生物类英语论文.docx
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生物类英语论文.docx
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生物类英语论文
Effectofdifferentcarbonsourceandnitrogensourceonthecoldresistanceofcitruscallusinduction
HuSiyuan
(CollegeofAgronomy,NorthwestA&FUniversity,Yangling,Shaanxi712100,China)
Abstract:
Citruscallusisidealmaterialforphysicalandchemicalmutagenesisandgenetictransformation,thecarbonsourceandnitrogensourceisanecessarycomponentintheinductionandsubcultureofcitruscallus,sotheexplorationofcarbon,nitrogensourcesisofgreatsignificancetotheeffectdegreeofcitruscallusinductionandeffectway.Thisstudychoosecoldresistanceofcitrusastestmaterial,throughculturevitrotissues,analysisoftheeffectsofcarbonsourceandconcentration,nitrogensourceandlevelontheinfluenceofcitruscallusinduction,ToprovidethebasisforthestudyoftheoccurrenceandthemechanismofcoldresistanceofCitrusgeneticimprovement,somaticembryos.
Keywords:
carbonsource;nitrogensource;citrus;callus
Introduction
Citrus(CitrusreticulataBlanco)RutaceaeCitrus,smallevergreentreeorshrub,aboutthreemetershigh,weakbranches,usuallywiththorns.Leavesarealternate,leathery,lanceolatetoovate-lanceolate,5.5-8cmlong,2.9-4cmwide,apexacuminate,basecuneate,entireorwithfinebluntteeth,petioleslender,wingedobvious.Flowerssmall,yellow-white,solitaryorclusteredinleafaxils,sepals5;petals5;stamens18-24,filaments3-5piecesoftenconnate,ovary9-15rooms.Citrusfruitflatspherical,5-7cmindiameter,orange-yelloworreddishyellow,peelloosefleshmeateasilyseparated.
Chenggucitruscultivationhasalonghistorydatingbackmorethan2,000yearsofhistory.ChengguCountyislocatedinSouthernShaanxi,CentralHanzhongBasin,northsubtropicaltotemperatetransitionregion,northofQinlingmountainblockingtheinvasionofcoldnorthwest,southwindsBashansowarmaircannotdrivestraight.InversioneffectQinlingbeltandfoothillsslopingwall,formingauniquemicro-climateregions,fewerwintercoldandsummerheatless.First,here'scitrushightemperaturehotwinddoesnotharmflowering,fruitsettingrate;Secondcoolearlyautumn,temperature,andisconducivetofruitcolor,ripe,citrusfruitsurfacehereisgorgeous,highsolublesolidscontent,moderatelysweetandsour.
Studies[1-4]haveshownthatbychangingthecarbonsourcetoinducesomaticembryogenesis.Asmostofthecitruscallushasnotregenerateembryoidbodyphenomena[5-7],whilethehigh-frequencyinductionofsomaticembryogenesisofcitruscellsforgenetictransformation,invitromutagenesis,invitroscreeningmutants,rapidpropagation,etc.workhasspecialsignificance[8-9].Inthispaper,sucrose,glucose,solublestarch,threekindsofcarbonandnitrogensourcesammoniumnitrateonekindofgrowthandtheabilityofsomaticembryogenesisofcitruscalluswerestudiedatdifferentlevels,inordertoselecttheoptimalcultureconditionshardycitrusstudiesprovideevidence.
1.Purposeandsignificance
1.1Purpose
1.Analysistheinfluenceofcarbontypesandcarbonlevelsonhardyofcitrusinvitroculturedexplantsdedifferentiation;
2.Explorethenitrogensourceforcitruscallusinductioneffectmannerandextentofeffect;
3.Screeningsuitablecarbon,nitrogenconditionsofcitrusdedifferentiationinvitrocallusinduction.
1.2Significance
CitrusCalliisanidealmaterialforthephysicalandchemicalmutagenesisandgenetictransformation,andcarbonandnitrogenisanecessaryingredientincitruscallusinductionandsubculture,thereforeitisimportanttoexploretheextentandtheeffectofthewayoncitruscallusinductionofcarbonsource,nitrogensource.
1.Explorethecarbonandnitrogensourcesforcitruscallusinductioneffect,especiallyontheinductionrateanddifferentiationspeedforin-depthstudyofcarbonandnitrogeninthemechanismoftissueculturecitrusfromthebody,andtoprovidesomedatatosupport;
2.Hanzhongcitrusvarietiesforthestudymaterial,trytofilteroutthebestofitscallusinductionofcarbonrequired,thetypeandcontentofnitrogenfortheproductionofvirus-freecitrusseedlingsthroughcallusidealwaytoproviderelativelybasiccultureconditions.
2.Materialsandmethods
2.1Materials
Seedsofthetestmaterialexplantsofthesamesize,healthyfullChengguhardycitrusvarieties.
2.2Preparation
2.2.1Instrument
HSserieshorizontalflowbench(SuzhouAntaiAirTechnologyLtd.);thedish;verticalcircularchamberpressuresteamsterilizer(ShanghaiMedicalNuclearInstrumentFactory);portableelectricpressuresteamsterilizer(ShandongXinhuaMedicalInstrument)electronicbalance(ShanghaibalanceInstrumentFactory);electricdistilledwater(ShanghaimedicalnuclearInstrumentFactory).
2.2.2Auxiliarymaterials
(1)Inaplannedwayofcleaningtheglassware(Erlenmeyerflasks,petridishes,etc.),andinnaturalconditionsorovendrying;
(2)TakeacertainamountofErlenmeyerflaskundersealmembrane,withcottonropeissecured;
(3)TakeacertainamountoftheErlenmeyerflaskwasaddedtotheflasktoapproximately2/3ofpurewater,sealing;
(4)Putthefilterpaperintoapetridish,wrap;
(5)PutthesealedErlenmeyerflasksandPetridishesintohigh-pressuresteamsterilizer,autoclavedat1.1kg/cm230min;
(6)Thesterilizationdish,flaskcoveredwithnewspaperspare,finishinglaboratory.
