西瓜(Citrullus lanatus (Thunb.)Matsum. & Nakai)是世界范围内种植的一种重要的瓜类作物。西瓜果实品质、育性、结实率与雄花发育密切相关。本研究通过细胞学观察,对西瓜新品种‘新特大郑抗9号’雄花发育的不同阶段进行了区分,并进行了转录组测序分析。对花药进行醋酸洋红染色,并测定未开放雄花的纵横径。对花药不同发育时期的细胞学观察表明,花药从四分体生长到成熟期,其纵横径逐渐增大,雄花的花蕾长度在发育过程中也发生了显著的变化。对花药发育的四分体期(A组)、单核期(B组)、双核期(C组)和成熟期(D组)四个发育时期进行转录组测序分析。结果表明,四个阶段总共发现了16288个差异表达基因(DEGs),随着发育阶段的延伸,各比较组间的DEGs数量逐渐增加,6个比较组(A-VS-B、A-VS-C、A-VS-D、B-VS-C、B-VS-D和C-VS-D)的DEGs分别为2014、3259、4628、1490、3495和1132个。GO和KEGG富集分析表明,DEGs主要富集于细胞组分、淀粉和蔗糖代谢、苯丙类生物合成和戊糖合成等途径。最后,我们在6个比较组中共筛选了59个DEGs,有趣的是,我们发现一个花粉特异表达蛋白(Cla001608)显著下调(log2Fold Change值高达17.32),这表明它可能在雄花发育中起重要作用。本研究为了解西瓜雄性花发育阶段的分子基础提供了依据,并有助于优势杂交育种。
In vitro gynogenesis is an important tool used in haploid or homozygous double-haploid plant breeding. However, because of low repeatability, embryoid induction rate and quality, the molecular mechanisms remain poorly understood. Heat shock treatment can promote the transformation of the gametophytic pathway into the sporophyte pathway, which induces the occurrence of haploid. In this study, unfertilized ovaries were heat shocked for 0 h (A0) before flowering and for 0 h (A1), 4 h (A3), 8 h (A5), 12 h (A7), and 24 h (A8), respectively, at 37°C at the first day of the flowering stage. The ovule enlargement rate was increased from 0% at 25°C to 96.8% at 37°C (24 h treatment). Thus, we aimed to investigate the gene expression patterns in unfertilized ovules of watermelon after different periods of heat shock by using RNA-Seq technology. The results showed that compared with A3, A5, A7, and A8, the biosynthesis of amino acid, glycine, serine and threonine metabolic pathways in A1 has changed significantly. This indicated that heat shock treatment affected the synthesis and transformation of amino acids during ovule expansion. The transcriptome data suggested gene expressions of ovule growth were significantly changed by heat-specific influences. The results provide new information on the complex relationship between in vitro gynogenesis and temperature. This provides a basis for further study of the mechanism of heat shock affecting the expansion of watermelon ovule.