Chinese Bulletin of Botany ›› 2013, Vol. 48 ›› Issue (1): 79-86.DOI: 10.3724/SP.J.1259.2013.00079

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In Vitro Culture of Pumpkin Unfertilized Ovules and Plant Regeneration

Shouru Sun, Peng Zhang, Jianbin Hu*, Liping Sun, Man Zhang, Zhiqiang Sun   

  1. College of Horticulture, Henan Agricultural University, Zhengzhou 450002, China
  • Received:2012-07-27 Revised:2012-09-30 Online:2013-01-01 Published:2012-11-01
  • Contact: Jianbin Hu

Abstract: We used unfertilized ovules of the pumpkin cultivar ‘Experiment 1’ as explants to examine the effect of combinations and concentrations of growth regulators, different developmental periods, high-temperature pretreatment, and AgNO3 concentrations on embryoid induction. The combination of 2,4-D, NAA, and 6-BA favoured embryoid induction; the medium with the best effect was Murishige and Skoog (MS)+1.0 mg·L–12,4-D+0.25 mg·L–1NAA+0.5 mg·L–1 6-BA, for 31.1% embryoid induction. The flowering ovules for the day showed the highest frequency of embryoid induction (26.7%) and the lowest frequency of callus formation (<5%). The explants treated with darkness and high temperature (35°C) for 5 days showed the best embryoid induction, 32.2%. However, the addition of AgNO3 remarkably inhibited embryoid induction. Transfer of embryoids to plant-regeneration medium led to the highest regeneration frequency, 64.3%. Plant regeneration expressed a typical pathway as for embryogenesis. Cytologic observations revealed that embryoids probably derived from cells close to the micropyle (i.e., ovums or synergids).