作物学报 ›› 2016, Vol. 42 ›› Issue (11): 1629-1637.
石磊,苗利娟,齐飞艳,张忠信,高伟,孙子淇,黄冰艳,董文召,汤丰收,张新友*
SHI Lei, MIAO Li-Juan, QI Fei-Yan, ZHANG Zhong-Xin, GAO Wei, SUN Zi-Qi, HUANG Bing-Yan, DONG Wen-Zhao, TANG Feng-Shou, and ZHANG Xin-You*
摘要:
Δ9-硬脂酰-ACP脱氢酶(SAD)是决定植物体内饱和脂肪酸与不饱和脂肪酸比值的关键酶。以花生品种豫花9326基因组DNA为模板,通过基因组步移技术,克隆到花生Δ9-硬脂酰-ACP脱氢酶基因(AhSAD)起始密码子ATG上游720 bp片段,利用5? RACE方法获得了该基因的5? UTR序列,通过序列比对确定720 bp片段为AhSAD启动子区域。PLACE在线启动子预测分析表明,该序列具有真核生物启动子必需的核心元件TATA-box和CAAT-box,含有多个与光诱导和激素响应相关顺式序列元件。将AhSAD启动子片段替换pBI121质粒中的CaMV35S启动子驱动下游GUS基因表达,构建植物表达载体pBI-PAhSAD。通过农杆菌介导法转化拟南芥和在花生不同组织中瞬时表达,利用GUS组织化学染色研究其表达特性。表明,在拟南芥和花生受体中,AhSAD启动子主要调控下游基因在根、茎、叶片和子叶中表达,在花生的果针中也检测到GUS活性;拟南芥的茎生叶只有叶脉中具有GUS活性,而花生整个叶片中都具有GUS活性。
| [1]SchwingshacklL,HoffmannG.Monounsaturatedfattyacidsandriskofcardiovasculardisease:synopsisoftheevidenceavailablefromsystematicreviewsandmeta-analyses.Nutrients,2012,4:1989–2007 [2]O’ByrneDJ,KnauftDA,ShiremanRB.Lowfat–monounsaturatedrichdietscontaininghigh-oleicpeanutsimproveserumlipoproteinprofiles.Lipids,1997,32:687–695 [3]CarrilloC,CaviaMdM,RoelofsH,WantenG,Alonso-TorreSR.Activationofhumanneutrophilsbyoleicacidinvolvestheproductionofreactiveoxygenspeciesandariseincytosoliccalciumconcentration:acomparisonwithN-6polyunsaturatedfattyacids.CellPhysiolBiochem,2011,28:329–338 [4]CarrilloC,CaviaMM,Alonso-TorreS.Roleofoleicacidinimmunesystem;mechanismofaction;areview.NutrHosp,2012,27:978–90 [5]DerbyshireEJ.Areviewofthenutritionalcomposition,organolepticcharacteristicsandbiologicaleffectsofthehigholeicpeanut.IntJFoodSciNutr,2014,65:1–10 [6]O’KeefeSF,WileyVA,KnauftDA.Comparisonofoxidativestabilityofhigh-andnormal-oleicpeanutoils.JAOCS,1993,70:489–492 [7]PatteeHE,IsleibTG,MooreKM,GorbetDW,GiesbrechtFG.Effectofhigh-oleictraitandpastestoragevariablesonsensoryattributestabilityofroastedpeanuts.JAgricFoodChem,2003,50:7366–7370 [8]BoltonGE,SandersTH.Effectofroastingoilcompositiononthestabilityofroastedhigh-oleicpeanuts.JAOCS,2002,79:129–132 [9]LindqvistY,HuangW,SchneiderG,ShanklinJ.CrystalstructureofΔ9stearoyl-acylcarrierproteindesaturasefromcastorseedanditsrelationshiptootherdi-ironproteins.EMBOJ,1996,15:4081–4092 [10]KachrooA,ShanklinJ,WhittleE,LapchykL,HildebrandD,KachrooP.TheArabidopsisstearoyl-acylcarrierprotein-desaturasefamilyandthecontributionofleafisoformstooleicacidsynthesis.PlantMolBiol,2007,63:257–271 [11]JungS,TatePL,HornR,KochertG,MooreK,AbbottAG.Thephylogeneticrelationshipofpossibleprogenitorsofthecultivatedpeanut.JHered,2003,94:334–340 [12]RuddlePII,WhettenR,CardinalA,UpchurchRG,MirandaL.EffectofanovelmutationinaΔ9-stearoyl-ACP-desaturaseonsoybeanseedoilcomposition.TheorApplGenet,2013,126:241–249 [13]SlocombeSP,PiffanelliP,FairbairnD,BowraS,HatzopoulosP,TsiantisM,MurphyDJ.Temporalandtissue-specificregulationofaBrassicanapusstearoyl-acylcarrierproteindesaturasegene.PlantPhysiol,1994,104:1167–1176 [14]LuoT,PengS,DengW,MaD,XuY,XiaoM,ChenF.Characterizationofanewstearoyl-acylcarrierproteindesaturasegenefromJatrophacurcas.BiotechnolLett,2006,28:657–662 [15]GuoZF,LiFZ,MaXG,LinF,MaH,ChenLJ,ZhongM,BaiLP,YiY.Molecularcloningoftwonovelstearoyl-acyldesaturasegenesfromwinternesswheat.GenesGenom,2011,33:583–589 [16]RameshAM,KesariV,RanganL.Characterizationofastearoyl-acylcarrierproteindesaturasegenefrompotentialbiofuelplant,PongamiapinnataL.Gene,2014,542:113–121 [17]LuoT,DengWY,ZengJ,ZhangFL.Cloningandcharacterizationofastearoyl–acylcarrierproteindesaturasegenefromCinnamomumlongepaniculatum.PlantMolBiolRep,2009,27:13–19 [18]WangH,CaoF,ZhangW,WangG,YuW.Cloningandexpressionofstearoyl-ACPdesaturaseandtwooleatedesaturasesgenesfromGinkgobilobaL.PlantMolBiolRep,2012,31:633–648 [19]GaoL,SunR,LiangY,ZhangM,ZhengY,LiD.CloningandfunctionalexpressionofacDNAencodingstearoyl-ACPΔ9-desaturasefromtheendospermofcoconut(CocosnuciferaL.).Gene,2014,549:70–76 [20]RajwadeAV,KadooNY,BorikarSP,HarsulkarAM,GhorpadePB,GuptaVS.DifferentialtranscriptionalactivityofSAD,FAD2andFAD3desaturasegenesindevelopingseedsoflinseedcontributestovarietalvariationinα-linolenicacidcontent.Phytochemistry,2014,98:41–53 [21]ThambugalaD,DuguidS,LoewenE,RowlandG,BookerH,YouFM,CloutierS.Geneticvariationofsixdesaturasegenesinflaxandtheirimpactonfattyacidcomposition.TheorApplGenet,2013,126:2627–2641 [22]ThambugalaD,CloutierS.Fattyacidcompositionanddesaturasegeneexpressioninflax(LinumusitatissimumL.).JApplGenet,2014,55:423–432 [23]ZhangY,MaximovaSN,GuiltinanMJ.Characterizationofastearoyl-acylcarrierproteindesaturasegenefamilyfromchocolatetree,TheobromacacaoL.FrontPlantSci,2015,6:239.doi:10.3389/fpls.2015.00239.eCollection2015 [24]ChiX,YangQ,PanL,ChenM,HeY,YangZ,YuS.Isolationandcharacterizationoffattyaciddesaturasegenesfrompeanut(ArachishypogaeaL.).PlantCellRep,2011,30:1393–1404 [25]KlinkenbergJ,FaistH,SaupeS,LambertzS,KrischkeM,StinglN,FeketeA,MuellerMJ,FeussnerI,HedrichR,DeekenR.Twofattyaciddesaturases,STEAROYL-ACYLCARRIERPROTEINΔ9-DESATURASE6andFATTYACIDDESATURASE3,areinvolvedindroughtandhypoxiastresssignalinginArabidopsiscrowngalls.PlantPhysiol,2014,164:570–583 [26]SlocombeSP,CumminsI,JarvisRP,MurphyDJ.Nucleotidesequenceandtemporalregulationofaseed-specificBrassicanapuscDNAencodingastearoyl-acylcarrierprotein(ACP)desaturase.PlantMolBiol,1992,20:151–155 [27]ZhaoN,ZhangY,LiQ,LiR,XiaX,QinX,GuoH.Identificationandexpressionofastearoyl-ACPdesaturasegeneresponsibleforoleicacidaccumulationinXanthocerassorbifoliaseeds.PlantPhysiolBiochem,2015,87:9–16 [28]LiuQ,SinghSP,GreenAG.High-stearicandhigh-oleiccottonseedoilsproducedbyhairpinRNA-mediatedpost-transcriptionalgenesilencing.PlantPhysiol,2002,129:1732–43 [29]KachrooA,LapchykL,FukushigeH,HildebrandD,KlessigD,KachrooP.Plastidialfattyacidsignalingmodulatessalicylicacid-andjasmonicacid-mediateddefensepathwaysintheArabidopsisssi2mutant.PlantCell,2003,15:2952–2965 [30]CraigW,LenziP,ScottiN,DePalmaM,SaggeseP,CarboneV,McGrathCurranN,MageeAM,MedgyesyP,KavanaghTA,DixPJ,GrilloS,CardiT.Transplastomictobaccoplantsexpressingafattyaciddesaturasegeneexhibitalteredfattyacidprofilesandimprovedcoldtolerance.TransgenicRes,2008,17:769–782 [31]单雷,唐桂英,徐平丽,赵学彬,柳展基.花生质体型酰基载体蛋白基因5′侧翼调控序列的克隆与分析.作物学报,2014,40:381–389 ShanL,TangGY,XuPL,ZhaoXB,LiuZJ.Cloningandanalysisof5′flankingregionsofArachisishypogaeaL.genesencodingplastidialacylcarrierprotein.ActaAgronSin,2014,40:381–389(inChinesewithEnglishabstract) [32]BrandY,HovavR.Identificationofsuitableinternalcontrolgenesforquantitativereal-timePCRexpressionanalysesinpeanut(Arachishypogaea).PeanutSci,2010,37:12–19 [33]ChuY,HolbrookCC,Ozias-AkinsP.TwoallelesofahFAD2Bcontrolthehigholeicacidtraitincultivatedpeanut.CropSci,2009,49:2029–2036 [34]KodamaH,HoriguchiG,NishiuchiT,NishimuraM,IbaK.Fattyaciddesaturationduringchillingacclimationisoneofthefactorsinvolvedinconferringlow-temperaturetolerancetoyoungtobaccoleaves.PlantPhysiol,1995,107:1177–1185 [35]GengL,DuanX,LiangC,ShuC,SongF,ZhangJ.Miningtissue-specificcontigsfrompeanut(ArachishypogaeaL.)forpromotercloningbydeeptranscriptomesequencing.PlantCellPhysiol,2014,55:1793–1801 [36]LiC,WuK,FuG,LiY,ZhongY,LinX,ZhouY,TianL,HuangS.Regulationofoleosinexpressionindevelopingpeanut(ArachishypogaeaL.)embryosthroughnucleosomelossandhistonemodifications.JExpBot,2009,60:4371–4382 [37]尹梦回,董静,李先碧,侯磊,罗明,李德谋,裴炎,肖月华.