Cotyledons Of Chickpea Research Articles

Impact of thermally induced wall breakage on the structural properties of water-soluble polysaccharides in chickpeas

The present work aimed to elucidate the influence of wall breakage induced by thermal processing on the molecular, structural, and antioxidant activities of water-soluble polysaccharides in chickpeas. Different extents of cell wall disruption were observed by fluorescence microscopy in chickpea cotyledons. Moreover, a decreasing fluorescence intensity of cell wall fragments was observed in the flour residues upon heat fluidization, autoclaving, and microwave heating, and the polysaccharide extraction rates were increased by 31.47%, 25.52%, and 9.79%, respectively. Furthermore, WPUCP, WPHCP, WPMCP, and WPACP (water-soluble polysaccharides from unprocessed, heat fluidized, microwaved, and autoclaved chickpeas, respectively) were RG-I (rhamnogalacturonan-I)-enriched pectic polysaccharides composed of galactose, arabinose, galacturonic acid, and rhamnose. After chickpea thermal processing, the degrees of branching decreased to 2.87, 3.79, and 2.53 in WPHCP, WPMCP, and WPACP, respectively, and the molecular weights were reduced by 46.46%, 24.83%, and 59.91%, respectively. Structural analysis showed that the semicrystalline regions of WPHCP, WPMCP, and WPACP were slightly damaged without changing the functional groups, but their thermal stability decreased. Interestingly, WPACP formed an ordered conformation (microporous network structure) through the formation of hydrogen bonds. Moreover, the antioxidant activities of WPHCP, WPMCP, and WPACP were enhanced, and the strongest radical scavenging activity was observed for WPHCP.

Genetic and environmental factors contribute to variation in cell wall composition in mature desi chickpea (Cicer arietinum L.) cotyledons.

Chickpea (Cicer arietinum L.) is an important nutritionally rich legume crop that is consumed worldwide. Prior to cooking, desi chickpea seeds are most oftendehulled and cleaved to release the split cotyledons, referred to as dhal. Compositional variation between desi genotypes has a significant impact on nutritional quality and downstream processing, and this has been investigated mainly in terms of starch and protein content. Studies in pulses such as bean and lupin have also implicated cell wall polysaccharides in cooking time variation, but the underlying relationship between desi chickpea cotyledon composition and cooking performance remains unclear. Here, we utilized a variety of chemical and immunohistological assays to examine details of polysaccharide composition, structure, abundance, and location within the desi chickpea cotyledon. Pectic polysaccharides were the most abundant cell wall components, and differences in monosaccharide and glycosidic linkage content suggest both environmental and genetic factors contribute to cotyledon composition. Genotype-specific differences were identified in arabinan structure, pectin methylesterification, and calcium-mediated pectin dimerization. These differences were replicated in distinct field sites and suggest a potentially important role for cell wall polysaccharides and their underlying regulatory machinery in the control of cooking time in chickpea.

Solid-state bioconversion of chickpea (Cicer arietinum L.) by Rhizopus oligosporus to improve total phenolic content, antioxidant activity and hypoglycemic functionality

The objective of this investigation was to study the effect of time during solid state bioconversion (SSB) on total phenolic content (TPC), antioxidant activity (AoxA), and inhibitory properties against α-amylase and α-glucosidase of chickpea. Chickpea cotyledons were inoculated with a suspension of Rhizopus oligosporus and incubated at 35 °C for 24, 36, 48, 60, 72, 84, 96 and 108 h. The best time to produce bioprocessed chickpea (added with seed coats) flour with the highest AoxA was 108 h. SSB substantially increased TPC and AoxA of chickpea extracts in 2.78 and 1.80–1.94 times, respectively. At 36 and 96 h of fermentation, the SSB process improved in vitro α-amylase and α-glucosidase inhibition (AI and GI indexes) activities of chickpea extracts in 83 and 370%, respectively. SSB is a good strategy to enhance health-linked functionality of chickpea, due to improved TPC, AoxA and content of strong natural inhibitors of enzymes associated with diabetes.

Optimization of regeneration and Agrobacterium-mediated transformation of immature cotyledons of chickpea (Cicer arietinum L.)

