Physiology and Molecular Biology of Plants

, Volume 23, Issue 2, pp 443–451 | Cite as

Identification of aroma volatiles and understanding 2-acetyl-1-pyrroline biosynthetic mechanism in aromatic mung bean (Vigna radiata (L.) Wilczek)

  • Usmangani Attar
  • Vidya Hinge
  • Rahul Zanan
  • Rahul Adhav
  • Altafhusain NadafEmail author
Research Article


Mung bean having high food value and easily digestible proteins, is one of the socioeconomically important crop of India. Among the varied cultivars, Sona mung is having aroma and hence popularly cultivated in the pockets of Ganga river basin at Bhutnir char village of Malda District in the West Bengal state. In the present study, aroma volatiles with special reference to 2-acetyl-1-pyrroline (2AP) were analyzed using HS–SPME–GCMS from Sona mung bean and compared with non-scented mung bean (PHULE M-9339). 26 volatiles in seeds of Sona mung and 20 in non-scented mung bean were identified, in which 3,7-dimethyl-6-octenal, (2E)-2-decen-1-ol, 2-ethyl-1-dodecanol and 3,5,5-trimethyl-2-cyclohexene-1-one are first time reported. 0.19 ± 0.001 ppm 2AP was recorded in Sona mung seeds whereas it was not detected in non-scented mung bean. PCA analysis indicated that 2AP, octanal, 1 pentanol, decanal, phenylmethanol and 2-nonen-1-ol were the major contributors in the aroma of Sona mung bean. The significantly higher level proline, methylglyoxal and lower level of BADH2 transcript were detected in Sona mung than non-scented mung, suggesting similar 2AP biosynthesis mechanism in Sona mung bean as reported in scented rice, sorghum and soybean.


Vigna radiata Sona mung bean Aroma volatiles 2-Acetyl-1-pyrroline biosynthesis 



Authors are thankful to Dr. P. Srinivas (Central Food and Technology Research Institute, Mysore, India) for generous gift of authentic 2AP.

Supplementary material

12298_2017_414_MOESM1_ESM.docx (596 kb)
Supplementary material 1 (DOCX 595 kb)
12298_2017_414_MOESM2_ESM.docx (59 kb)
Supplementary material 2 (DOCX 59 kb)
12298_2017_414_MOESM3_ESM.png (96 kb)
Supplementary material 3 (PNG 96 kb)


