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. 2019 Mar 8;56(4):1864–1876. doi: 10.1007/s13197-019-03646-4

Comparison of cultivated and wild chickpea genotypes for nutritional quality and antioxidant potential

Karamveer Kaur 1, Satvir Kaur Grewal 1,✉, Parmpal Singh Gill 2, Sarvjeet Singh 3
PMCID: PMC6443973  PMID: 30996422

Abstract

Fifteen cultivated (Ten desi, five kabuli) and fifteen wild species of chickpea (Cicer arietinum L.) were compared for nutritional traits, antinutritional factors and antioxidant potential. The average crude protein content in desi, kabuli and wild species was found to be 25.31%, 24.67% and 24.30%, respectively; total soluble sugars in these genotypes were 38.08, 43.75 and 33.20 mg/g, respectively and total starch content in these genotypes was 34.43, 33.43 and 28.77%. Wild species had higher antioxidant potential as compared to cultivated genotypes due to higher free radical scavenging activity, ferric reducing antioxidant power and reducing power. Kabuli genotypes had lower antioxidant potential than desi genotypes. Desi genotype, GL 12021 had high crude protein and total starch content, lower phytic acid and saponin content and higher antioxidant potential. GNG 2171 had high crude protein and total soluble sugar content and lower tannin and phytic acid content. Kabuli genotype L 552 possessed high total soluble sugar and total starch content, high Zn and Fe content and lower tannin, saponin and trypsin inhibitor content. Wild species C. pin ILWC 261 had high crude protein, lower phytic acid and trypsin inhibitor content and higher DPPH radical scavenging activity and hydroxyl radical scavenging activity. The observed diversity for quality traits in cultivated and wild genotypes can be further used.

Keywords: Antioxidant potential, Chickpea, Cultivated genotypes, Quality traits, Wild species

Introduction

Chickpea (Cicer arietinum L.) is an important food crop both for human food as well as animal feed. It is the world’s second largest grown pulse crop after beans. Chickpea is a cheap source of protein and ranked the fifth most valuable legume. India is the producer of 75% of world’s chickpea production. The chickpea production has increased globally by 56% and in India by 55% during the decade of 2004–2013. Other major chickpea producing countries are Australia, Pakistan, Myanmar, Ethiopia, Mexico, Canada, USA, Tanzania and Malawi (Gaur et al. 2016). Year 2016 was declared as the international year of pulses by the United Nations General Assembly. The production of pulses in India was 17.56 MT in 2016 and India emerged as the biggest chickpea producer in the world with a production of 7.8 MT in 2016 (Kumar et al. 2018).

Chickpea is an outstanding source of nutritional constituents such as proteins, carbohydrates, vitamins (niacin and thiamine), minerals and unsaturated fatty acids (linolenic and oleic acids) (Heiras-Palazuelos et al. 2013). Popularity of chickpea in human diet is due to the balanced seed nutrients composition and its low price. People who cannot afford animal protein and those who are vegetarians mainly consume chickpea as a protein source in semi-arid regions. Cereals are rich in thiol containing amino acids (methionine and cysteine) and deficient in lysine whereas pulses are rich in lysine and deficient in methionine and cysteine. Due to this pulses are taken with addition of cereals for proper intake of essential amino acids (Reinkensmeier et al. 2015).

Cultivated chickpea are categorized into Desi and Kabuli types. The Desi (microsperma) types have pink flowers, anthocyanin pigmentation on stems, and seed coat is thick and coloured. The Kabuli (macrosperma) types have white flowers, lack anthocyanin pigmentation on stems, seeds are white or beige coloured having ram’s head shape, seed coat is thin and a smooth seed surface. Geographical distribution of these chickpea types distinctly separate them as desi types which are mostly grown in Asia and Africa and account for up to 80–85% of the total chickpea area and kabuli chickpea types are grown mostly in West Asia, North Africa, North America and Europe (Gaur et al. 2016). Wild chickpea species are promising natural reservoir of potential genes for crop improvement due to their higher genetic variability and propagation in large range of habitats without human selection (Munoz et al. 2017).

Protein digestibility in legumes is affected by antinutritional factors such as phytate, raffinose family oligosaccharides, tannins, phenolic compounds, protease inhibitors (trypsin inhibitors and α amylase inhibitors) and saponins which are naturally occurring substances in edible seeds and impair absorption and nutritional utilization in humans by binding to proteins, minerals and vitamins (Soetan and Oyewole 2009).

Free radicals generated in many biochemical reactions cause functional and structural damage to nucleic acid, lipids, protein and cellular molecules and are mediators of many diseases in human body (Kumaran and Karunakaran 2007). Antioxidants in small quantities prevent the excessive collection of free radicals or reactive oxygen species (ROS) by retarding the oxidation of unsaturated fats which are easily oxidized. Pulses are also rich in natural antioxidants required for health and reduce the risk of diseases. Natural antioxidants are considered secure for the consumers than synthetic antioxidants such as butylated hydroytoluene (BHT), which have carcinogenic effects. Desi and kabuli chickpea significantly differ at nutritional level due to differences with respect to physiochemical properties, protein digestibility, phenolic content and antioxidant activity (Heiras-Palazuelos et al. 2013). The present investigation has been formulated to compare desi, kabuli and wild chickpea types for quality traits so as to identify cultivars with enhanced seed quality and this information can be further used to develop cultivars of desirable traits.

Materials and methods

Plant material

Thirty chickpea genotypes comprising of ten desi, five kabuli and fifteen wild species were procured from Pulses Section, Department of Plant Breeding and Genetics, Punjab Agricultural University, Ludhiana. Different accessions of wild species, Cicer judaicum (C. jud), Cicer pinnatifidum (C. pin), Cicer echinospermum (C. ech) were also evaluated for quality traits. Foreign material and damaged seeds were removed from the sample. The seeds were crushed into fine powder using grinder and the powdered chickpea seeds were used for estimation of quality traits.

Extraction and determination of nutritional and antinutritional traits

Crude protein was estimated by Kjeldahl method. Seed powder (0.2 g) wrapped in Whatman No. 1 filter paper was put in the kjeldahl digestion tube and 3 g of digestion mixture (copper sulphate and potassium sulphate (1:10) followed by addition of 10 ml of conc. sulphuric acid. The digestion was carried out for 2 h in kjeldahl digestor KELPLUS (KES 20L VADLS TS). After digestion, distillation was carried out in KELPLUS (CLASSIC-DX VATS (E) and then samples were titrated in titrator (Metrohm) using 0.1 N HCl. The nitrogen content determined was converted to protein content using conversion factor 6.25. Chickpea seed powder was extracted twice with 15 ml of 80% ethanol and 70% ethanol by keeping the tubes fitted with water condensers in boiling water bath for 20 min. The supernatants after each extraction were collected and pooled and total soluble sugars were estimated by using phenol sulphuric acid method as described previously by Sekhon et al. (2017). The collected dried soluble sugar free residue was used to extract and estimate starch using perchloric acid. Bound fructose of sucrose and raffinose family oligosaccharides was estimated by using resorcinol –HCl after destroying free fructose by boiling with 30% NaOH. Seed powder was digested with concentrated nitric acid and perchloric acid (4:1) till clear solution was obtained, volume was made to 25 ml with deionized distilled water and it was filtered using Whatman filter paper No. 1. Elements such as iron, zinc, calcium, magnesium, copper and manganese were estimated using Atomic Absorption Spectrophotometer (Perklin Elmer). The instrument was calibrated by using specific standards for each element.

Antinutritional factors such as phytic acid, tannins, trypsin inhibitor and saponins were determined from powdered chickpea seeds by the methods as described previously by Sekhon et al. (2017). Phytic acid was extracted with 1.2% HCl and precipitated with ferric chloride and inorganic phosphorus was estimated. Tannins were estimated from seed powder by Folin-Denis reagent. Saponins were extracted with acetone and later with methanol from the powdered seed powder and estimated by using reagent anisaldehyde (0.5 ml) mixed with (99.5 ml) ethyl acetate and concentrated sulphuric acid and concentration was calculated from standard curve prepared by using saponin (0–40 μg) as a standard. Trypsin inhibitor was extracted from powdered seeds with 0.1 M phosphate buffer (pH 7.5) containing 0.1 M NaCl and centrifuged at 10,000×g for 20 min. The supernatant was incubated at 80 °C for 20 min and then again centrifuged at 10,000×g for 20 min. The supernatant was used for assaying the bovine trypsin inhibition and Nα-benzoyl-dl-arginine p-nitroanilide (BAPNA) was used as a substrate. One inhibitor unit is defined as the quantity of inhibitor which causes 50% inhibition of trypsin activity.

