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Research Article

volume 40 issue 6 (december 2017) : 1004-1011,   Doi: 10.18805/lr.v40i04.9006

Identification of genomic Regions/genes for high iron and zinc content and cross transferability of SSR markers in mungbean (Vigna radiata L.)

V
Vijayata Singh
R
R.K. Yadav
N
N.R. Yadav
R
Rajesh Yadav
R
R.S. Malik
J
Jogendra Singh
1Chaudhary Charan Singh Haryana Agricultural University, Hisar-125004, Haryana, India.
Cite article:- Singh Vijayata, Yadav R.K., Yadav N.R., Yadav Rajesh, Malik R.S., Singh Jogendra (2017). Identification of genomic Regions/genes for high iron and zinc content and cross transferability of SSR markers in mungbean (Vigna radiata L.). Legume Research. 40(6): 1004-1011. doi: 10.18805/lr.v40i04.9006.

ABSTRACT

Among the legumes, mungbean has highest digestive protein but low micronutrient content like iron and zinc. Biofortification of mungbean has been undertaken to reduce micronutrient malnutrition. The objectives of this study were to identify QTLs for seed Fe and Zn content in F6 recombinant inbred line (RIL) population (ML776 x Sattya). A large genetic variation and transgressive segregation in RILs were observed for Fe and Zn content. Linkage map was developed which spanned 2919.7cM distance. 17 QTLs (2 for iron and 15 for zinc content) were mapped on four linkage groups; LG 4, LG 6, LG 7 and LG 11 in mungbean. The genomic regions qZn-4-3 and qFe-4-1 on chromosome 4 between PVBR82-BM210 markers; qZn-11-2 and qFe-11-1 on chromosome11 between BM141-BM184 markers, were co-located on the same chromosomal regions for Zn or Fe concentration, which probably were closely linked to each other, or were the same pleiotropic QTLs. The SSR markers associated with QTLs for both high iron and zinc content would also be useful in marker assisted breeding for biofortification in mungbean.  