  1. Arikit S, Yoshihashi T, Wanchana S, Tanya P, Juwattanasomran R, Srinives P, Vanavichit A (2011a) A PCR-based marker for a locus conferring aroma in vegetable soybean (Glycine max L.). Theor Appl Genet 122:311–316CrossRefPubMedGoogle Scholar
  2. Arikit S, Yoshihashi T, Wanchana S, Uyen TT, Huong NTT, Wongpornchai S, Vanavichit A (2011b) Deficiency in the amino aldehyde dehydrogenase encoded by GmAMADH2, the homologue of rice Os2AP, enhances 2-acetyl-1-pyrroline biosynthesis in soybeans (Glycine max L.). Plant Biotechnol J 9(1):75–87CrossRefPubMedGoogle Scholar
  3. Ayyanger GRR (1938) Studies in Sorghum. J Madras Univ 11:131–143Google Scholar
  4. Bates L, Waldren RP, Teare ID (1973) Rapid determination of free proline for water-stress studies. Plant Soil 39:205–207CrossRefGoogle Scholar
  5. Bradbury LMT, Henry RJ, Jin Q, Russell FRF, Waters DLE (2005) A perfect marker for fragrance genotyping in rice. Mol Breed 16:279–283CrossRefGoogle Scholar
  6. Bradbury LMT, Gillies SA, Brushett DJ, Waters DLE, Henry RJ (2008) Inactivation of an aminoaldehyde dehydrogenase is responsible for fragrance in rice. Plant Mol Biol 68:439–449CrossRefPubMedGoogle Scholar
  7. Brahmachary RL, Ghosh M (2002) Vaginal pheromone and other unusual compounds in mung bean aroma. J Sci Ind Res 61:625–629Google Scholar
  8. Bryant RJ, McClung AM (2011) Volatile profiles of aromatic and non-aromatic rice cultivars using SPME/GC–MS. Food Chem 124:501–513CrossRefGoogle Scholar
  9. Buttery RG, Ling LC (1995) Volatile flavor components of corn tortillas and related products. J Agric Food Chem 43:1878–1882CrossRefGoogle Scholar
  10. Buttery RG, Juliano BO, Ling LC (1983a) Identification of rice aroma compound 2-acetyl-1-pyrroline in Pandan leaves. Chem Ind 23:478Google Scholar
  11. Buttery RG, Ling LC, Juliano BO, Turnbaugh JG (1983b) Cooked rice aroma and 2-Acetyl- 1-pyrroline. J Agric Food Chem 31:823–826CrossRefGoogle Scholar
  12. Buttery RG, Turnbaugh JG, Ling LC (1988) Contributions of volatiles to rice aroma. J Agric Food Chem 36:1006–1009CrossRefGoogle Scholar
  13. Buttery RG, Orts WJ, Takeoka GR, Nam Y (1999) Volatile flavor components of rice cakes. J Agric Food Chem 47:4353–4356CrossRefPubMedGoogle Scholar
  14. Careri M, Mangia A, Barbieri G, Bolzoni L, Virgili R, Parolari G (1993) Sensory property relationships to chemical data of Italian-type dry-cured ham. J Food Sci 58:968–972CrossRefGoogle Scholar
  15. Chen S, Yang Y, Shi W, Ji Q, He F, Zhang Z, Cheng Z, Liu X, Xu M (2008) Badh2, encoding betaine aldehyde dehydrogenase, inhibits the biosynthesis of 2-acetyl-1-pyrroline, a major component in rice fragrance. Plant Cell 20(7):1850–1861CrossRefPubMedPubMedCentralGoogle Scholar
  16. Eyres G, Dufour JP, Hallifax G, Sotheeswaran S, Marriott PJ (2005) Identification of character-impact odorants in coriander and wild coriander leaves using gas chromatography-olfactometry (GCO) and comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry (GC x GC-TOFMS). J Sep Sci 28(9–10):1061–1074CrossRefPubMedGoogle Scholar
  17. Fan W, Xu Y, Qian MC (2012) Identification of aroma compounds in Chinese “Moutai” and “Langjiu” liquors by normal phase liquid chromatography fractionation followed by gas chromatography/olfactometry. In: Qian MC, Shellhammer TH (eds) Flavor chemistry of wine and other alcoholic beverages, American Chemical Society publication, Washington DC, 1104, pp 303–338Google Scholar
  18. Fery RL (2002) New opportunities in Vigna. In: Janick J, Whipkey A (eds) Trends in new crops and new uses. ASHS press, Alexandria, pp 424–428Google Scholar
  19. Fitzgerald TL, Waters DLE, Henry RJ (2008) The effect of salt on betaine aldehyde dehydrogenase transcript levels and 2-acetyl-1-pyrroline concentration in fragrant and non-fragrant rice (Oryza sativa). Plant Sci 175:539–546CrossRefGoogle Scholar
  20. Fushimi T, Masuda R (2001) 2-acetyl-1-pyrroline concentration of the aromatic vegetable soybean “Dadacha-Mame”. In: Lumpkin T, Shanmugasundaram S (eds) Proceeding of the 2nd international vegetable soybean conference. Washington State University, Pullman, p 39Google Scholar
  21. Ghosh M (2014) Preserving an indigenous cultivar. Curr Sci 107(12):1945Google Scholar
  22. Grimm CC, Bergman C, Delgado JT, Bryant R (2001) Screening for 2-Acetyl-1-pyrroline in the headspace of rice using SPME/GC-MS. J Agric Food Chem 49:245–249CrossRefPubMedGoogle Scholar