Immature cotyledons collected at different time intervals from four genotypes of chickpea (C 235, BG 256, P 362 and P 372) were cultured adaxially on Murashige and Skoog (MS) medium supplemented with 6-benzyladenine, thidiazuron, kinetin, zeatin and dimethylallylaminopurine (2-iP), either alone or in combination with indole-3-acetic acid (IAA) or α-napthoxyacetic acid (α-NOA) for dedifferentiation and regeneration of adventitious shoots. Morphogenesis was achieved with explants cultured adaxially on MS medium with 13.68 μM zeatin, 24.6 μM 2-iP, 0.29 μM IAA and 0.27 μM α-NOA. Explants prepared from pods of 21 days after pollination, responded favourably to plant growth regulator treatment in shoot differentiation. Histological studies of the regenerating explants, revealed the initiation of meristematic activity in the sub-epidermal region during the onset of morphogenesis, which can be correlated with elevated activity of cytokinin oxidase-dehydrogenase, for cytokinin metabolism. The regenerated shoots were efficiently rooted in MS medium supplemented with 2.46 μM indole-3-butyric acid and acclimatized under culture room and glasshouse conditions for normal plant development leading to 76–80 % survival of the rooted plantlets. The immature cotyledon explants were used for Agrobacterium-mediated transformation with critical manipulation of cultural conditions like age of explant, O.D. of Agrobacterium suspension, concentration of acetosyringone, duration of sonication and co-cultivation for successful genetic transformation and expression of the reporter gene uidA (GUS). Integration of transgene was confirmed by molecular analysis. Transformation frequency up to 2.08 % was achieved in chickpea, suggesting the feasibility of using immature cotyledon explants for Agrobacterium-mediated transformation.

Structural analysis of kappa-carrageenan isolated from Hypnea musciformis (red algae) and evaluation as an elicitor of plant defense mechanism

High molecular weight crude and purified polysaccharide fractions obtained from Hypnea musciformis, red algae were evaluated for their elicitor activity in terms of induced browning and phytoalexins production in the cotyledons of Chickpea and Peas. Intense browning was performed from purified fraction of algae that was extracted with water, dilute alkali and acid for elicitor preparations and maximum yield (30.2%) was obtained in aqueous extraction. Chemical composition of these extracted polysaccharides in terms of total sugar, protein, sulfate and uronic acid was performed and found that purified fractions contained sugar content (67.6%) and galactose as a major sugar component. using. Gel Permeation Chromatography of purified fraction identified molecular range ≥70,000Da. On the basis of FTIR, 1H and 13C NMR studies, the purified fraction was safely characterized as k-carrageenan.

English

Callus induction from cotyledon explants was studied in indigenous chick pea ( Cicer arietinum L.) cultivars KK-1 and Hassan-2K on MS and B 5 media containing different combinations and concentrations of growth regulators. Different MS and B 5 callusing media containing varying level of 2, 4-D (2 and 4 mg/l), NAA (0.50 and 1 mg/l), BAP (5 and 10 μM) and their combinations were tested for callus induction response. Percent callus and callus fresh weight (g) were recorded after two and four weeks of culture for both genotypes. For KK-1 cultivar, the maximum callus frequency (71 and 97%) followed by (65 and 96%) were observed on 4 mg/l 2,4-D+5 μM BAP in MS and 4 mg/l 2,4-D in B 5 media, respectively after two and four weeks of culture. Similarly, the highest callus fresh weight (0.411 and 0.787 g) were also recorded for MS+4 mg/l 2,4-D+5 μM BAP in contrast to B 5 where the highest callus weight (0.401 and 0.693 g) was achieved on 4 mg/l 2,4-D only. In Hassan -2K, the highest callus % (68 and 96) and fresh weight (0.572 and 0.821 g) were recorded on MS+4 mg/l 2,4-D+0.50 mg/l NAA after two and four weeks of culture, respectively. In B 5 medium, 2,4-D+BAP combination produced average callus induction response for both cultivars. Key words : Callus induction, chick pea cotyledons, growth regulators.

