College of Horticulture, South China Agricultural University, Guangzhou 510642, China.
Plant Cell Rep. 2013 Sep;32(9):1361-72. doi: 10.1007/s00299-013-1448-8. Epub 2013 May 1.
A suppression subtractive hybridization library was constructed using inflorescence primordia of 'Nuomici' litchi to identify EST clones responsive to MV-generated ROS. 93 ESTs could be aligned as unique gene sequences in the inflorescence primordia of litchi. Litchi is an evergreen woody tree widely cultivated in subtropical and tropical regions. However, defective flowering is a pending problem of litchi production. Our previous study indicated that reactive oxygen species (ROS) induced by methyl viologen dichloride hydrate (MV) promotes flowering in litchi. In the present study, a suppression subtractive hybridization (SSH) library was constructed using inflorescence primordia of 'Nuomici' with the aim to find out ROS responsive clones during floral differentiation. 1856 Expressed sequence tag (EST) clones were randomly selected. Clones carrying single exogenous fragments were screened by reverse northern analysis to identify those responsive to MV-generated ROS. A total of 783 differentially expressed EST clones were identified as MV responsive cDNA and were subjected to sequencing. Among them, 26 clones were represented more than three times. 783 clones were aligned to 93 unique gene sequences. The unique genes were classified into 9 categories. 16 % of them were involved in transport facilitation, 11 % in transcription regulation, 4 % in stress response, 9 % in carbohydrate metabolism, 1 % in secondary metabolism, 14 % in intracellular signaling, and 25 % in other metabolism, while 9 % were genes with unknown functions and 11 % were genes with no match in the database.
采用荔枝花序原基构建抑制性消减杂交文库，以鉴定对 MV 产生的 ROS 有反应的 EST 克隆。93 个 EST 可以在荔枝花序原基中被排列为独特的基因序列。荔枝是一种广泛种植在亚热带和热带地区的常绿木本植物。然而，开花不良是荔枝生产中的一个待解决的问题。我们之前的研究表明，由甲基紫精二水合物（MV）诱导的活性氧（ROS）促进荔枝开花。在本研究中，采用‘糯米糍’荔枝花序原基构建了抑制性消减杂交（SSH）文库，旨在寻找在花分化过程中对 ROS 有反应的克隆。随机选择了 1856 个表达序列标签（EST）克隆。通过反向Northern 分析筛选携带单个外源片段的克隆，以鉴定对 MV 产生的 ROS 有反应的克隆。共鉴定出 783 个差异表达的 EST 克隆为对 MV 有反应的 cDNA，并进行了测序。其中，26 个克隆的表达量超过 3 次。783 个克隆与 93 个独特的基因序列相匹配。这些独特的基因被分为 9 类。其中，16%参与运输促进，11%参与转录调控，4%参与应激反应，9%参与碳水化合物代谢，1%参与次生代谢，14%参与细胞内信号转导，25%参与其他代谢，9%是功能未知的基因，11%是数据库中没有匹配的基因。