REFERENCES

  1. Aneja, B., Yadav, N.R., Chawla, V. and Yadav, R.C. (2012). Sequence related amplified polymorphism (SRAP) molecular marker system and its application in crop improvement. Mol. Breeding, 1-14.
  2. Anuradha, K., Agarwal, S., Rao, Y.V., Rao, K.V., Viraktamath, B.C. and Sarla, N. (2012). Mapping QTLs and candidate genes for iron and zinc concentrations in unpolished rice of Madhukar x Swarna RILs. Gene, 508 (2): 233-40.
  3. Aubert, G., Morin, J., Jacquin, F., Loridon, K., Quillet, M.C., Petit, A., Rameau, C., Lejeune, H.I., Huguet, T. and Burstin, J. (2006). Functional mapping in pea, as an aid to the candidate gene selection and for investigating synteny with the model legume Medicago truncatula. Theor Appl. Genet., 112: 1024-1041.
  4. Beebe, S., Gonzalez, A.V. and Rengifo, J. (2000). Research on trace minerals in the common bean. Food. Nutr. Bull., 21 (4): 387-391.
  5. Blair, M.W. and Izquierdo, P. (2012). Use of the advanced backcross-QTL method to transfer seed mineral accumulation nutrition traits from wild to Andean cultivated common beans. Theor. Appl. Genet., 125:1015–1031.
  6. Blair, M.W., Astudillo, C., Grusak, M.A., Graham, R. and Beebe, S.E. (2009). Inheritance of seed iron and zinc concentrations in common bean (Phaseolus vulgaris L.). Mol. Breeding, 23:197-207.
  7. Blair, M.W., Buendia, H.F., Giraldo, M.C., Métais, I. and Peltier, D. (2008). Characterization of AT-rich microsatellites in common bean (Phaseolus vulgaris L.). Theor. Appl. Genet., 118: 91–103.
  8. Blair, M.W., Iriarte, G. and Beebe, S. (2006b). QTL analysis of yield traits in an advanced backcross population derived from a cultivated Andean x wild common bean (Phaseolus vulgaris L.) cross. Theor. Appl. Genet., 112:1149–    1163.
  9. Blair, M.W., Medina, J.I., Astudillo, C., Rengifo, J., Beebe, S.E., Machado, G. and Graham, R. (2010). QTL for seed iron and zinc concentration in a Mesoamerican common bean (Phaseolus vulgaris L.) population. Theor. Appl. Genet., 121: 1059-1070.
  10. Blair, M.W., Pedraza, F., Buendia, H.F., Gaitán-Solís, E., Beebe, S.E., Gepts, P. and Tohme, J. (2003). Development of a genome-wide anchored microsatellite map for common bean (Phaseolus vulgaris L.). Theor. Appl. Genet., 107(8):1362-74.
  11. Bouis, H.E. (2003). Micronutrient fortification of plants through plant breeding: can it improve nutrition in man at low cost? Proc. Nutr. Soc., 62: 403–411.
  12. Brown, K.H., Wuehler, S.E. and Peerson, J.M. (2001). The importance of zinc in human nutrition and estimation of the global prevalence of zinc deficiency. Food Nutr. Bull., 22: 113-125.
  13. Cakmak,    I., Torun, A., Millet, E., Feldman, M., Fahima, T., Korol, A., Nevo, E., Braun, H.J. and Ozkan, H. (2004). Triticum dicoccoides: An important genetic resource for increasing zinc and iron concentration in modern cultivated wheat. Soil Sci. Plant Nutri., 50:1047-1054.
  14. Chandel, G., Samuel, P., Dubey, M. and Meena, R. (2011). In silico expression analysis of QTL specific candidate genes for grain micronutrient (Fe/Zn) content using ESTs and MPSS signature analysis in rice (Oryza sativa L.). J. Plant. Genet. Transgenics, 1:11–22.
  15. Choudhary, S., Sethy, N.K., Shokeen, B. and Bhatia, S. (2008). Development of 3chickpea ESTSSR markers and analysis of allelic variation across related species. Theor. Appl. Genet., 118: 591-608.
