| China has carried out the research on conservation of the Empetrum nigrum L.var.japonicum K.Koch,since 1983.However,there is still no successful literature report on the artificial sexual reproduction.With the in-depth development of the medicinal value of northeastern crowberry.Its resources by wantonly destroyed.However,Northeastern crowberry in natural state seeds to germination and plant growth is extremely slow,as the Empetraceae only species in China,Northeastern crowberry has been in the brink of extinction.This paper mainly through three parts of the northeastern crowberry sexual reproduction and the establishment of northeastern crowberry rapid propagation system,for better protection and development resources to lay a solid foundation.1.Northeastern crowberry sexual reproduction of seed dormancy part:through full factorial experimental design method of pre experiment selected the six factors:water immersion processing(SS),concentrated sulfuric acid treatment processing(HS),different concentration of gibberellin(GA)(c),gibberellin in different temperature and(GA(T)),gibberellin deal with different time(GA(time)layer,product mode(CJway)experimental design,to the germination rate as response value.Through the analysis of selected concentrated sulfuric acid treatment(HS),normal atmospheric temperature stratification(WCJway)as the main influence factor.Combining the results of a full factorial experiment,according to the box Behnken central composite experimental design principles,through the response surface method to deal with the different concentrations of gibberellin(GA(c),gibberellin different temperature treatment(GA(T)of gibberellin deal with different time GA(time)were investigate and determine the best germination scheme:HS+WCJway+GA(c)600 mg/L+GA(T)18℃+GA(time)13h。2.Northeastern crowberry sexual reproduction study colonization:Northeastern crowberry seed germination and the survival rate of planting has been on 2014,7,17-2014,7,23 research group of northeastern crowberry natural habitats of field survey in climate chamber habitat simulation cultivation and determine the final colonization scheme:(1)The use of sand particle size less than 2.5mm and soil 1:3 ratio by mixing the tile to the bottom,the height of 10cm(the three layer first layer matrix).Sand particle size less than 5mm and more than 2.5mm mixed powder tile in the middle lacmus antlers,height 5cm(three layer second layer matrix).(2)Germinated seeds sown in three layered matrix layer 2,garden soil(three layered matrix layer 3)tile in the upper layer,2 cm in height,with 0.2%ferrous sulfate irrigation through the matrix,the top-level garden soil infiltration in the matrix,the surface residual about 1cm.(3)The germ has grown seedlings directly in accordance with the row spacing is 5cm,seedling spacing of 3cm plant three layer matrix in 0.2%ferrous sulfate is poured through the the upper layer.(4)The long width height:60cm×25cm×15cm rectangular pots for planting,planting 100 trees per pot Empetraceae seedlings.Planted a total of 6 basin,which germinated seeds planted three pots,has grown Seed Germ three pots,arranged in a climate chamber culture,daytime 10h simulation,simulation night 14h,box and climatic conditions for:humidity:simulate day 75%,60%of the night simulation;temperature:simulate day 25 DEG C,night 18 DEG C simulation;simulation daytime light intensity:4000Lx.In the process of seedling,keeping the substrate moist,every two weeks with a matrix of ferrous sulfate 0.2%irrigation.Using this method of colonization and field management,seedlings of Northeast High Rock Lanlu white survival was 94%,on average;a germ seedling survival rate was 87%.3.Northeastern crowberry micropropagation system part in the experimental group planting as test materials,discusses the choice of explants the best disinfection solution,explant,basic training and selecting,in northeastern crowberry stems as explants in different growth regulating substances in northeastern crowberry sterile short branches joggling in regeneration pathway(single bud induction,axillary shoot proliferation,rooting induction)and northeastern crowberry organ pathway(callus induction,callus proliferation and differentiation,rooting induction).The experimental results are as follows:(1)the best disinfection solution:75%alcohol after 30s of treatment with 10min 2%sodium hypochlorite solution.(2)the best explant selection:Empetraceae stem explant material is better.The rate of callus and germination rate were higher than the Empetraceae leaves.(3)basic training options:WPM(ML)is the best chose for Empetraceae tissue culture.(4)A pathway induced northeastern crowberry test of single bud regeneration.Empetrum nigrum var.japonicum sterile short cuttings.The results showed that 6-BA,NAA,2,4-D and KT difference on single bud induction rate greatly,which 6-BA and KT on induction rate effect is the most significant,the induction rate of effect sizes for 6-BA>KT>NAA>2,4-D.Single bud induction medium was the best WPM(ML)+2.0mg/L6-BA+2.0mg/L KT.(5)Pathways to induce Northeast High Rock Lancong bud and inducing cluster buds proliferation assay.Empetrum nigrum var.japonicum sterile short cuttings regeneration.The results showed that 6-BA,KT and IAA,the induction rate effect is most remarkable,the induction rate of effect sizes for 6-BA>IAA>kt>NAA.Induction of buds proliferation and the best medium for inducing WPM(ML)+2.0mg/L6-BA+0.03 mg/L IAA+2.0mg/L KT.(6)Empetrum nigrum var.japonicum sterile short cuttings regeneration pathways are induced in Empetrum nigrum var.japonicum rooting experiment.The results showed that IAA in the induction rate effect is most significant,different concentrations of plant growth regulator induction rate factor in the size of the order for IAA>NAA>IBA>KT.The best rooting medium was WPM(ML)+1.5mg/L IAA.(7)Empetrum nigrum var.japonicum organogenesis pathways to induce callus test.The results showed that 6-BA on callus induction effect is the most significant,and four kinds of hormones on Northeastern crowberry callus induction rate effect sizes for 6-BA>2,4-D>KT>IAA.The test results showed that the best medium for callus induction was WPM(ML)+1.0mg/L6-BA+0.9 mg/L2,4-D.(8)Empetrum nigrum var.japonicum organogenesis induced callus proliferation and differentiation test.The results showed that 6-BA and KT on callus induction effect is the most significant,and four kinds of hormones on Northeastern crowberry callus induction rate effect sizes for KT>6-BA>NAA>IBA.Induction of callus proliferation and differentiation of the best medium was WPM(ML)+1.5mg/L6-BA+1.5mg/LKT.(9)Empetrum nigrum var.japonicum organs induced pathways to induce rooting experiment.The results showed that IBA on root induction effect is the most significant,and four kinds of hormone of northeastern crowberry rooting induction rate of effect sizes for IBA>NAA>KT>IAA.The best rooting medium was WPM(ML)+0.9mg/LIBA. |
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