简介:Precisebaseeditingishighlydesiredinplantfunctionalgenomicresearchandcropmolecularbreeding.Inthisstudy,weconstructedarice-codonoptimizedadeninebaseeditor(ABE)-nCas9toolthatinducedtargetedA·TtoG·Cpointmutationofakeysinglenucleotidepolymorphismsiteinanimportantagriculturalgene.Combinedwiththemodifiedsingle-guideRNAvariant,ourplantABEtoolcanefficientlyachieveadeninebaseeditinginthericegenome.
简介:EliminationoftheCRISPR/Cas9constructsineditedplantsisaprerequisiteforassessinggeneticstability,conductingphenotypiccharacterization,andapplyingforcommercializationoftheplants.However,removaloftheCRISPR/Cas9transgenesbygeneticsegregationandbybackcrossislaboriousandtimeconsuming.WepreviouslyreportedthedevelopmentofthetransgenekillerCRISPR(TKC)technologythatusesapairofsuicidegenestotriggerself-eliminationofthetransgeneswithoutcompromisinggeneeditingefficiency.TheTKCtechnologyenablesisolationoftransgene-freeCRISPR-editedplantswithinasinglegeneration,greatlyacceleratingcropimprovements.Here,wepresentedtwonewTKCvectorsthatshowgreatefficiencyinbotheditingthetargetgeneandinundergoingself-eliminationofthetransgenes.ThenewvectorsreplacedtheCaMV35SpromoterusedinourpreviousTKCvectorwithtworicepromoterstodriveoneofthesuicidegenes,providingadvantagesoverourpreviousTKCvectorundercertainconditions.Thevectorsreportedhereofferedmoreoptionsandflexibilitytoconductgeneeditingexperimentsinrice.
简介:RiceisanimportantfoodcropinChina,andthedevelopmentofhybridriceisacrucialwaytoincreasegrainyield.Thecreationofdual-purposenuclear-sterilelinesfortwo-linehybridbreedinghasbecomevitalforcommercialricebreeding.WeconstructedthepC1300-2x35S::Cas9-sgRNAPTGMS2-1expressionvectorforeditingthemalefertilitygenePTGMS2-1intwowidelycompatiblericevarieties,93-11andHuazhan,byusingtheCRISPR/Cas9system.Weobtainedthemarker-freephotoperiod-/thermo-sensitivegenicmale-sterile(P/TGMS)linesinT1generation.Accordingtotheexperimentsinphytotronwithfourtemperatureandphotoperiodtreatments,wefoundthetemperatureisthemainfactorforrestoringthepollenfertilityofptgms2-1mutantsin93-11andHuazhan,andthephotoperiodalsohassomeeffectsonpollenfertilityintwodifferentricebackgrounds.Theapplicationofcultivatingnewmale-sterilelinesbygenomeeditingsystemwillsignificantlyacceleratethericebreedingprocess.