  16. Clemens, S. (2001). Molecular mechanisms of plant metal tolerance and homeostasis. Planta, 212: 475–486.
  17. Dikshit, H.K., Singh, D., Singh, A., Jain, N., Kumari, J. and Sharma, T.R. (2012). Utility of adzuki bean [Vigna angularis (Willd.) Ohwi & Ohashi] simple sequence repeats (SSR) markers in genetic analysis of mungbean and related Vigna spp. Afr. J. Biotechnol., 11 (69): 13261-13268. 
  18. Fletcher, R.J., Bell, I.P. and Lambert, J.P. (2004). Public health aspects of food fortification: A question of balance. Proc. Nutr. Soc., 63: 605-615.
  19. Garcia-Oliveira, A.L., Tan, L., Fu, Y. and Sun, C. (2009). Genetic identification of Quantitative Trait Loci for contents of mineral nutrients in rice grain. J. Integr. Plant. Biol., 51(1):84-92.
  20. Gelin, J.R., Forster, S., Grafton, K.F., McClean, P. and Rojas-Cifuentes, G.A. (2007). Analysis of seed-zinc and other nutrients in a recombinant inbred population of navy bean (Phaseolus vulgaris L.). Crop Sci., 47: 1361–1366.
  21. Ghandilyan, A., Vreugdenhil, D. and Aarts, M.G.M. (2006). Progress in the genetic understanding of plant iron and zinc nutrition. Physiol. Plant., 126: 407-417.
  22. Graham, R.D. and Welch, R.M. (1999). A new paradigm for world agriculture: productive, sustainable and nutritious food systems to meet human needs. Dev. Bull. (Canberra)., 49: 29–32.
  23. Gregorio, G.B., Senadhira, D., Htut, H. and Graham, R.D. (2000). Breeding for trace mineral density in rice. Food Nutr. Bull., 21:382–386.
  24. Grotz, N. and Guerinot, M.L. (2006). Molecular aspects of Cu, Fe and Zn homeostasis in plants. Biochim. Biophys. Acta., 7: 595–608.
  25. Hacisalihoglu, G., Osturk, L., Cakmak, I., Welch, R.M. and Kochian, L. (2004). Genotypic variation in common bean in response to zinc deficiency in calcareous soil. Plant and Soil, 259: 71-83.
  26. House, W.A., Welch, R.M., Beebe, S. and Cheng, Z. (2002). Potential for increasing the amounts of bioavailable zinc in dry beans through plant breeding. J. Sci. Food Agr., 82: 1452-1457. 
  27. Jeong, J. and Guerinot, M.L. (2009). Homing in on iron homeostasis in plants. Trends Plant Sci., 14: 280–285.
  28. Koornneef, M., Alonso-Blan, C. and Vreugdeuhil, D. (2004). Naturally occurring genetic variation. In: Arabidopsis thaliana. Annu. Rev. Plant Biol., 55:141-172.
  29. Kosambi, D.D. (1994). The estimation of a map distance from recombination values. Annals of Eugenics.12 (3): 172-175.
  30. Palmer, C.M. and Guerinot, M.L. (2009). Facing the challenges of Cu, Fe and Zn homeostasis in plants. Nat. Chem. Biol., 5 (5): 333–340.
  31. Pandian, A., Ford, R. and Taylor, W.J. (2000). Transferability of sequence tagged microsatellite sites (STMS) primers across major pulses. Plant Mol. Biol. Rep., 18: 395a-395h.
  32. Pfeiffer, W.H. and McClafferty, B. (2007). Harvest Plus: Breeding crops for better nutrition. Crop Sci., 47 (S3): S88-S105.
  33. Phan, H.T., Ellwood, S.R., Hane, J.K., Ford, R., Materne, M. and Oliver, R.P. (2007). Extensive macro synteny between Medicago truncatula and Lens culinaris ssp. culinaris. Theor. Appl. Genet., 114: 549-558.
  34. Roorkiwal, M. and Sharma, P. C. (2011). Mining functional microsatellite in legume unigenes. Bioinformation, 7 (5): 264-    270.
