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目的:建立适用于棉花的稳定可靠的MFLP技术体系,初步分析该技术在棉花应用中的适用性。方法:以冀棉169为材料对试验因素进行选择优化,建立适用于棉花分析体系。利用32对MFLP引物组合对33B、冀棉20、中棉所12、鲁棉1号4个品种进行扩增,初步分析该技术在棉花应用中的适用性。结果:确定的最佳酶切反应时间为2h,预扩增最适退火温度为54℃,选择性扩增使用的模板为预扩增产物的20倍稀释物。利用32对引物组合对4个棉花品种进行扩增分析,其中14对引物组合产生多态性条带,引物多态率为43.75%,每个组合的多态性条带为1~5条不等。结论:建立的MFLP技术体系稳定可靠,在棉花遗传多样性分析和分子标记辅助育种等方面具有良好的应用前景。
Objective: To establish a stable and reliable MFLP system suitable for cotton, and to analyze the applicability of the technology in cotton application. Methods: The experimental factors were selected and optimized by using Jimian 169 as material, and the suitable cotton analysis system was established. The 32 pairs of MFLP primer combinations were used to amplify 4 varieties of 33B, JiMian 20, ZhongmianShen 12 and LuMian 1, and the applicability of this technique in cotton was analyzed. Results: The optimum reaction time was 2h, the optimum temperature for pre-amplification was 54 ℃, and the template for selective amplification was 20-fold dilution of the pre-amplification product. Four cotton cultivars were amplified using 32 pairs of primers. Among them, 14 pairs of primers produced polymorphic bands with primer polymorphism of 43.75% and polymorphic bands of 1 ~ 5 for each combination Wait. Conclusion: The established MFLP technology system is stable and reliable, and has good application prospects in cotton genetic diversity analysis and molecular marker-assisted breeding.