红花(Carthamus tinctorius L.)SRAP反应体系建立与优化及对11个红花品种基因组的初步分析

Establishment and Optimization of SRAP-PCR and Preliminarily Analysis of Genomes in Carthamus tinctorius L.

摘要:

红花既是传统药用植物，也是一种新型油料经济作物。本研究利用SRAP技术对来源于不同地区的11个红花品种进行了条带分析。建立了红花种内SRAP-DNA的PCR扩增体系并进行了优化，进一步将最佳反应体系由25 μL减至10 μL，体系包括：Taq酶浓度0.04 u/μL，dNTP浓度为0.2 mM，正反引物浓度为0.3 mM，Mg2+浓度为2.0 mM，DNA模板为20 ng。为检测该优化体系的稳定性，选用了35对引物组合对11个分布于我国不同区域的红花品种进行PCR扩增和聚丙烯酰胺凝胶电泳，筛选出25对具有多态性条带的引物组合，获得了308条清晰的条带，其中109条具有多态性，比率为35%，说明用SRAP分子标记对于红花种内不同品种资源系统评估是完全可行的。红花SRAP反应体系的建立为深入评价红花的遗传多样性、分子辅助育种等研究奠定了基础。

Abstract: Safflower is a traditional medicinal plant, and also a new kind of oil economic crops. In this paper, the SRAP technology was used to analysis 11 safflower varieties. A SRAP-DNA PCR amplification system was established and optimized. The reaction mixture system was decreased from 25 μL to 10 μL, which contained: Taq DNA polymerase 0.04 u/μL, dNTP 0.20 mM, primers0.3 mM, MgCl2 2.0 mM, genomic DNA template 20 ng. In order to test the stability of SRAP-PCR system, the PCR amplification and polyacrylamide gel electrophoresis were used to analyse 11 safflower varieties with 35 primer pairs. 25 pairs of primers combination with polymorphism bands and 308 clear bands were obtained, 35 percent (109/308) bands were polymorphic. Therefore, the SRAP molecular marker system for safflower variety assessment was reliable. This research had put the foundation for genetic diversity research and molecular marker-assisted breeding of safflower.