  35. Saghai-Maroof, M.A., Soliman, K.M., Jorgensen, R.A. and Allard, R.W. (1984). Ribosomal DNA spacer-length polymorphisms in barly mendelian inheritance, chromosomal location and population dynamics. Proc. Natl. Acad. Sci. USA., 81: 8014-8018.
  36. Sarkar, A., Dikshit, H.K., Shrestha, R., Omar, M., Million, E. and Jalaluddin, M. (2009). Biofortification of grain legumes for quality improvement. International Conference on Grain Legumes, Kanpur, India. Pp. 5-6.
  37. Šimiæ, D., Mladenoviæ, D., Zduniæ, Z., Jambroviæ, A., Ledenèan, T., Brkiæ, J., Brkiæ, A. and Brkiæ, I. (2012). Quantitative trait loci for biofortification traits in maize grain. J. Hered., 103 (1): 47-54.
  38. Singh, R., VanHeusden, A.W., Kumar, R., Visser, R.G.F. and Yadav, R.C. (2013). Genetic diversity of Mungbean (Vigna radiata L.) in iron and zinc Content as Impacted by Farmer’s Varietal Selection in Northern India. Ecol. Food Nutr. 52 (2): 148-162.
  39. Singh,V., Yadav, R. K., Yadav, R., Malik, R.S., Yadav, N.R. and Singh, J. (2013). Stability analysis in Mungbean [Vigna Radiata (L.) Wilczek)] for nutritional quality and seed yield. Legume Res., 36(1): 56-61.
  40. Singh,V., Yadav, R. K., Yadav, R., Malik, R.S., Yadav, N.R., Singh, J. and Meena, M.D. (2013b). Effect of different Iron and Zinc application on growth, yield and quality parameters of Mungbean (Vigna radiata L.). Annals of Agri-Bio Res., 18 (2): 164-175.
  41. Stangoulis, J.C.R., Huynh, B.L., Welch, R.M., Choi, E.Y. and Graham, R.D. (2007). Quantitative trait loci for phytate in rice grain and their relationship with grain micronutrient content. Euphytica, 154: 289-294.
  42. Taunk, J., Yadav, N.R., Yadav, R.C. and Kumar, R. (2012). Genetic diversity among green gram [Vigna radiata (L.) Wilczek] genotypes varying in micronutrient (Fe and Zn) content using RAPD markers. Indian J. Biotechnol., 11: 48-53.
  43. Tiwari, V.K., Rawat, N., Chhuneja, P., Kumari, N., Aggarwal, R., Randhawa, G.S., Dhaliwal, H.S., Keller, B. and Singh, K. (2009). Mapping of Quantitative Trait Loci for Grain Iron and Zinc Concentration in Diploid A Genome. J. Hered., 100: 771-776.
  44. Vert, G., Grotz, N., De´dalde´champ, F., Gaymard, F., Guerinot, M.F., Briat, J.F. and Curie, C. (2002). IRT1, an Arabidopsis transporter essential for iron uptake from the soil and for plant growth. Plant Cell, 14: 1223–1233.
  45. Wang, L.X., Cheng, X.Z., Wang, S.H., Liu, C.Y. and Liang, H. (2009). Transferability of SSR markers from adzuki bean into mungbean. Acta Agron. Sinica, 35: 816-820.
  46. Weeden, N.F., Muehlbaur, F.J. and Ladizinsky, G. (1992). Extensive conservation of linkage relationships between pea and lentil genetic maps. J. Hered., 83: 123-129.
  47. Welch, R.M. (2002). Breeding strategies for biofortified staple plant foods to reduce micronutrient malnutrition globally. J. Nutri., 132: 495S-499S.
  48. Yu, K., Park, J., Poysa, V. and Gepts, P. (2000). Integration of simple sequence repeat (SSR) markers into a molecular linkage map of common bean (Phaseolus vulgaris L.). J. Hered., 91: 429-434.
  49. Zhu, C., Naqvi, S., Galera, S.G., Pelacho, A.M., Capell, T. and Christou, P. (2007). Transgenic strategies for the nutritional enhancement of plants. Trends Plant. Sci., 12: 548–555. 
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    Research Article

