甘蔗AFLP分析技术体系的建立

研究目的】为了建立甘蔗AFLP标记技术体系；【方法】本文通过对甘蔗抗、感黑穗病的杂交亲本CP84-1198和科5，以及根据其杂交后代的浸渍接种田间抗性鉴定结果所构建的抗、感黑穗病池的AFLP银染法分析；【结果】建立了甘蔗黑穗病抗性的AFLP分析实验体系，同时将AFLP-银染技术应用于甘蔗近缘植物斑茅抗旱胁迫及其对照材料的cDNA差别筛选，建立了一种mRNA差别显示技术体系。实验过程中还筛选出了四对能在抗、感亲本上扩增出特异条带的引物，以及一对在斑茅水分胁迫和对照中表现良好差异的引物，为下一步的实验工作的开展奠定了基础。【结论】通过本实验建立了甘蔗AFLP分析技术体系、变性聚丙烯酰胺凝胶的快速银染、凝胶简单保存以及聚丙烯酰胺凝胶中的特异条带的高效回收技术。

The System Establishment of AFLP Technique in Sugarcane

[OBJECTIVE] In order to establish the AFLP technique system in sugarcane; [METHOD] the hybrid parents CP84-1198 and Ke5 and correspondingly the resistance pool and susceptible pool were taken as plant materials and the AFLP(amplified fragment length polymorphism)-sliver stained experiment system was constructed accordingly. Moreover, the due system was used to analyze the drought-resistance character using the cDNA from E.arundinaceus as materials and a DD (differentially display) analysis method, which can also be named as cDNA-AFLP, was established. [RESULTS] During the experiment, four pairs of primers which can amplify differentially displayed bands between the resistant parent and susceptible parent, were selected. Among them, one pair of primer was found to produce favorable differentially displayed bands between the control and drought-menace E. arundinaceus. At the same time, both a quick method of silver staining of PAGE and a feasible gel conservation method were set up. And the recovery protocol of band on PAGE was also proved to be convenient and efficient. [CONCLUSION] In conclusion, the AFLP technique system in sugarcane analysis has been established in this paper.