluated for hepatoprotective activity.Results: Hydromethanolic seed extract exhibited hepatoprotective activity in post- and pre-treatment models of 20 mM APAP toxicity and restored the elevated levels of liver indices to normal values (p < 0.05). Post-treatment suppressed the generation of ROS by 58.37% and pre-treatment effectively prevented the generation of ROS by 90.5%. The mechanism of ROS suppression was further supported by antioxidant activity (IC50) data from DPPH (103.37 ± 4.2 µg AAE/mg), FRAP (83.26 ± 1.1 µg AAE/mg), ORAC (1115 µM GAE/ml), ABTS (83.05 µg GAE/ml), and superoxide (345.22 ± 5.15 µg AAE/mg) scavenging assays and by the restoration of cell cycle alterations. HPLC-DAD analysis revealed the presence quercetin, vitamin E, and catechin, which exhibited hepatoprotective activity.Discussion and conclusions: A phytoextract of mustard seeds acts by suppressing the generation of ROS in response to APAP toxicity.

Antioxidants from defatted Indian Mustard (Brassica Juncea) protect biomolecules against in vitro oxidation.

Indian mustard seeds were defatted by distillation with hexane and the residue extracted with methanol was analyzed for potential antioxidants; ascorbate, riboflavin, and polyphenols. Gallic acid (129.796μg), caffeic acid (753.455μg), quercetin (478.352μg) and kaempferol (48.060μg)/g dry seeds were identified by HPLC analysis of the extract. DPPH free radical scavenging activity and protection of lipids, proteins and DNA against metal induced oxidation was examined. Defatted mustard seed remnant had excellent free radical scavenging activity and protects biomolecules with IC50 value 2.0-2.25mg dry seed weight. Significant content of polyphenols in methanol extract of defatted seeds accounts for high antioxidant potential. We are the first to report the detailed analysis of antioxidant composition and protection of biomolecules against oxidative damage by methanol extract of mustard seed remnant after oil extraction.

Separation and Purification of Sinigrin and Gluconapin from Defatted Indian Mustard Seed Meals by Macroporous Anion Exchange Resin and Medium Pressure Liquid Chromatography

Defatted seed meals, the byproducts of Indian mustard seed oil industry, which are just used as animal feedstock and nitrogen fertilizers, possess various nutrients and phytochemical compounds, such as sinigrin and gluconapin. The present study demonstrated a novel, low-cost, and efficient method for the co-production of sinigrin and gluconapin from defatted Indian mustard seed meals by D261 strong basic anion-exchange macroporous resin and medium pressure liquid chromatography (MPLC). The adsorption/desorption capacities of eight adsorbents characterized by BET, IR, and EDS were screened. 26.4 g defatted Indian mustard seed meals, which contained about 1210.3 mg sinigrin and 696.6 mg gluconapin, could produce 2059.6 mg of glucosinolate-rich extract of 53.67% sinigrin and 31.30% gluconapin after separation by D261 resin. Then, 949.40 mg of 97% sinigrin and 568.40 mg of 95% gluconapin could be obtained after the extracts were purified by MPLC, with the recovery of 76.03% of sinigrin and 77.38% of gluconapin. The products were assessed by analytical HPLC and characterized by UV, MS, 1H NMR, and 13C NMR. In conclusion, this method is practical and environmentally friendly, and it is a low-cost process to make full use of Indian mustard seeds.

Relation between soil temperature and biophysical parameters in Indian mustard seeds

Abstract Temporal changes in surface soil temperature were studied in winter crop. Significant changes in bare and cropped soil temperature were revealed. Air temperature showed a statistically positive and strong relationship (R2 = 0.79** to 0.92**) with the soil temperature both at morning and afternoon hours. Linear regression analysis indicated that each unit increase in ambient temperature would lead to increase in minimum and maximum soil temperatures by 1.04 and 1.02 degree, respectively. Statistically positive correlation was revealed among biophysical variables with the cumulative surface soil temperature. Linear and non-linear regression analysis indicated 62-69, 72-86 and 72-80% variation in Leaf area index, dry matter production and heat use efficiency in Indian mustard crop as a function of soil degree days. Below 60% variation in yield in Indian mustard was revealed as a function of soil temperature. In contrast, non-significant relationship between oil content and soil temperature was found, which suggests that oil accumulation in oilseed crops was not affected significantly by the soil temperature as an independent variable.