烟草绒毡层特异启动子pTA29在棉花中的表达特性.作物学报,2008,34:2092–2098 YinMH,DongJ,LiXB,HuoL,LuoM,LiDM,PeiY,XiaoYH.Expressioncharacteristicsoftobaccotapetum-specificpromoterpTA29incotton.ActaAgronSin,2008,34:2092–2098(inChinesewithEnglishabstract) [38]BelostotskyDA,MeagherRB.Apollen-,ovule-,andearlyembryo-specificpoly(A)bindingproteinfromArabidopsiscomplementsessentialfunctionsinyeast.PlantCell,1996,8:1261–1275 [39]LuoK,DengW,XuS,PeiY.FunctionalanalysisoftheArabidopsisthalianapoly(A)bindingproteinPAB5genepromoterinNicotianatabacum.PlantCellRep,2008,27:1811–1819 |
| [1] | 郑玉珍, 齐飞艳, 孙子淇, 刘华, 秦利, 石磊, 王娟, 汪蒙蒙, 韩锁义, 徐静, 苗利娟, 黄冰艳, 董文召, 郑峥, 张新友. 花生籽仁总超长链脂肪酸和7种脂肪酸组分的QTL定位[J]. 作物学报, 2026, 52(6): 1646-1657. |
| [2] | 陆一楚, 李振影, 买春海, 赵晓蕊, 王利祥. 夜间复合体关键基因AhLUX1正向调控花生结瘤的功能研究[J]. 作物学报, 2026, 52(6): 1658-1668. |
| [3] | 谷春苗, 王润风, 黄璐, 刘浩, 鲁清, 李海芬, 李少雄, 何双呈, 洪彦彬, 陈小平, 谭斌, 余倩霞. 花生WOX基因家族的全基因组分析及不定芽再生候选基因的鉴定[J]. 作物学报, 2026, 52(5): 1326-1340. |
| [4] | 王懿涵, 李富昌, 刘意, 朱国鹏. 甘薯IbOPR2基因启动子克隆及调控因子的筛选[J]. 作物学报, 2026, 52(4): 1268-1276. |
| [5] | 于天一, 王春晓, 肖丽, 钟召迪, 王宣仓, 赵勇, 路亚, 吴月, 吴正锋. 不同结瘤特性花生品种氮素累积、产量及品质特性对氮肥用量的响应[J]. 作物学报, 2026, 52(3): 881-894. |
| [6] | 张胜忠, 李国卫, 戈立江, 王菲菲, 胡晓辉, 苗华荣, 李燕, 钟文, 陈静. 花生机械脱壳损伤相关农艺指标筛选与QTL定位[J]. 作物学报, 2026, 52(2): 644-652. |
| [7] | 王菲菲, 张胜忠, 杨贵华, 苗华荣, 胡晓辉, 张则林, 刘莎莎, 乔利仙, 单世华, 陈静. 331份花生种质苗期耐盐性综合评价和强耐盐种质鉴选[J]. 作物学报, 2026, 52(1): 279-294. |
| [8] | 金欣欣, 宋亚辉, 苏俏, 杨永庆, 王瑾. 高产高油高油酸花生品种的生长发育及干物质生产特征[J]. 作物学报, 2026, 52(1): 191-201. |
| [9] | 迟晓元, 刘庆, 张君, 赵旭红, 李美, 于天一, 潘丽娟, 许静, 姜骁, 殷祥贞, 马俊卿, 陈娜. 不同花生品种(系)耐盐碱性田间鉴定及各性状指标相关性研究[J]. 作物学报, 2026, 52(1): 85-98. |
| [10] | 孙辰硕, 张月, 田泽锴, 晏立英, 康彦平, 陈玉宁, 王欣, 淮东欣, 王前前, 姜慧芳, 罗怀勇, 黄莉, 廖伯寿, 王志慧, 雷永. 花生种质果柄强度的遗传分化与主要影响因子分析[J]. 作物学报, 2026, 52(1): 118-130. |
| [11] | 万书波, 张佳蕾, 高华鑫, 王才斌. 中国花生高产栽培研究进展与展望[J]. 作物学报, 2025, 51(7): 1703-1711. |
| [12] | 沈傲, 刘敏, 倪迪安, 刘炜. 谷子m6A甲基转移酶基因SiMTA1的启动子序列特征和基因表达模式分析[J]. 作物学报, 2025, 51(7): 1969-1978. |
| [13] | 郭腾达, 崔梦杰, 陈琳杰, 韩锁义, 郭敬坤, 吴晨迪, 付留洋, 黄冰艳, 董文召, 张新友. 花生磷脂酰肌醇转运蛋白基因AhSFH的克隆及其响应黄曲霉菌侵染的表达特征分析[J]. 作物学报, 2025, 51(6): 1489-1500. |
| [14] | 黄梦欣, 庄灵玲, 程佩佩, 李秦, 徐建堂, 陶爱芬, 方平平, 祁建民, 张立武. 黄麻U6启动子克隆及其转录活性分析[J]. 作物学报, 2025, 51(5): 1156-1165. |
| [15] | 李文佳, 廖泳俊, 黄璐, 鲁清, 李少雄, 陈小平, 金晶炜, 王润风. 花生开花时间的全基因组关联分析及候选基因筛选[J]. 作物学报, 2025, 51(5): 1400-1408. |
|
||