Preparation of Antioxidant Enzymatic Hydrolysates from Honeybee-Collected Pollen Using Plant Enzymes

Enzymatic hydrolysates of honeybee-collected pollen were prepared using food-grade proteinase and aminopeptidases entirely of plant origin. Bromelain from pineapple stem was applied (8 mAU/g substrate) in the first hydrolysis stage. Aminopeptidase (0.05 U/g substrate) and proline iminopeptidase (0.03 U/g substrate) from cabbage leaves (Brassica oleracea var. capitata), and aminopeptidase (0.2 U/g substrate) from chick-pea cotyledons (Cicer arietinum L.) were involved in the additional hydrolysis of the peptide mixtures. The degree of hydrolysis (DH), total phenolic contents, and protein contents of these hydrolysates were as follows: DH (about 20–28%), total phenolics (15.3–27.2 μg/mg sample powder), and proteins (162.7–242.8 μg/mg sample powder), respectively. The hydrolysates possessed high antiradical scavenging activity determined with DPPH (42–46% inhibition). The prepared hydrolysates of bee-collected flower pollen may be regarded as effective natural and functional dietary food supplements due to their remarkable content of polyphenol substances and significant radical-scavenging capacity with special regard to their nutritional-physiological implications.

Seed Size Is Associated with Sucrose Synthase Activity in Developing Cotyledons of Chickpea

Seed size, a key quality determinant in the marketing of the cool‐season food legume chickpea (Cicer arietinum L.), can be reduced under terminal drought. Sucrose synthase and invertase, important enzymes in sugar metabolism, play an important role in seed filling. This study aimed to determine whether these enzymes in the developing cotyledons are associated with the final seed size of chickpea by following their level of activity during seed filling in large‐ and small‐seeded chickpea genotypes. The level of invertase in the cotyledons was unchanged during seed filling, but sucrose synthase reached a maximum about 500 to 600 growing degree days (GDD) after flowering when cell numbers were no longer increasing, but the cotyledons were increasing rapidly in dry weight. In a range of genotypes differing in seed size, the level of sucrose synthase activity in the developing cotyledons at 500 to 600 GDD was correlated with the final seed size at maturity when measured at a rainfed site in two different years. The level of activity of sucrose synthase at the same stage of development in F2, F3, and F7 populations of crosses between a large‐seeded cultivar and two small‐seeded genotypes was also correlated with final seed size. It is concluded that sucrose synthase, but not invertase, in developing cotyledons is the key enzyme influencing seed size in chickpea.

Purification and Characterization of L-Phenylalanine Aminopeptidase from Chick-Pea Cotyledons (Cicer arietinum L.)

Chick-pea (Cicer arietinum L.) cotyledons are unique source of aminopeptidase - 8-9 U/g cotyledons was observed using L-leucine-p-nitroanilide as substrate. The aminopeptidase was purified (65 kDa, pI 4.8 ) reaching a specific activity of 220 U/mg at pH 7.0-7.2 and 35-40 degrees C. The determined constant of specificity k(cat)/K(m) during hydrolysis of N-unsubstituted amino acid-p-nitroanilides showed a decrease order: Phe>Leu>Pro>Ile>Val>Ala. The enzyme was strongly inhibited by p-chloromercuribenzoic acid as well as in a competitive rate by the antihypertensive peptides Ile-Pro-Pro and Val-Pro-Pro.

Enzymatic Hydrolysis of Soy Protein Isolate by Food Grade Proteinases and Aminopeptidases of Plant Origin

ABSTRACTEnzymatic hydrolysates of soy protein isolate were prepared using food grade proteinases and aminopeptidases entirely of plant origin. Papain from papaya fruit and bromelain from pineapple stem (both as catalog products and lyophilized crude extracts) were applied (20 mAU.g−1 substrate) in the first hydrolysis stage. Aminopeptidase (0.1 units.g−1 substrate) and proline iminopeptidase (0.08 units.g−1 substrate) from cabbage leaves (Brassica oleraceae var. capitata) and aminopeptidase (0.5 units. g−1 substrate) from chick-pea cotyledons (Cicer arietinum L.) were involved in the debittering process by additional hydrolysis of the peptide mixtures. The degree of hydrolysis was estimated from the amino nitrogen content and a level of36-38% degree of hydrolysis ensured an acceptable bland taste. Gel chromatography on BioGel P-2 of the obtained soy protein hydrolysates indicated a molecular weight of the all released peptides under 800 Da. Using entirely enzymes of plant origin the prepared hydrolysates are suitable for food supplements covering the requirements of vegetarians as well as the religious tolerance. The soy protein hydrolysates manifesting bland taste present a good substance for further design of food supplements with natural plant extracts from Vietnam.

