毛脉酸模无菌苗的制备及下胚轴愈伤组织诱导的研究.docVIP

毛脉酸模无菌苗的制备及下胚轴愈伤组织诱导的研究 ? 98? 中医药信息 InformationonTraditionalChineseMedicine 2012年第29卷第2期 Vo1.29,No.2,2012 毛脉酸模无菌苗的制备及下胚轴愈伤 组织诱导的研究 谢彦梅,许岩,常丰华,沈剑,王振月 (黑龙江中医药大学,黑龙江哈尔滨150040) 摘要:目的:建立制备毛脉酸模无菌苗及下胚轴愈伤组织诱导方法.方法:通过摩擦种皮,种子浸泡时 间,消毒次数,消毒剂种类,消毒时间以及种子贮存时间等因素研究对种子发芽的影响,探讨建立无茵苗 培养体系的最佳方法;以获得的无菌苗的下胚轴为外植体,通过在Ms基本培养基中附加6一BA和2, 4一D两种激素不同配比筛选诱导愈伤组织的最优培养基配方.结果:制备毛脉酸模无菌苗的方法为: 挑选粒大饱满的新种子,120目砂纸摩擦种皮,蒸馏水浸泡24h,75%酒精中浸2min,无菌水冲洗3次, 10%NaC10溶液处理10min,无菌水冲洗5次,将其放置在有两层湿润的无菌滤纸的培养皿中10h,再进 行第2次消毒,10%NaC10溶液处理8min,无菌水冲洗5次,在无激素的MS培养基中培养即可获得大 量无菌苗;下胚轴愈伤组织诱导的最优培养基为MS+3.0rag/L6一BA+1.7mg/L2,4一D.结论:建立 了毛脉酸模无菌苗培养体系,得到下胚轴愈伤组织诱导的最优培养基,为将来进一步研究奠定基础. 关键词:毛脉酸模;无菌苗;下胚轴;愈伤组织 中图分类号:R28文献标识码:A文章编号:1002—2406(2012)02—0098—03 AsepticSeedlingPreparationandInductionofCallusfrom HypocotylforRumexGmelini XIEYan—mei.XUYan,CHANGFeng—hua,SHENJian,WANGZhen—yue (HeilongfiangUniversityofChineseMedicine,Harbin150040,China) Abstract:0bjective:ToobtainthemethodsofasepticseedlingpreparationforRumexgmeliniandgetthecal— lusfromhypocotylforRumexgmelini.Methods:Frictionseedcoat,soakingtime,sterilizationtimes,disin— fectanttype,sterilizationtimeandstoragedurationwerestudiedtofindoutthebeststerilizationmethodfora— septicseedling.WiththehypocotylsoftheasepticseedlingsfromRumexgmeliniseedsasexplants,theopti— mummediumwasscreenedbyaddingdifferenthormone(6一BA,2,4一D)proportionsintoMSbasicmedi— um.Results:Thebestgermicidaltreatmentwasasfollows:TheseedcoatsofgoodandnewseedsfromRumex gmeliniwererubbedwith120eyesandpaper.Thetreatedseedswereduckedindistilledwaterfor24h,then weresoakedin75%alcoholfor2rain.washedbysterilewaterfor3times,thensoakedin10%sodiumhypo— chloritesolutionfor10min.washedbysterilewaterfor5times.Thetreatedseedswereplacedinasterileen— vironmentfor10h.thens0akedin10%sodiumhypochloritesolutionfor8min,washedbysterilewaterfor5 times.FinallytheseedswereculturedatMSbasicmediumtogettheasepticseedling.Theoptimummedium wasMS+3.0mg/L6一BA+1.7mL2,4一D.Conclusion:Thestudyhaslaidafoundationforthefurther researchonthattissueforactiveingredient