, 215:111 | Cite as

A unique strategy to improve the floral traits and seed yield of Brassica oleracea cytoplasmic male sterile lines through honey bee-mediated selection

  • S. S. DeyEmail author
  • R. Bhatia
  • Achintya Pramanik
  • Kanika Sharma
  • Chander Parkash
Original Article


Improvement in flower structure and seed yield is one of the main objectives when developing cytoplasmic male sterile lines (CMS) in vegetable Brassicas for their successful use in the development of F1 hybrids. However, it is found that the introgression of alien cytoplasm alters the reproductive structure of Brassica oleracea genotypes and the altered floral structure in these lines can be improved only marginally even after several generations of manual selection. We have tested an alternative selection strategy using honey bee (Apis cerena indica) as the selection agent in five selection cycles in three CMS lines of cauliflower (Ogu1A, Ogu2A and Ogu3A). Selected plants with the highest seed yield, determined through honey bee selection, were bulked to raise the plants for next generation. The reproductive structures of the CMS plants improved significantly after five cycles of honey bee-mediated selection. Seed yield/plant, nectar quantity, size of nectary and flower size improved significantly in all three CMS lines at the end of the selection cycles. Quantity of nectar increased from 0.45 to 1.45 µl and from 0.37 to 1.49 µl per flower in CMS lines Ogu1A and Ogu3A, respectively, after five cycles of selection; similarly, quantity of seeds per plant improved from 13.48 to 20.71 g and from 12.77 to 17.57 g in the same CMS lines, respectively. The extent of improvement in nectar quantity and seed yield was comparatively lower in CMS line Ogu2A. The occurrence of different floral deformities was also reduced in all three CMS lines after five cycles of honey bee-mediated selection. CMS lines Ogu1A and Ogu3A responded better to honey bee-mediated selection than did CMS line Ogu2A. This novel strategy of honey bee-mediated selection can be used to develop CMS lines with higher seed yield in different crops where cytoplasmic male sterility is commercially used in hybrid seed production.


CMS lines Brassica oleracea Seed yield Floral traits Honey bee Selection 



The authors are grateful to ICAR–Indian Agricultural Research Institute, New Delhi for providing financial assistance and other logistic support that enabled this long-term experiment to be conducted.

Author’s contributions

SSD has planned and supervised the entire research program. RB collected the data in the laboratory and was involved in conducting the field experiments. AP arranged the suitable bee colony and recorded data on nectar quantity and nectar development. KS, CP and RK were involved in the field experiments and recorded data on different occasions. SSD compiled the entire data set and carried out the statistical analysis.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict(s) of interest.