SURVIVAL TO EARLY TOXIC COPPER EXCESS: BIOCHEMICAL AND ANATOMICAL CHANGES DURING GERMINATION OF INDIAN MUSTARD

It is well-known that essential heavy metals like copper (Cu), mainly at higher concentrations usually cause overproduction of reactive oxygen species (ROS) resulting in oxidative stress in plants. Till date many experiments were carried out to evaluate how Cu toxicity influences in adult plants but only a few reports are available about the effects during germination. Since this is a very sensitive period and the effects of heavy metal stress are more serious. The aims of our study were to investigate potential oxidative stress and antioxidative defense mechanisms beside potential morphological and/or anatomical alterations in germinating seeds of Indian mustard ( Brassica juncea L. ) exposed to excess Cu. The following parameters were evaluated to describe oxidative stress: FRAP (ferric reducing ability of plasma), lipid peroxidation (LP), reduced glutathione content (GSH), total protein content and the activity of glutathione-S-transferase (GST), superoxide dismutase (SOD), catalase (CAT), guaiacol peroxidase (GPOX) and glutathione reductase (GR). We also made an assessement of histochemically LP and the loss of plasma membrane integrity in the root tips, the production of callose and the lignification of cell walls. Our results showed that Cu treatments were followed by notable GSH-depletion. We could detect LP histochemically in the root tips. The application of Cu increased the activity of SOD in time and dose-dependent manner. The activity of CAT and GPOX increased after 48-96h Cu excess. Morphological symptoms of metal toxicity occurred such as stunted, hooked-formed and brownish root tips. Production of callose and lignification of cell walls could be visualized, too.

Separation of sinigrin from Indian mustard (Brassica juncea L.) seed using macroporous ion-exchange resin

Sinigrin is a major glucosinolate present in Indian mustard (Brassica juncea L.) seeds as the precursor of the anticancer compound allyl isothiocyanate. In the present study, the adsorption and desorption characteristics of six macroporous ion-exchange resins for the separation of sinigrin from crude aqueous extracts have been compared. The results indicated that D261 resin showed the best adsorption and desorption capacity to sinigrin, and its adsorption data fit best to the Freundlich isotherm. The dynamic adsorption/desorption experiments were carried out to optimize the separation process. After treatment with D261 resin in one run, the purity of sinigrin in the product was increased 15.57-fold from 3.75% to 58.37% with the recovery of 79.82%. Meanwhile, the separation effect of D261 resin was also supported by UV and IR. The separation process using macroporous ion-exchange resin in our paper provides a novel, rapid and economical method for separation of sinigrin.

A novel technique to collect root exudates from mustard (Brassica juncea)

A very simple, novel, cost effective, easy to use technique has been developed for the collection of root exudates from small seeded plants, under laboratory conditions. 200-1000 μl micro tips (Tarsons), kept in 100 ml glass beakers, were used as holders for the small seeds of Indian mustard (Brassica juncea) and the exudates were trapped in liquid culture medium. The exudates, so obtained, were authenticated and analyzed for organic compounds such as sugars, amino acids and organic acids, as well as chemotactic response towards rhizobacteria. Method was found to be suitable and easy to handle for small seeds.

Optimization of ultrasonic‐stimulated solvent extraction of sinigrin from Indian mustard seed (Brassica Juncea L.) using response surface methodology

Sinigrin, a major glucosinolate present in Indian mustard (Brassica juncea L.) seeds as the precursor of the anticancer compound allyl isothiocyanate, shows a wide range of biological activities. It's necessary to optimize the extraction methods and conditions, in order to improve the extraction productivity and save raw material. To systemically investigate and optimize the most important factors affected the productivity of sinigrin in the process of extraction using response surface methodology. The ranges of three main factors including the ethanol concentration, extraction time and extraction temperature were selected by the one-factor-at-a-time method. The conditions of ultrasonic-stimulated extraction of sinigrin from defatted Indian mustard seed powder were optimized by Box-Behnken design to obtain the maximum productivity. The predicted productivity (3.81%) was obtained using 57% ethanol concentration at 81 °C for 60 min, with the coefficient of the model R² > 0.96 (n = 17). The actual productivity (3.84 ± 0.02%) of sinigrin under the optimized condition was increased by 70.67% compared with the result of conventional extraction. Meanwhile, HPLC, UV and IR were applied to examine if there is a difference between the ultrasonic-stimulated solvent extraction and conventional extraction, and the improvement of productivity of sinigrin depended on the destruction of cell wall caused by the elimination of outer pectinous material was explained by SEM and composition content analysis. The ultrasonic-stimulated solvent extraction was suggested to be a promising method to improve the productivity of sinigrin. And the results demonstrated that sinigrin productivity may be related to pectinous materials existed in the seeds.