Identification of useful recombinants from interspecific hybrids of Citrullus lanatus and C. colocynthis

, , , ,

Research Articles | Published:

Print ISSN : 0970-4078.
Online ISSN : 2229-4473.
Pub Email:
Doi: 10.1007/s42535-020-00131-8
First Page: 475
Last Page: 482
Views: 2324

Keywords: Citrullus, Bitter apple, SSR, Phytochemicals, Water melon


Watermelon (Citrullus lanatus) is an important fruit crop in the arid region of western India. The modern watermelon cultivars share a narrow genetic base and are susceptible to various stresses. Thus, there is a need to expand its genetic base through distant hybridization by interspecific crossing with C. colocynthis. An attempt was made to develop interspecific hybrids between lanatus and colocynthis followed by hybridity testing of F1 through SSR markers and biochemical analysis of F2. Out of nine primers, eight primers could successfully show amplification in parents and hybrids out of which two markers could correctly discriminate the true hybrid. The morphological analysis demonstrated that F1 progeny resembles much with C. colocynthis for most of the characters. Biochemical analysis of pulp of selected F2 plants and parents indicated that in many F2 plants biochemicals were higher than parents which suggest that the blending of genes from both species made a favourable effect. Diterpene was only present in watermelon and not detected in C. colocynthis or any of the F2 plants. GC–MS based fatty acid profiling of nine identified fatty acids namely palmitic acid, stearic acid, oleic acid, linoleic acid and linolenic acid was done. It can be concluded that the interspecific hybrids between water melon and C. colocynthis as also confirmed by molecular markers and superior F2 plants can be used in watermelon breeding program for enhancing vital phytochemicals.

Citrullus, Bitter apple, SSR, Phytochemicals, Water melon

*Get Access

(*Only SPR Members can get full access. Click Here to Apply and get access)



  1. Anghel I (1969) Studies on the cytology and evolution of the Citrullus vulgaris. Commun Bot 11:49–55

  2. Anuragi H, Dhaduk HL, Kumar S, Dhruve JJ, Parekh MJ, Sakure AA (2016) Molecular diversity of Annona species and proximate fruit composition of selected genotypes. 3 Biotech 6(2):204

  3. AOAC (Association of Analytical Chemists) (1984) Standard official methods of analysis of the association of analytical chemists, 14th edn. Williams SW, Washington DC, p 121

  4. Baboli MZ, Kordi SAA (2010) Characteristics and composition of watermelon seed oil and solvent extraction parameters effects. J Am Oil Chem Soc 87(6):667–671

  5. Biles CL (1989) Isozymes and general proteins from various watermelon cultivars and tissue types. Hortic Sci 24:810–812

  6. Chaturvedi RK, Sankar K (2006) Laboratory manual for the physico-chemical analysis of soil, water and plant. Wildlife Institute of India, Dehradun, p 97

  7. Dane F, Liu J, Zhang C (2007) Phylogeography of the bitter apple, Citrullus colocynthis. Genet Resour Crop Evol 54(2):327–336

  8. Demirel G, Wachira AM, Sinclair LA, Wilkinson RG, Wood JD, Enser M (2004) Effects of dietary n-3 polyunsaturated fatty acids, breed and dietary vitamin E on the fatty acids of lamb muscle, liver and adipose tissue. Br J Nutr 91(4):551–565

  9. Denwick PM (2002) Natural products: a biosynthetic approach, 2nd edn. Wiley, Chichester, pp 241–243

  10. Dhingra DR, Biswas AK (1945) Component fatty acids of oil of Citrullus vulgaris Schrad. (watermelon) seed. J Indian Chem Soc 22:119–122

  11. Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12(13):39–40

  12. Ekins-Daukes S, Helms PJ, Taylor MW, Simpson CR, McLay JS (2005) Paediatric homoeopathy in general practice: where, when and why? Br J Clin Pharmacol 59(6):743–749

  13. El-Adawy TA, Taha KM (2001) Characteristics and composition of watermelon, pumpkin, and paprika seed oils and flours. J Agric Food Chem 49(3):1253–1259