Determination of antioxidant potential

The seed powder was refluxed with 80% methanol for 1 h at 80 °C. The mixture was vortexed for two hours and then centrifuged at 10,000 rpm for 10 min. The supernatant was collected and the pellet was re-extracted twice with 80% methanol. The pooled supernatants were used to measure antioxidant capacity of chickpea genotypes. Total phenols, flavonols, total reducing power, ferric reducing antioxidant power and free radical scavenging activities such as DPPH (2,2-diphenyl-1-picryl hydrazyl) free radical, hydroxyl radical, superoxide anion radical and nitric oxide radical were estimated in seeds of cultivated and wild species by the methods as described previously by Sekhon et al. (2017).

Statistical analysis

Mean and standard deviation were calculated. The results were analyzed by Tukey’s test. Software SPSS pc version 20.0 was used to identify correlation between the nutritional, antinutritional and antioxidant potential in chickpea genotypes. Equal class distribution method was used to categorize chickpea genotypes into three equal groups (low, medium and high) for quality traits.

Results and discussion

Nutritional constituents

Cultivated chickpea comprises two distinct seed types, desi and kabuli: desi seeds are generally brown or blackish brown while kabuli are of cream or beige colour. Due to these differences both desi and kabuli have different uses. We evaluated desi and kabuli chickpea genotypes to assess differences for nutritional quality and for antioxidant components so that potential donors can be identified for their use in breeding programmes to improve both desi and kabuli types for nutritional quality and antioxidant traits by transfer of desirable traits from potential donors. Similarly, it was assumed that wild species may possess some desirable nutritional and antioxidants which can be introgressed into cultivated chickpea. The variation in seed weight in desi, kabuli and wild species are presented in (Table 1). Seed weight of desi genotypes ranged from 7.19 g/100 seeds (GL 29095) to 13.69 g/100 seeds (GL 15017) with an average value of 9.07 g/100 seeds and in kabuli genotypes ranged from 15.16 g/100 seeds (L 552) to 22.97 g/100 seeds (GLK 14313) with an average value of 18.0 g/100 seeds. Wild species are smaller in size and their seed weight ranged from 0.76 g/100 seeds to 1.59 g/100 seeds with mean value of 0.93 g/100seeds. Kabuli genotype (GLK 14313) has the highest seed weight and wild species (C. jud 17148) has the lowest seed weight. Gaur et al. (2016) reported that seed weight of desi chickpea genotypes was found to be 10.2 g/100seeds and that of kabuli chickpea was 54.5 g/100 seeds. Seed weight, an extrinsic seed quality trait, also affects the reserve accumulation. Cooking time depends on seed size (weight and volume) and seed size is related to hydration and swelling capacity (Iqbal et al. 2006).

Table 1.

Distribution of seed weight (g/100 seeds), crude protein (%), total soluble sugar (mg/g), total starch (%), zinc (mg/100 g), iron (mg/100 g), calcium (mg/100 g), magnesium (mg/100 g), copper (mg/100 g) and manganese (mg/100 g) content in cultivated and wild chickpea genotypes

