贵州21份玉米优质种质资源的ssr标记分析

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1、2013I旧五田Ⅱ田E匹日j匿茁a1579bridvarieties，andprovidingmolecularMethodsincubatedat65℃for1h：thenequiv．-basisforbreedinghighyieldingmaizelsolationofmaizegenomicDNIAalentvolumeofphenol：chloroform：hybridcombinations．AbIout0．5-0．8afreshIeaveswasisoamy。~alcohoI(25：24：1)wasaddedtogroundinliquidnitrogenintopo

2、wder．extractfor2Omin．andthemixedso．-MaterialsandMethodstransferredtoa2．0mlcentrifugelutionwascentrifuged12minat8000ExperimentaImaterialstube，followedby700uIofpreheatedr／minfortransferringthesupernatantTwentyonesharesofmaizeCTABlysisbufer(2％CTAB．1O0toanotherfreshtube；theequivalentgermplasmswere

3、collectedfrOmmmOI／LTris-HCI，1．4mol／LNaCq1．20volumeofchloroform：isoamylalcohoIGuizhouInstituteofUplandCropsmmoI／LEDTA，1％13-mercapt0ethanoI，(24：11wasaddedtothesupernatant(Table1)．PVP1％，pH8．0)，welImixedandfor20minofextraction．thencen—Table1TheexperimentalmaizegermplasmsusedinthisstudyNo．NameSourc

4、eNo．NameSource1QB44Suwanderivatives12HuangC(HuangXiaol62x330／O2)xTuxpeno2QB48Suwanderivatives13Qiandanl6cml171xHuangC3QB18178599xsuwan14Qiandan18QB44xHuangC4QB5069046x919515Qiandan918QB181xJ1O65QB516F06(Suwanpopulation)xF1g(Brazilpopulation)16Jindan999QB48xJ1066QB552Dika007(PA212xPA31)17Jindan

5、506$273xQB5067QB576QB48×T3218Jindan516QB576x(B5168cml171Tropicalderivatives19Jindan518QB48xQB5169J106U．S．hybrid324320Jindan519QB552xQB51610$273Suwan+unknown21Jindan818QB506xT3211T32Linesselectedfr0mSuwan3populationtrifugedat12minat8000r／minforPCRamplificationwasconductedinannealing,．andwithadr

6、opof0．5oCfortransferringthesupernatanttoanotheranEppendorf5331PCRsystemac--eachcycle；d，72oC30sforextension；freshtube；KAc(5moVL)equalto1O％cordingtothefollowingscheme：a，95e．repeatofb-dfor9cycles；f，5℃30volumeofsupernatantwasaddedandoC5minforpredenaturation；b，95ocsfordenaturation；g．55℃30sforan—mix

7、ed，followedbyprecooledisopropyl30sfordenaturation；c，60oC30sfornealing；h_72℃30sforextension；i．alcoholequaIto0．8-1multipleofsu-Table2Allelesandpolymorphisminformationcontentof21maizeinbredlinesby20SSRpernatant：theresultantsolutionwasprime