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1、園藝學(xué)報，2017，44(1)：1–10.ActaHorticulturaeSinicadoi：10.16420/j.issn.0513-353x.2016-0489；http：//www.ahs.ac.cn1蘋果MdCBL3基因的克隆與功能鑒定*劉亞靜，馬齊軍，李浩浩，路靜，郝玉金，由春香（山東農(nóng)業(yè)大學(xué)園藝科學(xué)與工程學(xué)院，作物生物學(xué)國家重點實驗室，農(nóng)業(yè)部黃淮地區(qū)園藝作物生物學(xué)與種質(zhì)創(chuàng)制重點實驗室，山東泰安271018）摘要：從‘嘎拉’蘋果（Malus×domesticaBorkh.）中克隆MdCBL3（序列號：MDP0000155124），長852bp。系統(tǒng)進化樹分析表明，
2、蘋果MdCBL3與白梨親緣關(guān)系最近。利用PlantCare數(shù)據(jù)庫進行啟動子順式作用元件預(yù)測分析，MdCBL3啟動子序列中存在干旱、低溫、光、生長素、防御等響應(yīng)元件。構(gòu)建MdCBL3表達載體并利用農(nóng)桿菌介導(dǎo)法侵染蘋果愈傷組織和擬南芥。半定量RT-PCR證明MdCBL3在轉(zhuǎn)基因愈傷組織中過量表達，在擬南芥中得到異源表達。表型分析發(fā)現(xiàn)，在鹽脅迫中，與野生型相比，轉(zhuǎn)基因愈傷組織和擬南芥成齡苗生長更好，鹽脅迫的抗性更強。在擬南芥中異源表達MdCBL3，能提高擬南芥對干旱和低溫的抗性。關(guān)鍵詞：蘋果；MdCBL3；基因克隆；功能鑒定；鹽脅迫中圖分類號：S661.1文獻標(biāo)志碼：A文章編號：0
3、513-353X（2017）01-0001-10MolecularCloningandFunctionalCharacterizationofMdCBL3inApple*LIUYajing，MAQijun，LIHaohao，LUJing，HAOYujin，andYOUChunxiang（StateKeyLaboratoryofCropBiology，MOAKeyLaboratoryofHorticulturalCropBiology（HuanghuaiRegion）andGermplasmInnovation，CollegeofHorticultureScienceandEn
4、gineering，ShandongAgriculturalUniversity，Tai’an，Shandong271018，China）Abstract：AgenenamedMdCBL3（MDP0000155124）wasclonedfrom‘Gala’apple（Malus×domesticaBorkh.）.ThelengthofMdCBL3was852bp.AphylogenetictreeanalysisindicatedthatthePyrusbretschneideriexhibitedthehighestsequencesimilaritytoMdCBL3.Th
5、epromoter’scis-actingelementswereanalyzedandpredictedusingPlantCareDatabases，analysisshowedthattheMdCBL3promotersequenceincludeddrought，lowtemperature，light，auxin，defenseresponsivenessetc.ConstructexpressionvectorofMdCBL3anduseAgrobacteriummediatedmethodtoinfectapplecallusandArabidopsisseed
6、.SemiRT-PCRindicatedthatMdCBL3wasgeneticallytransformedintoapplecallusandArabidopsis.PhenotypicanalysisfoundthattransgeniccallusandArabidopsisseedlinggrowbetterandhavestrongerresistanceunderNaClstresscomparedwithcontrast.DemonstratedthatoverexpressionofMdCBL3，heterogenousexpressionofMdCBL3r
7、emarkablyincreasedthetoleranceoftransgenicapplecallusandArabidopsistohighsalinity.ThestudyfoundthatheterogenousexpressionofMdCBL3inArabidopsisenhencedthetoleranceofdroughtandlowtemperature.Keywords：apple；MdCBL3；molecularclone；functionalidentification；sal