  23. Hanelt P (2001) Institute of plant genetics and crop plant research, mansfeld’s encyclopedia of agricultural and horticultural crops, 1-5. Springer, BerlinCrossRefGoogle Scholar
  24. Hinge V, Patil H, Nadaf A (2015) Comparative characterization of aroma volatiles and related gene expression analysis at vegetative and mature stages in basmati and non-basmati rice (Oryza sativa L.) cultivars. Appl Biochem Biotechnol 178(4):619–639CrossRefPubMedGoogle Scholar
  25. Huang TC, Huang YW, Hung HJ, Ho CT, Wu ML (2007) Δ1-Pyrroline-5-carboxylic acid formed by proline dehydrogenase from the Bacillus subtilis ssp. natto expressed in Escherichia coli as a precursor for 2-acetyl-1-pyrroline. J Agric Food Chem 55:5097–5102CrossRefPubMedGoogle Scholar
  26. Huang TC, Teng CS, Chang JL, Chuang HS, Ho CT, Wu ML (2008) Biosynthetic mechanism of 2-acetyl-1-pyrroline and its relationship with Δ1-pyrroline-5-carboxylic acid and methylglyoxal in aromatic rice (Oryza sativa L.) callus. J Agric Food Chem 56:7399–7404CrossRefPubMedGoogle Scholar
  27. ICAR (2012). All India coordinated research project on MULLaRP. In: Proceedings and recommendations, annual group meet on mungbean and urdbean, Indian council of agricultural research. Accessed 25 Mar 2016
  28. Kaikavoosi K, Kad TD, Zanan RL, Nadaf AB (2015) 2-Acetyl-1-Pyrroline augmentation in scented indica rice (Oryza sativa L.) varieties through Δ1-Pyrroline-5-Carboxylate Synthetase (P5CS) gene transformation. Appl Biochem Biotech 177(7):1466–1479CrossRefGoogle Scholar
  29. Kosuge T, Kamiya H (1962) Discovery of a pyrazine in a natural product: tetramethylpyrazine from cultures of a strain of Bacillus subtilis. Nature 193:776CrossRefPubMedGoogle Scholar
  30. Lee KG, Shibamoto T (2000) Antioxidant properties of aroma compounds isolated from soybeans and mung beans. J Agric Food Chem 48:4290–4293CrossRefPubMedGoogle Scholar
  31. Liu T, Fan W, Xu Y (2008) Characterization of volatile and semi-volatile compounds in Chinese rice wines by headspace solid phase microextraction followed by gas chromatography—mass spectrometry. J Inst Brew 114(2):172–179CrossRefGoogle Scholar
  32. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2-∆∆ct method. Methods 25:402–408CrossRefPubMedGoogle Scholar
  33. Lovergren NV, Fisher GS, Legendre MG, Schuller WH (1979) Volatile constituents of dried legumes. J Agric Food Chem 27(4):851CrossRefGoogle Scholar
  34. Mahatheeranont S, Keawsaard S, Dumri K (2001) Quantification of the rice aroma compound, 2-Acetyl-1-pyrroline, in uncooked Khao Dawk Mali 105 brown rice. J Agric Food Chem 49(2):773–779CrossRefPubMedGoogle Scholar
  35. Mathure SV, Wakte KV, Jawali N, Nadaf AB (2011) Quantification of 2-Acetyl-1-pyrroline and other rice aroma volatiles among Indian scented rice cultivars by HS-SPME/GC-FID. Food Anal Method 4(3):326–333CrossRefGoogle Scholar
  36. Mathure SV, Jawali N, Thengane RJ, Nadaf AB (2014) Comparative quantitative analysis of headspace volatiles and their association with BADH2 marker in non-basmati scented, basmati and non-scented rice (Oryza sativa L.) cultivars of India. Food Chem 142:383–391CrossRefPubMedGoogle Scholar
  37. Midya S, Brahmachary R (1996) The aroma of Bassia flower. Curr Sci 71:430Google Scholar
  38. Oomah BD, Razafindrainibe M, Drover JC (2014) Headspace volatile components of Canadian grown low-tannin faba bean (Vicia faba L.) genotypes. J Sci Food Agric 94(3):473–481CrossRefPubMedGoogle Scholar
  39. Pal M, Brahmachary RL, Ghosh M (2010) Comparative studies on physicochemical and biochemical characteristics of scented and non-scented strains of mung Beans (Vigna radiata) of Indian origin. Legum Res 33(1):1–9Google Scholar
  40. Phillips SA, Thornalley PJ (1993) The formation of methylglyoxal from triose phosphates. Investigation using a specific assay for methylglyoxal. Eur J Biochem 212:101–105CrossRefPubMedGoogle Scholar
  41. Poehlman JM (1991) The mungbean. Oxford and IBH Publishing Co. Pvt. Ltd., New DelhiGoogle Scholar
  42. Pramnoi P, Somta P, Chankaew S, Juwattanasomran R, Srinives P (2013) A single recessive gene controls fragrance in cucumber (Cucumis sativus L). J Genet 92:147–149CrossRefPubMedGoogle Scholar
  43. Shanmugasundaram S (2007) Exploit mungbean with value added products. Acta Hortic 752:99–102CrossRefGoogle Scholar
  44. Sharma OP, Bambawale OM, Gopali JB, Bhagat S, Yelshetty S, Singh SK, Anand R, Singh OP (2011) Field guide mungbean and urdbean, National Centre for Integrated Pest Management 2011. Assessed 20 Dec 2015