Genotypes Seed weight Crude protein Total soluble sugar Total starch Zinc Iron Calcium Magnesium Copper Manganese
GL 12021 8.42 ± 0.25gh 27.69 ± 0.5ab 39.06 ± 1.23cd 35.62 ± 9.13bcde 3.12 ± 0.21abc 7.99 ± 0.12abcd 62.01 ± 5.23e 83.86 ± 6.05fg 0.35 ± 0.13bcd 2.41 ± 0.13bcdefgh
GL 13037 11.83 ± 0.42e 25.03 ± 0.28efgh 52.73 ± 1.95ab 34.71 ± 9.14cdef 1.63 ± 0.18cd 9.29 ± 0.70abc 82.51 ± 7.66e 87.03 ± 5.82efg 0.30 ± 0.05bcd 1.96 ± 0.11efgh
GL 13042 9.11 ± 0.17fg 24.09 ± 0.72ijkl 31.25 ± 1.54fgh 39.27 ± 9.14a 2.00 ± 0.21bcd 7.83 ± 0.13abcd 102.36 ± 15.94de 84.91 ± 3.45fg 0.40 ± 0.01bcd 2.28 ± 0.13cdefgh
GL 14015 10.01 ± 0.28f 23.19 ± 0.25m 33.20 ± 1.95efg 36.99 ± 4.57b 2.12 ± 0.18abcd 9.83 ± 0.19a 40.83 ± 3.93e 71.96 ± ±11.39g 0.21 ± 0.14bcd 2.73 ± 0.12bcdefg
GL 15003 7.38 ± 0.07h 25.94 ± 0.09cd 27.34 ± 1.75h 36.08 ± 4.57bcd 2.93 ± 0.07abcd 5.53 ± 0.15cd 56.95 ± 8.85e 76.42 ± 7.09fg 0.26 ± 0.04bcd 3.49 ± 0.08abc
GL 15017 13.69 ± 0.28d 23.97 ± 0.16ijklm 31.25 ± 1.66fgh 33.79 ± 9.14efgh 2.42 ± 0.25abcd 4.56 ± 1.75d 55.11 ± 4.92e 73.23 ± 7.23fg 0.13 ± 0.05d 2.06 ± 0.17defgh
GL 29078 7.74 ± 0.14h 23.72 ± 0.09klm 37.10 ± 1.95cde 31.51 ± 4.57ijk 3.76 ± 0.12ab 8.33 ± 0.13abcd 53.78 ± 9.49e 80.32 ± ±2.22fg 0.29 ± 0.01bcd 2.60 ± 0.04bcdefgh
GL 29095 7.19 ± 0.03h 25.66 ± 0.22def 35.15 ± 1.45efd 31.51 ± 4.57ijk 3.75 ± 0.10ab 8.92 ± 0.54abc 76.69 ± 6.56e 92.86 ± 8.08efg 0.33 ± 0.15bcd 2.26 ± 0.17cdefgh
GNG 2171 8.02 ± 0.43gh 28.00 ± 0.38a 54.68 ± 1.34a 30.60 ± 4.57jk 2.81 ± 0.29abcd 5.80 ± 0.61abcd 140.38 ± 14.46abcde 145.12 ± 4.18abcdefg 0.27 ± 0.06bcd 1.80 ± 0.16fgh
PBG 7 7.30 ± 0.14h 25.85 ± 0.1de 39.06 ± 1.27cd 34.25 ± 22.84defg 2.24 ± 0.24abcd 7.24 ± 0.52abcd 242.60 ± 13.57abc 119.09 ± 4.23abcdefg 0.18 ± 0.06cd 2.12 ± 0.13defgh
Mean 9.07 ± 0.22 25.31 ± 0.28 38.08 ± 1.61 34.43 ± 6.19 2.68 ± 0.18 7.53 ± 0.38 91.32 ± 8.06 91.48 ± 5.98 0.27 ± 0.07 2.37 ± 0.12
L 552 15.16 ± 0.26c 24.13 ± 0.22ijkl 52.73 ± 1.95ab 36.53 ± 18.27bc 4.05 ± 0.41a 9.87 ± 0.94a 56.44 ± 3.90e 132.27 ± 6.82abcdefg 0.38 ± 0.10bcd 2.88 ± 0.19abcdefg
GLK 14313 22.97 ± 1.66a 24.63 ± 0.13ghij 41.02 ± 1.95c 35.62 ± 9.13bcde 1.23 ± 0.14cd 4.59 ± 0.23d 79.64 ± 6.99e 73.71 ± 4.24fg 0.21 ± 0.07bcd 1.64 ± 0.23gh
GLK 28127 18.22 ± 0.16b 25.47 ± 0.03defg 37.11 ± 1.95cde 34.71 ± 9.14cdef 2.05 ± 0.11abcd 9.69 ± 0.61abc 61.66 ± 5.40e 101.68 ± 3.44bcdefg 0.26 ± 0.05bcd 3.28 ± 0.13abcd
GLK 07-42 15.51 ± 0.82c 23.15 ± 0.1m 39.06 ± 1.77cd 32.42 ± 21.11ghij 0.96 ± 0.30d 5.49 ± 0.23cd 122.80 ± 3.00bcde 131.32 ± 5.85abcdefg 0.43 ± 0.08bcd 1.31 ± 0.16h
GLK 08-134 18.12 ± 0.39b 25.97 ± 0.16cd 48.83 ± 1.95b 27.86 ± 4.57m 2.47 ± 0.20abcd 5.59 ± 0.63bcd 64.63 ± 5.19e 84.13 ± 1.79fg 0.26 ± 0.08bcd 1.71 ± 0.12fgh
Mean 18.00 ± 0.66 24.67 ± 0.13 43.75 ± 1.91 33.43 ± 5.17 2.15 ± 0.23 7.05 ± 0.53 77.03 ± 4.90 104.62 ± 4.42 0.31 ± 0.07 2.16 ± 0.17
IC 525199 0.87 ± 0.02i 24.32 ± 0.19hijk 39.06 ± 1.27cd 30.14 ± 9.14kl 1.46 ± 0.11cd 9.15 ± 0.25abc 238.73 ± 7.29abc 167.91 ± 11.01abcde 0.42 ± 0.15bcd 3.16 ± 0.22abcde
IC 525202 0.84 ± 0.02i 23.38 ± 0.13lm 35.16 ± 1.75def 31.97 ± 0.10hijk 1.48 ± 0.21cd 8.66 ± 0.23abcd 257.63 ± 21.54a 118.19 ± 10.84abcdefg 0.41 ± 0.03bcd 3.31 ± 0.19abcd
IC 525691 0.77 ± 0.03i 23.85 ± 0.41jklm 35.15 ± 1.63def 28.31 ± 9.14lm 1.55 ± 0.18cd 9.39 ± 0.18abc 250.83 ± 16.15abc 153.33 ± 6.65abcdef 0.47 ± 0.07bc 2.72 ± 0.06bcdefg
C. jud 95 0.86 ± 0.03i 24.75 ± 0.11ghi 29.30 ± 1.95gh 27.40 ± 1.20m 1.51 ± 0.12cd 9.45 ± 0.34abc 217.52 ± 20.65abcd 93.26 ± 3.81defg 1.07 ± 0.28a 4.07 ± 0.34a
C. jud 185 0.91 ± 0.02i 25.82 ± 0.38de 39.06 ± 1.76cd 27.86 ± 4.57m 1.41 ± 0.06cd 7.65 ± 0.17abcd 238.57 ± 10.00abc 187.86 ± 8.32a 0.28 ± 0.01bcd 2.86 ± 0.02abcdefg
C. jud 185 B 0.87 ± 0.08i 24.32 ± 0.19hijk 27.34 ± 1.95h 28.31 ± 9.14lm 3.18 ± 0.15abc 8.98 ± 0.37abc 237.96 ± 3.18abc 174.19 ± 4.57abcd 0.54 ± 0.32b 3.01 ± 0.06abcdef
C. jud 17148 0.76 ± 0.03i 23.78 ± 0.03jklm 29.30 ± 1.95gh 27.40 ± 1.00m 1.70 ± 0.01cd 6.97 ± 0.08abcd 256.53 ± 9.49ab 180.06 ± 4.61ab 0.31 ± 0.00bcd 2.96 ± 0.04abcdef
C. jud 17150 0.88 ± 0.01i 23.03 ± 0.59efgh 35.15 ± 1.95def 30.14 ± 9.14kl 1.81 ± 0.13bcd 8.45 ± 0.46abcd 259.58 ± 5.66a 137.22 ± 7.71abcdefg 0.36 ± 0.05bcd 2.87 ± 0.16abcdefg
C. jud ILWC 30 1.30 ± 0.05i 24.22 ± 0.28hijkl 39.06 ± 1.66cd 33.34 ± 13.7fghi 2.07 ± 0.17abcd 8.39 ± 0.58abcd 240.80 ± 12.06abc 179.46 ± 12.51abc 0.25 ± 0.03bcd 3.56 ± 0.28abc
C. pin ILWC 0 0.99 ± 0.05i 23.76 ± 0.13jklm 27.34 ± 1.6h 30.14 ± 0.09kl 2.65 ± 0.19abcd 9.77 ± 0.08ab 260.95 ± 3.44a 111.03 ± 3.52abcdefg 0.27 ± 0.04bcd 2.96 ± 0.15abcdef
C. pin ILWC 261 1.59 ± 0.04i 26.82 ± 0.32bc 39.06 ± 1.88cd 27.40 ± 0.09m 2.53 ± 0.15abcd 7.79 ± 0.38abcd 150.49 ± 10.90abcde 99.00 ± 3.78cdefg 0.44 ± 0.02bcd 3.69 ± 0.18ab
EC 366338 0.84 ± 0.02i 23.82 ± 0.13jklm 31.25 ± 1.95fgh 26.49 ± 9.14m 1.91 ± 0.10bcd 8.63 ± 0.26abcd 224.02 ± 22.38abcd 119.72 ± 13.08abcdefg 0.36 ± 0.04bcd 3.30 ± 0.39abcd
EC 366342 0.84 ± 0.01i 24.78 ± 0.28fghi 33.20 ± 1.95efg 27.40 ± 18.27m 1.84 ± 0.05bcd 7.80 ± 0.19abcd 120.31 ± 10.47cde 90.65 ± 3.37efg 0.40 ± 0.01bcd 3.29 ± 0.25abcd
EC 556270 R 0.81 ± 0.03i 23.38 ± 0.07hijk 31.25 ± 1.8fgh 27.86 ± 4.57m 1.43 ± 0.17cd 8.58 ± 0.66abcd 141.39 ± 11.89abcde 85.42 ± 7.62fg 0.20 ± 0.05cd 2.94 ± 0.27abcdefg
C. ech 17159 0.83 ± 0.03i 24.53 ± 0.03hkij 27.34 ± 1.75h 27.40 ± 1.75m 1.56 ± 0.21cd 9.24 ± 0.29abc 147.33 ± 12.07abcde 89.36 ± 6.12efg 0.27 ± 0.02bcd 2.70 ± 0.19bcdefg
Mean 0.93 ± 0.03 24.30 ± 0.22 33.20 ± 1.79 28.77 ± 5.23 1.87 ± 0.13 8.59 ± 0.30 216.18 ± 9.81 132.44 ± 7.17 0.40 ± 0.07 3.67 ± 0.19
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Data represent the mean ± SD of triplicates and values with different letters in the same column are significantly different (p < 0.05)

Legumes are known as ‘meat of the poor people’ because of their high protein content and are considered as staple food for those who cannot afford animal proteins or vegetarian by choice and for people affected by nutrition related health problems such as diabetes, obesity and over weight. Chickpea has protein quality better than other legumes and is a good source of dietary protein (Kaur and Singh 2007). The crude protein content in chickpea genotypes (Table 1) ranged from 23.03% (C. jud 17150) to 28.00% (GNG 2171). Crude protein content in desi genotypes was found in the range of 23.19 to 28.00% with the mean value of 25.31% and in kabuli genotypes it lied in the range of 23.15 to 25.97% with the mean value of 24.67%. In wild species, crude protein ranged from 23.03 to 26.82% with an average value of 24.30%. The chickpea genotypes with respect to crude protein were characterized into high, low and medium group using equal class distribution method. Crude protein content higher than 26.35% is categorized under higher crude protein group, value between 24.69 and 26.35% is placed under medium crude protein content and lower than 24.69% is placed under low crude protein group (H ≥ 26.35 ≤ 24.69). Three genotypes namely GL 12021 (Desi), GNG 2171 (Desi) and C. pin ILWC 261 (Wild) possessed higher protein content in the range of 26.35 to 28.00%. Ten genotypes possessed medium range of crude protein content ranging from 24.69 to 26.35%. Seventeen genotypes had lower crude protein content ranging from 23.03 to 24.69%. Differences of crude protein content between desi and kabuli types have been reported to be inconsistent. Our results are in accordance with Gaur et al. (2008) who reported that crude protein content in desi chickpea (29.2%) was found to be higher than kabuli chickpea (20.5%). However Ghribi et al. (2015) reported that crude protein content in desi and kabuli chickpea cultivars differ significantly as desi chickpea genotypes possessed lower amount of crude protein (20.29%) and kabuli possessed the higher amount of crude protein (24.51%). Singh et al. (2010) reported that protein content in chickpea genotypes ranged between 15.7 and 31.5%.

