[1]尹光晶,宋廷宇,吴春燕,等.豇豆ISSR-PCR反应体系的建立[J].北方园艺,2012,36(12):141-143.
　YINGuang-jing,SONGTing-yu,WUChun-yan,et al.EstablishmentofInterSimpleSequenceRepeatPCRReactionSysteminAsparagusBean[J].Northern Horticulture,2012,36(12):141-143.
点击复制

豇豆ISSR-PCR反应体系的建立

《北方园艺》[ISSN:1001-0009/CN:23-1247/S] 卷: 第36卷 期数: 2012年12期 页码: 141-143 栏目: 出版日期: 2012-12-04
Title:
EstablishmentofInterSimpleSequenceRepeatPCRReactionSysteminAsparagusBean
文章编号:
1001-0009(2012)12-0141-03
作者:
尹光晶1; 2; 宋廷宇2; 吴春燕2; 李静2; 宋述尧2
1.松原职业技术学院，吉林松原138005;
2.吉林农业大学，吉林长春130118
Author(s):
YINGuang-jing 1; 2; SONGTing-yu2; WUChun-yan2; LIJing2; SONGShu-yao2
1.SongyuanTechnicalCollege,Songyuan,Jilin138005；
2.JilinAgriculturalUniversity，Changchun,Jilin130118
关键词:
豇豆; ISSR标记; 反应体系
Keywords:
asparagusbean; ISSR; reactionsystem
分类号:
S643.403.6
文献标志码:
A
摘要:
以豇豆为试材，用新型植物基因组DNA提取试剂盒提取豇豆DNA，选取Mg2+、dNTPs、TaqDNA聚合酶、引物及模板DNA为主因素进行正交优化试验，从3个不同循环次数中筛选出最佳效果循环次数。结果表明：优化的反应体系为25μL的反应体系中Mg2+1.0mmol/L、dNTP0.2μmol/L、TaqDNA聚合酶1.5U、引物0.3μmol/L、模板DNA45ng，循环次数为40次。
Abstract:
GenomicDNAofasparagusbeanwasextractedbythemethordofDNAsecurePlantkit.FivefactorssuchasconcentrationofMg2+,dNTPs,TaqDNA,primer,andtemplateDNAwhichwereprimaryfactorfortheorthogonaldesignwasselected,whilethebestcyclesofPCRreactionwasalsoscreenedfromthreedifferenttrements.TheresultsshowedthattheoptimizationforISSR-PCRreactionsystemonasparagusbeanwasasfollows：25μLreactionsystemcontaining1.0mmol/LMg2+，0.2μmol/LdNTP，1.5UTaqDNApolymerase，0.3μmol/Lprimerand45ngDNAtemplet,thewholePCRreactionranfor40cycles.

参考文献/References:

[1]朱德蔚,王德槟,李锡香.中国作物及其野生近缘植物(蔬菜作物卷下)[M].北京：中国农业出版社,2008.
[2]ZietkiewiczE,RafalskiA,LabudaD.Genomefingerprintingbysimplesequencerepeat（SSR）-anchoredpolymerasechainreactionamplification[J].Genomics,1994,20：176-183.
[3]BlairmW,PanaudO,MccouchSR.Inter-simplesequencerepeat（ISSR）amplificationforanalysisofmicrosatellitemotiffrequencyandfingerprintinginrice（OryzasativaL.）[J].Theor.ApplGenet，1999，98：780-792.
[4]KojimaT,NagaokaT,NodaK.GeneticlinkagemapofISSRandRAPDmarkersinEinkornwheatinrelationtothatofRFLPmarkers[J].Theor.ApplGenet,1998,96:37-45.
[5]RatnaparkhemB，SanTradK，TulluA，etal.Inheritanceofinter-simple-sequece-repeatpolymorphismsandlinkagewithafusariumwiltresistancegeneinchickpea[J].Theor.Appl.Genet,1998,96:348-353.
[6]JoshisP，GuPtavS，AggarwalrK，etal.Geneticdiversityandphylogeneticrelationshipasrevealedbyintersimplesequencerepeat（ISSR）polymorphisminthegenusOryza[J].Theor.Appl.Genet，2000，100：1311-1320.
[7]王建波.ISSR分子标记及其在植物遗传学研究中的应用[J].遗传,2002,24(5):613-616.
[8]DoyleJJ,DoyleJL.ArapidDNAisolationprocedureforsmallquantitiesoffreshleaftissue[J].PhytochenBul,1987,19:11-15.

