MENG Fanjuan,XING Chun,XU Fuling,et al.Optimization of ISSR on Prunus mira koehne Kov et.Kpst[J].Northern Horticulture,2015,39(20):95-98.[doi:10.11937/bfyy.201520023]
光核桃ISSR扩增体系的优化
- Title:
- Optimization of ISSR on Prunus mira koehne Kov et.Kpst
- Keywords:
- :Prunus mira koehne Kov et.Kpst; ISSR; system optimazation
- 文献标志码:
- A
- 摘要:
- 为确保光核桃ISSR反应条件的稳定性,以改良的CTAB法提取光核桃基因组DNA,对模板DNA浓度、MgCl2浓度、dNTPs浓度、Taq酶用量、引物浓度等因素进行了优化,确立了光核桃ISSR反应体系。结果表明:最佳反应体系为模板浓度40 ng/μL、引物浓度0.4 μmol/L、Mg2+的浓度2.5 mmol/L、酶浓度0.5 U、dNTPs浓度0.5 mmol/L,为光核桃这一优质的种质资源的遗传多样性研究奠定了理论基础。
- Abstract:
- To ensure the stability of ISSR for Prunus mira koehne Kov et.Kpst,total genomic DNA was extracted by improved CTAB method,the factors including template DNA concentration,MgCl2 concentration,dNTPs concentration,Taq polymorphism dose and primer concentration were studied to establish optimized ISSR reaction system.The result showed that template concentration 40 ng/μL,primer concentration 0.4 μmol/L,Mg2+ concentration 2.5 mmol/L,enzyme concentration 0.5 U,dNTPs concentration 0.5 mmol/L were the optimized reaction system.The study would provide the foundation for the genetic diversity on germplasm resources of Prunus mira koehne Kov et.Kpst.
参考文献/References:
[1]陈巍,王力荣,张绍铃,等.利用SSR研究不同国家桃育成品种的遗传多样性[J].果树学报,2007,24(5):580-584. [2]罗冉,吴委林,张旸,等.SSR分子标记在作物遗传育种中的应用[J].基因组学与应用生物学,2010,29(1):137-143. [3]罗海燕,陈业渊.ISSR分子标记及其应用[J].安徽农学通报,2008(19):45-46,27. [4]朱岩芳,祝水金,李永平,等.ISSR分子标记技术在植物种质资源研究中的应用[J].种子,2010(2):55-59. [5]韩宏.分子标记在果树种质资源研究中的应用[J].陕西农业科学,2010(4):74-76. [6]余贤美,艾呈祥.杧果野生居群遗传多样性ISSR 分析[J].果树学报,2007(3):329-333. [7]叶春海,王耀辉,李映志,等.菠萝蜜遗传多样性的ISSR 分析[J].果树学报,2009(5):659-665. [8]刘威生,冯晨静,杨建民,等.杏ISSR 反应体系的优化和指纹图谱的构建[J].果树学报,2005(6):626-629. [9]陈少瑜,杨恩,习学良,等.云南主要核桃品种的ISSR分子鉴别[J].经济林研究,2006(4):41-45. [10]姜静,杨传平,刘桂丰,等.桦树ISSR-PCR反应体系的优化[J].生态学杂志,2003(3):91-93. [11]谢运海,夏德安,姜静,等.利用正交设计优化水曲柳ISSR-PCR反应体系[J].分子植物育种,2005(3):445-450. [12]邹枚伶,夏志强,王文泉.白木香基因组DNA提取与ISSR反应体系的优化[J].中国农学通报,2009(2):250-254. [13]刘威生,冯晨静,杨建民,等.杏ISSR 反应体系的优化和指纹图谱的构建[J].果树学报,2005(6):626-629.
相似文献/References:
[1]肖志娟,翟梅枝,许静,等.不同核桃品种的ISSR标记分析[J].北方园艺,2013,37(04):103.
[2]刘志曦,李志强,魏海莲,等.平菇ISSR遗传多样性分析[J].北方园艺,2014,38(12):94.
LIU Zhi-xi,LI Zhi-qiang,WEI Hai-lian,et al. Phylogenetic Analysis of Oyster Mushrooms by ISSR Marker[J].Northern Horticulture,2014,38(20):94.
[3]李 娜,张存旭,崔晓燕,等.栓皮栎ISSR反应体系的建立[J].北方园艺,2014,38(13):93.
LI Na,ZHANG Cun-xu,CUI Xiao-yan,et al.Establishment of ISSR Reaction System in Quercus variabilis [J].Northern Horticulture,2014,38(20):93.
[4]王鸿磊,丁 强,吕 蔚,等.我国双孢菇菌株与国外引进菌株间亲缘关系的ISSR分析[J].北方园艺,2014,38(11):96.
WANG Hong-lei,DING qiang,LV Wei,et al.Genetic Relationship Based on ISSR Markers Among Agaricus bisporus Strains From China and Overseas[J].Northern Horticulture,2014,38(20):96.
[5]李 娜,张存旭,刘 伟,等.毛梾ISSR-PCR反应体系的建立[J].北方园艺,2013,37(22):114.
LI Na,ZHANG Cun-xu,LIU Wei,et al.Establishment of ISSRPCR Reaction System of Cornus walteri[J].Northern Horticulture,2013,37(20):114.
[6]周淑香,符亚茹,王 超,等.西藏光核桃果实及种子表型性状变异研究[J].北方园艺,2013,37(23):38.
ZHOU Shu-xiang,FU Ya-ru,WANG Chao,et al.Study on Phenotypic Variation of Seed and Fruit on Prunus miakoehne in Tibet[J].Northern Horticulture,2013,37(20):38.
[7]黄萌,陈尚钘,杨光耀,等.古樟ISSR扩增条件的优化[J].北方园艺,2013,37(07):116.
HUANG Meng,CHEN Shang-xing,YANG Guang-yao,et al.Optimization of ISSR Amplification Conditions in Ancient Camphor-trees[J].Northern Horticulture,2013,37(20):116.
[8]吴春燕,高义,宋廷宇,等.正交设计优化大白菜ISSR-PCR 反应体系[J].北方园艺,2012,36(05):127.
WU Chun-yan,GAO Yi,SONG Ting-yu,et al.Optimization of an ISSR-PCR Reaction System of Chinese Cabbage by Orthogonal Design[J].Northern Horticulture,2012,36(20):127.
[9]陈名红,李 玉,刘 多,等.适于ISSR-PCR扩增的百合基因组DNA的提取[J].北方园艺,2012,36(24):108.
CHEN Ming-hong,LI Yu,LIU Duo,et al.Extraction of Genomic DNA for ISSR-PCR Reaction in Lilium[J].Northern Horticulture,2012,36(20):108.
[10]张运峰,胡芬.红掌组培再生植株中表型突变体的ISSR分析[J].北方园艺,2014,38(21):101.
ZHANG Yun-feng,HU Fen.ISSR Analysis of Phenotypic Variants in Tissue Culturing Regeneration Plants of Anthurium andraeanum[J].Northern Horticulture,2014,38(20):101.
备注/Memo
第一作者简介:孟凡娟(1975-),女,黑龙江兰西人,博士,副教授,现主要从事植物资源等研究工作。E-mail:mfj19751@163.com.责任作者:李荣钦(1968-),女,四川成都人,硕士,副教授,现主要从事植物资源等研究工作。E-mail:mfj19751@126.com.基金项目:国家自然科学基金资助项目(31160386)。