The total carbohydrates in chickpea seeds are up to 62% and the major proportion is of starch. One distinguish feature of legumes is that legume carbohydrates are digested slowly and due to this reason legumes are considered as low glycemic index foods that prevent diabetes, heart disease and obesity (Jenkins et al. 2012). Total soluble sugar content in chickpea genotypes varied from 27.34 to 54.68 mg/g with mean value of 36.59 mg/g (Table 1). Among cultivated genotypes, desi genotypes possessed total soluble sugar content in the range of 27.34 to 54.68 mg/g with mean value of 38.08 mg/g and kabuli genotypes possessed total soluble sugar content in the range of 37.11 to 52.73 mg/g with an average value of 43.75 mg/g. Sugar content in wild species ranged from 27.34 to 39.06 mg/g with mean value of 33.20 mg/g. Maheri-Sis et al. (2008) reported that total soluble sugar content in kabuli genotypes was higher as compared to desi genotypes.

Total starch content in chickpea genotypes ranged from 26.49 to 39.27% with mean value of 31.43% (Table 1). Wild species (EC 366338) had the lowest starch content and desi genotype (GL 13042) had the highest starch content. In desi genotypes, starch content varied from 30.60 to 39.27% and in kabuli genotypes it ranged from 27.86 to 36.53%. Wild species possessed total starch content in the range of 26.49 to 33.34% with mean value of 28.77%. Six genotypes comprising four desi GL 12021, GL 13042, GL 14015, GL 15003 and two kabuli L 552, GLK 14313 possessed higher total starch content that ranged from 35.62 to 39.27%. Nine genotypes possessed medium range of starch content from 30.75 to 35.01% and fifteen genotypes had lower starch content in the range of 26.49 to 30.60%. Starch content among desi and kabuli genotypes varied significantly from 30.2 to 55.2% and 38.3 to 51.3% respectively (Wang et al. 2017). Amylose content of starches from kabuli and desi chickpea types ranged from 43.24 to 47.22 and 28.26 to 52.82%, respectively (Singh et al. 2010). Sekhon et al. (2017) reported that starch content in pigeonpea cultivars ranged from 27.27 to 52.12% and among wild species it ranged from 9.04 to 19.12%.

Mineral elements are involved in all defense mechanisms as they are important components of cells and enzymes. They also act as inhibitors, activators and regulators of metabolism. Chickpea can overcome mineral malnutrition as it is a good source of iron and zinc. As compared to other pulses, manganese, zinc and phosphorous are present in higher amount in chickpea (Raes et al. 2014). Zinc content in chickpea genotypes varied from 0.96 mg/100 g to 4.05 mg/100 g (Table 1). Content of iron in chickpea genotypes ranged from 4.56 mg/100 g to 9.87 mg/100 g (Table 1). Zinc content in desi, kabuli and wild species ranged from 1.63 to 3.76; 0.96 to 4.05; 1.41 to 3.18 mg/100 g and the corresponding iron content ranged from 4.56 to 9.83; 4.59 to 9.87 and 6.97 to 9.77 mg/100 g respectively. Thavarajah and Thavarajah (2012) reported that Zn and Fe content in chickpea cultivars ranged from 3.7 mg/100 g to 7.4 mg/100 g and 4.6 mg/100 g to 6.7 mg/100 g.

Calcium content in chickpea genotypes ranged from 40.83 mg/100 g to 260.95 mg/100 g (Table 1). Magnesium content ranged from 71.96 to 187.86 mg/100 g (Table 1). Copper content in chickpea genotypes was found in the range of 0.13 mg/100 g to 1.07 mg/100 g (Table 1). Manganese content ranged from 1.31 mg/100 g to 3.69 mg/100 g (Table 1). Calcium, magnesium, copper and manganese content in desi, kabuli and wild genotypes ranged from 40.83 to 246.60, 56.44 to 122.80, 217.52 to 260.95 and 1.96 to 145.12 mg/100 g; 73.71 to 132.27, 85.42 to 187.86, 0.13 to 0.40 and 0.21 to 0.43 mg/100 g and 0.20 to 1.07 and 1.80 to 3.49, 1.31 to 3.28 and 2.70 to 4.07 mg/100 g. Wild species had higher calcium, magnesium and manganese content as compared to cultivated genotypes. Thavarajah and Thavarajah (2012) reported that calcium content ranged from 93 to 197 mg/100 g and magnesium content ranged from 125 to 159 mg/100 g in chickpea cultivars. Khan et al. (2015) reported that copper and manganese content of twenty-eight chickpea genotypes ranged from (0.66 to 1.04 mg/100 g) and (1.3 to 2.1 mg/100 g). Parmar et al. (2017) reported that hard to cook kidney bean grains had significantly higher Ca and Zn while Cu, Mn and Fe were lower than easy to cook kidney bean grains. They found that most of the minerals were correlated with the colour of the grains as hard to cook grains from light coloured accessions had lower Zn content.

Bound fructose of sucrose and raffinose series oligosaccharides

Raffinose oligosaccharides (RFO’s) are galactosyl derivative of sucrose and characterized by the presence of α (1 → 6) linkage between the galactose residue and the C-6 of the glucose moiety of sucrose (Gangola et al. 2013). The highest content of raffinose oligosaccharides was found in wild species (EC 366342) 10.33 mg/g and the lowest content was found in kabuli genotypes (GLK 14313) 0.66 mg/g (Table 2). In kabuli genotypes, bound fructose content ranged from 3.66 to 4.33 mg/g except in GLK 08-134 which has the lowest bound fructose content (0.66 mg/g). Bound fructose content in desi chickpea varieties found to be in range of 2.0 to 5.33 mg/g with the mean value of 3.80 mg/g. In wild chickpea species it ranged from 5.0 to 10.33 mg/g with an average value of 7.73 mg/g. Eight genotypes from wild species had higher bound fructose content (8.33 to 10.33 mg/g).

Table 2.

Distribution of total phenol (mg/100 g), flavonol (mg/100 g), bound fructose (mg/g), tannin (mg/g), phytic acid (mg/g), trypsin inhibitor (IU/g) and saponin (mg/g) content in cultivated and wild chickpea genotypes

