[1]张云秀,杨美玲,于玮玮,等.新疆野苹果SSR-PCR反应体系的优化[J].北方园艺,2017,41(15):29-35.[doi:10.11937/bfyy.20165044]
　ZHANG Yunxiu,YANG Meiling,YU Weiwei,et al.Optimization of SSR-PCR Reaction System forMalus sieversii (Lebed.) Roem.[J].Northern Horticulture,2017,41(15):29-35.[doi:10.11937/bfyy.20165044]
点击复制

新疆野苹果SSR-PCR反应体系的优化

《北方园艺》[ISSN:1001-0009/CN:23-1247/S] 卷: 第41卷 期数: 2017年15 页码: 29-35 栏目: 出版日期: 2017-08-15
Title:
Optimization of SSR-PCR Reaction System forMalus sieversii (Lebed.) Roem.
作者:
张云秀1; 杨美玲2; 于玮玮1; 邹家辉1; 龙鸿1; 阎国荣1
（1.天津农学院 园艺园林学院，天津 300384;2.南开大学 生命科学学院,天津 300071）
Author(s):
ZHANG Yunxiu1; YANG Meiling2; YU Weiwei1; ZOU Jiahui1; LONG Hong1; YAN Guorong1
(1.Garden and Horticulture Institute,Tianjin Agricultural University,Tianjin 300384;2.College of Life Sciences,Nankai University,Tianjin 300071)
关键词:
新疆野苹果; 基因组DNA; SSR-PCR; 优化
Keywords:
Malus sieversii; extract genomic DNA; SSR-PCR; optimization
DOI:
10.11937/bfyy.20165044
文献标志码:
A
摘要:
以新疆野苹果叶片为试材，采取改良CTAB法提取基因组,参照基本PCR反应体系,采用控制变量的方法，研究了各成分不同的浓度梯度对新疆野苹果SSR-PCR扩增的影响，为新疆野苹果的遗传育种奠定基础。结果表明：新疆野苹果SSR扩增的最佳体系为10×PCR buffer 2.5 μL，dNTP 0.2 mmol?L-1，0.2 mmol?L-1引物，0.5 U Taq DNA聚合酶，100 ng DNA模板。并用此优化体系对154份新疆野苹果进行了稳定性、普遍性验证，扩增效果较好。
Abstract:
CTAB method was adopted to extract genomic DNA from the dried leaves of Malus sieversii (Lebed.) Roem.The effect of different concentrations of components on SSR-PCR was studied by using control variate method referring to basic PCR system in order to lay the foundation of genetic breeding for Malus sieversii (Lebed.) Roem.The results showed that the optimal SSR-PCR system was 10×PCR buffer 2.5 μL，dNTP 0.2 mmol?L-1，0.2 mmol?L-1 primers，0.5 U Taq DNA polymerase，100 ng DNA template;154 kinds of Malus sieversii germplasm were used to test the stability and universality of system,amplification effect was good.

参考文献/References:

 

[1]李育农.苹果属植物种质资源研究[M].北京:中国农业出版社,2001:20-134.

[2]阎国荣,许正.中国新疆野生果树研究[M].北京:中国林业出版社,2010:62-114.

[3]林培钧,崔乃然.天山野果林资源伊犁野果林综合研究[M].北京:中国林业出版社,2000:6-140.

[4]SOUSA A G R,SOUZA M M,MELO C A F,et al.ISSR markers in wild species of Passiflora L.(Passifloraceae) as a tool for taxon selection in ornamental breeding[J].Genetics and Molecular Research,2015,14 (4):18534-18545.

[5]YU H P,CHENG F Y,ZHONG Y,et al.Development of simple sequence repeat (SSR) markers from Paeonia ostii to study the genetic relationships among tree peonies (Paeoniaceae)[J].Scientia Horticulturae,2013,164:58-64.

[6]温景辉.基于SSR分子标记的山葡萄种质遗传多样性研究与核心种质构建[D].长春:吉林农业大学,2011.

[7]巴巧瑞,赵政阳,高华,等.基于SSR和SRAP标记的苹果品种亲缘关系分析[J].西北农林科技大学学报,2011,39(9):124-128.

