利用cDNA-AFLP技术分析全膜覆盖对玉米苗期基因表达的影响

发布时间：2018-01-26 01:21

本文关键词： 玉米全膜覆盖 cDNA-AFLP 差异表达　出处：《内蒙古大学》2017年硕士论文　论文类型：学位论文

【摘要】：为了研究玉米苗期在全膜覆盖条件下,基因的差异表达。本实验利用cDNA-AFLP技术对全膜覆盖条件下种植的玉米与不覆膜条件下种植的玉米,进行差异表达基因片段的筛选、分离、测序和功能性分析,结果如下:1.建立适合玉米苗期根系和叶片总RNA提取的方法,并将总RNA经过反转录形成双链cDNA。使用PstI和MseI两种内切酶对双链cDNA进行酶切,设计与两种内切酶相对应的接头以及9对选择性扩增引物,进行特异片段扩增及测序。2.运用cDNA-AFLP技术,共扩增出条带1481条,336条差异表达片段,约占总条带数的22.7%。随后将其中的60条进行测序得到58条测序结果。3.利用NCBI系统对58条测序结果进行同源性分析和功能预测。结果如下:与植物同源性较高的有13条并且大多具有蛋白功能;与动物同源性较高的有6条;与微生物同源性较高的有20条;另有,19条差异表达片段未能找到同源性序列。4.获得的与植物同源性较高的13条基因片段中,有3条与小麦、高粱、柑桔植物体中相关功能基因高度相似,其余10条均与玉米体内某些克隆基因高度相似。以上这些差异条带的获得为更深层次研究玉米基因在覆膜条件下的差异表达打下了良好的基础。5.测序结果中获得了 20条与微生物同源性高度相似的基因片段,这些基因片段的获得以及在玉米生长过程中的作用还需进一步研究确定。
[Abstract]:In order to study the differential expression of genes in seedling stage of maize under the condition of full film mulching, cDNA-AFLP technique was used to treat maize planted under the condition of full film mulching and maize without film mulching. Screening, isolation, sequencing and functional analysis of differentially expressed gene fragments were carried out. The results were as follows: 1. A suitable method for extracting total RNA from roots and leaves of maize at seedling stage was established. The total RNA was reversed to form double-stranded cDNA.The double strand cDNA was digested by PstI and MseI endonuclease. The specific fragments were amplified and sequenced by 9 pairs of primers. 1 481 bands were amplified by cDNA-AFLP technique. 336 differentially expressed fragments. Then 60 of them were sequenced to get 58 sequencing results .3.The homology analysis and functional prediction of 58 sequencing results were carried out by using NCBI system. The results were as follows:. Below:. 13 of them had high homology with plants and most of them had protein function. Six of them had high homology with animals. There were 20 high homology with microorganisms. Another 19 differentially expressed fragments could not find the homologous sequence. 4. Of the 13 genes with high homology with plants, 3 were with wheat and sorghum. The related functional genes in citrus plants are highly similar. The other 10 genes were highly similar to some cloned genes in maize. The above results laid a good foundation for further research on the differential expression of maize genes under the condition of film mulching. 5. Sequencing results. In the. There were 20 gene fragments with high homology with microorganism. The acquisition of these gene fragments and their roles in maize growth need to be further studied.
【学位授予单位】：内蒙古大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S513

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