    volume 40 issue 6 (december 2017) : 1004-1011,   Doi: 10.18805/lr.v40i04.9006

    Identification of genomic Regions/genes for high iron and zinc content and cross transferability of SSR markers in mungbean (Vigna radiata L.)

    V
    Vijayata Singh
    R
    R.K. Yadav
    N
    N.R. Yadav
    R
    Rajesh Yadav
    R
    R.S. Malik
    J
    Jogendra Singh
    1Chaudhary Charan Singh Haryana Agricultural University, Hisar-125004, Haryana, India.
    Cite article:- Singh Vijayata, Yadav R.K., Yadav N.R., Yadav Rajesh, Malik R.S., Singh Jogendra (2017). Identification of genomic Regions/genes for high iron and zinc content and cross transferability of SSR markers in mungbean (Vigna radiata L.). Legume Research. 40(6): 1004-1011. doi: 10.18805/lr.v40i04.9006.

    ABSTRACT

    Among the legumes, mungbean has highest digestive protein but low micronutrient content like iron and zinc. Biofortification of mungbean has been undertaken to reduce micronutrient malnutrition. The objectives of this study were to identify QTLs for seed Fe and Zn content in F6 recombinant inbred line (RIL) population (ML776 x Sattya). A large genetic variation and transgressive segregation in RILs were observed for Fe and Zn content. Linkage map was developed which spanned 2919.7cM distance. 17 QTLs (2 for iron and 15 for zinc content) were mapped on four linkage groups; LG 4, LG 6, LG 7 and LG 11 in mungbean. The genomic regions qZn-4-3 and qFe-4-1 on chromosome 4 between PVBR82-BM210 markers; qZn-11-2 and qFe-11-1 on chromosome11 between BM141-BM184 markers, were co-located on the same chromosomal regions for Zn or Fe concentration, which probably were closely linked to each other, or were the same pleiotropic QTLs. The SSR markers associated with QTLs for both high iron and zinc content would also be useful in marker assisted breeding for biofortification in mungbean.  

