Evaluation of Iron, Zinc, and Protein Contents of Common Bean (Phaseolus vulgaris L.) Genotypes: A Collection from Jammu & Kashmir, India  

Reetika Mahajan1 , Sajad Majeed Zargar1 , Aziz Mujtaba Aezum2 , Sufia Farhat1 , Mudasir Gani3 , Aftab Hussain I.S4 , Ganesh Kumar Agrawal5 , Randeep Rakwal5,6
1. School of Biotechnology, S K University of Agricultural Sciences and Technology of Jammu, Chatha, Jammu, J&K-180009, India
2. Department of Soil Science, MRCFC, Khudwani, S K University of Agricultural Sciences and Technology of Kashmir, Srinagar, J&K-192101, India
3. Division of Entomology, S K University of Agricultural Sciences and Technology of Jammu, Chatha, Jammu, J&K-180009, India
4. Department of Crop Physiology, University of Agricultural Sciences, GKVK, Bangalore-65, India
5. Research Laboratory for Biotechnology and Biochemistry (RLABB), Kathmandu, Nepal
6. Organization for Educational Initiatives, University of Tsukuba, Tsukuba, Ibaraki 305-8577, Japan
Author    Correspondence author
Legume Genomics and Genetics, 2015, Vol. 6, No. 2   doi: 10.5376/lgg.2015.06.0002
Received: 04 Jan., 2015    Accepted: 15 Feb., 2015    Published: 26 Feb., 2015
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Mahajan et al., 2015, Evaluation of Iron, Zinc, and Protein Contents of Common Bean (Phaseolus vulgaris L.) Genotypes: A collection from Jammu & Kashmir, India, Legume Genomics and Genetics, Vol.6, No.2, 1-7 (doi: 10.5376/lgg.2015.06.0002)


Common bean (Phaseolus vulgaris. L), is the most important legume crop, rich in high-quality protein, energy, fiber and micronutrients especially iron, zinc, and pro-vitamin A. High genetic diversity is known to exist among bean genotypes for micronutrient densities. Considering these facts and the status of micronutrient malnutrition in the developing countries, we have evaluated the seeds of 51 common bean genotypes collected from different locations of Jammu and Kashmir, with the major objective of identifying genotypes with high protein and micronutrient contents. Results revealed that the variation in the iron (Fe) content was very high ranging from 0.71 mg to 7.22 mg 100g-1, and the zinc (Zn) content varied from 0.43 mg to 1.93 mg 100g-1. The variation in protein content was also very high ranging from 7.2% to 31.6%. No correlation was found for Fe, Zn, and protein contents. This variability implies that, the screened genotypes could serve as a source for breeding new varieties with improved biochemical and nutritional traits and could be highly suited to meet specific dietary requirements.

