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1、河南農(nóng)業(yè)科學(xué)離子束介導(dǎo)遺傳轉(zhuǎn)化小麥突變體的遺傳分析111231王鐵固,楊天佑,趙新亮,馮偉森,黃群策,陳士林(1.河南科技學(xué)院生命科技學(xué)院,河南新鄉(xiāng)453003;2.洛陽市農(nóng)業(yè)科學(xué)院,河南洛陽471022;3.鄭州大學(xué)河南省離子束生物工程重點(diǎn)實(shí)驗(yàn)室,河南鄭州450052)摘要:以離子束介導(dǎo)后代的9個(gè)小麥突變體和4個(gè)受體作供試材料,對(duì)其進(jìn)行了RAPD和SSR分子標(biāo)記,在此基礎(chǔ)上獲得了矮稈突變株系1042的2個(gè)特異片斷,并進(jìn)行了測(cè)序和同源序列分析。13個(gè)材料間分子標(biāo)記遺傳差異比較結(jié)果表明,離子束介導(dǎo)遺傳轉(zhuǎn)化后的突變明顯小于小麥品種間的遺傳差
2、異。矮稈突變株系1042的SSR標(biāo)記特異片斷序列BLAST分析結(jié)果表明,從1042中擴(kuò)增到的特異片斷與小麥高稈基因RhtD1a中的部分序列高度一致,1042的株高明顯降低可能是由于該株系遺傳突變,產(chǎn)生了2個(gè)矮稈基因所致;該特異片斷與小麥BAC克隆中LTRretrotransposonAngela3s轉(zhuǎn)座子的部分片斷同源性最高,說明離子束介導(dǎo)遺傳轉(zhuǎn)化誘發(fā)變異可能與小麥基因組中反轉(zhuǎn)錄轉(zhuǎn)座子的激活有關(guān)。關(guān)鍵詞:小麥;離子束;遺傳轉(zhuǎn)化;RAPD;SSR;序列分析中圖分類號(hào):S512.1文獻(xiàn)標(biāo)識(shí)碼:A文章編號(hào):10043268(
3、2010)05001105GeneticAnalysisofWheatMutantsbyIonBeammediatedTransformation1112WANGTiegu,YANGTianyou,ZHAOXinliang,FENGWeisen,31HUANGQunce,CHENShilin(1.SchoolofLifeScienceandTechnology,HenanInstituteofScienceandTechnology,Xinxiang,453003,China;2.AgriculturalScience
4、InstituteofLuoyang,Luoyang471022,China;3.HenanProvincialKeyLaboratoryofIonBeamBioengineering,ZhengzhouUniversity,Zhengzhou450052,China)Abstract:Ninewheatmutantsfromgenetictransformationmediatedbyionbeamandtheir4recipientvarietiesweretakenastestmaterialstoconductRAPDand
5、SSRmarkeranalysis,andthen2specialfragmentsfromthedwarfmutant1042weresequenced,homologyanalysisofwhichwasdone.Theresultsshowedthatthevarianceofmutantsfromgenetictransformationbytionbeamwassmallerthanthegeneticvariancebetweendifferentrecipientvarieties.AfterBLASTanalysis,
6、thespecialfragmentsofSSRmarkerfromthedwarfmutant1042werehighlyidenticaltopartialsequenceofhighstrawgeneRhtD1a,suggestingthatthedwarfingmightbecausedbythetwomutationdwarfgenes;whilethespecialfragmentsofRAPDmarkerhadhighhomologywithpartialsequenceofLTRretrotransposonAng
7、ela3sinwheatBACclones,whichsuggestedthatthemechanismofmutagenesisbyionbeamispartiallyattributedtothereactivationoftheretrotransposons.Keywords:Wheat;Ionbeam;Genetictransformation;RAPD;SSR;SequenceanalysisRAPD是以基因組DNA為模板,以一個(gè)隨機(jī)PCR擴(kuò)增反應(yīng)產(chǎn)生不連續(xù)的DNA產(chǎn)物,用以檢測(cè)[1]的寡核苷酸序列(通常為10個(gè)堿基
8、對(duì))作引物,通過DNA序列的多態(tài)性。目前該方法已廣泛用于種收稿日期:20091012基金項(xiàng)目:農(nóng)業(yè)部農(nóng)業(yè)公益性行業(yè)科研專項(xiàng)經(jīng)費(fèi)項(xiàng)目(200803034)作者簡(jiǎn)介:王鐵固(1971),男,河南伊川人,博
