论文部分内容阅读
以黄瓜高抗霜霉病自交系为材料,构建了两个抑制差减文库,经反向Northern杂交筛选、测序和序列比对,筛选到3个未知功能抗病相关基因。荧光实时定量RT-PCR分析结果表明,未知功能抗病相关基因2I15(GD254229)、3C19(GD254243)和1O08(GD254207)在抗病自交系接种霜霉病病原菌后,可早期高丰度特异上调表达,而在感病自交系中特异上调表达丰度较低或被特异抑制表达。水杨酸(SA)可诱导1O08特异上调表达,抑制2I15和3C19的表达;茉莉酸(JA)处理可诱导1O08特异上调表达,而抑制2I15的表达。ABA处理可显著诱导3C19和1O08特异上调表达,而抑制2I15的表达。机械伤害、高盐、冷害和热激等其它非生物胁迫可显著抑制或诱导这些基因的表达,因此推测,所筛选到的功能未知基因可能具有参与生物和非生物胁迫反应的作用。
Two inbred subtractive libraries were constructed from inbred lines with high resistance to downy mildew in cucumber. Three unknown resistance genes were screened by reverse Northern hybridization, sequencing and sequence alignment. Real-time quantitative RT-PCR analysis showed that the resistance genes 2I15 (GD254229), 3C19 (GD254243) and 1O08 (GD254207) with unknown function could be specifically upregulated early after inoculated with downy mildew pathogen in resistant inbred lines Expression, while in susceptible inbred lines specifically upregulate the expression of low or specifically inhibited expression. Salicylic acid (SA) induced a specific up-regulation of 1O08 and inhibited the expression of 2I15 and 3C19. JA treatment induced up-regulation of 1O08 and down-regulation of 2I15 expression. ABA treatment significantly induced 3C19 and 1O08 specific up-regulated expression, while inhibiting 2I15 expression. Other abiotic stress such as mechanical injury, high salt, chilling injury and heat shock can significantly inhibit or induce the expression of these genes, so it is speculated that the functionally unknown genes screened may be involved in the biotic and abiotic stress responses.