  14. Ethiraj S, Balasundaram J (2016) Phytochemical and biological activity of Cucurbita seed extract. J Adv Biotechnol 6(1):813–821

  15. FAOSTAT (2018) Watermelon production in 2016: crops/regions (world list)/production quantity (from pick lists). FAOSTAT, Rome

  16. Fougat RS, Joshi CG, Kulkarni K, Kumar S, Patel A, Sakure AA, Mistry J (2014) Rapid development of microsatellite markers for Plantago ovata Forsk.: using next generation sequencing and their cross-species transferability. Agriculture 4:199–216

  17. Girgis P, Said F (1968) Lesser known Nigerian edible oils and fats 1: characteristics of melon seeds. J Food Agric 19:615–616

  18. Goldman D, Merril CR (1982) Silver staining of DNA in polyacrylamide gels: linearity and effect of fragment size. Electrophoresis 3(1):24–26

  19. Gurudeeban S, Satyavani K, Ramanathan T (2010) Bitter Apple (Citrullus colocynthis): an overview of chemical composition and biomedical potentials. Asian J Plant Sci 1:1–8

  20. Harborne JB (1973) Phytochemical methods. Chapman and Hall, London, pp 49–188

  21. Hussain AI, Rathore HA, Sattar MZ, Chatha SA, Sarker SD, Gilani AH (2014) Citrulluscolocynthis (L.) Schrad (bitter apple fruit): a review of its phytochemistry, pharmacology, traditional uses and nutritional potential. J Ethnopharmacol 155(1):54–66

  22. Jamatia J, Choudhary H, Basavaraj B (2018) Morphological studies on interspecific hybrids between Citrulluslanatus L. and its wild species. Int J Chem Stud 6(4):199–202

  23. Jarret RL, Levy IJ (2012) Oil and fatty acid contents in seed of Citrullus lanatus Schrad. J Agric Food Chem 60:5199–5204

  24. Joobeur T, Gusmini G, Zhang X, Levi A, Xu Y, Wehner TC, Oliver M, Dean RA (2006) Construction of a watermelon BAC library and identification of SSRs anchored to melon or Arabidopsis genomes. Theor Appl Genet 112(8):1553–1562

  25. Kumar V, Rathee P, Kohli K, Chaudhary H, Rathee S (2009) Phytochemical and biological potential of Indrayan: an overview. Pharmacogn Rev 3(5):193

  26. Kumar M, Fougat RS, Sharma AK, Kulkarni K, Ramesh Mistry JG, Sakure AA, Kumar S (2014) Phenotypic and molecular characterization of selected species of Plantago with emphasis on Plantago ovata. Aust J Crop Sci 8:1639–1647

  27. Kumar S, Parekh MJ, Patel CB, Zala HN, Sharma R, Kulkarni KS, Fougat RS, Bhatt RK, Sakure AA (2016) Development and validation of EST-derived SSR markers and diversity analysis in cluster bean (Cyamopsis tetragonoloba). J Plant Biochem Biotechnol 25(3):263–269

  28. Mahajan S, Kumawat RN (2013) Study of seed dormancy in colocynth (Citrulluscolocynthis L.) with after-ripening of fruits, seed extraction procedures and period of seed storage. Natl Acad Sci Lett 36(4):373–378

  29. Malik EP, Singh MB (1980) Plant enzymology and histo-enzymology, 1st edn. Kalyani Publishers, New Delhi, p 286

  30. Mehta SL, Lodha ML, Sane PV (1993) Recent advances in plant biochemistry. Publication and Information Division, New Delhi

  31. Navot N, Zamir D (1987) Isozyme and seed protein phylogeny of the genus Citrullus (Cucurbitaceae). Plant Syst Evol 156(1–2):61–67

  32. Nelson N (1944) A photometric adaptation of the Somogyi method for the determination of sugar. J Biol Chem 153:375

  33. Onimawo IA (2002) Proximate composition and selected physicochemical properties of seed, pulp and oil of soursop (Annonamuricata L.). J Plant Foods Hum Nutr 57(2):155–171

