| 均匀设计优化新疆主要葡萄品种ISSR-PCR反应体系 |
| |
| 引用本文： | 彭媛媛,高展,艾克拜尔·毛拉,周龙.均匀设计优化新疆主要葡萄品种ISSR-PCR反应体系[J].分子植物育种,2021,19(2):539-545. |
| |
| 作者姓名： | 彭媛媛 高展 艾克拜尔·毛拉 周龙 |
| |
| 作者单位： | 新疆农业大学林学与园艺学院,乌鲁木齐, 830052;新疆农业大学林学与园艺学院,乌鲁木齐, 830052;新疆农业大学林学与园艺学院,乌鲁木齐, 830052;新疆农业大学林学与园艺学院,乌鲁木齐, 830052 |
| |
| 基金项目： | 新疆维吾尔自治区科技计划重大专项(2017A01001);自治区级研究生创新项目(XJ2019G161);新疆农业大学2612创新团队(XJAU201528);新疆维吾尔自治区园艺学重点学科项目(2016-10758-3)共同资助。 |
| |
| 摘 要： | 为了建立适用于新疆主要葡萄品种的ISSR-PCR反应体系,本研究以新疆不同葡萄品种DNA为模板,采用均匀设计法,对影响ISSR-PCR体系的Mg^2+、dNTPs、引物、Taq DNA聚合酶等进行了U20(54)和U12(34)两轮优化,并在此基础上对退火温度及循环数进行了摸索。结果表明:在一定的浓度范围之内,Mg2+、d NTPs和引物浓度对体系的影响相对较小,而Taq DNA酶浓度对于整个ISSR-PCR体系的影响较大。优化后的25μL反应体系包含2.5 mmol/L Mg^2+,2.0 mmol/L dNTPs,1.0μmol/L引物,0.25 U Taq DNA聚合酶及1.0μmol/L模板DNA。最后运用优化体系对24份不同葡萄品种的DNA进行扩增验证,结果获得的DNA条带清晰,多态性比较丰富。说明优化后的ISSR-PCR体系可用于新疆不同葡萄种间亲缘关系和遗传多样性等领域的研究。
|
| 关 键 词： | 葡萄 ISSR-PCR 体系优化 均匀设计 |
Uniform Design Optimizes ISSR-PCR Reaction System for Major Grape Varieties in Xinjiang |
| |
| Peng Yuanyuan,Gao Zhan,Maola Aikebaier,Zhou Long.Uniform Design Optimizes ISSR-PCR Reaction System for Major Grape Varieties in Xinjiang[J].Molecular Plant Breeding,2021,19(2):539-545. |
| |
| Authors: | Peng Yuanyuan Gao Zhan Maola Aikebaier Zhou Long |
| |
| Institution: | (College of Forestry and Horticulture,Xinjiang Agricultural University,Urumqi,830052) |
| |
| Abstract: | In order to establish the ISSR-PCR reaction system suitable for the main grape varieties in Xinjiang.The DNA of different grape varieties in Xinjiang was used as the template,homogeneous design method was adopted to optimize the Mg2+,dNTPs,primers,Taq DNA polymerase,etc.in two rounds of U20(54)and U12(34),which affected the ISSR-PCR system.On this basis,the annealing temperature and cycle number are explored.The results showed that Mg^2+,dNTPs and primer concentration had little effect on the system,while Taq DNA polymerase concentration had greater effect on the whole ISSR-PCR system.The optimized 25μL reaction system contained 2.5 mmol/L Mg^2+,2.0 mmol/L dNTPs,1.0μmol/L primer,0.25 U Taq DNA polymerase and 1.0μmol/L template DNA.Finally,the optimized system was used to amplify and verify the DNA of 24 different grape varieties,and the obtained DNA bands were clear and polymorphic.It was indicated that the optimized ISSR-PCR system can be used to study the genetic relationship and genetic diversity of different grape species in Xinjiang. |
| |
| Keywords: | Grape ISSR-PCR System optimization Uniform design |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
|