Application of unreacted-core model to in situ gelatinization of chickpea starch

Unreacted-core model for reaction-controlled systems was tested on modeling of starch gelatinization in whole chickpea (in situ) during cooking. Experiments were conducted in deionized water at 50, 60, 70, 80, 90, and 100 °C. The process was followed through images of the flat sides of the chickpea cotyledons. During cooking between 60 and 100 °C, a white core in the original color of the cotyledons and a surrounding opaque yellow zone were observed on the cotyledons. According to birefringence studies starch granules in the yellow zone were gelatinized, and in the white core they remained ungelatinized. The formation of the yellow color was connected to the gelatinization in the peripheral zone. During cooking at 50 °C the color change was not observed because of working below the gelatinization temperature of chickpea starch. The area of the gelatinized zone increased at the expense of the area of the ungelatinized core with the progress of the cooking. The unreacted-core model fitted the process very well, and the estimated gelatinization times were in good agreement with the experimental gelatinization times. The kinetic data for the gelatinization reaction estimated after verifying the unreacted-core model were in agreement with the literature. These findings indicated that the in situ gelatinization of chickpea starch can be modeled using the unreacted-core model, and the process is effectively gelatinization-controlled under the given conditions.

Ethylene regulates the expression of a cysteine proteinase gene during germination of chickpea (Cicer arietinum L.).

Synthetic oligonucleotides corresponding to conserved regions of cysteine proteinases were used as primers in the RT-PCR amplification of a fragment of cDNA corresponding to a region of a cysteine proteinase gene expressed during germination of chickpea (cac for Cicer arietinum cysteine proteinase). The identity of the PCR-amplified fragment was confirmed by sequencing and the fragment used as a probe to investigate the pattern of cac gene expression during germination and its hormonal regulation. The corresponding transcript is undetected in the seed during embryogenesis and before imbibition, being detected 24 h after imbibition. Ablation of the embryonic axis before imbibition results in a dramatic decrease in the amount of transcript detected. Expression of the cac transcript in excised cotyledons is restored in the presence of aqueous extracts from embryonic axes and also by incubating the excised cotyledons in 1 mM ethephon. Experiments with various known inhibitors of ethylene action indicate that ethylene activates the expression of cac gene in the cotyledons of chickpea during normal germination.

Effect of dehulling on nutrient losses in chickpea ( Cicer arietinum L.)

Chickpea cotyledons were dehulled (scarified) for different lengths of times (0, 2, 4, 8, and 12 min) using a Tangential Abrasive Dehulling Device (TADD). The resulting decorticated dry split cotyledons (dhal) and powder fractions were then analyzed for protein, sugar, starch, fiber, minerals, and amino acids. Protein, soluble sugar, and ash of dhal (scarified cotyledons) fraction decreased with increasing dehulling time, but starch content increased. Considerable amounts of calcium, iron, and zinc were removed by dehulling for 4 min. This time is assumed to be an equivalent of traditional dehulling in terms of quantitative losses of powder fraction. No notable differences were observed in the amino acids of dhal and powder fractions of cotyledons dehulled for different intervals. Albumin and glutelin fractions were considerably reduced due to dehulling, while the globulin fraction increased. It is concluded that the dehulling process (4 min) would incur considerable losses in protein, calcium, iron, and zinc, whereas it would not adversely affect the protein quality in terms of amino acids.

Construction and Screening of Chickpea cDNA Library with Heterologous Legumin cDNA Probe

A cDNA library was constructed from developing chickpea cotyledons in the expression vector λ gt 11. The library when plated on X-gal and IPTG showed about 35% recombinants. These recombinants were screened using pea legumin cDNA probe. Of a total of 4 × 104 cDNA clones screened, 20 clones showed homology to legumin probe. The presence of cDNA insert in these clones was further confirmed by Southern blot hybridization.

Regulatory role of phytochrome in the induction of α-amylase activity in excised chick-pea cotyledons

During the germination and primary growth of chick-pea seedlings two clearly differentiable phases are observed in the development of α-amylase activity. The activity detected is thermolabile and is negatively affected by the absence of the embryonic axis. A short pulse of red light is able to substitute the effect of the axis in inducing α-amylase activity, the main stimulatory effect being obtained when red light is applied at 25 h after the start of incubation of the cotyledons in darkness. Phytochrome seems to be involved in the stimulatory effect of red light since, in general, this effect is reverted by the application of far red light, 5 min of this type of irradiation being the most effective treatment.

