Haploid Embryogenesis in Tea

  • R. Bajpai
  • R. ChaturvediEmail author
Part of the Forestry Sciences book series (FOSC, volume 85)


History unveils decades of strenuous efforts made by mankind to strengthen agricultural productivity and attain sustainable food security at global level. Traditional breeding practices have been followed by plant breeders to attain genetic improvement in plants and to ease down the challenges faced by local farmers. However, there are limitations to implement these conventional breeding practices to improve woody perennials due to their long reproductive cycle and out breeding nature. Plant tissue culture techniques has emanated as a potential branch of biotechnology to help plant breeders in addressing long existing problems of inbreeding depression, prolong juvenile period and extreme heterozygosity encountered in majority of perennial crops and recalcitrant woody trees. Gamete biology, involving in vitro haploid plant production via embryogenesis from immature explants of gametic origin in anther or ovary cultures, routes easier, single step procedure of obtaining doubled haploid (pure breeding) lines within reduced time duration. This chapter accentuates the progress achieved in the field of haploid plant induction using in vivo and in vitro means and brings together the research updates on recent strategies deployed to induce diploidization of haploids and cytological and molecular basis of their identification.


  1. Akula A, Becker D, Bateson M (2000) High-yielding repetitive somatic embryogenesis and plant recovery in a selected tea clone, ‘TRI-2025’, by temporary immersion. Plant Cell Rep 19:1140–1145CrossRefGoogle Scholar
  2. Alan AR, Mutschler MA, Brants A, Cobb E, Earle ED (2003) Production of gynogenic plants from hybrids of Allium cepa L. and A. roylei Stearn. Plant Sci 165:1201–1211CrossRefGoogle Scholar
  3. Bartošová Z, Obert B, Takacomá Š, Kormutak A, Pretová A (2005) Using enzyme polymorphism to identify the gametic origin of flax regenerants. Acta Biol Cracov Bot 47:173–178Google Scholar
  4. Bhatt JG, Murthy HN (2007) Factors affecting in vitro gynogenic haploid production in niger (Guizotia abyssinica (L. f.) Cass.). Plant Growth Regul 52:241–248CrossRefGoogle Scholar
  5. Bhojwani SS, Razdan MK (1996) Haploid Production. Plant tissue culture: theory and practice. Elsevier Scientific Publishers, Amsterdam, The Netherlands, pp 113–142Google Scholar
  6. Blakeslee AF, Belling J, Farnham ME, Berner AD (1922) A haploid mutant in Jimson weed, Dhatura stramonium. Science 55:646–647CrossRefPubMedGoogle Scholar
  7. Bohanec B (2009) Doubled haploids via gynogenesis. In: Touraev A, Forster BP, Jain SM (eds) Advances in haploid production in higher plants. Springer, Dordrecht, Netherlands, pp 35–46CrossRefGoogle Scholar
  8. Bueno MA, Gómez A, Boscaiu M, Manzanera JA, Vicente O (1997) Stress-induced formation of haploid plants through anther culture in cork oak (Quercus suber). Physiol Plant 99:335–341CrossRefGoogle Scholar
  9. Chaturvedi R, Razdan MK, Bhojwani SS (2003) Production of haploids of neem (Azadirachta indica A. Juss.) by anther culture. Plant Cell Rep 21:531–537PubMedGoogle Scholar
  10. Chen JF, Cui L, Malik AA, Mbira KG (2010) In vitro haploid and dihaploid production via unfertilized ovule culture. Plant Cell Tissue Organ Cult 104:311–319CrossRefGoogle Scholar
  11. Chen Y, Kenaschuk E, Dribnenki P (1998) High frequency of plant regeneration from anther culture in flax, Linum usitatissimum L. Plant Breed 117:463–467CrossRefGoogle Scholar
  12. Clapham D (1971) In vitro development of callus from the pollen of Lolium and Hordeum. Z Pflanzenziichtg. 65:285–292Google Scholar
