学科分类
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37 个结果
  • 简介:Precisebaseeditingishighlydesiredinplantfunctionalgenomicresearchandcropmolecularbreeding.Inthisstudy,weconstructedarice-codonoptimizedadeninebaseeditor(ABE)-nCas9toolthatinducedtargetedA·TtoG·Cpointmutationofakeysinglenucleotidepolymorphismsiteinanimportantagriculturalgene.Combinedwiththemodifiedsingle-guideRNAvariant,ourplantABEtoolcanefficientlyachieveadeninebaseeditinginthericegenome.

  • 标签: PRECISE optimized ADENINE AGRICULTURAL GENE
  • 简介:Homeoboxtranscriptionfactorsparticipateinthegrowthanddevelopmentofplantsbyregulatingcelldifferentiation,morphogenesisandenvironmentalsignalresponse.Torevealthefunctionsofthesetranscriptionfactorsinrice,weconstructedtheRNAivectorsofOsHox9,amemberofhomeoboxfamily,andanalyzedthefunctionofOsHox9usingreversegenetics.TheplantheightandtilleringnumberofRNAitransgenicplantsdecreasedcomparedwiththoseofwild-typeplants.Reversetranscription-polymerasechainreactionanalysisshowedthatOsHox9expressionreducedinthetransgenicplantswithphenotypicvariance,whereasthatinthetransgenicplantswithoutphenotypicvariancewassimilartothatinthewild-typeplants.ThisresultsuggeststhatthephenotypesofthetransgenicplantswerecausedbyRNAieffects.Thetissue-specificityofOsHox9expressionindicatedthatitwasexpressedindifferentorgans,withhighexpressioninstemapicalmeristemandyoungpanicles.SubcellularlocationofOsHox9demonstratedthatitwaslocalizedonthecellmembrane.

  • 标签: 转录因子 同源盒 遗传分析 家族 水稻 转基因植物
  • 简介:EliminationoftheCRISPR/Cas9constructsineditedplantsisaprerequisiteforassessinggeneticstability,conductingphenotypiccharacterization,andapplyingforcommercializationoftheplants.However,removaloftheCRISPR/Cas9transgenesbygeneticsegregationandbybackcrossislaboriousandtimeconsuming.WepreviouslyreportedthedevelopmentofthetransgenekillerCRISPR(TKC)technologythatusesapairofsuicidegenestotriggerself-eliminationofthetransgeneswithoutcompromisinggeneeditingefficiency.TheTKCtechnologyenablesisolationoftransgene-freeCRISPR-editedplantswithinasinglegeneration,greatlyacceleratingcropimprovements.Here,wepresentedtwonewTKCvectorsthatshowgreatefficiencyinbotheditingthetargetgeneandinundergoingself-eliminationofthetransgenes.ThenewvectorsreplacedtheCaMV35SpromoterusedinourpreviousTKCvectorwithtworicepromoterstodriveoneofthesuicidegenes,providingadvantagesoverourpreviousTKCvectorundercertainconditions.Thevectorsreportedhereofferedmoreoptionsandflexibilitytoconductgeneeditingexperimentsinrice.

  • 标签: genome EDITING SUICIDE gene TRANSGENE KILLER
  • 简介:有低glutelin内容的瑞斯作为为肾失败影响的病人的功能的食物合适。在米饭的低glutelin内容基因Lgc1有在二高度类似的glutelin基因GluB4和GluB5之间的3.5-kb删除,它在染色体2的短手臂上定位。在低glutelin内容米饭改进选择效率繁殖,指定为InDel-Lgc1-1和InDel-Lgc1-2的二个分子的标记被开发检测低glutelin内容基因Lgc1。双PCR察觉显示二个标记的联合使用能容易把Lgc1的遗传型与不同米饭变化区分开来。作为一种简单、便宜的技术,因此,分子的标记能广泛地被用来与Lgc1基因识别不同变化并且在低glutelin内容米饭的帮助标记的选择适用。

  • 标签: 蛋白基因 水稻育种 分子标记 蛋白含量 ORYZA PCR检测
  • 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.

  • 标签: RICE CREATION MARKER-FREE
  • 简介:Rice(Oryzasativa)issensitivetosalinity,butthesalttoleranceleveldiffersamongcultivars,whichmightresultfromnaturalvariationsinthegenesthatareresponsibleforsalttolerance.High-affinitypotassiumtransporter(HKTs)hasbeenproventobeinvolvedinsalttoleranceinplants.Therefore,wescreenedfornaturalnucleotidepolymorphisminthecodingsequenceofOsHKT1,whichencodestheHKTproteinineightVietnamesericecultivarsdifferinginsalttolerancelevel.Intotal,sevennucleotidesubstitutionsincodingsequenceofOsHKT1werefound,includingtwonon-synonymousandfivesynonymoussubstitutions.Furtheranalysisrevealedthatthesetwonon-synonymousnucleotidesubstitutions(G50TandT1209A)causedchangesinaminoacids(Gly17ValandAsp403Glu)atsignalpeptideandtheloopofthesixthtransmembranedomain,respectively.Toassessthepotentialeffectofthesesubstitutionsontheproteinfunction,the3DstructureofHKTproteinvariantswasmodelledbyusingPHYRE2webserver.Theresultsshowedthatnodifferencewasobservedwhencomparedthosepredicted3DstructureofHKTproteinvariantswitheachother.Inaddition,thecodonbiasofsynonymoussubstitutionscannotclearlyshowcorrelationwithsalttolerancelevel.Itmightbeinterestingtofurtherinvestigatethefunctionalrolesofdetectednon-synonymoussubstitutionsasitmightcorrelatetosalttoleranceinrice.