Scanning Electron Microscopy Study of Starch Granule Degradation in Chick‐pea Cotyledons

AbstractThe embryonic axis plays an important role in the processes of starch degradation. Before germination starts, the starch granules of chickpea cotyledons have an almost smooth surface. As germination progresses, membrane‐like material is deposited on the granules from the adaxial to the abaxial face of the cotyledon. At 10 days after the start of germination, the starch granules disappear, giving rise to numerous polyhaedral starch bodies. When excised cotyledons are cultured, a pronounced temporal delay occurs in these events.

Accumulation of Phenolic Compounds and Phytoalexins in Sliced and Elicitor-Treated Cotyledons of Cicer arietinum L.

Upon slicing cotyledons of chickpea, Cicer arietinum L.. accumulate the pterocarpan phyto­alexins medicarpin and maackiain. Treatment of this tissue with an elicitor from the phytopathogcnic deuteromyccte Ascochyta rabiei (Pass.) Lab. greatly enhances accumulation of the pterocarpans and of other isoflavones and flavonoids. Isolation, chromatographic purification and structural elucidation by spectroscopic techniques of 16 phenolic compounds is described. Cotyledons induced for phytoalexin biosynthesis readily accumulate the isoflavones daidzein, formononetin, calycosin and pseudobaptigenin which are thought to be intermediates in pterocarpan formation.

Elicitation and Suppression of Phytoalexin and Isoflavone Accumulation in Cotyledons of Cicer arietinum L. as Caused by Wounding and by Polymeric Components from the Fungus Ascochyta rabiei

AbstractShortly after sowing cotyledons of chickpea (Cicer arietinum) start to accumulate the isoflavones biochanin A and formononetin together with their 7‐0‐glucosides and their 7‐0‐glucoside‐6″‐malonates. The additional accumulation of the pterocarpan phytoalexins medicarpin and maackiain can be induced by wounding of the cotyledons. Treatment of sliced cotyledons with a crude elicitor fraction obtained from the growth medium or the mycelium of the chickpea pathogenic fungus Ascochyta rabiei (Pass.) Lab. leads to a dramatic increase in the level of numerous aromatic compounds, especially of the isoflavone aglyca and the phytoalexins. Accumulation of isoflavone conjugates is not altered by elicitor treatment as shown by time course studies, and dose‐response curves.A protein preparation (“suppressor”) isolated from the culture filtrate of the same fungus was shown to inhibit the accumulation of isoflavone aglyca, isoflavone conjugates and phytoalexins in the sliced cotyledons.The possible relevance of elicitor‐suppressor counteraction with regard to the defence mechanisms of the host plant is discussed.

Imbibitional Stress and Transverse Cracking of Bean, Pea, and Chickpea Cotyledons

Abstract Transverse cracking was examined in cotyledon tissue of snap bean (Phaseolus vulgaris L. ‘Apollo’), pea (Pisum sativum L. ‘Garfield 81’), and chickpea (Cicer arietinum L. ‘Surutato 77’). The hypothesis that imbibitional cracks develop along lines of least resistance or along preexisting cracks was evaluated. Columns of tissue were cut from cotyledons. Time from the start of imbibition to detection of the first crack was compared between whole bean cotyledons and columns. Bean columns cracked transversely with respect to the long axes of the columns. They did not crack longitudinally, even if the long axis of a column was originally perpendicular to the long axis of the cotyledon. These results do not support the hypothesis that imbibitional cracks form along preexisting cracks or along lines of weakness. Imbibitional stresses induced new cracks to form. The mean time to first crack and SE associated with its measurement were both significantly reduced in columns relative to the corresponding values for whole cotyledons (40 ± 2.6 and 100 ± 8.0 min, respectively) at an imbibition temperature of 19.3°C. Pea and chickpea cotyledon tissues also cracked transversely when they were cut into long, narrow columns.

Alcohol dehydrogenase isoenzymes in chickpea cotyledons

Alcohol dehydrogenase (ADH) (EC 1.1.1.1) in the cotyledons of chickpea consists of three isoenzymes, ADH-1, ADH-2 and ADH-3, in order of decreasing ele