Genotypes Total Phenol Flavonol Bound fructose Tannin Phytic acid Trypsin inhibitor Saponin
GL 12021 83.52 ± 2.12fg 12.67 ± 0.92efg 4.33 ± 0.33ghij 13.28 ± 0.15ijk 8.48 ± 0.95fghi 72.37 ± 0.25k 6.49 ± 0.11ij
GL 13037 62.02 ± 0.71ijk 9.77 ± 0.61ijkl 3.33 ± 0.02ijk 11.75 ± 0.30klmn 9.90 ± 0.47f 32.91 ± 1.76o 6.54 ± 0.05hij
GL 13042 56.56 ± 5.11jkl 13.18 ± 1.34e 4.66 ± 1.33ghi 13.96 ± 0.38ghi 12.73 ± 0.42e 50.45 ± 5.04mn 6.70 ± 0.22ghij
GL 14015 55.85 ± 1.94jkl 12.88 ± 0.65ef 3.99 ± 0.66ghi 9.08 ± 0.07p 15.56 ± 0.47bcd 98.72 ± 4.28hi 7.25 ± 0.11fghi
GL 15003 48.28 ± 2.47lm 6.39 ± 0.27no 2.66 ± 0.07jk 11.98 ± 0.53jklm 18.39 ± 0.47a 89.35 ± 2.55ij 5.51 ± 0.11l
GL 15017 38.59 ± 4.06mno 5.04 ± 0.15o 2.02 ± 0.09kl 10.61 ± 0.07mnop 13.20 ± 0.95de 61.53 ± 1.34klm 8.97 ± 0.11abcd
GL 29078 42.46 ± 4.41mn 10.74 ± 0.19hij 3.66 ± 0.33hijk 13.43 ± 0.15hij 15.09 ± 0.94cde 91.08 ± 8.41ij 7.51 ± 0.05fg
GL 29095 60.44 ± 1.24jk 9.47 ± 0.08ijklm 5.33 ± 0.21efgh 12.44 ± 0.07ijkl 17.45 ± 0.47abc 112.32 ± 3.61fg 9.51 ± 0.22ab
GNG 2171 60.61 ± 1.02jk 9.13 ± 0.19jklm 3.33 ± 0.14ijk 11.67 ± 0.22lmn 8.49 ± 0.48fghi 107.03 ± 1.51gh 7.41 ± 0.05fg
PBG 7 53.74 ± 1.94kl 8.40 ± 0.08lm 4.66 ± 0.66ghi 11.06 ± 0.53lmno 9.90 ± 0.47f 85.04 ± 1.18j 6.27 ± 0.86jkl
Mean 56.21 ± 2.50 9.77 ± 0.45 3.80 ± 0.38 11.93 ± 0.25 12.92 ± 0.70 80.08 ± 1.99 7.22 ± 0.19
L 552 61.14 ± 5.46jk 10.23 ± 0.08ijk 4.33 ± 0.33ghij 9.84 ± 0.22op 12.73 ± 0.47e 38.78 ± 0.59no 5.57 ± 0.05kl
GLK 14313 32.24 ± 2.29op 0.80 ± 0.04p 0.66 ± 0.19l 10.30 ± 0.38nop 13.67 ± 0.48de 38.53 ± 1.85o 6.38 ± 0.54jk
GLK 28127 27.48 ± 1.06p 1.37 ± 0.23p 3.66 ± 0.33hijk 11.67 ± 0.38lmn 9.43 ± 0.94fg 58.09 ± 3.27lm 7.46 ± 0.11fg
GLK 07-42 48.01 ± 1.59lm 5.50 ± 0.99o 3.99 ± 0.66hij 10.76 ± 0.38mno 9.90 ± 0.47f 39.29 ± 1.09no 7.84 ± 0.27ef
GLK 08-134 33.30 ± 1.94nop 1.22 ± 0.38p 4.33 ± 0.33ghij 10.53 ± 0.15mnop 17.92 ± 0.94ab 64.47 ± 1.09kl 7.82 ± 0.17ef
Mean 40.45 ± 2.47 3.82 ± 0.34 3.40 ± 0.37 10.63 ± 0.31 12.73 ± 0.66 47.83 ± 1.58 7.02 ± 0.23
IC 525199 97.44 ± 4.05cd 11.04 ± 0.42hij 5.99 ± 0.66efg 14.96 ± 1.98egh 5.65 ± 0.95jk 127.01 ± 2.35de 7.24 ± 0.32fghi
IC 525202 98.50 ± 0.53cd 16.26 ± 0.92d 6.66 ± 0.05def 17.40 ± 0.30abcd 6.13 ± 0.47ijk 138.94 ± 2.85abc 7.35 ± 0.22fgh
IC 525691 104.49 ± 1.59abc 24.20 ± 1.45a 5.02 ± 1fghi 16.41 ± 0.07bcdef 6.6 ± 0.47hijk 135.91 ± 3.86bcd 8.38 ± 0.27de
C. jud 95 84.22 ± 0.71efg 18.17 ± 0.53c 6.65 ± 0.05def 16.87 ± 0.99bcde 9.90 ± 0.47f 122.73 ± 2.94ef 8.49 ± 0.16cde
C. jud 185 93.92 ± 2.65de 8.93 ± 0.08jklm 6.68 ± 0.05def 17.70 ± 0.30abc 5.18 ± 0.48jk 129.87 ± 1.18cde 8.70 ± 0.05bcd
C. jud 185 B 84.75 ± 2.65efg 11.11 ± 0.11fhij 6.01 ± 0.06efg 17.93 ± 0.38ab 5.18 ± 0.42jk 141.03 ± 0.59abc 9.68 ± 0.05a
C. jud 17148 63.08 ± 1.41ijk 8.59 ± 0.19klm 9.05 ± 0.55ab 15.72 ± 0.15ef 4.24 ± 0.47k 139.86 ± 1.09abc 8.81 ± 0.16bcd
C. jud 17150 71.36 ± 0.18hi 7.94 ± 0.31mn 6.99 ± 0.33cde 15.72 ± 0.30ef 8.96 ± 0.47fgh 150.18 ± 2.35a 8.92 ± 0.27abcd
C. jud ILWC 30 109.07 ± 4.05ab 20.31 ± 0.53b 9.33 ± 0.66ab 18.47 ± 0.15a 8.48 ± 0.45fghi 15.95 ± 0.59p 9.24 ± 0.05abc
C. pin ILWC 0 79.47 ± 4.76gh 9.85 ± 0.31ijkl 8.33 ± 1.66bcd 16.10 ± 0.22def 5.66 ± 0.95jk 137.34 ± 0.42bcd 6.54 ± 0.05hij
C. pin ILWC 261 63.96 ± 1.94ij 12.75 ± 0.69efg 8.33 ± 0.33bcd 16.41 ± 0.07bcdef 8.48 ± 0.95fghi 8.23 ± 0.59p 8.76 ± 0.11bcd
EC 366338 93.03 ± 4.23def 12.06 ± 0.23efgh 8.66 ± 0.66abc 15.49 ± 0.07efg 6.13 ± 0.47ijk 142.38 ± 12.17ab 7.84 ± 0.27ef
EC 366342 102.73 ± 6.52bcd 10.31 ± 0.46hijk 10.33 ± 0.33a 15.19 ± 0.07fg 9.90 ± 1.42f 131.13 ± 3.78bcde 9.30 ± 0.11abc
EC 556270 R 113.30 ± 2.64a 10.36 ± 0.38hijk 8.33 ± 0.33bcd 16.33 ± 0.30cdef 7.07 ± 1.42fghi 137.26 ± 0.34bcd 9.46 ± 0.27ab
C. ech 17159 76.82 ± 0.71gh 8.97 ± 0.19jklm 9.66 ± 0.33ab 14.96 ± 0.15fgh 5.65 ± 0.95kj 126.43 ± 0.08de 9.08 ± 0.43abcd
Mean 89.07 ± 2.57 12.72 ± 0.45 7.73 ± 0.47 16.38 ± 0.37 6.88 ± 0.81 118.0.95 ± 1.68 8.52 ± 0.19
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Data represent the mean ± SD of triplicates and values with different letters in the same column are significantly different (p < 0.05). One inhibitor unit is the quantity of inhibitor that inhibits 50% of bovine trypsin activity

Tannins

Tannins constitute low molecular weight (0.5–3 kDa) naturally occurring anti-nutritional compounds. They precipitate with proteins and form complexes with proline rich proteins, polysaccharides and alkaloids, some of these are water soluble while some are insoluble. Binding of tannin with enzyme proteins or minerals cause inactivation of digestive enzymes and reduced protein digestion. Tannin content in chickpea genotypes ranged from 9.08 mg/g (GL 14015) to 18.47 mg/g (C. jud ILWC 30) with mean value of 13.94 mg/g (Table 2). The average tannin content in desi, kabuli and wild species was found to be 11.93, 10.63 and 16.38 mg/g respectively. Genotypes with tannin content higher than 15.34 mg/g are high tannin group; between 12.21 and 15.34 mg/g are medium tannin group and lower than 12.21 mg/g is placed under low tannin group (H ≥ 15.34 ≤ 12.21). Kaur et al. (2014) reported that tannin content in chickpea seeds ranged from 5.44 to 10.87 mg/g. Bulbula and Urga (2018) reported that tannin content in chickpea seeds was found to be 17.52 mg/g.

Phytic acid

Phytic acid (myo-inositol hexaphosphate) is the main inhibitor of iron and zinc absorption and form protein complexes. In gastrointestinal tract, absorption of minerals like calcium, copper, magnesium, iron and zinc are adversely affected by formation of complexes between minerals and phytic acid (Tiwari and Singh 2012). Due to the lack of sufficient level of phytate degrading enzyme activity in monogastric animals, metabolization of phytic acid is impossible and excreted in manure. Kabuli genotypes had phytic acid content in medium range i.e. from 9.43 to 13.67 mg/g (Table 2) except in GLK 08-134 which had higher content of phytic acid (17.92 mg/g). Phytic acid content in desi genotypes ranged from 8.48 to 18.39 mg/g with an average value of 12.92 mg/g. All wild species had lower phytic acid content that ranged from 4.24 to 8.48 mg/g. The chickpea genotypes were categorized into high, low and medium groups on the basis of phytic acid content (H ≥ 13.68 ≤ 8.96). Mondor et al. (2009) reported that phytic acid content ranged from 3.49 to 11.52 mg/g in desi genotypes and it ranged from 3.45 to 12.35 mg/g in kabuli genotypes. Vadivel and Biesalski (2010) reported that phytic acid content in wild type legume seeds was found to be 0.98 to 3.14 g/100 g. In our present investigation phytic acid content in desi genotypes was found to be higher than kabuli genotypes and wild species had phytic acid content in lower range. Shi et al. (2018) reported that phytic acid content in whole chickpea, split chickpea and desi chickpea was found to be 11.33 mg/g, 11.53 mg/g and 14.00 mg/g respectively.

