| 新疆野生欧洲李ISSR-PCR反应体系的建立与优化 |
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| 引用本文： | 经建永,陈曦,马百强,耿文娟. 新疆野生欧洲李ISSR-PCR反应体系的建立与优化[J]. 分子植物育种, 2021, 19(4): 1223-1231 |
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| 作者姓名： | 经建永 陈曦 马百强 耿文娟 |
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| 作者单位： | 新疆农业大学林学与园艺学院,乌鲁木齐,830052;新疆园艺作物种质资源与高效生产重点实验室,乌鲁木齐,830052;新疆农业大学林学与园艺学院,乌鲁木齐,830052;新疆园艺作物种质资源与高效生产重点实验室,乌鲁木齐,830052;新疆农业大学林学与园艺学院,乌鲁木齐,830052;新疆园艺作物种质资源与高效生产重点实验室,乌鲁木齐,830052;新疆农业大学林学与园艺学院,乌鲁木齐,830052;新疆园艺作物种质资源与高效生产重点实验室,乌鲁木齐,830052 |
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| 基金项目： | 国家自然科学基金项目(31560545);新疆维吾尔自治区园艺重点学科(2016-10758-3)共同资助。 |
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| 摘 要： | 为建立适合新疆野生欧洲李的ISSR-PCR反应体系,本研究以野生欧洲李为供试材料,采用L16(45)正交试验设计和单因素试验设计相结合的方法,对影响野生欧洲李ISSR-PCR反应体系的5个因素(DNA模板,Taq酶,dNTPs,引物,Mg2+)进行筛选与优化,对16条引物的退火温度进行筛选.结果表明,Taq酶对PCR扩...
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| 关 键 词： | 野生欧洲李 ISSR-PCR 正交试验 单因素试验 体系优化 |
Establishment and Optimization of ISSR-PCR Reaction System in Xinjiang Prunus domestica L. |
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| Jing Jianyong,Chen Xi,Ma Baiqiang,GengWenjuan. Establishment and Optimization of ISSR-PCR Reaction System in Xinjiang Prunus domestica L.[J]. Molecular Plant Breeding, 2021, 19(4): 1223-1231 |
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| Authors: | Jing Jianyong Chen Xi Ma Baiqiang GengWenjuan |
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| Affiliation: | (College of Forestry and Horticulture,Xinjiang Agricultural University,Urumqi,830052;Laboratory of Germplasm Resources and Efficient Production of Horticultural Crops in Xinjiang,Urumqi,830052) |
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| Abstract: | In order to establish an ISSR-PCR reaction system suitable for Xinjiang Prunus domestica L.,the Prunus domestica was used as the test material.In this research,the ISSR-PCR reaction system for Prunus domestica was optimized by using the L16(45)orthogonal experimental design combined with signal factor analysis to screen five main factors(DNA template,Taq polymerase,dNTPs,primer,Mg2+)of PCR reaction.Based on the optimal ISSR-PCR reaction system of Prunus domestica,the optimal annealing temperature of 16 primers was determined.The results showed that Taq polymerase had the greatest influence on PCR amplification reaction.The optimal reaction system of Prunus domestica ISSR-PCR was:total volume 20μL,5 U Taq polymerase 0.10μL,10 mmol/L primer 1.00μL,2.5 mmol/L dNTPs 1.50μL,10×Buffer(containing Mg2+)2.00μL,50 ng/μL DNA template 1.00μL,and double distilled water 14.4μL.The established ISSR-PCR reaction system was verified by 22 Prunus domestica samples,which indicated that the reaction system was stable and reliable.The ISSR-PCR reaction system could be used for subsequent genetic diversity research,providing a theoretical reference for the protection and utilization of Prunus domestica germplasm resources. |
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| Keywords: | Wild European plum ISSR-PCR Orthogonal test Single factor test System optimization |
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