[8]左力辉,张文林,邱彤,等.新疆野苹果叶形性状变异及其与 SSR 标记关联分析[J].园艺学报,2015,42(4):759-768.

[9]BUDAK H,SHEARMAN R C,PARMAKSIZ I,et al.Comparative analysis of seeded and vegetative biotype buffalo grasses based on phylogenetic relationship using ISSRs,SSRs,RAPDs,and SRAPs[J].Theor Appl Genet,2004,109(1):280-288.

[10]LITT M,LUTY J A.A hypervariable microsatellite revealed by in vitro amplification of a dinucleotide repeat within the cardiac muscle actin gene[J].Am J Hum Genet,1989,44:397-401.

[11]TAUTZ D.Hyper variability of simple sequence as a general source for polymorphic DNA marker[J].Nucleic Acids Search,1989,17(16):6463-6471.

[12]SMEETS A J M,BRUNNER H G,ROPERS H H,et al.Use of variable simple sequence motifs as genetic markers:Application to study of myotonic dystrophy[J].Human Genetics,1989,83:245-251.

[13]WEBER J L,MAY P E.Abundant class of human DNA polymorphisms which can be typed using the polymerase chain reaction[J].American Journal of Human Genetics,1989,44:388-396.

[14]高源,田路明,刘凤之.利用SSR荧光标记构建92个梨品种指纹图谱[J].园艺学报,2012(8):1437-1446.

[15]张萌.基于SSR分子标记的葡萄种质资源的遗传多样性分析及品种鉴定[D].南京:南京农业大学,2012.

[16]岳娜.枇杷(Eriobotrya Lindl)种质资源SSR标记指纹图谱构建与遗传连锁分析[D].苏州:苏州大学,2012.

[17]王爱德,李天忠,许雪峰,等.苹果品种的SSR分析[J].园艺学报,2005,32(5):721-726.

[18]COTA-SANCHEZ′ J H,REMARCHUK K,UBAYASENA K.Ready-to-use DNA extracted with a CTAB method adapted for herbarium specimens and mucilaginous plant tissue[R].Canada:International Socieiy for Plant Molecular Biology,2006.

[19]LISTON A,RIESEBERG L H,ADAMAR P,et al.A method for collecting dried plant specimens for DNA and isozyme analyses,and the results of a field test in Xinjiang,China[J].Annals of Missouri Botanic Garden,1990,77(4):859-863.

[20]张春雨,陈学森,何天明,等.中国新疆野苹果（Malus sieversii （Lebed.） Roem.）群体遗传结构的SSR 分析[J].遗传学报,2007,34(10):947-955.

[21]高源,刘风之,曹玉芬,等.苹果属种质资源亲缘关系的SSR分析[J].果树学报,2007,23(2):129-134.

[22]徐兴兴,梁海永,甄志先,等.苹果SSR反应体系的建立[J].果树学报,2006,23(2):161-164.

[23]王云,秦伟,王永丰.新疆野苹果S基因特异性PCR体系优化[J].新疆农业科学 2013,50(11):2023-2030.

[24]俞明亮,马瑞娟,沈志军,等.应用SSR 标记进行部分黄肉桃种质鉴定和亲缘关系分析[J].园艺学报,2010,37(12):1909-1918.

[25]张文娜,王忆,孔瑾,等.甜樱桃品种SSR-PCR反应体系的优化[J].生物技术通报,2009(11):103-107.

[26]张靖国.湖北海棠遗传多样性的SSR分析[D].武汉:华中农业大学,2007:33-44.

[27]张淑静.葡萄SSR反应体系的建立及遗传多样性分析[D].保定:河北农业大学,2008:48-50.