  13. Comai L (2014) Genome elimination: translating basic research into a future tool for plant breeding. PLoS Biol 12:e1001876CrossRefPubMedPubMedCentralGoogle Scholar
  14. Custers JB, Cordewener JH, Nöllen Y, Dons HJ, Campagne MM (1994) Temperature controls both gametophytic and sporophytic development in microspore cultures of Brassica napus. Plant Cell Rep 13:267–271CrossRefPubMedGoogle Scholar
  15. Datta SK (2005) Androgenic haploids: factors controlling development and its application in crop improvement Curr Sci 1870–1878Google Scholar
  16. Da Silva DJC (2001) Effect of incubation temperature regimes and culture medium on broccoli microspore culture embryogenesis. Euphytica 119:389–394CrossRefGoogle Scholar
  17. Dunwell JM, Cornish M, De Courcel AGL (1985) Influence of genotype, plant growth temperature and anther incubation temperature on microspore embryo production in Brassica napus ssp. oleifera. J Exp Bot 36:679–689CrossRefGoogle Scholar
  18. Dunwell JM (2010) Haploids in flowering plants: origins and exploitation. Plant Biotechnol J 8:377–424CrossRefPubMedGoogle Scholar
  19. Dwivedi SL, Britt AB, Tripathi L, Sharma S, Upadhyaya HD, Ortiz R (2015) Haploids: constraints and opportunities in plant breeding. Biotechnol Adv 33:812–829CrossRefPubMedGoogle Scholar
  20. Ferrie AMR, Caswell KL (2011) Isolated microspore culture techniques and recent progress for haploid and doubled haploid plant production. Plant Cell Tissue Organ Cult 104:301–309CrossRefGoogle Scholar
  21. Forster BP, Heberle-Bors E, Kasha KJ, Touraev A (2007) The resurgence of haploids in higher plants. Trends Plant Sci 12:368–375CrossRefPubMedGoogle Scholar
  22. Gaillard A, Vergne P, Beckert M (1991) Optimization of maize microspore isolation and culture conditions for reliable plant regeneration. Plant Cell Rep 10:55–58CrossRefPubMedGoogle Scholar
  23. Germanà MA (2005) Protocol of somatic embryogenesis from Citrus spp. anther culture. In: Jain SM, Gupta PK (eds) Protocol for somatic embryogenesis in woody plants. Springer, Netherlands, pp 191–207CrossRefGoogle Scholar
  24. Germana MA (2006) Doubled haploid production in fruit crops. Plant Cell Tissue Organ Cult 86:131–146CrossRefGoogle Scholar
  25. Germanà MA (2011) Anther culture for haploid and doubled haploid production. Plant Cell Tissue Organ Cult 104:283–300CrossRefGoogle Scholar
  26. Gharyal PK, Maheshwari SC (1982) In vitro differentiation of plantlets from tissue cultures of Albizzia lebbeck L. Plant Cell Tissue Organ Cult 2(1):49–53CrossRefGoogle Scholar
  27. Gharyal PK, Rashid A, Maheshwari SC (1983) Androgenic response from cultured anthers of a leguminous tree Cassia siamea Lam. Protoplasma 118:91–93CrossRefGoogle Scholar
  28. Gresshoff PM, Doy CH (1972) Haploid Arabidopsis thaliana callus and plants from anther culture. Aust J Biol Sci 25:259–264CrossRefGoogle Scholar
  29. Guha S, Maheswari SC (1964) In vitro production of embryos from anthers of Datura. Nature 204:497CrossRefGoogle Scholar
  30. Guha S, Maheswari SC (1966) Cell division and differentiation of embryos in the pollen grains of Datura in vitro. Natur 212:97–98CrossRefGoogle Scholar
  31. Hazarika RR, Chaturvedi R (2013) Establishment of dedifferentiated callus of haploid origin from unfertilized ovaries of tea (Camellia sinensis (L.) O. Kuntze) as a potential source of total phenolics and antioxidant activity. In Vitro Cell Dev Biol Plant 49:60–69CrossRefGoogle Scholar
  32. Hazarika R, Mishra V K, Chaturvedi R (2013) In Vitro Haploid Production—A Fast and Reliable Approach for Crop Improvement. In: Tuteja N, Gill S (eds) Crop improvement under adverse conditions. Springer, New York, pp 171–212Google Scholar