  1. Abrol DP (2007) Honeybees and rapeseed: a pollinator–plant interaction. Adv Bot Res 45:337–367CrossRefGoogle Scholar
  2. Bhatia R, Dey SS, Sood S, Sharma K, Sharma VK, Parkash C, Kumar R (2016) Optimizing protocol for efficient microspore embryogenesis and doubled haploid development in different maturity groups of cauliflower (B. oleracea var. botrytis L.) in India. Euphytica 212:439–454CrossRefGoogle Scholar
  3. Bhatia R, Dey SS, Sood S, Sharma K, Parkash C, Kumar R (2017) Efficient microspore embryogenesis in cauliflower (Brassica oleracea var. botrytis L.) for development of plants with different ploidy level and their use in breeding programme. Sci Hortic 216:83–92CrossRefGoogle Scholar
  4. Bhatia R, Dey SS, Parkash C, Sharma K, Sood S, Kumar R (2018) Modification of important factors for efficient microspore embryogenesis and doubled haploid production in field grown white cabbage (Brassica oleracea var. capitata L.) genotypes in India. Sci Hortic 233:178–187CrossRefGoogle Scholar
  5. Brunet J (2009) Pollinators of the rocky mountain columbine: temporal variation, functional groups and associations with floral traits. Ann Bot 103:1567–1578CrossRefGoogle Scholar
  6. Brunet J, Thairu MW, Henss JM, Link RI, Kluever JA (2015) The effects of flower, floral display and reward sizes on bumble bee foraging behaviour when pollen is the reward and plants are dichogamous. Int J Plant Sci 176:811–819CrossRefGoogle Scholar
  7. Conner JK, Rush S (1996) Effects of flower size and number on pollinator visitation to wild radish, Raphanus raphanistrum. Oecologia 105:509–516CrossRefGoogle Scholar
  8. Dey SS, Sharma SR, Parkash C, Kumar PR, Bhatia R (2011) Development and characterization of “Ogura” based improved CMS lines of cauliflower (Brassica oleracea var. botrytis L.). Indian J Genet Plant Breed 71:37–42Google Scholar
  9. Dey SS, Bhatia R, Sharma SR, Parkash C, Sureja AK (2013a) Effects of chloroplast substituted Ogura male sterile cytoplasm on the performance of cauliflower (Brassica oleracea var. botrytis L.) F1 hybrids. Sci Hortic 157:45–51CrossRefGoogle Scholar
  10. Dey SS, Singh N, Bhatia R, Parkash C, Chandel C (2013b) Genetic combing ability and heterosis for important vitamins and antioxidant pigments in cauliflower (Brassica oleracea var. botrytis L.). Euphytica 195:169–181CrossRefGoogle Scholar
  11. Dey SS, Bhatia R, Bhardwaj I, Mishra V, Sharma K, Parkash C, Kumar S, Sharma VK, Kumar R (2017a) Molecular-agronomic characterization and genetic study reveals usefulness of refined Ogura cytoplasm based CMS lines in hybrid breeding of cauliflower (Brassica oleracea var. botrytis L.). Sci Hortic 224:27–36CrossRefGoogle Scholar
  12. Dey SS, Bhatia R, Parkash C, Kumar R (2017b) Heterosis and combining ability analysis in snowball cauliflower using indigenously developed CMS lines. Indian J Hortic 74:374–381CrossRefGoogle Scholar
  13. Dey SS, Bhatia R, Sharma SR, Sharma K, Parkash C, Kumar R (2018) Population dynamics in introgression of wild male sterile cytoplasm into Brassica oleracea: a small population based breeding model. Sci Hortic 232:231–239CrossRefGoogle Scholar
  14. Eckhart VM (1991) The effects of floral display on pollinator visitation vary among populations of Phacelia linearis (Hydrophyllaceae). Evol Ecol 5:370–384CrossRefGoogle Scholar
  15. Engel EC, Irwin R (2003) Linking pollinator visitation rate and pollen receipt. Am J Bot 90:1612–1618CrossRefGoogle Scholar
  16. Golubov J, Eguiarte LE, Mandujano MC, Lopez-Portillo J, Montana C (1999) Why be a honeyless honey mesquite? Reproduction and mating system of nectarful and nectarless individuals. Am J Bot 86:955–963CrossRefGoogle Scholar
  17. Heinrich B (1975) Energetics of pollination. Annu Rev Ecol Syst 6:139–170CrossRefGoogle Scholar
  18. Heinrich B (1979) Majoring and minoring by foraging bumblebees, Bombus vagans: an experimental analysis. Ecology 60:245–255CrossRefGoogle Scholar
