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1、園藝學(xué)報,2016,43(7):1402–1410.ActaHorticulturaeSinica1402doi:10.16420/j.issn.0513-353x.2015-0872;http://www.ahs.ac.cn紅掌查爾酮異構(gòu)酶基因的克隆與表達分析121,31,3111,*楊哲,劉克林,彭佳佳,秦?fù)P,鄭高言,明軍,劉春123(中國農(nóng)業(yè)科學(xué)院蔬菜花卉研究所,北京100081:北京市大東流苗圃,北京102211;寧夏大學(xué)農(nóng)學(xué)院,銀川750021)摘要:以紅掌栽培品種‘Dakota’為材料,采用RA
2、CE技術(shù)克隆獲得AnCHI基因的cDNA序列,分析花青素含量與該基因表達的關(guān)系,進一步完善紅掌花青素合成途徑。結(jié)果表明:AnCHI序列全長為1117bp,其中開放閱讀框為777bp,編碼258個氨基酸;AnCHI在不同時期紅掌佛焰苞以及根、莖、葉、肉穗組織中均有表達,且在佛焰苞發(fā)育前期表達量明顯高于后期,測得花青素含量與基因表達量變化基本一致;構(gòu)建的pGEX-4T1-AnCHI重組表達載體在原核中正確表達。初步驗證了AnCHI的表達可能對紅掌佛焰苞花青素的合成有一定促進作用。關(guān)鍵詞:紅掌;AnCHI;花青素
3、;表達分析中圖分類號:S682文獻標(biāo)志碼:A文章編號:0513-353X(2016)07-1402-09CloningandExpressionAnalysisofChalconeIsomeraseGenefromAnthuriumandraeanum121,31,3111,*YANGZhe,LIUKe-lin,PENGJia-jia,QINYang,ZHENGGao-yan,MINGJun,andLIUChun12(InstituteofVegetablesandFlowers,ChineseAcadem
4、yofAgriculturalSciences,Beijing100081,China;Nurseryof3Dadongliu,Beijing102211,China;CollegeofAgricuture,NingxiaUniversity,Yinchuan750021,China)Abstract:AfulllengthcDNAofchalconeisomerase(CHI)wasamplifiedfromAnthuriumandraeanumL.‘Dakota’byusingRACEtechnolog
5、y.Therelationshipbetweenitsexpressionpatternsandanthocyanincontentweredissectedforthepurposeoffurtherimprovingtheanthocyaninbiosyntheticpathwayofanthurium.TheresultsshowedthattheAnCHIcDNAwas1117bpwitha777bpopenreadingframeinfull-lengthwhichencoding258amino
6、acids.AnCHIwasexpressedindifferentdevelopingperiodsofspathesaswellasinroots,stems,leavesandspadixesandtheexpressionlevelsofAnCHIintheearlystages(1–4)ofspathesweresignificantlyhigherthanthatinmaturestages(5–8).Thecontentofanthocyaninwaschangedconsistentlywi
7、ththegeneexpression,whiletherecombinantexpressionvectors—pGEX-4T1-AnCHIwascorrectlyexpressedinprokaryotes.TheresearchsuggestedthatAnCHImayplayanimportantroleinpromotingtheanthocyaninsynthesisofAnthuriumandraeanum.Keywords:Anthuriumandraeanum;AnCHI;anthocya
8、nin;expressionanalysis收稿日期:2016–02–01;修回日期:2016–07–07基金項目:北京市科技計劃研發(fā)攻關(guān)項目(YLHH201200109);中國農(nóng)業(yè)科學(xué)院與北京市大興區(qū)科技合作項目;農(nóng)業(yè)部園藝作物生物學(xué)與種質(zhì)創(chuàng)制重點實驗室項目*通信作者Authorforcorrespondence(E-mail:liuchun@caas.cn)楊哲,劉克林,彭佳佳,秦?fù)P,鄭高言,明軍,劉春.紅掌