适合于胞质基因组扩增的红麻成熟叶片DNA提取改良方法.pdf

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1、植物遗传资源学报2013，14(2)：347．351Journa1ofPlantGeneticResources适合于胞质基因组扩增的红麻成熟叶片DNA提取改良方法徐建堂，祁建民，陈涛，陈美霞，方平平一，陶爱芬，。(福建农林大学作物遗传育种与综合利用教育部重点实验室，福州350002；福建农林大学农业部东南黄红麻科学观测站，福州350002；3宁德师范学院，宁德352100)摘要：为了从成熟红麻叶片中提取高质量、高产量的基因组DNA，针对红麻成熟叶片中多糖、多酚含量较高的特性，利用改良CTAB法及改良SDS法分别提取红麻品种福红952成熟叶基因组DN

2、A，并通过琼脂糖凝胶电泳和紫外分光光度计测定进行DNA质量检测。结果表明：改良CTAB法提取的基因组DNA电泳时点样孔干净，条带整齐无拖带，0D6o／OD：。。为1．9左右，产率可达1．84g／g，其质量、产量都高于改良SDS法，所提取的DNA可用于红麻RAPD分子标记、线粒体DNA、叶绿体DNA通用引物PCR扩增。改良CTAB法是提取成熟红麻叶片DNA的有效方法，并且可用于红麻分子标记及胞质基因组学研究。关键词：红麻；DNA提取；CTAB法；线粒体DNA；叶绿体DNATheImprovementMethodofExtractionDNAinMatu

3、reKenafLeavesSuitableforCytoplasmicGenomeAmplifyingXUJian．tang’，QIJian—min'，CHENTao，，CHENMei—xia，FANGPing-ping’．TAOAi-fen’(KeyLaboratoryofMinistryofEducationforGenetics，BreedingandMultipleUtilizationofCrops／FujianAgricultureandForestryUniversity，Fuzhou350002；MinistryofAgricultu

4、reofSoutheastJute&KenafScientifeObservationExperimentalStation，Fuzhou350002；NingdeNormalUniversity，Ningde352100)Abstract：TheresearchaimedtoextractthehighqualityandyieldgenomicDNAfrommaturekenaf(HibiscuscannabinusL．)leaves．Accordingtothecharacterofmaturekenafleavescontainingmore

5、amylaseandpolyphenolthanyoungleaves，thekenafvarietiesFuhong952matureleaveswereextractedbyimprovedCTABandSDSmeth-ods，andtheywereidentifiedbyagarosege1electrophoresisandultravioletspectroph0tometermethods．Theresultsshowedthatthesampleholewascleanwithnopulling，OD260／OD280wasabout1

6、．9，theyieldratewasupto1．84g／g，anditsqualityandyieldwashigherthanthatextractedbyimprovedSDSmethod．GenomicDNAextractedbyimprovedCTABalsocanbeusedinkenafRAPD(RandomAmplifiedPolymorphicDNA)molecularmarkers，mito-chondrialDNA，andchloroplastDNAuniversalprimerPCRamplification．Therefore

7、，theimprovedCTABmethodwasaneffectivemethodforextractingthehighqualityandyieldgenomicDNAfrommaturekenafleaves，anditcouldbeusedforthekenafmolecularmarkersandcytoplasmicgenomicsresearch．Keywords：Kenaf(HibiscuscannabinusL．)；genomicDNAextraction；CTABmethod；mitochondrialDNA；chloropla

8、stDNA收稿日期：2012—04—19修回日期：2012．07．10网络出版日期：2012．12．19UR