LaPCBER1基因在‘Tresor’百合鳞茎发育过程中的功能解析

    Functional Analysis of LaPCBER1 Gene During the Development of Bulbs of Lilium 'Tresor'

    • 摘要:
      目的 分析苯基香豆满苄基醚还原酶基因LaPCBER1在‘Tresor’百合鳞茎发育过程中的表达模式,并探究其对百合鳞茎膨大的影响以及在此过程中对木质素含量的调控作用。
      方法 基于‘Tresor’百合鳞茎膨大过程的转录组数据,筛选到1个注释为PCBER1的差异基因,以‘Tresor’百合鳞茎cDNA为模板克隆LaPCBER1基因,并利用生物信息学软件分析其编码蛋白的理化性质;通过荧光定量PCR(qRT-PCR)检测LaPCBER1在百合不同组织及不同发育时期的表达模式;利用瞬时过表达技术探究其对鳞茎膨大的影响,并利用试剂盒检测不同时期以及过表达LaPCBER1后百合鳞茎的木质素含量。
      结果 LaPCBER1的DNA序列全长为930 bp,编码309个氨基酸,共预测到51个磷酸化位点,编码蛋白为非跨膜且不稳定亲水性蛋白。qRT-PCR结果显示,LaPCBER1在茎中表达量最高,在鳞茎发育S4时期(快速膨大期)表达量达峰值。瞬时过表达LaPCBER1后,转基因鳞茎木质素含量较空载体对照提高49.7%,鲜质量、干质量及直径相对增长率分别提高29.0%、29.7%和37.2个百分点;而沉默LaPCBER1后,鳞茎鲜质量、干质量及直径相对增长率分别降低35.9%、38.7%和32.2个百分点,木质素含量降低24.3%。
      结论 LaPCBER1基因在鳞茎发育早期高表达,且能促进木质素积累与鳞茎膨大,推测LaPCBER1参与‘Tresor’百合鳞茎的膨大过程,并在该过程中发挥正向调控作用。

       

      Abstract:
      Objective To analyze the expression pattern of the phenylcoumaran benzylic ether reductase gene LaPCBER1 during bulb development in Lilium 'Tresor', and to investigate its effects on bulb enlargement as well as its role in regulating lignin content during this process.
      Method By analyzing transcriptomic data from the bulb enlargement process of Lilium 'Tresor', a differentially expressed gene annotated as PCBER1 was screened. The LaPCBER1 gene was cloned using cDNA from Lilium 'Tresor' bulbs as the template, and the physicochemical properties of the encoded protein were analyze using bioinformatics software.The expression patterns of LaPCBER1 in different tissues and different developmental stages of Lilium 'Tresor' were detected by quantitative real-time PCR (qRT-PCR). The effects of LaPCBER1 on bulb enlargement were investigated using transient overexpression techniques, and the lignin content of lily bulbs at different stages and following overexpression was measured using assay kits.
      Result The full-length DNA sequenceof LaPCBER1 is 930 bp, encoding 309 amino acids. The protein is predicted to have 51 phosphorylation sites and is characterized as a non-transmembrane, unstable, hydrophilic protein. qRT-PCR results showed that LaPCBER1 was highly expressed in the stem, with expression peaking during the S4 stage (rapid expansion stage) of bulb development. Following transient overexpression of LaPCBER1, the lignin content in transgenic bulbs increased by 49.7% compared to the vector. The fresh weight, dry weight, and relative growth rates of circumference of the bulbs increased by 29.0%, 29.7%, and 37.2 percentage points, respectively; Conversely, after silencing LaPCBER1, the fresh weight, dry weight, and relative growth rates of circumference of the bulbs decreased by 35.9%, 38.7%, and 32.2 percentage points, respectively, and the lignin content decreased by 24.3%.
      Conclusion Expression pattern and functional analysis of LaPCBER1 suggest that this gene is highly expressed during the early stages of bulb development and promotes lignin accumulation and bulb expansion. It is therefore hypothesized that LaPCBER1 is involved in the bulb expansion process of Lilium 'Tresor' and plays a positive regulatory role in this process.