Trypsin inhibitor activity

Protease inhibitor interfere with digestion by irreversibly binding to digestive enzymes trypsin and chymotrypsin and also due to their resistance to pepsin and acidic pH of human digestive tract. Presence of trypsin inhibitor in human foods led to impaired growth, pancreatic hyperplasia and metabolic disturbance of sulphur and amino acid utilization. The trypsin inhibitor content in chickpea genotypes varied significantly from 8.23 to 150.18 IU/g (Table 2). Trypsin inhibitor content in desi genotypes ranged from 32.91 to 112.32 IU/g with mean value of 80.08 IU/g. Trypsin inhibitor in kabuli genotypes ranged from 38.53 to 64.47 IU/g with mean value of 47.83 IU/g. All the wild species had higher trypsin inhibitor content ranging from 122.73 to 150.18 IU/g except in C. jud ILWC 30 and C. pin ILWC 261. Genotypes with trypsin inhibitor content higher than 102.87 IU/g are categorized under high trypsin inhibitor content group; values between 55.55 and 102.87 IU/g are medium group and lower than 55.55 IU/g are low trypsin inhibitor group (H ≥ 102.87 ≤ 55.55). Gupta et al. (2017) reported that trypsin inhibitor content in chickpea genotypes ranged from 111.5 to 218.4 TIU/g.

Saponins

Saponins are naturally occurring surface active glycosides that reduce absorption of nutrients and cause systemic toxicity by directly binding and inactivating enzymes. Saponins causes leakage of cells by creating holes, disturb the fluidity and causes lysis of red blood cells. Saponin affects metabolism in different ways like reduced cholesterol, depressed growth rate, erythrocyte haemolysis and reduced nutrient absorption. The average content of saponin in desi and kabuli genotypes was found to be 7.22 and 7.02 mg/g, respectively (Table 2). However all the wild species had higher saponin content that ranged from 8.38 to 9.68 mg/g except IC 525199, IC 525202 and EC 366338 which had medium saponin content in the range of 7.24 to 7.84 mg/g and C. pin ILWC 0 had lower saponin content (6.54 mg/g). Choudhary et al. (2015) reported that saponin content in kabuli chickpea genotypes varied from 4.98 to 12.23 mg/g. Sekhon et al. (2017) reported that saponin content ranged from 4.73 to 17.98 mg/g in pigeonpea genotypes.

Total phenol and flavonol in chickpea genotypes

Phenols bind to positively charged proteins, amino acids or multivalent cations and minerals such as iron, zinc and calcium in foods and decrease their bioavailability. On the other hand prevention of reactive oxygen species and removal of existing reactive oxygen species from blood are beneficial effect of phenols. Singh et al. (2017) reported that primary phenolic compounds present in legume seeds are phenolic acids, flavonoids and condensed tannins which play significant roles in many physiological and metabolic processes. These phenolic compounds had numerous health benefits as they act as anticarcinogenic, anti-thrombotic, anti-ulcer, anti-artherogenic, anti-allergenic, anti-inflammatory, antioxidant, immunemodulating, anti-microbial, cardioprotective and analgesic agents. Seed coat of legumes has high concentration of phenolic acids and acts as protective barrier for the cotyledon. The total phenol content in chickpea genotypes lied in the range of 27.48 mg/100 g to 113.30 mg/100 g (Table 2). All kabuli genotypes have lower total phenol content that ranged from 27.48 mg/100 g to 48.01 mg/100 g except in L 552 which has medium range of total phenol content. Wild species possessed total phenol content in higher and medium range that ranged from 63.08 mg/100 g to 113.30 mg/100 g with an average value of 89.08 mg/100 g. Total phenol content varied from 38.59 mg/100 g to 83.52 mg/100 g in desi genotypes. Higher total phenolic content in desi and wild genotypes in comparison to kabuli genotypes might be due to their dark coloured seed coat. Xu et al. (2007) compared different legumes for total phenol content and reported that soybeans (1.57–5.57 mg GAE/g), chickpeas (0.98 mgGAE/g), yellow peas (0.85–1.14 mg GAE/g) and green peas (0.65–0.99 mg GAE/g) have lower total phenol content than lentils (4.86–9.60 mg GAE/g). Sekhon et al. (2017) studied reported that total phenol content in pigeonpea wild species C. Scarabaeoides ranged from 69 mg/100 g to 129 mg/100 g. Flavonol content in chickpea genotypes ranged from 0.80 mg/100 g (GLK 14313) to 24.20 mg/100 g (IC 525691) with an average value of 10.25 mg/100 g (Table 2). Kabuli genotypes possessed lower flavonol content ranging from 0.08 mg/100 g to 0.55 mg/100 g. Flavonol content in desi genotypes varied from 5.04 mg/100 g to 13.18 mg/100 g. Flavonol content varied from 7.94 mg/100 g to 24.20 mg/100 g in wild species with an average value of 12.72 mg/100 g. Sekhon et al. (2017) reported that flavonol content in pigeonpea genotypes ranged from 5.32 mg/100 g to 16.02 mg/100 g. In present investigation, the higher total phenol and flavonol content in desi and wild species as compared to kabuli cultivars could be due to their dark coloured seeds as dark coloured legume had higher total phenol content as compared to light coloured seeds.

Free radical scavenging activity (DPPH)

DPPH is a stable free radical having maximum absorbance at 517 nm in methanol. It is used to determine antioxidant activity in natural compounds and its assay is mainly based on an electron transfer reaction and hydrogen-atom abstraction. DPPH free radical scavenging activity in chickpea genotypes ranged from 10.57 to 89.61% with mean value of 56.72% (Table 3). All kabuli genotypes possessed DPPH activity in the lower range (10.57 to 25.64%) and all wild species possessed DPPH activity in higher range (64.74 to 89.61%) except C. jud 17148, C. jud 17150 and C. pin ILWC 0 which had DPPH activity in medium range. DPPH scavenging activity in desi genotypes ranged from 21.02 to 75.57% with an average value of 49.40%. DPPH scavenging activity in wild chickpea species was higher than desi and kabuli genotypes indicating their higher antioxidative potential which might help to reduce oxidative stress in them. Gupta et al. (2017) reported that DPPH radical scavenging activity in forty chickpea genotypes ranged from 32.6 to 58.9%.

Table 3.

Distribution of DPPH radical scavenging activity (%), FRAP (mg/g), reducing paper (mg/g), hydroxyl radical scavenging activity (%), superoxide anion radical scavenging activity (%) and nitric oxide radical scavenging activity (%)