2.2.3Thebasicculturemedium
Table2.1 thebasicculturemedium
Motherliquorcategory
Composition
Workingliquidconcentration(mg/L)
Avarietyofpracticalquantity(mg)
Constantvolumestandard(ml)
Alargenumberofelementclass
KNO3
1900
19000
1000
NH4NO3
1650
16500
MgSO4·7H2O
370
3700
KH2PO4
170
1700
CaCl2
332
3319
Traceelementclass
MnSO4·H2O
22.3
1115
500
ZnSO4·7H2O
8.6
430
H3BO3
6.2
310
KI
0.83
41.5
Na2MoO4·2H2O
0.25
12.5
CuSO4·5H2O
0.025
1.25
CoCl2·6H2O
0.025
1.25
Lronclass
Na2EDTA·2H2O
37.25
1862.5
500
FeSO4·7H2O
27.85
1392.5
Organicmaterialclass
glycine
2.0
100
500
Thiaminehydrochloride
0.4
20
Hydrochloricacidpyrazolecompoundelement
10
500
niacin
5
250
inositol
100
5000
2.3Experimental design
2.3.1Variableselection
1.Argument:
(1)Typesofcarbonsource:
sucrose;glucose;solublestarch;
(2)Thecontentofcarbonsource:
fourlevelsofsucrose15g/L;30g/L;45g/L;60g/L;
(3)Thecontentofnitrogensource:
KNO3/NH4NO3=1:
1andKNO3/NH4NO3=1:
2.
2.Thedependentvariable:
(1)Inductionrate:
thenumberofcallusformationslices/thenumberofinoculationslices*100%;
(2)Expansionrate:
thenumberofinvitrotissueexpansionslices/thenumberofinoculationslices*100%;
(3)Expansionspeed:
expansionspeed=(NK-N0)/K*numberofslices.N0representsthenumberofinvitrotissueexpansionslicesatfirsttime,NKrepresentsthenumberofinvitrotissueexpansionslicesatKthdayafterthefirsttime;
(4)Inductionspeed:
inductionspeed=(NK-N0)/K*numberofslices.N0representsthenumberofinvitrotissueinductionslicesatfirsttime,NKrepresentsthenumberofinvitrotissueinductionslicesatKthdayafterthefirsttime.
2.3.2Designscheme
Inthisexperiment,threekindsofcarbonsources,namelysucrose,glucoseandsolublestarch,sucrosedensitygradientset4,glucoseandsolublestarchconcentrationgradientisnotset;nitrogenconcentrationgradientset2werenormal1.65gMTmedium/L(KNO3/NH4NO3=1:
1)anddoublingthevalueof3.30g/L(KNO3/NH4NO3=1:
2),thespecificcontentisshowninTable2.2table2.3:
Table2.2 threekindsofcarbon source materials inculture
Composition
Serialnumber
Sucrose(g/L)
Glucose(g/L)
Starch(g/L)
NH4NO3(g/L)
1
30
0
0
1.65
2
30
0
0
3.30
3
0
30
0
1.65
4
0
30
0
3.30
5
0
0
12
1.65
6
0
0
12
3.30
Table2.3 fourdifferentcarbon source level medium materialcontent
Composition
Serialnumber
Sucrose(g/L)
Glucose(g/L)
Starch(g/L)
NH4NO3(g/L)
1
15
0
0
1.65
2
30
0
0
1.65
3
45
0
0
1.65
4
60
0
0
1.65
2.4 ResearchonTechnologyRoadmap
Accesstostudymaterial,asepticinoculation,invitrocultureobservation,analysisofcarbonandnitrogensourceontheeffectofcitrusinvitrotissueculture
2.4.1 Asepticinoculation
Inthecleanbenchcutcitrusseedcotyledonaboutthesizeof3mm×3mmrandomexplantsinoculatedintoeachculturemedium,thebodynumberofeachmediumwasinoculatedexplantisnotlessthan30.
2.4.2Culture observation
Materialsplacedinatemperature25±2℃,relativehumidityof50-60%underconditionsofdarknesstraining,periodicstatisticalswellingnumberofexplantswithoutafewmore,andsocalculatethecorrespondingsecondaryvariables.
2.4.3Statisticalanalysis
Regularobservationofstatistics,dataobtainedentryExceldatabaseandthenconvertthedatainSPSSprofessionalplatformforsingle-factoranalysisofvarianceandtwofactorfactorialanalysis.
3.results andanalysis
3.1Theeffectofcarbonspeciesoncitruscallusinduction
Inviewofthethreetypesofcarbonintheexperimentalgroupwereprovidedwithtwonitrogenlevels,andthereforecannotexcludetheimpactofnitrogenratioofdifferentcarbonsources,itshouldtaketwo-factorfactorialanalysisandtheresultsareshowninTable3.1andFigure3.1:
Table3.1 Effect ofdifferentcarbonsourceson citrus callus
Two-factorfactorialanalysis
Sumofsquares
Degreesoffreedom
Themeansquare
TheFvalue
Significant
Expansionrate
0.070
2
0.035
9.422
0.008
Inductionrate
0.047
2
0.023
5.626
0.030
Expansionspeed
0.000
2
0.000
2.107
0.184
Inductionspeed
2.04E-005
2
1.02E-005
4.711
0.044
Earlyexpansionspeed
0.003
2
0.002
6.629
0.020
Lateexpansi
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