简介:CRISPR/Cas9(Clusteredregulatoryinterspacedshortpalindromicrepeat/Cas9)基因编辑技术在生命科学领域掀起了一场重大的技术革命,它比锌指核酸酶(ZFNs)和转录激活因子效应物核酸酶(TALENs)技术更易于操作,而且更高效。CRISPR/Cas9系统已经被广泛应用到植物科学研究领域中。本实验选取拟南芥MS2为目的基因,构建植物基因编辑系统的表达载体,并通过农杆菌介导的方法转化拟南芥,从而利用CRISPR/Cas9系统靶向敲除拟南芥MS2基因。对转基因后代的MS2测序结果分析表明,在获得的12个阳性转化植株中,发现了5个阳性植株存在基因序列上的新碱基插入,由此形成CRISPR/Cas9介导的拟南芥MS2突变体。这一工作为后续分析其他物种中MS2同源基因的功能研究提供参考依据。
简介:本刊讯:首批6台于“3·15”日启用、能独立快速完成12类检测是否是注水肉、假酒、含碘盐,甚至禽类有无禽流感病毒的流动监测车,近日亮相广州。广州市工商局局长陈斯达在广州市政府召开的新闻发布会上表示,3月15日,广州市将举行食品安全监测车交接启用仪式,首批6台监测车将在活动当日向公众亮相。据悉,今后,市工商部门将按照日常质量检查与快速检测相结合的原则,用流动监测车对全市流通领域的食品进行检测,对检测不合格的食品,将依法进行处理。据了解,该车的检验仪器标准配备有7套检验仪器和7种速测工具,检验仪器分别是:禽流感检测仪;酶标仪,可检测猪肉、瘦肉精及兽药残毒;农药残毒速测仪,可检测各类蔬菜瓜果的农药残毒;二氧化硫速测仪,可对腐竹、粉丝、白糖、黄花菜等商品的食用漂白剂进行检测;亚硝酸盐速测仪,可对腊味、火腿肉、各类肉类罐头食品的亚硝酸盐进行检测;甲醛速测仪,可对冰鲜鱼等水产品的甲醛含量进行检测;全自动气相色谱仪,可迅速检测出食品中的甲醇、甲醛和二氧化硫等30多种常见的禁止使用的化学成分。7种速测工具分别是注水肉快速测速盒、碘盐含碘量速测液等,能在2分钟内对食品安全检...
简介:Leafshapesarenotonlytheusefulindicatorsinplanttaxonomy,butalsotheimportantfactorsaffectingenergyandmaterialexchangeinleaves.Inthispaper,wecollectedandscannedtheleavesofNitrariatangutoruminDengkouofInnerMongoliaAutonomousRegion(themeanannualprecipitation145mm)andMinqinofGansuProvince(themeanannualprecipitation115mm)andN.sphaerocarpainDunhuang,andthenanalyzedleafshapeparameterswithImage-ProPlus6.0imageprocessingsoftwareandleafδ13CvaluesintheisotopelaboratoryoftheChineseAcademyofForestry.Theresultshowedthat:1)asleafareaincreasedwithincreasingwateravailabilitytheincreasesintheleaflengthandwidthwereasynchronously;2)withthesameleafwidth,the1eavesofN.tangutorumandN.sphaerocarpaweresignificantlylongerinhighwateravailableconditions;and3)althoughthereweresignificantlydifferencesinwateravailabilitybetweenDengkouandMinqin,aswellasbetweenthebottomandmiddleofthealluvialfanneartheEastLakeinDunhuang,theleafδ13CvaluesofN.tangutorumorN.sphaerocarpaweresimilarindifferentwaterconditions(P>0.05).Ourresultssuggestedthattheratioofleafperimetertoareawouldbeanimportantfactorwhichlinkedleafshapetoplantwaterphysiology.Duringgrowingprocedureofleafarea,leaflengthincreasewaspriortoitswidthtoalleviatethereductioninratioofperimetertoareaandmaintainwateruseefficiencyoftheplant.
简介:为解决竹子种子长期保存问题,以马来龙竹、云南龙竹、巨龙竹、金平龙竹、麻竹、长节勒竹、云南箭竹、棉花竹、爪哇巨竹、大泰竹、笻竹、方竹和毛竹13种竹子的成熟种子为材料,通过研究不同低温、低温下不同储存时间、不同含水量、不同发芽设施和不同基质对种子发芽率的影响,来判断竹子种子保存的适合条件。结果表明:1)除笻竹和方竹的种子外,大多数竹种的种子在低温储存5个月内,随着温度由10℃逐渐降低到-16℃,种子发芽率会有不同程度的增高;2)在-16℃条件下,随着储存时间的延长,种子发芽率会有所下降;3)种子含水量会在很大程度上影响种子的发芽率,含水量超过在一定临界值,随着含水量的增加,种子发芽率会显著下降;4)选择不同的种子发芽设施也会影响种子发芽率,生产中宜选用具有微喷灌条件的温室进行育苗;5)在无菌红土、黑土和蛭石这3种基质中,蛭石是竹子播种育苗的适宜基质。