    REFERENCES

    1. Aneja, B., Yadav, N.R., Chawla, V. and Yadav, R.C. (2012). Sequence related amplified polymorphism (SRAP) molecular marker system and its application in crop improvement. Mol. Breeding, 1-14.
    2. Anuradha, K., Agarwal, S., Rao, Y.V., Rao, K.V., Viraktamath, B.C. and Sarla, N. (2012). Mapping QTLs and candidate genes for iron and zinc concentrations in unpolished rice of Madhukar x Swarna RILs. Gene, 508 (2): 233-40.
    3. Aubert, G., Morin, J., Jacquin, F., Loridon, K., Quillet, M.C., Petit, A., Rameau, C., Lejeune, H.I., Huguet, T. and Burstin, J. (2006). Functional mapping in pea, as an aid to the candidate gene selection and for investigating synteny with the model legume Medicago truncatula. Theor Appl. Genet., 112: 1024-1041.
    4. Beebe, S., Gonzalez, A.V. and Rengifo, J. (2000). Research on trace minerals in the common bean. Food. Nutr. Bull., 21 (4): 387-391.
    5. Blair, M.W. and Izquierdo, P. (2012). Use of the advanced backcross-QTL method to transfer seed mineral accumulation nutrition traits from wild to Andean cultivated common beans. Theor. Appl. Genet., 125:1015–1031.
    6. Blair, M.W., Astudillo, C., Grusak, M.A., Graham, R. and Beebe, S.E. (2009). Inheritance of seed iron and zinc concentrations in common bean (Phaseolus vulgaris L.). Mol. Breeding, 23:197-207.
    7. Blair, M.W., Buendia, H.F., Giraldo, M.C., Métais, I. and Peltier, D. (2008). Characterization of AT-rich microsatellites in common bean (Phaseolus vulgaris L.). Theor. Appl. Genet., 118: 91–103.
    8. Blair, M.W., Iriarte, G. and Beebe, S. (2006b). QTL analysis of yield traits in an advanced backcross population derived from a cultivated Andean x wild common bean (Phaseolus vulgaris L.) cross. Theor. Appl. Genet., 112:1149–    1163.
    9. Blair, M.W., Medina, J.I., Astudillo, C., Rengifo, J., Beebe, S.E., Machado, G. and Graham, R. (2010). QTL for seed iron and zinc concentration in a Mesoamerican common bean (Phaseolus vulgaris L.) population. Theor. Appl. Genet., 121: 1059-1070.
    10. Blair, M.W., Pedraza, F., Buendia, H.F., Gaitán-Solís, E., Beebe, S.E., Gepts, P. and Tohme, J. (2003). Development of a genome-wide anchored microsatellite map for common bean (Phaseolus vulgaris L.). Theor. Appl. Genet., 107(8):1362-74.
    11. Bouis, H.E. (2003). Micronutrient fortification of plants through plant breeding: can it improve nutrition in man at low cost? Proc. Nutr. Soc., 62: 403–411.
    12. Brown, K.H., Wuehler, S.E. and Peerson, J.M. (2001). The importance of zinc in human nutrition and estimation of the global prevalence of zinc deficiency. Food Nutr. Bull., 22: 113-125.
    13. Cakmak,    I., Torun, A., Millet, E., Feldman, M., Fahima, T., Korol, A., Nevo, E., Braun, H.J. and Ozkan, H. (2004). Triticum dicoccoides: An important genetic resource for increasing zinc and iron concentration in modern cultivated wheat. Soil Sci. Plant Nutri., 50:1047-1054.
    14. Chandel, G., Samuel, P., Dubey, M. and Meena, R. (2011). In silico expression analysis of QTL specific candidate genes for grain micronutrient (Fe/Zn) content using ESTs and MPSS signature analysis in rice (Oryza sativa L.). J. Plant. Genet. Transgenics, 1:11–22.
    15. Choudhary, S., Sethy, N.K., Shokeen, B. and Bhatia, S. (2008). Development of 3chickpea ESTSSR markers and analysis of allelic variation across related species. Theor. Appl. Genet., 118: 591-608.
    16. Clemens, S. (2001). Molecular mechanisms of plant metal tolerance and homeostasis. Planta, 212: 475–486.
    17. Dikshit, H.K., Singh, D., Singh, A., Jain, N., Kumari, J. and Sharma, T.R. (2012). Utility of adzuki bean [Vigna angularis (Willd.) Ohwi & Ohashi] simple sequence repeats (SSR) markers in genetic analysis of mungbean and related Vigna spp. Afr. J. Biotechnol., 11 (69): 13261-13268. 
    18. Fletcher, R.J., Bell, I.P. and Lambert, J.P. (2004). Public health aspects of food fortification: A question of balance. Proc. Nutr. Soc., 63: 605-615.
    19. Garcia-Oliveira, A.L., Tan, L., Fu, Y. and Sun, C. (2009). Genetic identification of Quantitative Trait Loci for contents of mineral nutrients in rice grain. J. Integr. Plant. Biol., 51(1):84-92.
    20. Gelin, J.R., Forster, S., Grafton, K.F., McClean, P. and Rojas-Cifuentes, G.A. (2007). Analysis of seed-zinc and other nutrients in a recombinant inbred population of navy bean (Phaseolus vulgaris L.). Crop Sci., 47: 1361–1366.
    21. Ghandilyan, A., Vreugdenhil, D. and Aarts, M.G.M. (2006). Progress in the genetic understanding of plant iron and zinc nutrition. Physiol. Plant., 126: 407-417.
    22. Graham, R.D. and Welch, R.M. (1999). A new paradigm for world agriculture: productive, sustainable and nutritious food systems to meet human needs. Dev. Bull. (Canberra)., 49: 29–32.
    23. Gregorio, G.B., Senadhira, D., Htut, H. and Graham, R.D. (2000). Breeding for trace mineral density in rice. Food Nutr. Bull., 21:382–386.
    24. Grotz, N. and Guerinot, M.L. (2006). Molecular aspects of Cu, Fe and Zn homeostasis in plants. Biochim. Biophys. Acta., 7: 595–608.