  45. Singh N, Kaul VK, Megeji NW, Singh V, Ahuja PS (2008) Essential oil composition of three accessions of Dracocephalum heterophyllum Benth. Cultivated at Palampur, India. Nat Prod Res 22(11):927–936CrossRefPubMedGoogle Scholar
  46. Sood BC, Siddiq EA (1978) A rapid technique for scent determination in rice. Ind J Genet Plant Breed 38(2):268–275Google Scholar
  47. Sun W, Zhao Q, Zhao H, Zhao M, Yang B (2010) Volatile compounds of Cantonese sausage released at different stages of processing and storage. Food Chem 121(2):319–325CrossRefGoogle Scholar
  48. Van Den Dool H, Kratz PD (1963) A generalization of the retention index system including linear temperature programmed gas-liquid partition chromatography. J Chromatogr 11:463–471CrossRefGoogle Scholar
  49. Vavilov NI (1926) Studies on the origin of cultivated plants. Institute of Applied Botany and Plant Breeding, LeningradGoogle Scholar
  50. Virgili R, Parolari G, Bolzoni L, Barbieri G, Mangia A, Careri M, Spagnoli S, Panari G, Zannoni M (1994) Sensory-chemical relationships in Parmigiano-Reggiano cheese. LWT Food Sci Technol 27:491–495CrossRefGoogle Scholar
  51. Wakte KV, Thengane RJ, Jawali N, Nadaf AB (2010) Optimization of HS-SPME conditions for quantification of 2-acetyl-1-pyrroline and study of other volatiles in Pandanus amaryllifolius Roxb. Food Chem 121:595–600CrossRefGoogle Scholar
  52. Widjaja R, Craske JD, Wootton M (1996) Comparative studies on volatile components of non-fragrant and fragrant rices. J Sci Food Agric 70(2):151–161CrossRefGoogle Scholar
  53. Wild R, Ooi L, Srikanth V, Münch G (2012) A quick, convenient and economical method for the reliable determination of methylglyoxal in millimolar concentrations: the N-acetyl-l-cysteine assay. Anal Bioanal Chem 403:2577–2581CrossRefPubMedGoogle Scholar
  54. Wongpornchai S, Sriseadka T, Choonvisase S (2003) Identification and quantitation of the rice aroma compound, 2-acetyl-1-pyrroline, in bread flowers (Vallaris glabra Ktze). J Agric Food Chem 51:457–462CrossRefPubMedGoogle Scholar
  55. Wu ML, Chou KL, Wu CR, Chen JK, Huang TC (2009) Characterization and the possible formation mechanism of 2-Acetyl-1-Pyrroline in aromatic vegetable soybean (Glycine max L.). J Food Sci 74(5):192–197CrossRefGoogle Scholar
  56. Xiao Z, Hou X, Lyu X, Xi L, Zhao J (2014) Accelerated green process of tetramethylpyrazine production from glucose and diammonium phosphate. Biotech Biofuels 7:106CrossRefGoogle Scholar
  57. Yadav SK, Singla-Pareek SL, Ray M, Sopory SK (2005) Methylglyoxal levels in plants under salinity stress are dependent on glyoxalase I and glutathione. Biochem Biophys Res Commun 337:61–67CrossRefPubMedGoogle Scholar
  58. Yang DS, Shewfelt RL, Lee KS, Kays SJ (2008) Comparison of odor-active compounds from six distinctly different rice flavor types. J Agric Food Chem 56(8):2780–2787CrossRefPubMedGoogle Scholar
  59. Yoshihashi T (2002) Quantitative analysis on 2-acetyl-1-pyrroline of aromatic rice by stable isotope dilution method and model studies on its formation during cooking. J Food Sci 67(2):619–622CrossRefGoogle Scholar
  60. Yoshihashi T, Huong NTT, Inatomi H (2002) Precursors of 2-acetyl-1-pyrroline, a potent flavor compound of an aromatic rice variety. J Agric Food Chem 50:2001–2004CrossRefPubMedGoogle Scholar
  61. Yundaeng C, Somta P, Tangphatsornruang S, Wongpornchai S, Srinives P (2013) Gene discovery and functional marker development for fragrance in sorghum (Sorghum bicolor (L.) Moench). Theor Appl Genet 26(11):2897–2906CrossRefGoogle Scholar
  62. Zanan R, Khandagale K, Hinge V, Elangovan M, Henry FJ, Nadaf A (2016) Characterization of fragrance in sorghum (Sorghum bicolor (L.) Moench) grain and development of a gene-based marker for selection in breeding. Mol Breed 36:146. doi: 10.1007/s11032-016-0582-8 CrossRefGoogle Scholar
  63. Zukovskij PM (1962) Cultivated plants and their wild relatives. Commonwealth Agriculture Bureau, LondonGoogle Scholar

Copyright information

© Prof. H.S. Srivastava Foundation for Science and Society 2017

Authors and Affiliations

  • Usmangani Attar
    • 1
  • Vidya Hinge
    • 1
  • Rahul Zanan
    • 1
  • Rahul Adhav
    • 1
  • Altafhusain Nadaf
    • 1
    Email author
  1. 1.Department of BotanySavitribai Phule Pune UniversityPuneIndia

Personalised recommendations