Genotypes DPPH FRAP Reducing Power Hydroxyl radical scavenging power Superoxide anion radical scavenging activity Nitric oxide radical scavenging activity
GL 12021 63.97 ± 1.41efg 1.71 ± 0.12efg 33.09 ± 1.34fg 28.89 ± 0.47ab 35.91 ± 5.26cde 61.18 ± 0.93bcd
GL 13037 46.28 ± 0.09kl 1.56 ± 0.10g 26.13 ± 0.82jk 3.63 ± 3.15kl 48.43 ± 0.50a 64.80 ± 0.41ab
GL 13042 46.02 ± 2.69kl 1.97 ± 0.10def 30.21 ± 0.17h 2.77 ± 1.33l 40.67 ± 2.00bcd 58.49 ± 0.31de
GL 14015 52.82 ± 3.20ij 1.66 ± 0.27fg 32.36 ± 0.61g 7.65 ± 2.00jkl 37.04 ± 0.87cde 56.25 ± 0.41ef
GL 15003 48.07 ± 2.05jk 1.49 ± 0.19g 25.06 ± 0.26jk 15.02 ± 1.72fghi 25.03 ± 0.12f 61.28 ± 1.86bcd
GL 15017 21.02 ± 3.07no 1.39 ± 0.01ghi 23.00 ± 0.17lm 7.94 ± 1.14jkl 41.17 ± 3.75abcd 62.94 ± 2.07abcd
GL 29078 39.03 ± 0.70m 1.40 ± 0.22gh 24.59 ± 0.47kl 9.76 ± 8.51ijkl 48.06 ± 0.87ab 53.00 ± 0.62fg
GL 29095 42.37 ± 0.32lm 1.55 ± 0.12g 25.32 ± 0.60jk 12.05 ± 2.00ghij 32.79 ± 0.37e 65.22 ± 1.45ab
GNG 2171 75.57 ± 0.70d 0.92 ± 0.03jk 24.63 ± 0.17kl 21.62 ± 0.09cdef 42.67 ± 1.25abc 61.59 ± 0.72bcd
PBG 7 58.78 ± 0.32gm 0.86 ± 0.02jkl 22.66 ± 0.35lm 6.41 ± 1.14jkl 12.64 ± 0.25gh 58.80 ± 1.04cde
Mean 49.40 ± 1.63 1.46 ± 0.12 26.70 ± 0.50 11.58 ± 2.16 36.45 ± 1.39 60.38 ± 0.98
L 552 24.93 ± 1.73n 0.74 ± 0.01kl 16.09 ± 0.64o 12.24 ± 2.00ghij 17.07 ± 0.06g 63.25 ± 0.52abc
GLK 14313 13.52 ± 2.11p 1.09 ± 0.11hij 15.62 ± 0.17o 13.30 ± 0.19ghij 25.28 ± 0.87f 67.18 ± 1.55a
GLK 28127 10.57 ± 1.60p 0.94 ± 0.05jk 17.25 ± 0.52o 10.62 ± 4.21hijk 40.67 ± 0.25bcd 59.42 ± 0.41cde
GLK 07-42 25.64 ± 0.76n 1.06 ± 0.04ijk 19.48 ± 0.43n 12.34 ± 1.14ghij 39.29 ± 0.62cde 56.52 ± 2.07ef
GLK 08-134 19.55 ± 2.24o 0.52 ± 0.08l 22.18 ± 0.91m 11.67 ± 0.47ghij 34.79 ± 0.87de 61.59 ± 0.93bcd
Mean 18.85 ± 1.69 0.88 ± 0.06 18.12 ± 0.53 12.04 ± 1.61 31.43 ± 0.54 61.59 ± 1.10
IC 525199 89.61 ± 0.51a 2.26 ± 0.05cd 35.87 ± 0.52de 15.31 ± 2.58fghi 11.51 ± 1.37gh 49.07 ± 4.35gh
IC 525202 77.62 ± 1.98d 2.09 ± 0.05cd 40.81 ± 0.56b 21.53 ± 1.72cdef 10.38 ± 0.50gh 42.03 ± 1.24jkl
IC 525691 84.80 ± 0.19ab 2.41 ± 0.11abc 34.42 ± 0.09ef 31.29 ± 0.38a 10.26 ± 2.62gh 41.82 ± 0.41jkl
C. jud 95 74.10 ± 0.51d 2.00 ± 0.08de 34.03 ± 0.13efg 26.98 ± 0.28abc 8.13 ± 2.25h 49.17 ± 1.55gh
C. jud 185 78.58 ± 0.76cd 2.63 ± 0.08ab 37.46 ± 1.16cd 9.18 ± 1.62ijkl 16.14 ± 0.50g 43.37 ± 0.31ijk
C. jud 185 B 77.75 ± 2.62d 2.22 ± 0.02cd 40.12 ± 0.22b 22.77 ± 0.86bcde 10.01 ± 0.12gh 49.59 ± 0.93gh
C. jud 17148 61.85 ± 0.32efgh 2.02 ± 0.05de 28.58 ± 0.26hi 26.88 ± 0.19abc 10.63 ± 1.50gh 38.20 ± 0.31lmno
C. jud 17150 59.48 ± 3.46fgh 2.66 ± 0.02ab 33.26 ± 0.22fg 11.86 ± 0.47ghij 10.51 ± 0.37gh 47.10 ± 1.14hi
C. jud ILWC 30 73.71 ± 1.02d 2.11 ± 0.08cd 38.96 ± 0.95bc 26.02 ± 0.47abc 10.26 ± 2.62gh 35.61 ± 0.62no
C. pin ILWC 0 57.17 ± 1.15hi 2.74 ± 0.09a 34.03 ± 0.39efg 26.12 ± 0.95abc 7.38 ± 0.25h 41.30 ± 1.14jklm
C. pin ILWC 261 66.66 ± 1.02e 1.64 ± 0.02fg 26.73 ± 0.13ij 24.78 ± 0.38abcd 9.76 ± 2.62gh 45.34 ± 2.28hij
EC 366338 83.20 ± 0.38bc 2.36 ± 0.13bc 37.33 ± 1.21cd 21.05 ± 0.66defg 10.26 ± 0.12gh 33.95 ± 0.21o
EC 366342 75.38 ± 2.30d 2.62 ± 0.10ab 37.89 ± 0.90cd 17.99 ± 2.00defg 6.25 ± 0.87h 36.75 ± 1.35mno
EC 556270 R 88.78 ± 0.57a 2.38 ± 0.04bc 43.60 ± 0.69a 7.84 ± 0.47jkl 5.75 ± 0.62h 39.44 ± 2.59klmn
C. ech 17159 64.74 ± 0.76ef 2.17 ± 0.03cd 32.40 ± 0.82fg 17.22 ± 1.43efgh 7.25 ± 2.12h 33.64 ± 0.52o
Mean 74.23 ± 1.18 2.29 ± 0.07 35.70 ± 0.55 20.46 ± 0.97 9.64 ± 1.10 41.76 ± 1.26
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Data represent the mean ± SD of triplicates and values with different letters in the same column are significantly different (p < 0.05)

Ferric reducing antioxidant power (FRAP)

The reaction measures reduction of ferric-2,4,6-tripyridyltriozine (TPTZ) complex to ferrous form and an intense blue colour with an absorption maximum at 593 nm develops. FRAP is a reasonable screening method for the ability to maintain redox status in cells or tissues because reaction detects compounds with redox potentials of < 0.7 V. FRAP activity in chickpea genotypes varied from 0.52 mg/g (GLK 08-134) to 2.74 mg/g (C. pin ILWC 0) (Table 3). All the kabuli genotypes had lower FRAP activity and all desi genotypes had medium FRAP activity except GNG 2171 and PBG7 which had FRAP activity in the range of (0.92 and 0.86 mg/g), respectively. All wild species possessed higher FRAP activity except C. pin ILWC 261 which had FRAP activity in the medium range (1.64 mg/g). Quintero-Soto et al. (2018) reported that FRAP values in desi chickpeas were found to be higher than those of kabuli chickpeas.

Hydroxyl radical scavenging activity

Hydroxyl radical is the most reactive among reactive oxygen radicals. It causes cell damage and other diseases by readily reacting with other groups or substances in body. Hydroxyl radicals react with polypeptides, DNA (thymine and guanosine) and proteins. Scavenging of hydroxyl radical is very important for protection against various metabolic disorders. The scavenging rate of hydroxyl radical in chickpea cultivars ranged from 2.77 (GL 13042) to 31.29% (IC 525691) with an average value of 16.10% (Table 3). Kabuli genotypes have hydroxyl radical scavenging activity ranging from 10.62 to 13.30% with an average value of 12.04%. In desi genotypes, it ranged from (2.77 to 28.89%) with an average value of 11.58%. Wild species have the average value of 20.46% with the scavenging activity ranging from 7.84 to 31.29%. Zhao et al. (2013) reported that hydroxyl radical scavenging activity in cowpea, kidney bean, chickpea and lentil was found to be 58.64%, 85.68%, 66.22% and 60.09%, respectively.

Superoxide anion radical scavenging activity

Superoxide radicals are highly toxic species which are generated by numerous biological reactions such as oxidation of haemoglobin and normal catalytic function of a number of metabolic enzymes. The concentration of superoxide free radicals increases under condition of oxidative stress and these are also generated by auto-oxidation processes or by enzymes. These free radicals are potential precursor of highly reactive species, powerful and dangerous hydroxyl radical. Superoxide anion radical scavenging activity in wild species was found to be lower (5.75 to 16.14%) as compared to desi (12.64 to 48.43%) and kabuli (17.07 to 40.67%), respectively (Table 3). Sekhon et al. (2017) investigated cultivated and wild species of pigeonpea and found that superoxide radical scavenging activity of pigeonpea ranged from 13.33 to 52.45% and in wild species it ranged from 15.5 to 24.5%.

Total reducing power

The presence of reductones is associated with reducing properties which can break the free radical chain with the donation of hydrogen atom. Reductones avert the peroxide formation by reacting with precursors of peroxides (Kumaran and Karunakaran 2007). Reducing power content in desi, kabuli and wild chickpea genotypes ranged from 22.66 to 33.09; 15.62 to 22.18 and 26.73 to 43.60 mg/100 g, respectively (Table 3).