  • 标签: 水稻科学 农业
  • 简介:ThecompleteopenreadingframeofOsPIN1awasamplifiedthroughreversetranscriptase-polymerasechainreaction(RT-PCR)basedonthesequencedepositedinGenBanktoexploretherelationshipbetweentheauxineffluxproteinOsPIN1aandthenegativephototropismofriceroots.SequencingresultsshowedthattheGCcontentofOsPIN1awas65.49%.ThefusionexpressionvectorpCAMBIA-1301-OsPIN1a::GFPcontainingtheOsPIN1ageneandacodinggreenfluorescentprotein(gfp)genewasconstructed.ThefusionvectorwastransferredintoonionepidermalcellsbyAgrobacteriumtumefacienstransformation.ThetransientexpressionofOsPIN1a-GFPwasmainlylocatedinthenucleusandcellmembrane.Moreover,thetransgenicplantswereobtainedbyAgrobacterium-mediatedgenetictransformation.MoleculardetectionperformedbyusingPCRandβ-glucuronidasestainingshowedthatthetargetconstructwasintegratedintothegenomeofrice.Thenegativephototropiccurvaturesofthetransgenicricerootswerehigherthanthoseofthewildtype.Similarly,theexpressionlevelsofOsPIN1ainthetransgenicplantswereconsiderablyhigherthanthoseinthewild-typeplants.TheseresultssuggestthatOsPIN1aiscrucialinthenegativephototropiccurvatureofriceroots.

  • 标签: RICE OsPIN1a green FLUORESCENT protein TRANSIENT
  • 简介:Membersoftheactivityofbc1complex(ABC1)familyareproteinkinasesthatarewidelyfoundinprokaryotesandeukaryotes.PreviousstudiesshowedthatseveralplantABC1genesparticipatedintheabioticstressresponse.Here,wepresentthesystematicidentificationofriceandArabidopsisABC1genesandtheexpressionanalysisofriceABC1genes.Atotalof15and17ABC1genesfromthericeandArabidopsisgenomes,respectively,wereidentifiedusingabioinformaticsapproach.Phylogeneticanalyseso...

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  • 简介:米饭镉(Cd)敏感变异的cadB-1用Agrobacteriumtumefaciens被获得调停的系统。在cadB-1和野类型(WT)的暴露以后米饭幼苗到为有增加外部Cd集中的cadB-1和WT的10d,在根积累到高水平的Cd,茎和叶子的Cd集中的一个范围,并且在cadB-1的幼苗生长的抑制比在WT更严肃。氢过氧化物累积在cadB-1的叶子和根是更高的。减少的谷胱甘肽(GSH)的比率/oxidized谷胱甘肽(GSSG),ascorbate(ASC)/dehydroascorbate(DHA)和减少的菸碱腺嘌dinucleotide磷酸盐(NADPH)/oxidized菸碱腺嘌dinucleotide磷酸盐(NADP+)在高Cd层次下面在叶子和根两个都比在WT在cadB-1是更低的。ascorbateperoxidase(APX)的活动,谷胱甘肽peroxidase(GR),dehydroascorbatereductase(DHAR)和monodehydroascorbatereductase(MDHAR)在Cd的高水平的处理下面在叶子和根两个都比在WT在cadB-1是也更低的。我们的结果建议在Cd应力下面,ASC-GSH周期更严重比在WT在cadB-1被禁止,显示变异的cadB-1不太能清除反应的氧种类并且对Cd敏感。

  • 标签: 脱氢抗坏血酸还原酶 还原型谷胱甘肽 镉(CD) 水稻幼苗 突变体 还原型烟酰胺腺嘌呤二核苷酸
  • 简介:一个实验被进行用荧光灯的微分显示器(软式磁碟机)方法在干旱应激和正常条件下面在米饭叶子和根比较信使rna表达式差别。一积极碎片被H.A的联合孤立黄页(contained0.1%H.A。黄)分离和宏数组屏蔽方法。比作ArabidopsisthalianaNADPH氧化还原酶基因,它有96%身份。cDNA是1423bp,并且包含了与345氨基酸残余编码蛋白质的1048bp的一个完全的开的读物框架。而且,基因表示水平在正常条件下面比那在干旱应激下面是更高的。在干旱反应下面的NADPH氧化还原酶基因的可能的角色也被讨论。

  • 标签: 水稻 干旱压力 还原型辅酶II类氧化还原酶基因 克隆化 基因表达
  • 简介:到在米饭germplasm91-1A2的米饭胆量小蚊的抵抗被识别并且遗传上分析了。米饭人口的F1s从作为一个男父母与米饭材料Jinggui,TN1,W1263(Gm1),IET2911(Gm2),BG404-1(gm3),OB677(Gm4),ARC5984(Gm5)和Duokang1(Gm6)交叉的91-1A2被导出。到米饭胆量小蚊的所有父母线和F1,BC1F1和F2人口的抵抗被识别。结果证明91-1A2和所有F1s对中国米饭胆量小蚊遗传因子型IV抵抗。到在BC1F1和F2的易受影响的抵抗植物的分离比率被X2测试与1:3和9:7规则给予,建议到中国米饭胆量小蚊遗传因子型IV的91-1A2的抵抗被是新抵抗基因的二主导的基因控制,对已知的米饭胆量小蚊抵抗基因非突变而产生之遗传因子。

  • 标签: 水稻种质 遗传分析 稻瘿蚊 抗性鉴定 显性基因控制 F2群体
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