不同生长时期罗汉果果实转录因子的转录组分析及酵母单杂交文库的构建

Transcriptome Analysis on Transcription Factors of Siraitia grosvenorii Fruits at Different Growing Stages and Construction of Yeast One-Hybrid Library

目的:研究罗汉果果实中的转录因子(transcription factors,TFs)。方法:在授粉后3、50、70 d(days after flowering,DAF)的罗汉果果实转录组中查询注释为转录因子的所有非重复Unigene,并在表达谱中查找其表达情况;应用Matchmaker Yeast One-Hybrid Library Construction系统构建酵母单杂交c DNA文库。结果:罗汉果表达谱数据中共有38个家族的119个转录因子Unigene具有表达差异,其中以70 DAF/3 DAF和50 DAF/3 DAF组中具有差异表达的转录因子数目较多,且上调和下调的转录因子Unigene数目相近。此外,初步筛选到4个可能参与罗汉果甜苷合成调控的b HLH(basic helix-loop-helix,b HLH)转录因子Unigene(b HLH014、b HLH025、b HLH093、b HLH096)及8个可能参与黄酮类化合物合成调控的MYB转录因子Unigene;成功构建了酵母单杂交文库p GADT7-Rec2-Lib,其库容量为2.3×106 cfu,插入片段的平均长度约为1.5 kbp。结论:转录组数据分析和酵母单杂交实验有助于筛选罗汉果果实中具有调控作用的转录因子。

Objective： To study transcription factors （TFs） in the fruits of Siraitia grosvenorii. Methods： Non repeating TFs unigenes were selected in transcriptome of S. grosvenorii fruits at 3, 50 and 70 DAF and their expressions were investigated in gene expression profiles; Matchmaker Yeast One-Hybrid Library Construction system was applied to construct yeast one-hybrid cDNA library. Results： 119 differentially expressed TFs unigenes which belong to 38 TFs families were found out. The numbers of TFs unigenes in 70 DAF/3 DAF and 50 DAF/3 DAF groups were greater and the numbers of the up-regulated and down-regulated TFs unigenes were similar. What＇ s more, 4 bHLH （basic helix-loop-helix, bHLH ） TFs unigenes （bHLHO14, bHLH025, bHLH093, bHLH096 ） which might involve in the regulation of mogrosides synthesis and 8 MYB TFs unigenes which might involve in the regulation of flavonoids synthesis were screened. The yeast one-hybrid library pGADT7-Rec2-Lib was successfully constructed with 2. 3 × 10^6 cfu storage capacity. And the inserted PCR fragments sizes are about 1.5 kbp. Conclusion ： Transcriptome analysis and yeast one-hybrid system are able to help the selection of transcription factors with regulatory effect in S. grosvenorii fruits.