杜仲est-ssr引物开发及遗传多样性研究

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1、AbstractEucommiaulmoidesOliv．istheonlyspeciesofgenus(Eucommia)intheEucommiaceaefamily．Itisatreespeciesofimportanteconomicandmedicinalvalueswiths打ongadaptabilitywidelydistributedin27provinces(areas)ofChina,italsohasstrongresistancetodiseasesandinsectpests．Wild

2、resourcesofE．ulmoidesareonthevergeofextinction．RelatedstudieshaveshownthatE．ulmoideshasabundantvariationtypes，i．evariationinshapes，structuresandeventheembryonictypes．Thereforetheresearchesareurgentlyneededonitsgeneticsfromtheperspectiveofmolecularbiology．Inth

3、isstudy,weusedthetranscriptomesequencingdataofleavesandfi'uitsofE．ulmo，idesfromdifferentgrowingperiodtodevelopprimerssuitablefortheSSRmarkers．Afterthat，systematicanalysiswasconductedonitsgeneticdiversity,geneticrelationship，thedegreeofdifferentiationbetweenan

4、dwithin,geneticstructureofgrouppopulationofexperimentalmaterialsincluded136samplesfrom20groupsofE．ulmoides．Theresultsofthepresentstudycouldprovidetheoreticalbasisforgermplasmresourcescollection，conservationandforgeneticbreedingofE．ulmoides．Themainresultswere-

5、asfollows：1．TheSSRprimersofE．ulmoideswereindependentlydeveloped．SSRlociof2to6repeatedunitsincluding1334stripsdistributedin150kindswereseekedoutthroughbioinformaticsmethodsfrom49610Unigenesequencesoftranscriptomedataextractedfromeucommialeavesandfruits，showing

6、thatfrequencyofSSRlociinthetranscriptomedataWaslowbyonly2．9％．mostrepeatedunitswerefromdinucleotideandtrinucleotide，accountsfor92．72％ofallSSRloci．Atotalof114pairsofPrimerweredesignedtobesynthesizedusingPrimer5softwareaccordingtoUnigenesequencesofranscriptome，i

7、nwhich13primersofbj曲polymorphismandstabilitywereselectedthroughPAGEelectrophoresistest．ThelengthofthetargetfragmentWasbetween120bpto250bp．2．ThegeneticdivisityofE．ulmoidesSSRsitesweredetected．73alleleshavebeendetectedfrom10SSRsites．Numberofalleles(瑚)Wasinthera

8、ngeof2to13，、^，ith7．3allelesIIIdetectedonallaveragenumberofpersite．Effectivenumberofalleles(he)was20．18，anaveragenumberwas2．018．Expectedheterozygosity(He)andobservedheterozygosity(Ho)range