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1、作物学报ACTAAGRONOMICASINICA2010,36(3):517525http://www.chinacrops.org/zwxb/ISSN0496-3490;CODENTSHPA9E-mail:xbzw@chinajournal.net.cnDOI:10.3724/SP.J.1006.2010.00517甘蓝和白菜紫色酸性磷酸酶17基因家族的克隆和比较分析1,21,2,**1,21,21,21,2,*卢坤张凯柴友荣陆俊杏唐章林李加纳12西南大学农学与生物科技学院,重庆400716;重庆市油菜工程技术
2、研究中心,重庆400716摘要:从甘蓝和白菜中分别克隆到2个PAP17,根据序列相似性,将PAP17分为类型I(BoPAP17-1和BrPAP17-1)和类型II(BoPAP17-2和BrPAP17-2)。Southern杂交表明,白菜和甘蓝中均只存在2个PAP17成员,与克隆结果吻合。系统发育和分子进化分析表明,PAP17基因家族受纯化选择,编码低分子量PAP蛋白。荧光定量PCR结果表明,PAP17在所检测的9个组织器官中均有表达,尤以花和蕾中的表达量特别高,种子中也有一定程度的表达,表明其与体内储备态磷的动用有
3、关,尤其在花和蕾的发育阶段。磷饥饿诱导表达结果表明,除BrPAP17-2在幼苗叶片中表达受抑制外,BrPAP17和BoPAP17在幼苗根部和叶片中均受磷饥饿诱导,表达量先降后升,诱导4d或8d后达到峰值;恢复供磷4d后,表达量迅速降低至基础表达水平以下。与幼苗叶片相比,白菜和甘蓝幼苗根部的PAP17磷饥饿诱导表达似乎更为强烈。这些结果表明PAP17在白菜和甘蓝根部可能参与了根外磷的活化与吸收及无机磷由根部向其他组织器官的转运。关键词:白菜;甘蓝;紫色酸性磷酸酶;基因家族;磷饥饿CloningandComparati
4、veAnalysisofPURPLEACIDPHOSPHATASE17GeneFamiliesinBrassicaoleraceaandBrassicarapa1,21,2,**1,21,21,21,2,*LUKun,ZHANGKai,CHAIYou-Rong,LUJun-Xing,TANGZhang-Lin,andLIJia-Na12CollegeofAgronomyandLifeSciences,SouthwestUniversity,Chongqing400716,China;ChongqingRapesee
5、dTechnologyResearchCenter,Chongqing400716,ChinaAbstract:TwoPAP17geneswereclonedfromparentspeciesofBrassicanapus,B.oleracea,andB.rapa,respectively.Accord-ingtosequencesimilarity,PAP17genescouldbedividedintotwotypes,typeI(BoPAP17-1andBrPAP17-1)andtypeII(BoPAP17-
6、2andBrPAP17-2).SouthernhybridizationresultedintwobandsbothinB.oleraceaandB.rapa,thisisaccordancewithformercloningresults.PhytogeneticandmolecularevolutionanalysisindicatedthatPAP17genesinBrassicaspeciesun-derwentpurifyingselection,andtheirdeducedproteinsaretyp
7、icallowmolecularweightPAPproteins.ExpressionpatternsofBoPAP17andBrPAP17geneswereassayedbyfluorescentquantitativePCR.TheresultsrevealedthatPAP17genesexpressedinallninetestedtissuesandorgans,withtheextremelyhighexpressioninflowerandbud,andcertainexpressioninseed
8、satdiffer-entstages,implyingthesePAP17genesmostlikelymobilizephosphorusreservesinplants,particularlyduringflowerandbuddevelopmentstages.Underphosphatestarvationconditions,expressio