皇竹草是一种高产、优质牧草,但因耐寒性较差而极大限制了其在我国大面积种植和推广.选择在冬季平均温度接近皇竹草耐寒极限温度的四川地区进行了5年自然驯化,筛选在最低温度-5℃下能够自然过冬的样本,并在冬季最低温度下和夏季最高温度下分别取样进行转录组测序,挖掘皇竹草本身具有的耐寒、抗冻基因.结果显示:皇竹草转录组共有转录本125 467条,注释转录本65 633条,注释信息及邻近分析显示皇竹草与小米、玉米高度同源.冬夏两季皇竹草比较转录组分析共计筛选差异表达基因11 443个,其中冬季相对于夏季表达量上调基因6 943个,下调基因4 500个,差异显著.在以上差异表达基因中,共计挖掘在冬季低温条件下高度表达的抗冻基因112个,且有17个抗冻基因在冬季样品中特异表达.随机选取6个差异表达的抗冻基因进行荧光定量PCR实验验证,结果显示与预测转录本表达量一致.本研究对皇竹草进行了转录组测序并通过比较转录组的方法挖掘其在自然驯化条件下成功越冬样本中的抗冻基因,有利于揭示皇竹草冷适应下的抗性机制,同时也有助于热带引种植物分子育种的开展. Huangzhu grass is a kind of high-yield and high-quality forage grass, but due to poor cold tolerance and greatly restricted its large-scale planting and popularization in our country.Selected in the winter average temperature close to the imperial bamboo cold tolerance temperature in Sichuan for 5 years Natural acclimation, screened at the lowest temperature -5 ℃ can survive the winter samples, and in the lowest temperature in winter and the summer maximum temperature samples were taken for transcriptome sequencing, dig Emperor Xian grass has its own cold, antifreeze genes.The results showed : There were 125 467 transcripts and 65 633 annotated transcripts in Huangzhuban transcriptome. The annotation information and the adjacent analysis showed that Huangzhucao was highly homologous to millet and maize. Total difference in transcriptome analysis of imperial bamboo between winter and summer 11 443 genes were expressed, of which 6 943 genes were up-regulated in summer and 4 500 genes were down-regulated in summer, of which 112 genes were highly expressed in winter under low temperature , And 17 antifreeze genes were specifically expressed in winter samples.Five antifreeze genes that were differentially expressed were randomly selected and verified by real-time quantitative PCR. The results showed that compared with the predicted transcription The same amount of expression.We carried out the transcriptome sequencing of Pseudostellaria barbata and compared it with the transcriptome to find its antifreeze genes in the successful overwintering samples under natural acclimation conditions to reveal the resistance of Pseudostellaria barbata under cold acclimation Mechanism, but also contribute to tropical introduction of plant molecular breeding.