西瓜离体雌核发育诱导单倍体植株再生

Haploid Plants Regeneration Induced by Watermelon Gynogenesis in Vitro

为建立完善的西瓜未授粉子房离体培养技术体系,本研究以西瓜未授粉子房为外植体,在离体条件下研究了激素种类、基因型、取样时间和热激处理时间对胚状体诱导效果的影响,并对再生植株倍性进行了鉴定。结果表明:外植体在MS+3.0 mg/L 2,4-D+1.5 mg/L 6-BA+0.5 mg/L NAA或MS+0.04 mg/L TDZ两种培养基上,胚状体诱导率分别为23%和22.3%,二者之间差异不显著;不同西瓜品种胚状体诱导率差异显著,‘早佳’最高可达23.3%,‘QL-2’最低只有2.3%;开花前12 h取样接种,胚状体诱导率最高可达22.3%;在35℃黑暗条件下热激5 d有利于子房的膨大和出胚,平均每瓶出胚5.3个;流式细胞术是鉴定再生植株倍性快速准确的方法。本研究结果为西瓜单倍体育种技术在育种实践中的应用提供了技术保障。

In order to establish a complete watermelon unfertilized ovary in vitro culture technological system, In this study, using unfertilized ovaries of watermelon as explants, we studied the effects of hormone types, genotypes, sampling time and heat shock treatment time on embryoid induction via in vitro culture, and we identified the ploidy of the regenerated plants. The results showed that media MS+3.0 mg/L 2,4-D+1.5 mg/L 6-BA +0.5 mg/L NAA and MS+0.04 mg/L TDZ achieved different embryoid induction rates, 23% and 22.3%, respectively, without significant difference. There were remarkable differences on embryoid induction rates of different watermelon cultivars. The ’zao jia’ was up to 23.3%, and the ’QL-2’ was only 2.3%. Sampling and inoculating 12 hours before flowering, and embryoid induction rate was up to 22.3%. Treatments with darkness and heat shock(35℃) for 5 days could help ovaries swelling and induce embryoids. The average of embryo was 5.3 per bottle. Flow cytometry was a rapid and accurate method for identifying the ploidy of regenerated plants The results of this study provide a technical guarantee for the application of watermelon haploid breeding technology in breeding practice.