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  • 简介:Thehighgrowth-stimulatingeffectofplantextracthasurgedtheplantbiotechnologiststousenaturalsupplementsintheculturemediainsteadofsyntheticphytohormones.Weadvocatedtheeffectofsproutedsorghumextract(SSE)onemergence,invitroacclimatization,andgeneticfidelityincoleoptilederivedcallusofindicaricevarietyADT36.TheuseofSSEwithMurashigeSkoogmediumefficientlyacclimatizedtherootandshootapicalsystems.Ahighermatandseminalroots(3.4gbiomass)withanefficientshootprimordiumelongationwereobservedwithanincreaseintheconcentrationofSSE.SeedstreatedwithSSEmediumshowedhighergerminationandearliercoleoptilematurationabout48hcomparedtountreatedseeds,andtherewasahigherexpressionofeEF-1αwithanincreaseincoleoptilelength.B5mediumwaseffectiveoninducingembryogenicandnodularcallusfrom3-day-oldcoleoptilewith3.0mg/L2,4-dichlorophenoxyaceticacidandfurtherproliferatedeffectivelywith0.8mg/Lkinetinwithafreshweightof180mg.Highlysignificantregenerationwasobservedwithcombinationof2.5mg/L6-benzylaminopurineand3.0mg/Lα-naphthaleneaceticacid.Themetabolicandgeneticprofilesofinvitroanddirectlycultivatedplantswerethesame,examinedthroughFourier-transforminfraredspectroscopy,randomamplifiedpolymorphicDNA(RAPD),inter-simplesequencerepeat(ISSR)andR-ISSR(combinationofRAPDandISSR)markers,respectively,andthusconfirmingthesignificantefficacyoftheSSEincorporatedmedium.DisarmedT-DNAwastransformedtocoleoptilederivedcallusthroughAgrobacteriumtumefaciensLBA4404andconfirmedbyGUSassay.TheT-DNAintegrationwasconfirmedbyDNAblotanalysisusingDNAfromtransientGUS-expressedexplants.Thus,SSEcanbeusedasanaturalandorganicsupplementfororganogenesisandefficientacclimatizationsofshootandrootapicalmeristemsinregeneratedplants.

  • 标签: CALLUS induction COLEOPTILE Oryza sativa random