相似文献/References:

[1]董学卫,李有志,何庆芳,等.聚球藻7002基因组DNA提取方法比较[J].北方园艺,2013,37(20):95.
　DONG Xue-wei,LI You-zhi,HE Qing-fang,et al.Comparison of Different Methods for Extracting Genomic DNA of Synechococcus sp.PCC 7002[J].Northern Horticulture,2013,37(15):95.
[2]刘锴栋,袁长春,陈燕,等.不同方法提取野生荔枝基因组DNA效果的比较[J].北方园艺,2012,36(23):105.
　LIU Kai-dong,YUAN Chang-chun,CHEN Yan,et al.Comparison on Effects of Different Methods on Extract Genomic DNA of Wild Litchi Leaves[J].Northern Horticulture,2012,36(15):105.
[3]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(15):108.
[4]左力辉,张 军,董 研,等. 新疆野苹果自然群体叶形遗传多样性分析[J].北方园艺,2015,39(11):1.[doi:10.11937/bfyy.201511001]
　ZUO Li-hui,ZHANG Jun,DONG Yan,et al.Genetic Diversity of Malus sieversii Natural Population from Xinjiang Analyzed by Leaf Traits [J].Northern Horticulture,2015,39(15):1.[doi:10.11937/bfyy.201511001]
[5]刘航空,王安柱,赵彩平,等.高通量提取桃树叶片组织基因组DNA的研究[J].北方园艺,2015,39(13):120.[doi:10.11937/bfyy.201513034]
　LIU Hangkong,WANG Anzhu,ZHAO Caiping,et al.Study on Extracting DNA From Peach Leaves Efficiently[J].Northern Horticulture,2015,39(15):120.[doi:10.11937/bfyy.201513034]
[6]成晓静,周楚奇,周丽英,等.三种黑籽南瓜RAPD分析[J].北方园艺,2015,39(17):75.[doi:10.11937/bfyy.201517019]
　CHENG Xiaojing,ZHOU Chuqi,ZHOU Liying,et al.RAPD Analysis on Three Cucurbita ficifotia[J].Northern Horticulture,2015,39(15):75.[doi:10.11937/bfyy.201517019]
[7]杨 亚 亚,麻 冬 梅,许 　 兴.改良CTAB 法提取冷季型草坪草基因组DNA 的研[J].北方园艺,2009,33(05):0.[doi:10.11937/bfyy.200905012]
　YANG Ya - ya,MA Dong - mei,XU Xing,et al.Studies on Genomic DNA Extraction of Cool - season T urf Grass with Improved CT AB Method[J].Northern Horticulture,2009,33(15):0.[doi:10.11937/bfyy.200905012]
[8]袁云香,王志平.猕猴桃基因组DNA提取方法的研究进展[J].北方园艺,2011,35(08):0.[doi:10.11937/bfyy.201108070]
　YUAN Yurrxiang,WANG Zhi-ping.The Progress of Extract Method of Kiwifruit Genomic DNA[J].Northern Horticulture,2011,35(15):0.[doi:10.11937/bfyy.201108070]
[9]于立洋,左力辉,徐卫华,等.套袋对五个新疆野苹果优系果实品质的影响[J].北方园艺,2017,41(21):42.[doi:10.11937/bfyy.20171387]
　YU Liyang,ZUO Lihui,XU Weihua,et al.Effect of Bagging on Fruit Quality of Five Malus sieversii Clones[J].Northern Horticulture,2017,41(15):42.[doi:10.11937/bfyy.20171387]
[10]王怡玢,陈锡龙,张东,等.新疆野苹果与栽培苹果挥发性香气物质分析[J].北方园艺,2018,42(03):46.[doi:10.11937/bfyy.20172800]
　WANG Yibin,CHEN Xilong,ZHANG Dong,et al.Comparison Study of Volatile Components in Malus sieversii and Malus pumila[J].Northern Horticulture,2018,42(15):46.[doi:10.11937/bfyy.20172800]

备注/Memo

第一作者简介：张云秀(1991-)，女，山东章丘人，硕士研究生，研究方向为新疆野苹果种质资源利用与保护。E-mail：13116012750@163.com.责任作者：阎国荣(1957-)，男，甘肃敦煌人，博士，教授，现主要从事果树种质资源与生理生态等研究工作。E-mail：yanguorong@eyou.com.基金项目：新疆维吾尔自治区财政林业科技专项资助项目（400474）。收稿日期：2017-02-16

更新日期/Last Update: 2017-08-03