  34. Palevitch D, Yaniv Z (1991) Medicinal plants of the Holyland. Tamus Modan Press, Tel Aviv

  35. Pareek OP, Vashishtha BB (1980) Variability in Citrullus colocynthis in the Thar Desert [plant of Cucubitaceae family]. Ann Arid Zone (India)

  36. Perkins-Veazie P, Collins JK, Pair SD, Roberts W (2001) Lycopene content differs among red-fleshed watermelon cultivars. J Sci Food Agric 81(10):983–987

  37. Ren Y, McGregor C, Zhang Y, Gong G, Zhang H, Guo S, Sun H, Cai W, Zhang J, Xu Y (2014) An integrated genetic map based on four mapping populations and quantitative trait loci associated with economically important traits in watermelon (Citrullus lanatus). BMC Plant Biol 14(1):33

  38. Rickman JC, Barrett DM, Bruhn CM (2007) Nutritional comparison of fresh, frozen and canned fruits and vegetables. Part 1: Vitamins C and B and phenolic compounds. J Sci Food Agric 87(6):930–944

  39. Rubatzky VE, Yamaguchi M (1997) World vegetables, 2nd edn. Chapman and Hall, New York

  40. Sadasivam S, Manickam A (1992) Biochemical methods for agricultural sciences. Wiley Eastern Ltd., New Delhi

  41. Sao TCM, Potts WM (1952) The analysis and characterization of the oil from the seed of Citrullus vulgaris. J Am Oil Chem Soc 29:444–445

  42. Shimotsuma M (1960) Cytogenetical studies in the genus Citrullus. IV. Intra-and interspecific hybrids between C. colocynthis Schrad. and C. vulgaris Schrad. Jpn J Genet 35(10):303–312

  43. Singh AK, Yadava KS (1977) Cytomorphological studies in Citrullus L. Biol Contemp 4:168–172

  44. Singh D, Singh R, Sandhu JS, Chunneja P (2017) Morphological and genetic diversity analysis of Citrullus landraces from India and their genetic inter relationship with continental watermelons. Sci Hortic 218:240–248

  45. Sofowara A (1993) Medicinal plants and traditional medicine in Africa. Spectrum Books Ltd, Ibadan, p 289

  46. Takashi I, Keiichi U, Hisayuki R (1986) Gourd saponins as antioxidants in oils, foods, cosmetics and pharmaceuticals. Chem Abstr 105:151–181

  47. Horticultural Statistics at a Glance (2017) Government of India Ministry of Agriculture and Farmers’ Welfare Department of Agriculture, Cooperation and Farmers’ Welfare Horticulture Statistics Division

  48. Whitaker TW (1933) Cytological and phylogenetic studies in the Cucurbitaceae. Bot Gaz 94(4):780–790

  49. Yin YL, McEvoy JDG, Schulze H, Hennig U, Souffrant WB, McCracken KJ (2000) Apparent digestibility (ileal and overall) of nutrients and endogenous nitrogen losses in growing pigs fed wheat (var. Soissons) or its by-products without or with xylanase supplementation. Livest Prod Sci 62(2):119–132

  50. Yoshikawa M, Morikawa T, Kobayashi H, Nakamura A, Matsuhira K, Nakamura S, Matsuda H (2007) Bioactive saponins and glycosides. XXVII. Structures of new cucurbitane-type triterpene glycosides and antiallergic constituents from Citrulluscolocynthis. Chem Pharm Bull 55(3):428–434

  51. Zamir D, Navot N, Rudich J (1984) Enzyme polymorphism in Citrullus lanatus and C. colocynthis in Israel and Sinai. Plant Syst Evol 146(3–4):163–170



Authors acknowledge Anand Agricultural University, Gujarat, India, for providing facilities to experiment.

Author Information

Parihar Akarsh
Department of Agricultural Biotechnology, Anand Agricultural University, Anand, India
Vaja M. B.
Department of Agricultural Biotechnology, Anand Agricultural University, Anand, India

Dhruve J. J.
Department of Agricultural Biochemistry, BACA Anand Agricultural University, Anand, India

Department of Agricultural Biotechnology, Anand Agricultural University, Anand, India

Kumar Sushil
Department of Agricultural Biotechnology, Anand Agricultural University, Anand, India