Micronutrient; Malnutrition; Fe; Zn; Protein; Common bean

Common bean (Phaseolus vulgaris L.) is predominantly a self-pollinated crop originating in Latin America and mostly cultivated in the tropics and subtropics as well as in the temperate regions of the world (Gepts and Bliss 1988; Zeven, 1997; Zeven et al., 1999). Common bean represents 50% of the grain legume consumed worldwide, and is thus considered as the most important legume crop (McConnell et al., 2010). This crop is grown for its dry beans, and immature green pods. Being a good and cheap source of protein (2028%), minerals like Fe (70 mg/kg) and Zn (33 mg/kg), energy (32%), and fiber (56%) can solve malnutrition and hunger-related problems. Large variability among bean genotypes for micronutrient densities is known to exist (Beebe et al., 2000; Tryphone and Nichumbi-Msolla, 2010). In countries where daily bean consumption is high, it provides significant amount of proteins, calories and micronutrients helping to avoid the consequences of malnutrition and hunger (Valdemiro and Whitaker, 1982). Common bean is considered as an important functional food because of its high levels of chemically diverse components (phenols, starch, vitamins, fructo-oligosaccharides) which give protection against oxidative stresses, cardiovascular diseases, diabetes, metabolic syndrome and many types of cancers (Camara et al., 2013). Loss of crop diversity and extinction of genetic resources have led to a deterioration of nutritional quality of food crops (Gouveia et al., 2011; Singh, 2001). Micronutrients like Fe and Zn play an important role in the metabolism of both plants and animals. Iron deficiency anemia (IDA) and other micronutrient deficiencies affect large number of people worldwide and in many parts of Africa and Latin America it is the top health concern especially among the poor (Graham et al., 2001). Worldwide, over three billion people are affected by IDA while zinc deficiency in the human diet is probably almost as widespread as iron deficiency (Frossard et al., 2000). IDA causes loss in work productivity and complications in child birth while zinc deficiency causes stunting and reduces immunity against disease causing organisms (Welch and Graham 1999, 2004). Keeping these facts in view, in this study we evaluated the common bean genotypes for seed Fe, Zn & protein contents in order to identify superior genotypes that can act as a genetic resource for mining alleles/QTLs contributing for higher amounts of Fe and Zn. These genes/QTLs can further be introgressed in desired background through molecular breeding for enhancing nutritional value of common bean varieties.

1 Results and Discussion
1.1 Evaluation of common bean genotypes for Fe, Zn, & protein contents in seed material
Fe, Zn, and protein contents in the seeds of 51 diverse genotypes of common bean were estimated to determine the variation among them. Wide variation was found in Fe concentration ranging from 0.71 to 7.22 mg 100g-1 with an average of 1.81 mg 100g-1. Genotype R2 possesses highest seed Fe content (7.22 mg 100g-1) whereas genotype K12 has lowest Fe content (0.71 mg 100g-1). The Zn concentration in seed varied from 0.43 to 1.93 mg 100g-1 with an average of 0.78 mg 100g-1. Genotype K15 possesses high Zn content in seed (1.93 mg 100g-1) whereas genotype KS6 has lowest Zn content (0.43 mg 100g-1). Table 1 and Figure 1 represent the mean values of Fe and Zn content observed in seeds of 51 diverse common bean genotypes. In earlier reports, we observed a similar pattern of variation in Fe & Zn content in common bean seeds. Silva et al. (2010) observed a wide variation in Fe & Zn contents among 100 diverse common bean lines ranging from 54.20 to 161.50 mg kg-1 and 29.33 to 65.50 mg kg-1, Fe and Zn respectively. Similarly, another study also revealed a variation in Fe and Zn content from 34 to >100 mg kg-1 and 21 to 54 mg kg-1, respectively among 2000 common bean accessions of CIAT (Beebe et al., 2000). In a recent report, 117 genotypes of common bean collected from Uganda showed variation in Fe and Zn contents ranging from 45 to 87mg kg-1 and 22 to 40mg kg-1, respectively (Mukamuhirwa et al., 2012). Since, there is a huge variation in Fe and Zn contents in our material, we, suggest that, genotypes with highest Fe and Zn can be used as genetic resource for improving nutritional quality of adopted common bean cultivars.
We further analyzed seeds for their protein content, which ranged from 7.2% to 31.6% with an average content of 20.30%. Highest protein content was observed in KS6 (31.6%) whereas genotype K12 had the least protein content of 7.2%. Table 1 and Figure 1 represent the mean values of protein content observed in seeds of 51 diverse common bean genotypes. Similar results have been reported in previous studies, although variation between the content may be due to the environmental factors, geographical location, and growing season. In earlier reports, the variation in the seed protein content was observed ranging from 17.4% to 29% (Sood et al., 2003; Silva and Iachan, 1975; Sgarbieri et al., 1979; Márquez and Lajolo, 1981, Durigan & Sgarbieri, 1985; Durigan et al., 1987 & Tezoto & Sgarbieri, 1990). Protein content of 36 North American bean cultivars evaluated by Koehler et al., (1987) also ranged from 19.6 to 32.2%. A wide range in micronutrient and protein contents among 51 genotypes in this study indicates the existence of extensive genetic variation which can be explored for enhancing nutritional value of common bean varieties. Further, we grouped these 51 genotypes on the basis of their mean values (Table 2) to cluster them based on particular range of micronutrient (Fe and Zn) and protein content.