  33. Höfer M (2004) In vitro androgenesis in apple—improvement of the induction phase. Plant Cell Rep 22:365–370CrossRefPubMedGoogle Scholar
  34. Hu T, Kasha KJ (1997) Performance of isolated microspore-derived doubled haploids of wheat (Triticum aestivum L.). Can J Plant Sci 77:549–554CrossRefGoogle Scholar
  35. Islam SS, Tuteja N (2012) Enhancement of androgenesis by abiotic stress and other pretreatments in major crop species. Plant Sci 182:134–144CrossRefPubMedGoogle Scholar
  36. Jacquard C, Asakaviciute R, Hamalian AM, Sangwan RS, Devaux P, Clement C (2006) Barley anther culture: effects of annual cycle and spike position on microspore embryogenesis and albinism. Plant Cell Rep 25:375–381CrossRefPubMedGoogle Scholar
  37. Karasawa MMG, Chiancone B, Gianguzzi V, Abdelgalel AM, Botta R, Sartor C, Germanà MA (2016) Gametic embryogenesis through isolated microspore culture in Corylus avellana L. Plant Cell Tissue Organ Cult 124:635–647CrossRefGoogle Scholar
  38. Kasha KJ, Kao KN (1970) High frequency haploid production in barley (Hordeum vulgare L.). Natur 225:874–876CrossRefGoogle Scholar
  39. Kasha KJ (1974) Haploids in higher plants: advances and potential. In: Proceedings of the 1st international symposium, University of Guelph, GuelphGoogle Scholar
  40. Katiyar S, Mukhtar HA (1996) Tea in chemoprevention of cancer. Int J Oncol 8:221–238PubMedGoogle Scholar
  41. Keller WA, Armstrong KC (1979) Stimulation of embryogenesis and haploid production in Brassica campestris anther cultures by elevated temperature treatments. Theor Appl Genet 55:65–67CrossRefPubMedGoogle Scholar
  42. Khush GS, Virmani SS (1996) Haploids in plant breeding. In: Jain SM, Sopory SK, Veilleux R (eds) In Vitro haploid production in higher plants. Springer, Netherlands, pp 11–33CrossRefGoogle Scholar
  43. Larsen ET, Tuvesson IK, Andersen SB (1991) Nuclear genes affecting percentage of green plants in barley (Hordeum vulgare L.) anther culture. Theor Appl Genet 82:417–420CrossRefPubMedGoogle Scholar
  44. Laurie DA, Bennett MD (1988) The production of haploid wheat plants from wheat x maize crosses. Theor Appl Genet 76:393–397CrossRefPubMedGoogle Scholar
  45. Maluszynska J (2003) Cytogenetic tests for ploidy level analyses—chromosome counting. In: Maluszynski M, Kasha K.J, Forster BP, Szarejko I (eds) Doubled haploid production in crop plants. Springer, Dordrecht, pp 391–395CrossRefGoogle Scholar
  46. Magoon ML, Khanna KR (1963) Haploids Caryologia 16:191–235CrossRefGoogle Scholar
  47. Maheshwari SC, Rashid A, Tyagi AK (1982) Haploids from pollen grains–retrospect and prospect. Am J Bot 69:865–879CrossRefGoogle Scholar
  48. Maluszynski M, Kasha KJ, Szarejko I (2003) Published doubled haploid protocols in plant species. Doubled haploid production in crop plants. Springer, Netherlands, pp 309–335CrossRefGoogle Scholar
  49. Mishra VK, Bajpai R, Chaturvedi R (2017) An efficient and reproducible method for development of androgenic haploid plants from in vitro anther cultures of Camellia assamica ssp. assamica (Masters). In Vitro Cell Dev Biol Plant 53:239–248CrossRefGoogle Scholar
  50. Mishra VK, Goswami R (2014) Haploid production in higher plant. Int J Chem Biol Sci 1:26–45Google Scholar
  51. Mondal BA, Laxmikumaran M, Ahuja PS (2004) Recent advances of tea (Camellia sinensis) biotechnology. Plant Cell Tissue Organ Cult 76:195–254CrossRefGoogle Scholar
  52. Murashiage T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  53. Nair S, Gupta PK, Mascarenhas AF (1983) Haploid plants from in vitro anther culture of Annona squamosa Linn. Plant Cell Rep 2:198–200CrossRefPubMedGoogle Scholar
  54. Nakajima I, Kobayashi S, Nakamura Y (2000) Embryogenic callus induction and plant regeneration from unfertilized ovule of ‘Kyoho’grape. J Jpn Soc Hortic Sci 69:186–188CrossRefGoogle Scholar
