【作者】 王莉；

【导师】 石大兴；

【作者基本信息】 四川农业大学 ， 园林植物与观赏园艺， 2008， 硕士

【摘要】 本试验以“草原”美国薄荷（Monarda didyma‘Praricnacht’）带芽茎段、叶片和叶柄为材料,进行茎段培养和愈伤组织培养同时,利用GC—MS技术分析“草原”美国薄荷实生苗和试管苗的叶片挥发油成份,以期为美国薄荷离体培养建立有效途径,为美国薄荷工厂化试管育苗及种质资源保存提供理论依据。运用正交试验设计方法和Excel、SPSS软件进行数据处理,主要研究结果如下:（1）通过组织培养技术研究得出:①种子发芽及无菌苗的获得的最佳途径为:先将“草原”美国薄荷种子在发芽盒中萌发出苗,再用0.1%HgCl₂灭菌2min,接入MS空白培养基中,效果最好。发芽率为45%,污染率为10%,死亡率为45%。②初代培养:带芽茎段培养以MS+6-BA1.0 mg·L^-1+NAA0.05 mg·L^-1为最适宜培养基配方组合,启动率达98.33%,平均新梢长为4.23cm。叶片、叶柄、和茎段的愈伤组织培养以MS+6-BA3.0 mg·L^-1+NAA0.05 mg·L^-1+2,4-D0.1 mg·L^-1为最适宜培养基配方组合,出愈率为100%。③继代培养:适宜于茎段芽苗增殖培养的最适宜培养基配方组合为:MS+6-BA2.0 mg·L^-1+NAA0.01 mg·L^-1+IBA0.5 mg·L^-1,芽苗增殖倍数达3.9,腋芽萌发早,有效苗多,且健壮。愈伤增殖的最适宜培养基配方组合为:MS+6-BA2.0mg·L^-1+NAA0.1 mg·L^-1+2,4-D 0.1 mg·L^-1,愈伤的增殖率为81.6%。④愈伤组织分化培养:愈伤组织分化的最适宜培养基配方组合为:MS+BA2.0mg·L^-1+TDZ1.0 mg·L^-1+ZT0.25 mg·L^-1+NAA 0.05mg·L^-1分化培养最好,分化率为40%。⑤壮苗与生根培养:以空白培养基MS为最适宜的壮苗培养基配方。生根培养的最适宜培养基配方为:1/2MS+蔗糖30g·L^-1+IBA0.05mg·L^-1,生根率达100%,平均根长达4.16cm,平均根数为4.5条。⑥炼苗与移栽:最适宜的移栽基质为珍珠岩:蛭石:河沙（1:1:2）,成活率为92.5%。（2）通过挥发油成份的气相色谱-质质谱（GC—MS）分析研究得出:以“草原”美国薄荷实生苗与组培苗的叶片为材料,利用GC-MS技术共分离鉴定出28种化学成分。其中实生苗挥发油鉴别出25种化学成份,而组培苗鉴别出有23种。发现两者的叶片挥发油的主要化学成份无明显的差异,其主要的化学成分为:麝香草酚,γ-松油烯,邻-聚伞花素,1-辛烯-3-醇,2-甲氧基-5-甲基-异丙苯,芳樟醇,4-蒈烯,3-崖柏烯,月桂烯。更多还原

【Abstract】 In this experiment,seeds,stems,leaves and petioles of Monarda didyma ’Praricnacht’ were used as explants for the studies on tissue culture of stem segment and callus.At the same time,the essential oil components of the leaves of Monarda didyrna ’Praricnacht’in vitro and seedling were analysised by GC-MS.An effective way on tissue culture of Monarda didyma would be established,which could procide a theoretical basis on industrial producing of plantet and protection.The main results of experiment were as follows:（1）The study of tissue culture technique and the better medium for various stages are as follows:①Put seeds of Monarda didyma’Praricnacht’ into the germination boxes and germinated,and then sterilized 2 rain with 0.1%HgCl₂ and Put them on MS medium,which is the best method to germinate and get no vaccine seedlings. Thegermination rate was 45%,the pollution rate was 10%and mortality was 45%.②Initial cultureMS+6-BA1.0 mg·L^-1+NAA0.05 mg·L^-1was the best ignition medium,in which the germination rate reach 98.33%and the average shoot length reach 4.23 cm.The bese medium for induction of calluses from leaves,petioles and stems was MS+6-BA3.0 mg·L^-1+NAA0.05 mg·L^-1+2,4-D0.1 mg·L^-1.The induction rate reach 100%.③Multiplication cultureMS+6-BA2.0 mg·L^-1+NAA0.01mg·L^-1+IBA0.5 mg·L^-1was best proliferation medium for stems,in which the multiplication rate was 3.9.The most suitable medium for the proliferation of the callus was MS+6-BA 2.0 mg·L^-1+NAA0.1 mg·L^-1+2,4-D 0.1 mg·L^-1,the proliferation rate was 81.6%.④Callus differentiation cultureThe most suitable differentiation medium was MS+BA2.0 mg·L^-1+TDZ1.0 mg·L^-1+zT 0.25 mg·L^-1+NAA 0.05 mg·L^-1,in which differentiation rate was40%.⑤Stanger seedling and rooting cultureBlack medium（MS）was the best stanger seedling medium.1/2MS + sucrose 30g·L^-1+ IBA 0.05mg·L^-1was the best rooting medium,in which the rooting rate reach 100%,the average length was 4.16cm and the average numbers was 4.5.⑥TransplantingThe transplant medium in vermiculite,perlite and riversand by 1:1:2 was optimal,in which the survival rate was 92.5%.（2）The study of Essention oil constituents by GC-MS are as follows:seedlings and tissue culture of Monarda didyma ’Praricnacht’were used as the materials,using GC-MS separate 28 kinds of chemical components.The seedling volatile oil identify 25 kinds of chemical composition,and in vitro have indentified 23 spacies.found that there was no significant differences in the chemical composition of its major between seedlings ang in vitro.The major chemical composition were Thymol,γ-terpinen-o - Poly-parachutes,1 - octene -3 - alcohol,2 - methoxy-5 -methyl - Cumene,linalool,4 - carene,3 - Cliff Parker ene,Myrcene.更多还原