Table 1 Variation among Fe, Zn, and protein content in diverse common bean genotypes

Table 2 Distribution of 51 common bean genotypes on the bases of their mean value range of Fe, Zn, & protein

Figure 1 Mean±SE of mineral (Fe & Zn) and protein concentrations in seeds of common bean and their correlation with each other


1.2 Variation in Fe, Zn, and protein among the genotypes and within the genotype
The comparison of Fe, Zn, and protein contents revealed non-significant differences among most of the genotypes, whereas the comparison of the Fe & Zn, Zn & Protein, and Fe & protein contents within the genotype indicated that, these traits are significantly different except three genotypes detailed in (Table 1).
1.3 Correlation among micronutrient (Fe & Zn) and protein contents
Negative but non-significant correlation was found between Fe and Zn (r=-0.022; p>0.05), Fe and protein (r=-0.037; p>0.05), and Zn and protein (r=-0.037; p>0.05) (Figure 1, Table 3). In an earlier study, Zn & Fe content in common bean seeds were found inversely correlated (r=-0.11; p<0.05) (Akond et al., 2011). Thus, it can be interpreted that accumulation of one micronutrient (in this case Fe or Zn) has negative impact on concentration of the other, as such there is a genetic regulation governing the transport and accumulation of these micronutrients.

Table 3 Correlation among Fe, Zn, & protein

2 Conclusion
In the present study, we fulfilled the objective of the study by evaluating the micronutrient (Fe, and Zn) and protein contents of the common bean genotypes collected from different regions of Jammu & Kashmir, India. The findings from this study suggest that there is a wide variability in Fe, Zn, and protein content in this genetic stock. As such it can be used as a genetic resource for further improvement of this crop for enhancing its nutritional value.
3 Material and Methods
3.1 Plant material
The purified seed material of 51 genotypes of common bean, collected from different locations of Jammu & Kashmir, India were used in the present study. The details for these genotypes are previously described in Zargar et al. (2014).
3.2 Chemical analysis
Dry seeds (10~20) from each genotype were finely powdered in a grinder. 250 mg of this powder was cold digested in 5 mL concentrated nitric acid over-night, followed by digestion in 5 ml of diacid mixture containing nitric acid: perchloric acid, in 10:4 proportion (Jackson 1973). The resulting clear solution was diluted to 25 mL using double distilled water. The concentration of Fe and Zn was determined using ICPOES (Thermo Fischer 600). The concentration values were further converted and expressed in mg100g-1 using the following formula.

Protein content was determined by using Lowry's method (Lowry et al., 1951). 500 mg of powdered sample was extracted in sodium buffer followed by centrifugation (5000 rpm for 10 minutes). Protein content was estimated by adding alkaline copper sulphate and folin ciocalteau solution to the supernatant. Absorbance was recorded at 660 nm using UV visible spectrophotometer (UV-1601, Shimadzu, Japan). Protein content was calculated and expressed in percentage.
3.3 Statistical analysis
All the observations were taken in three replicates and values were then averaged. One-way ANOVA was applied to evaluate the variance of protein and mineral contents (Fe & Zn) among the genotype. Paired-t test was used to determine variance of Fe, Zn & protein within the genotype. The Pearson’s correlation coefficient between Fe, Zn, and protein was determined by Pearson's correlation analysis using SPSS (version 16).
SMZ is grateful to SERB, DST New Delhi for financial support of this work (Project sanction order No. SR/FT/LS-27/2011).
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