【作者】 郝玉立；

【导师】 石大兴；

【作者基本信息】 四川农业大学 ， 森林培育， 2010， 硕士

【摘要】 本研究以红花羊蹄甲(Bauhinia blakeana)当年生幼嫩带芽茎段、叶片和叶柄为试验材料,通过正交实验设计结合方差分析,探讨了红花羊蹄甲离体培养的关键影响因素,存在问题及解决方法,对红花羊蹄甲无菌苗诱导、茎段和叶片、叶柄愈伤组织诱导及分化、壮苗生根、炼苗与移栽等方面进行了研究,以得到最佳培养配方及培养程序,从而为建立红花羊蹄甲的快繁体系、工厂化生产提供理论与技术依据。通过组织培养技术研究得出：①红花羊蹄甲外植体组织培养最佳取材时间为3月,启动率高,污染率相对其它时期较低,茎段、叶片和叶柄的启动率分别为91.6%、81.8%和89.5%,污染率分别为21.6%、19.3%和19.6%。②用0.1%的升汞灭菌,茎段的最佳灭菌时间为8 min,叶片为4 min,叶柄为6 min。③初代培养：茎段培养以MS+6-BA 2.0 mg·L-1+NAA 0.01 mg·L-1+IBA 0.5mg·L-1为最适宜培养基配方,启动率高达91.4%,愈伤组织诱导率81.2%,出芽指数1.8；叶片愈伤组织诱导培养以MS+6-BA 1.0 mg·L-1+NAA 0.1 mg·L-1+2,4-D0.1 mg·L-1为最适宜培养基配方,愈伤组织诱导率为75.6%；叶柄愈伤组织诱导培养以MS+6-BA 0.5 mg·L-1+NAA 0.1 mg·L-1+2,4-D 0.05 mg·L-1为最适宜培养基配方,愈伤组织诱导率为71.8%。④继代培养：适宜于芽继代增殖培养的最优组合是MS+6-BA 1.0 mg·L-1+NAA 0.1 mg·L-1+IBA 0.5 mg·L-1,增殖倍数达3.7,有效苗数2.7；适宜于茎段愈伤组织继代增殖培养的最优组合是MS+6-BA 0.5 mg·L-1+NAA 0.1 mg·L-1+2,4-D 0.5mg·L-1,增殖倍数达4.4；适宜于茎段愈伤组织分化培养的最优组合是MS+6-BA 1.0 mg·L-1+NAA 1.0 mg·L-1,分化率达85.9%。⑤有效苗的培养：以MS培养基为最适宜培养基配方,植株生长健壮,有效苗率为85.9%。⑥生根培养：以MS+IBA 1.0 mg·L-1为最适宜培养基配方,植株生长健壮,生根率最高,达76.8%,平均根数最多,为3.3条,平均根长为5.1 cm。⑦移栽与炼苗：炼苗基质以蛭石、珍珠岩、河沙以1：1：2的比例效果最好,成活率较高,达到73.5%,且植株长势良好。更多还原

【Abstract】 The study was mainly to probe an efficient way through tissue and rapid propagation of Bauhinia blakeana. The stems with shoot, the leaves, petiole were taken as explants. For know the better medium and proceeding of culturing Bauhinia blakeana., we used orthogonal experimental design and analysis of variance to establish the technique system of tissue culture. The affected factors of propagation, existing problems in this technique and the corresponding solutions was also described, which provide the theoretical base and technical measures for industrial producing of plantlet.The study of tissue culture technique and the better medium for various stages are as follows:①March is the best season to draw materials, materials, in which the germination rate are 91.6%,81.8% and 89.5% by stem, laminae and petiole respectively, the contaminated rate only are 21.6%,19.3% and 19.6% by stem, laminae and petiole respectively.②During the trial:8 minutes is good to treat the stem by HgC12 (0.1%),4 minutes is good to treat the laminae, and 6 minutes is good to treat the petiole.③Initial medium of stem:MS+6-BA 2.0 mg.L-1+NAA 0.01 mg.L-1+IBA 0.5 mg.L-1 in which the germination rate is 91.4%, the callus induction rate is 81.2%, and the shoot forming index is 1.8; Initial medium of aminae:MS+6-BA 1.0 mg.L-1+NAA 0.1 mg.L-1+2,4-D 0.1 mg.L-1,in which the callus induction rate is 75.6%; Initial medium of petiole:MS+6-BA 0.5 mg·L-1+NAA 0.1 mg·L-1+2,4-D 0.05 mg·L-1,in which the callus induction rate is 71.8%.④Proliferating of stems cultivation:The optional proliferating buds culture medium stems and shoots was:MS+6-BA 1.0 mg·L-1+NAA 0.1 mg·L-1+IBA 0.5 mg·L-1,in which the multiplication rate is 3.7 and the multiplication of efficiency seedings is 2.7.Highly proliferating callus was obtained following cultivation on MS+6-BA 0.5 mg.L-1+NAA 0.1 mg.L-1+2,4-D 0.5 mg.L-1.The proliferating rates could be up to 4.4.The best culture medium for callus of stems differentiating was MS+6-BA 1.0 mg.L-1+NAA 1.0 mg.L-1.The regeneration rate was 85.9%. ⑤Efficiency seeding culture:MS, in which the rate of efficiency seedings is 85.9%.⑥Rooting culture:MS+IBA 1.0 mg.L-1 in which the rate of rooting is 76.8%, the quantity of roots is 3.3, and the length is 5.1 cm.⑦Forging seeding transplanting:The transplant medium in vermiculite, perlite and riversand by 1:1:2 is the optimal for Bauhinia blakeana., which the rate of survival is 73.5%.更多还原