Nitric oxide radical scavenging activity

Nitric oxide is a free radical and has an unpaired electron. Free radicals like nitrate, nitrite and s-nitrosothiols are formed when nitric oxide react rapidly in the intracellular environment. These metabolites play a key role in mediating many xenotoxic effects such as DNA damage. All kabuli genotypes possessed higher nitric oxide radical scavenging activity ranging from 59.42 to 67.18% with mean value of 61.59% (Table 3). Desi genotypes also possessed higher nitric oxide radical scavenging activity ranging from 56.52 to 65.22% except in GL29078 which had nitric oxide scavenging activity of 53.00%. In wild species, nitric oxide radical scavenging activity is in the lower range from 33.64 to 43.37% except in IC 525199, C. jud 95, C. jud 185 B, C. jud 17150 and C. pin ILWC 261 which had nitric oxide radical scavenging activity ranging from 45.34 to 49.59%. Nithiyanantham et al. (2012) reported that raw chickpea seed extract had 54% nitric oxide radical scavenging activity and green pea had 45% nitric oxide radical scavenging activity.

Correlation analysis

Correlation analysis was carried out between all the parameters and has been represented in (Table 4) using the software SPSS. The Pearson correlation indicated the positive and negative relationships between different parameters. Bound fructose (V6) has negative correlation with seed weight (r = − 0.782). Therefore, wild species with lower seed weight has higher bound fructose of sucrose and raffinose oligosaccharides. Tannin has significant negative correlation with seed weight (r = − 0.846**) and total sugars (r = − 0.437**). Trypsin inhibitor has negative correlation with seed weight (r = − 0.657**) and as from our results kabuli genotypes having higher seed weight had lower trypsin inhibitor content whereas wild species having lower seed weight had higher trypsin inhibitor content. Total phenol has significant and positive correlation with DPPH, FRAP, hydroxyl radical scavenging activity and reducing power. Verma et al. (2008) reported that antioxidant ability of seeds is positively correlated with phenolic content. Most of the genotypes possessing higher content of total phenol had higher level of DPPH radical scavenging activity (Tables 2, 3). Positively strong and significant correlation was observed in total phenol content and total antioxidant content in parameters FRAP, reducing power and DPPH (Yeo and Shahidi 2015). Total soluble sugar has positive correlation with total starch content (r = 0.225*). Phytic acid has negative correlation with minerals (iron, calcium, magnesium, copper and manganese) except zinc which showed positive correlation with phytic acid. Negative correlation of minerals with phytic acid showed that as the phytic acid content increases, reduction in minerals take place because phytic acid affect the absorption of minerals (Tiwari and Singh 2012).

Table 4.

Correlation of biochemical parameters related to nutritional and antioxidant potential

V0 V1 V2 V3 V4 V5 V6 V7 V8 V9 V10 V11 V12
V1 1
V2 NS 1
V3 0.486** 0.403** 1
V4 0.608** NS 0.225* 1
V5 − 0.782** NS − 0.381** − 0.635** 1
V6 − 0.846** NS − 0.437** − 0.598** 0.723** 1
V7 0.621** NS NS 0.497** − 0.586** − 0.672** 1
V8 − 0.481** NS − 0.340** − 0.660** 0.542** 0.543** − 0.294** 1
V9 − 0.657** NS − 0.492** − 0.563** 0.431** 0.468** − 0.428** 0.337** 1
V10 − 0.809** NS − 0.249* − 0.474** 0.678** 0.780** − 0.634** 0.395** 0.505** 1
V11 − 0.643** NS NS NS 0.392** 0.587** − 0.348** NS NS 0.711** 1
V12 − 0.897** NS − 0.265* − 0.542** 0.636** 0.749** − 0.647** 0.360** 0.605** 0.881** 0.676** 1
V13 − 0.822** − 0.304** − 0.588** − 0.461** 0.671** 0.799** − 0.592** 0.446** 0.637** 0.778** 0.505** 0.715**
V14 0.709** NS 0.412** 0.582** − 0.728** − 0.684** 0.569** − 0.345** − 0.495** − 0.689** − 0.381** − 0.632**
V15 − 0.482** NS NS − 0.444** 0.424** 0.539** − 0.463** NS 0.252* 0.456** 0.509** 0.471**
V16 − 0.841** NS − 0.462** − 0.465** 0.673** 0.802** − 0.562** 0.474** 0.616** 0.888** 0.615** 0.862**
V17 0.810** 0.339** 0.482** 0.673** − 0.880** − 0.785** 0.677** − 0.527** − 0.522** − 0.726** − 0.476** − 0.694**
V18 NS 0.241* NS 0.230* NS − 0.225* 0.343** NS NS − 0.249* NS − 0.216*
V19 − 0.376** NS NS NS 0.346** 0.317** − 0.243** NS NS 0.386** 0.435** 0.306**
V20 − 0.681** NS − 0.260* − 0.497** 0.529** 0.690** − 0.682** 0.311** 0.500** 0.574** 0.431** 0.631**
V21 − 0.381** NS NS − 0.292** 0.320*8 0.442** − 0.549** 0.215* 0.244* 0.385*8 0.270** 0.409**
V22 − 0.291** NS NS − 0.269* NS 0.332** NS NS NS 0.259* 0.418** 0.279**
V23 − 0.447** NS NS − 0.214* 0.485** 0.536** − 0.220** 0.281** − 0.212* 0.367** 0.445** 0.346**
V0 V13 V14 V15 V16 V17 V18 V19 V20 V21 V22 V23
V1
V2
V3
V4
V5
V6
V7
V8
V9
V10
V11
V12
V13 1
V14 − 0.613** 1
V15 0.322** − 0.491** 1
V16 0.850** − 0.582** 0.351** 1
V17 − 0.758** 0.744** − 0.467** − 0.740** 1
V18 − 0.294** 0.225* NS − 0.249* 0.331** 1
V19 0.347** − 0.253* NS 0.364** NS NS 1
V20 0.570** − 0.676** 0.439** 0.552** − 0.638** − 0.375*8 0.241** 1
V21 0.273** − 0.356** 0.320** 0.270** − 0.355** NS NS 0.521** 1
V22 NS − 0.274** 0.382** 0.213* NS NS 0.298** NS NS 1
V23 0.279** − 0.442** − 0.420** 0.315** − 0.464** NS NS NS NS NS 1
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NS represent non significant coefficients. V1, Seed Weight; V2, Crude Protein; V3, Total Soluble Sugar; V4-Total Starch; V5, Bound Fructose; V6, Tannin; V7, Phytic Acid; V8, Saponin; V9, Trypsin Inhibitor; V10, Total Phenol; V11, Flavonol; V12, DPPH (2;2-diphenyl-1-picryl hydrazyl) activity; V13, Ferric Reducing Antioxidant Power; V14, Superoxide anion radical scavenging activity; V15, Hydroxyl radical scavenging activity; V16, Reducing Power; V17, Nitric Oxide radical scavenging activity; V18, Zinc; V19, Iron; V20-Calcium; V21, Magnesium; V22, Copper; V23, Manganese

**Correlation is significant at the 0.01 level (2-tailed), *Correlation is significant at the 0.05 level (2-tailed)

Conclusion

Thirty chickpea genotypes (ten desi, five kabuli and fifteen wild) compared for nutritional, antinutritional and antioxidant potential showed a lot of diversity which can be further used. In desi genotypes, GL 12021 had high crude protein content, high soluble protein content and high starch content, lower phytic acid and saponin and medium trypsin inhibitor and tannin content. GNG 2171 possessed higher crude protein content and soluble sugar content and GLK 28127 possessed crude protein and soluble sugar content in medium range whereas both possessed lower content of tannin and phytic acid. Kabuli genotype (L 552) contained higher content of total starch, soluble sugar, Zn and Fe and lower content of tannin, saponin and trypsin inhibitor. Wild species C. pin ILWC 261 had higher crude protein content and lower trypsin inhibitor and phytic acid content. Kabuli genotypes possessed the lowest antioxidant activity.

Abbreviations

BAPNA

Nα-benzoyl-dl-arginine p-nitroanilide

DPPH

(2,2-Diphenyl-1-picryl hydrazyl)

FRAP

Ferric reducing antioxidant power

ROS

Reactive oxygen species

Footnotes

Publisher's Note

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