    25. Hacisalihoglu, G., Osturk, L., Cakmak, I., Welch, R.M. and Kochian, L. (2004). Genotypic variation in common bean in response to zinc deficiency in calcareous soil. Plant and Soil, 259: 71-83.
    26. House, W.A., Welch, R.M., Beebe, S. and Cheng, Z. (2002). Potential for increasing the amounts of bioavailable zinc in dry beans through plant breeding. J. Sci. Food Agr., 82: 1452-1457. 
    27. Jeong, J. and Guerinot, M.L. (2009). Homing in on iron homeostasis in plants. Trends Plant Sci., 14: 280–285.
    28. Koornneef, M., Alonso-Blan, C. and Vreugdeuhil, D. (2004). Naturally occurring genetic variation. In: Arabidopsis thaliana. Annu. Rev. Plant Biol., 55:141-172.
    29. Kosambi, D.D. (1994). The estimation of a map distance from recombination values. Annals of Eugenics.12 (3): 172-175.
    30. Palmer, C.M. and Guerinot, M.L. (2009). Facing the challenges of Cu, Fe and Zn homeostasis in plants. Nat. Chem. Biol., 5 (5): 333–340.
    31. Pandian, A., Ford, R. and Taylor, W.J. (2000). Transferability of sequence tagged microsatellite sites (STMS) primers across major pulses. Plant Mol. Biol. Rep., 18: 395a-395h.
    32. Pfeiffer, W.H. and McClafferty, B. (2007). Harvest Plus: Breeding crops for better nutrition. Crop Sci., 47 (S3): S88-S105.
    33. Phan, H.T., Ellwood, S.R., Hane, J.K., Ford, R., Materne, M. and Oliver, R.P. (2007). Extensive macro synteny between Medicago truncatula and Lens culinaris ssp. culinaris. Theor. Appl. Genet., 114: 549-558.
    34. Roorkiwal, M. and Sharma, P. C. (2011). Mining functional microsatellite in legume unigenes. Bioinformation, 7 (5): 264-    270.
    35. Saghai-Maroof, M.A., Soliman, K.M., Jorgensen, R.A. and Allard, R.W. (1984). Ribosomal DNA spacer-length polymorphisms in barly mendelian inheritance, chromosomal location and population dynamics. Proc. Natl. Acad. Sci. USA., 81: 8014-8018.
    36. Sarkar, A., Dikshit, H.K., Shrestha, R., Omar, M., Million, E. and Jalaluddin, M. (2009). Biofortification of grain legumes for quality improvement. International Conference on Grain Legumes, Kanpur, India. Pp. 5-6.
    37. Šimiæ, D., Mladenoviæ, D., Zduniæ, Z., Jambroviæ, A., Ledenèan, T., Brkiæ, J., Brkiæ, A. and Brkiæ, I. (2012). Quantitative trait loci for biofortification traits in maize grain. J. Hered., 103 (1): 47-54.
    38. Singh, R., VanHeusden, A.W., Kumar, R., Visser, R.G.F. and Yadav, R.C. (2013). Genetic diversity of Mungbean (Vigna radiata L.) in iron and zinc Content as Impacted by Farmer’s Varietal Selection in Northern India. Ecol. Food Nutr. 52 (2): 148-162.
    39. Singh,V., Yadav, R. K., Yadav, R., Malik, R.S., Yadav, N.R. and Singh, J. (2013). Stability analysis in Mungbean [Vigna Radiata (L.) Wilczek)] for nutritional quality and seed yield. Legume Res., 36(1): 56-61.
    40. Singh,V., Yadav, R. K., Yadav, R., Malik, R.S., Yadav, N.R., Singh, J. and Meena, M.D. (2013b). Effect of different Iron and Zinc application on growth, yield and quality parameters of Mungbean (Vigna radiata L.). Annals of Agri-Bio Res., 18 (2): 164-175.
    41. Stangoulis, J.C.R., Huynh, B.L., Welch, R.M., Choi, E.Y. and Graham, R.D. (2007). Quantitative trait loci for phytate in rice grain and their relationship with grain micronutrient content. Euphytica, 154: 289-294.
    42. Taunk, J., Yadav, N.R., Yadav, R.C. and Kumar, R. (2012). Genetic diversity among green gram [Vigna radiata (L.) Wilczek] genotypes varying in micronutrient (Fe and Zn) content using RAPD markers. Indian J. Biotechnol., 11: 48-53.
    43. Tiwari, V.K., Rawat, N., Chhuneja, P., Kumari, N., Aggarwal, R., Randhawa, G.S., Dhaliwal, H.S., Keller, B. and Singh, K. (2009). Mapping of Quantitative Trait Loci for Grain Iron and Zinc Concentration in Diploid A Genome. J. Hered., 100: 771-776.
    44. Vert, G., Grotz, N., De´dalde´champ, F., Gaymard, F., Guerinot, M.F., Briat, J.F. and Curie, C. (2002). IRT1, an Arabidopsis transporter essential for iron uptake from the soil and for plant growth. Plant Cell, 14: 1223–1233.
    45. Wang, L.X., Cheng, X.Z., Wang, S.H., Liu, C.Y. and Liang, H. (2009). Transferability of SSR markers from adzuki bean into mungbean. Acta Agron. Sinica, 35: 816-820.
    46. Weeden, N.F., Muehlbaur, F.J. and Ladizinsky, G. (1992). Extensive conservation of linkage relationships between pea and lentil genetic maps. J. Hered., 83: 123-129.
    47. Welch, R.M. (2002). Breeding strategies for biofortified staple plant foods to reduce micronutrient malnutrition globally. J. Nutri., 132: 495S-499S.
    48. Yu, K., Park, J., Poysa, V. and Gepts, P. (2000). Integration of simple sequence repeat (SSR) markers into a molecular linkage map of common bean (Phaseolus vulgaris L.). J. Hered., 91: 429-434.
    49. Zhu, C., Naqvi, S., Galera, S.G., Pelacho, A.M., Capell, T. and Christou, P. (2007). Transgenic strategies for the nutritional enhancement of plants. Trends Plant. Sci., 12: 548–555. 
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