相似文献/References:

[1]饶贵珍,鲁红学,王香萍,等.豇豆无公害安全高效生产技术及效益分析[J].北方园艺,2014,38(14):230.
[2]贾利元,郭秀英,崔保伟.黄淮海地区豇豆品种筛选试验[J].北方园艺,2013,37(17):44.
　JIA Li-yuan,GUO Xiu-ying,CUI Bao-wei.Screening Test on Cowpea Breeds in Huanghuaihai Region[J].Northern Horticulture,2013,37(12):44.
[3]常文静,张学超,杨明花.伊犁河谷豇豆品种比较试验[J].北方园艺,2013,37(17):46.
　CHANG Wen-jing,ZHANG Xue-chao,YANG Ming-hua.Comparative Experiment of Cowpea Varieties in Yili Valley[J].Northern Horticulture,2013,37(12):46.
[4]康利平,张禄.水分胁迫对豇豆幼苗光合特性的影响[J].北方园艺,2012,36(08):17.
　KANG Li-ping,ZHANG Lu.Effect of Water Stress on Photosynthetic Characteristics of Cowpea Seedlings[J].Northern Horticulture,2012,36(12):17.
[5]周娟,邹礼平.壳聚糖对盐胁迫下豇豆幼苗生长发育的影响[J].北方园艺,2014,38(23):30.
　ZHOU Juan,ZOU Li-ping.Effect of Chitosan on Seedling Growth of Cowpea Under Salt Stress[J].Northern Horticulture,2014,38(12):30.
[6]张金梦,江洪,吴丹娜,等.三种有机蔬菜结果期光合及荧光特性研究[J].北方园艺,2015,39(12):10.[doi:10.11937/bfyy.201512003]
　ZHANG Jinmeng,JIANG Hong,WU Danna,et al.Study on Photosynthetic and Chlorophyll Fluorescence Characteristics of Three Kinds of Organic Vegetables[J].Northern Horticulture,2015,39(12):10.[doi:10.11937/bfyy.201512003]
[7]颜秀娟,何鑫,王学梅,等.宁夏羊角椒雄性不育基因ISSR的分子标记[J].北方园艺,2015,39(17):82.[doi:10.11937/bfyy.201517021]
　YAN Xiujuan,HE Xin,WANG Xuemei,et al.ISSR Marker of Male Sterility in Pepper of Ningxia[J].Northern Horticulture,2015,39(12):82.[doi:10.11937/bfyy.201517021]
[8]范林林,高丽朴,王清,等.预冷方式对豇豆采后生理特性的影响[J].北方园艺,2015,39(17):115.[doi:10.11937/bfyy.201517030]
　FAN Linlin,GAO Lipu,WANG Qing,et al.Research on the Pre-cooling Treatment on Physiological Characteristics of Postharvest Cowpea[J].Northern Horticulture,2015,39(12):115.[doi:10.11937/bfyy.201517030]
[9]范林林,王清,高丽朴,等.二氧化氯对豇豆采后生理特性的影响[J].北方园艺,2015,39(24):122.[doi:10.11937/bfyy.201524034]
　FAN Linlin,WANG Qing,GAO Lipu,et al.Effect of ClO2 Treatment on Quality of Postharvest Cowpea During Cold Storage[J].Northern Horticulture,2015,39(12):122.[doi:10.11937/bfyy.201524034]
[10]代鹏,曹岩坡,戴素英,等.早熟豇豆新品种“长青101”的选育[J].北方园艺,2016,40(02):156.[doi:10.11937/bfyy.201602042]
　DAI Peng,CAO Yanpo,DAI Suying,et al.A New Early Maturing Cowpea Variety ‘Changqing 101’[J].Northern Horticulture,2016,40(12):156.[doi:10.11937/bfyy.201602042]

备注/Memo

第一作者简介：尹光晶（1987-），女，吉林松原人，现主要从事果树生理研究工作。E-mail：yinguangying@yahoo.com.cn。
责任作者：宋廷宇（1977-），男，吉林德惠人，博士，现主要从事蔬菜种质资源创新与利用方面的研究工作。E-mail：tysong422@163.com。
收稿日期：2012-04-17

更新日期/Last Update: 2014-09-25