  55. Namita P, Mukesh R, Vijay KJ (2012) Camellia sinensis (green tea): a review. Global J Pharmacol 6:52–59Google Scholar
  56. Ochatt S, Pech C, Grewal R, Conreux C, Lulsdorf M, Jacas L (2009) Abiotic stress enhances androgenesis from isolated microspores of some legume species (Fabaceae). Plant Physiol 166:1314–1328CrossRefGoogle Scholar
  57. Olmedilla A (2010) Microspore embryogenesis In: Plant developmental biology-biotechnological perspectives. Springer, Berlin Heidelberg, pp 27–44CrossRefGoogle Scholar
  58. Palmer CD, Keller WA (2005) Overview of haploidy. In: Haploids in crop improvement II. Springer, Berlin Heidelberg, pp 3–9Google Scholar
  59. San Noeum LH (1976) Haploïdes d’ Hordeum vulgare L. par culture in vitro d’ovaires non fécondés. Ann Amelior Plant 26:751–754Google Scholar
  60. Scott P, Lyne RL (1994) The effect of different carbohydrate sources upon the initiation of embryogenesis from barley microspores. Plant Cell Tissue Organ Cult 36:129–133CrossRefGoogle Scholar
  61. Scott P, Lyne RL, Rees T (1995) Metabolism of maltose and sucrose by microspores isolated from barley (Hordeum vulgare L). Planta 197:435–441CrossRefGoogle Scholar
  62. Seguí-Simarro JM (2010) Androgenesis revisited. Bot Rev 76:377–404CrossRefGoogle Scholar
  63. Seran TH (2007) Anther culture in tea improvement. Med Aromat Plant Sci Biotechnol 1:234–239Google Scholar
  64. Shen Y, Pan G, Lübberstedt T (2015) Haploid strategies for functional validation of plant genes. Trends Biotechnol 33:611–620CrossRefPubMedGoogle Scholar
  65. Sita GL (1996) Gynogenic haploids in vitro. In: In vitro haploid production in higher plants. Springer, Netherlands, pp 175–193CrossRefGoogle Scholar
  66. Sopory SK, Maheswari SC (1976) Development of Pollen Embryoids in Anther Cultures of Datura innoxia: I. General observations and effects of physical factors. J Exp Bot 27:49–57CrossRefGoogle Scholar
  67. Soriano M, Li H, Boutilier K (2013) Microspore embryogenesis: establishment of embryo identity and pattern in culture. Plant Reprod 206:181–196CrossRefGoogle Scholar
  68. Srivastava P, Chaturvedi R (2008) In vitro androgenesis in tree species: an update and prospect for further research. Biotechnol Adv 26:482–491CrossRefPubMedGoogle Scholar
  69. Srivastava P, Chaturvedi R (2011) Increased production of azadirachtin from an improved method of androgenic cultures of a medicinal tree Azadirachta indica A. Juss Plant Signal Behav 6:974–981CrossRefPubMedGoogle Scholar
  70. Thomas WTB, Newton AC, Wilson A, Booth A, Macaulay M, Keith R (2000) Development of recombinant chromosome substitution lines-a barley resource. SCRI annual report, pp 99–100Google Scholar
  71. Touraev A, Ilham A, Vicente O, Heberle-Bors E (1996) Stress-induced microspore embryogenesis in tobacco: an optimized system for molecular studies. Plant Cell Rep 15:561–565CrossRefPubMedGoogle Scholar
  72. Touraev A, Vicente O, Heberle-Bors E (1997) Initiation of microspore embryogenesis by stress. Trends Plant Sci 2:297–302CrossRefGoogle Scholar
  73. Touraev A, Forster BP, Jain SM (2009) (eds) Advances in haploid production in higher plants. Berlin, Springer, pp 1–208Google Scholar
  74. Xu L, Najeeb U, Tang GX, Gu HH, Zhang GQ, He Y, Zhou WJ (2007) Haploid and doubled haploid technology. Adv Bot Res 45:181–216CrossRefGoogle Scholar
  75. Zenkteler M, Nitzsche W (1984) Wide hybridization experiments in cereals. Theor Appl Genet 68:311–315CrossRefPubMedGoogle Scholar

Copyright information

© Springer International Publishing AG, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Biosciences and BioengineeringIndian Institute of Technology GuwahatiGuwahatiIndia

Personalised recommendations