  19. Karron JD, Mitchell RJ (2012) Effects of floral display size on male and female peproductive success in Mimulus ringens. Ann Bot 109:563–570CrossRefGoogle Scholar
  20. Kucera V, Chytilova V, Vyvadilova M, Klima M (2006) Hybrid breeding of cauliflower using self-incompatibility and cytoplasmic male sterility. Hortic Sci Prague 33:148–152CrossRefGoogle Scholar
  21. Makino TT, Sakai S (2007) Experience changes pollinator responses to fl oral display size: from size-based to reward-based foraging. Funct Ecol 21:854–863CrossRefGoogle Scholar
  22. Mesquida J, Renard M (1978) The entomophilous pollination of male sterile strains of Winter rapeseed (Brassica napus L.) and a preliminary study of alternating devices. In: Caron TA (ed) Fourth Int Symp on Pollination. Washington DC, pp 49–57Google Scholar
  23. Mitchell RJ (1994) Effects of fl oral traits, pollinator visitation, and plant size on Ipomopsis aggregata fruit production. Am Nat 143:870–889CrossRefGoogle Scholar
  24. Mitchell RJ, Karron JD, Holmquist KG, Bell JM (2000) The influence of Mimulus ringens floral display size on pollinator visitation patterns. Funct Ecol 18:116–124CrossRefGoogle Scholar
  25. Nieuwhof M (1963) Pollination and contamination of Brassica oleracea L. Euphytica 12:17–26CrossRefGoogle Scholar
  26. Ogura H (1968) Studies on the new male-sterility in Japanese radish, with special reference to the utilization of this sterility towards the practical raising of hybrid seeds. Mem Fac Agric Kagoshima Univ 6:36–78Google Scholar
  27. Pelletier G, Primard G, Vedel F, Chetrit P, Remy R, Rousselle P, Renard M (1983) Intergeneric cytoplasmic hybridization in Cruciferae by protoplast fusion. Mol Genes Genet 191:244–250CrossRefGoogle Scholar
  28. Pyke GH, Day LP, Wale KA (1998) Pollination ecology of Christmas Bells (Blanfordia nobilis Sm): effects of adding artificial nectar on pollen removal and seed-set. Aust J Ecol 13:279–284CrossRefGoogle Scholar
  29. Real LA, Rathcke BJ (1991) Individual variation in nectar production and its effect on fitness in Kalmia latifolia. Ecology 72:149–155CrossRefGoogle Scholar
  30. Sharma SR, Vinod (2002) Breeding for cytoplasmic male sterility in broccoli (Brassica oleracea L. var. italica Plenk). Indian J Genet 62:65–166Google Scholar
  31. Sharma SR, Singh PK, Chable V, Tripathy SK (2004) A review in hybrid cauliflower development. In: Singh PK, Dasgupta SK, Tripathi SK (eds) Hybrid vegetable development. Haworth Press, New York, pp 217–221Google Scholar
  32. Singh BK, Singh B (2016) Development of hybrids and hybrid seed production of cole crops. In: Singh B, Pandey S, Singh N, Manimurugan C, Devi J, Singh PM (eds) Principles and production techniques of hybrid seeds in vegetables. TM No. 67. Indian Council of Agricultural Research–Indian Institute of Vegetable Research, Varanasi, pp 112–125Google Scholar
  33. Singh KH, Srivastava KK (2006) Characterization of different cytoplsmic male sterility systems in Indian mustard (Brassica juncea L. Czern & Coss). Plant Breed 125:72–76CrossRefGoogle Scholar
  34. Thomson JD, Plowright RC (1980) Pollen carryover, nectar rewards and pollinator behaviour with special reference to Diervilla lonicera. Oecologia 46:68–74CrossRefGoogle Scholar
  35. Yamagishi H, Bhat SR (2014) Cytoplasmic male sterility in Brassicaceae crops. Breed Sci 64:38–47CrossRefGoogle Scholar
  36. Zimmerman M (1988) Nectar production, fowering phenology and strategies for pollination. In: Lovett Doust J, Lovett Doust L (eds) Plant reproductive ecology. Oxford University Press, New York, pp 157–178Google Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.ICAR-Indian Agricultural Research InstituteRegional StationKatrain, KulluIndia
  2. 2.Division of Vegetable ScienceICAR–Indian Agricultural Research InstituteNew DelhiIndia
  3. 3.Division of Floriculture and LandscapingICAR–Indian Agricultural Research InstituteNew DelhiIndia

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