含有母性影响遗传基因的连锁定位系的构建

HEREDITAS(Beijing)200711,29(11):1393―1398ISSN0253-9772研究报告收稿日期:2007−05−22;修回日期:2007−08−18基金项目:(973)(2005CB121000);(30671591);(20060635016);(CSTC2006BB1051)(2006052)[SupportedbytheNationalBasicResearchProgramofChina(No.2005CB121000),theNationalNaturalScienceFoundationofChina(No.30671591),ScienceFoundationofMinistryofEducationofChina(No.20060635016),theNationalNaturalScienceFoundationofChongqing(No.CSTC2006BB1051),TechnologicalInnovativeFoundationProjectforpostgraduatesofSouthwestUniversity(No.2006052)]作者简介:(1969−),,,,,,E-mail:[e-mail],Tel:[phone]通讯作者:(1957−),,,E-mail:[e-mail]:10.1360/yc-007-1393代方银,谭端,童晓玲,胡海,鲁成,向仲怀,400716:母性影响遗传基因由于其杂交后代的表型受母本基因型的影响,而不能直接反映当代个体的基因型,这给连锁定位测交亲本(三隐性或双隐性系统)的培育带来困难,从而影响这类基因的定位研究。设计了一套杂交培育方案,其核心是使母性影响遗传基因先纯合,再使非母性影响遗传基因纯合。采用所设计的方案,成功培育了家蚕第13连锁群的赤蚁(ch)、无鳞毛翅(nlw,新突变)和褐色卵t(b-t,母性影响遗传)的三隐性系统,并培育了第19连锁群的狭胸(nb)和第二肾形卵(ki-2,母性影响遗传,待定位突变基因)的双隐性系统。:母性影响遗传基因;三隐性系统;双隐性系统;培育方案;家蚕Constructionoflinkagelocalizationlinescontaininggeneswithma-ternaleffectsDAIFang-Yin,TANDuan,TONGXiao-Ling,HUHai,LUCheng,XIANGZhong-HuaiTheKeySericulturalLaboratoryofAgriculturalMinistry,SouthwestUniversity,Chongqing400716,ChinaAbstract:Sincethephenotypesofhybridprogeniesinvolvinggeneswithmaternaleffectsareaffectedbythegeno-typeoffemaleparent,theycannotreflectthegenotypesofindividuals.Thismakesitdifficulttodeveloptestcrossparents(tripleordoublerecessivelines)forlinkagelocalizationand,consequently,hinderstheprogressinlocaliza-tionresearchesforthesetypesofgenes.Inthisstudy,wedesignedasetofhybridizationschemes,thekeyofwhichwastomakethe“maternal-effect”geneshomozygousatfirst,andthenthenon-maternal-effectgenes.Usingthisscheme,wesuccessfullyproducedatriplerecessivelineforgenesch(chocolate),nlw(non-lepiswing)andb-t(ma-ternalbrowneggofTsujitan)onthe13thlinkagegroupandadoublerecessivelineforgenesnb(narrowbreast)andki-2(kidney-shapedegg2)onthe19thlinkagegroupofBombyxmori.Keywords:geneswithmaternaleffects;triplerecessiveline;doublerecessiveline;hybridizationscheme;Bombyxmori[1],,1900[2],,[3],,(),1394HEREDITAS(Beijing)200729,,,32F1,(BC1)F1,,,,,,,[2,4],,,,,,(),,[5],[4],F1,F2,,11.11.1.113(1)(ch:13-9.6),;(2)(nlw:13-),,,;(3)t(b-t),,[6]1.1.219(1)(nb:19-),,,,;(2)2(ki-2:19-),,,,1.21.2.1chnlwb-t/chnlwb-t(1),(ch)(nlw),,F2,F2,chnlw/chnlwF2nlwch,(ch),ch+/ch+(,)ch+/chnlw(,)+nlw/+nlwF2,,,nlw,ch,,chnlw/chnlw(2)chb-t(1)chb-tF2,b-t,,b-t,F2chb-t/+b-t,F2+b-t/+b-t,,F2+b-t/+b-tch+/+b-t,F2(4:+b-t/ch++b-t/+b-t++/ch+++/+b-tchb-t/+b-t),b-t(b-t+/b-t+),b-t,+b-t/+b-tchb-t/+b-