資源描述:
《利用基因芯片鑒定梨品種自交不親和基因型》由會(huì)員上傳分享,免費(fèi)在線閱讀,更多相關(guān)內(nèi)容在學(xué)術(shù)論文-天天文庫。
1、園藝學(xué)報(bào)2014,41(10):1983–1992http://www.ahs.ac.cnActaHorticulturaeSinicaE-mail:yuanyixuebao@126.com利用基因芯片鑒定梨品種自交不親和基因型1,2,*221江南,譚曉風(fēng),張琳,鄧靖12(湖南工業(yè)大學(xué)包裝與材料工程學(xué)院,湖南株洲412007;中南林業(yè)科技大學(xué)經(jīng)濟(jì)林培育與保護(hù)教育部重點(diǎn)實(shí)驗(yàn)室,經(jīng)濟(jì)林育種與栽培國家林業(yè)局重點(diǎn)實(shí)驗(yàn)室,長沙410004)摘要:根據(jù)東方梨中已鑒定的46個(gè)S基因序列和S基因的結(jié)構(gòu)特點(diǎn),設(shè)計(jì)了86條寡核苷酸探針并制備成S基因寡核苷酸檢測芯片,采
2、用Cy3熒光修飾引物標(biāo)記被檢測品種的PCR產(chǎn)物并與芯片雜交,以檢測不同品種的S基因型。結(jié)果表明:利用芯片與華梨2號(hào)、秀玉和德勝香等已知S基因型的品種雜交,雜交信號(hào)顯示與各品種已知基因型相符合。利用芯片鑒定了麗江黃酸梨等27個(gè)未知S基因型的梨品種,獲得了各品種的S基因型,隨機(jī)選取部分品種進(jìn)行DNA測序和序列分析,結(jié)果與芯片雜交結(jié)果完全一致,證明利用S基因寡核苷酸芯片鑒定梨品種S基因型結(jié)果準(zhǔn)確可靠。關(guān)鍵詞:梨;自交不親和;S基因型;寡核苷酸;芯片中圖分類號(hào):S661.2文獻(xiàn)標(biāo)志碼:A文章編號(hào):0513-353X(2014)10-1983-10Ident
3、ifyingPearCultivarsS-genotypesUsingGeneChips1,2,*221JIANGNan,TANXiao-feng,ZHANGLin,andDENGJing12(SchoolofPackingandMaterialEngineering,HunanUniversityofTechnology,Zhuzhou,Hunan412007,China;KeyLaboratoryofCultivationandProtectionforNon-woodForestofMinistryofEducation,theKeyLabo
4、ratoryofNon-woodForestProductofForestryMinistry,Central-southUniversityofForestryandTechnology,Changsha410004,China)Abstract:Basedonthestructurecharacteristicofself-incompatibilitygeneandthegenegroupsequencesof46SgenesinOrientalpearcultivars,86probesforS-RNasedetectionsweredes
5、ignedandoligonucleotidegenechipwasestablished.InordertodetecttheS-genotypeofpearcultivars,thePCRproductsofS-RNasewereamplifiedbyCy3-labeledprimersandhybridizedwiththemicroarray.ThecultivarswithknownS-genotypesuchasHuali2werehybridizedwiththegenechipand27cultivarswithunknownS-g
6、enotypewereS-genotypedusingthegenechip.TheDNAsequencedresultsofHuali2,XiuyuandDeshengxiangpearsandrandomlyselectedunknownS-genotypecultivarswereconsistentwiththemicroarrayanalyses.Inconclusion,theoligonucleotidegenechipprovidedanaccurateandreliablemethodfordetectingS-genotypeo
7、fpearcultivars.Keywords:pear;self-incompatibility;S-genotype;oligonucleotide;genechip近20年,在梨S自交不親和基因的研究中,利用PCR-RFLP技術(shù)、S基因DNA序列分析、cDNA克隆、雜交授粉試驗(yàn)、花粉管生長檢測及蛋白質(zhì)電泳分析等方法已分離鑒定了53個(gè)梨S等位基因(GenBank已登錄),鑒定了部分梨品種的S基因型。日本學(xué)者建立了鑒別梨S基因的PCR-RFLP收稿日期:2014–06–26;修回日期:2014–08–11基金項(xiàng)目:國家自然科學(xué)基金面上項(xiàng)目(312
8、72124);湖南省自然科學(xué)基金項(xiàng)目(12JJ6017)*E-mail:namijiangnan@126.com1984園