t,,,,b-t,,ch,chb-t/+b-t,chb-t/chb-t(3)chnlw/chnlwchb-t/chb-t,ch,(2),nlwb-t,chnlwb-t1.2.2(nb)(ki-2)ki-2,,,,,(F3)1.2.1(2)ki-2nb2002200622.1,211:1395F29331,1/16;2,,F2,,,,,F21/16,,,(),,,,F12,22,F213(ch)(nlw),1F2,,,,,,chnlw,,F2(ch+nlw/ch+nlwchnlw/ch+nlw)nlw(+chnlw/+chnlw),nlw,,chnlw/chnlw2:2F2,580,140;140,,123,,nlw(),,118;,,,,102,16nlw,chnlw/+chnlw,ch,:(chnlw/chnlw),2:(1):a.F2(ch),ch+nlw/ch+nlwchnlw/ch+nlw,,,,nlw,,;b.F2(nlw),a(2)图1家蚕2个隐性连锁基因的F2代基因型Fig.1GenotypesofF2generationoftworecessivelinkagegenesinsilkworm1396HEREDITAS(Beijing)200729图2利用对F2代进行测交的方法构建家蚕连锁基因的双隐性纯合系统Fig.2ConstructionofthehomozygosisdoublerecessivelineofsilkwormlinkagegenesbytestcrosswithF2generation2.2,,F2,,,F2,,,,,,(ch)t(b-t),3:F21000,b-t,,978(22),,231b-t;b-t1,2,,230,231,,223,8,,,,,8(b-t),:t(chb-t/chb-t)2.32.12.2,(ch)(nlw)(ch)t(b-t),,(ch),2.2,(nlw)t(b-t),chnlwbt:A(chnlw/chnlw)()B(chb-t/chb-t)()F2800(),B,,783,,184b-t;b-t1,2,,183,184,,,,142,42,10,,,10(b-t),:t(chb-tnlw/chb-tnlw)2.419(nb)(ki-2),(ki-2)11:1397图3含有一个母性影响遗传基因的家蚕双隐性连锁系统的育成方案Fig.3Hybridizationschemeofsilkwormdoublerecessivelinkagelinecontainingagenewithmaternaleffect19[7]ki-2,,,,[4,8],,ki-2nbbtch(3):ki-2nb,F2800,ki-2,761,(ki-2)170;170,,9(nb),(ki-2nb/ki-2nb)3,,,,,,,(),,,,,,,,,,(1%),,1398HEREDITAS(Beijing)200729,450[6,9~13],[4](ki-2)2080,199319[7],ki-2nb(nbki-2/nbki-2),,19(Gl)[10],nbki-2/nbki-2,nbki-2/nbki-2ki-2ki-2,,nb,,,,,nbki-2/++nbki-2/nbki-2♂(),ki-2()(nlw)[15],(TP01),nlw(,2001,)(),,[16,17],nlw13(ch),13(),t(b-t)chnlwb-t/chnlwb-t,(References):[1]Mendel(TranslatedbyLiangHong).TheExperimentofPlantHybridization.In:ClassicPapersinGenetics.Bei-jing:SciencePress,1984.():.:,1984.[2]ZHUJun.Genetics.3rdedition.Beijing:ChinaAgricul-turalPublishingPress,2002.():,2002.[3]Morgan(TranslatedbyLuHui-lin).GeneTheory.Beijing:SciencePress,1959.,:,1959.[4]XIANGZhong-Huai.SilkwormGeneticsandBreeding.Beijing:AgriculturePress,1994.:,1994.[5]ZHANGWen-Qing.InsectGenetics.Beijing:SciencePress,2000.:,2000.[6]XIANGZhong-Huai.SilkwormBiology.Beijing:ChinaForestryPress,2005.:,2005.[7]LUCheng,XIANGZhong-Huai,HUANGYing.Linkagestudiesonthe“kidneyegg2”(ki-2)mutantinBombyxmori.JournalofSouthwestAgriculturalUniversity,1993,15(6):557−559.,,2(ki-2),1993,15(6):557−559.[8]LUCheng,XIANGZhong-Huai.Geneticalstudiesonkidney2mutationsinBombyxmori.ScienceofSericulture,1990,16(4):234−236.,2,l990,l6(4):234−236.[9]TazimaY.TheRecentAdvancesinGene-analysisoftheSlikwom.Tokyo:Gihodo,1951,67−144.:,1951,67−144.[10]FujiiH.GeneticstocksandmutationsofBombyxmori:importantgeneticre