墨兰论文墨兰组培体系的优化及根状茎在生物反应器培养的研究.doc

墨兰论文：墨兰组培体系的优化及根状茎在生物反应器培养的研究 【中文摘要】墨兰是一种极具观赏价值的高档兰花,但因其繁殖系数低,规模化生产受到阻碍,‘组织培养是解决这一问题的重要途径。本研究为了建立墨兰组培快繁体系,以墨兰根状茎为外植体,调查了影响墨兰根状茎增殖,芽分化和生根的若干因素；此外,利用气升式生物反应器影响根状茎增殖培养的几种理化参数进行了研究,为墨兰种苗的规模化生产提供理论依据。通过研究影响墨兰根状茎增殖的因素,本研究结果表明当花宝一号(H1)和花宝二号(H2)单独使用时,墨兰根状茎的生长及增殖效果不及MS培养基,但当H1和H2混合使用(H12)时明显好于MS培养基。BA和NAA的混合使用有利于根状茎的增殖生长,当H12培养基中加入BA 2mg·L-1+NAA 0.2 mg·L-1时,根状茎分化数多且鲜重和干重良好；糖种类对根状茎增殖生长影响较大,在H12+BA 2 mg·L-1+NAA 0.2 mg·L-1培养基中加入30 g·L-1的蔗糖或食用白糖时增殖效果最佳,其次是葡萄糖,果糖不利于根状茎增殖培养,因根状茎增殖在蔗糖和食用白糖无明显差异,所以可用白糖代替蔗糖。为了从组培根状茎中诱导绿芽,本文对BA和NAA浓度的影响进行了研究,NAA 0.2 mg·L-1和BA4 mg·L-1+食用白糖30 g·L-1+琼脂7.0 g·L-1的培养基中芽分化数最多,生长状态最好。此外,研究墨兰试管苗适宜生根的培养基,结果发现H12培养基中加入NAA1.0 mg·L-1可促进试管苗生根。利用气升式生物反应器可大量增殖墨兰根状茎,其增殖效果显著好于固态培养基培养和液态培养基振荡培养。墨兰根状茎在完全浸没式反应器中增殖效果好于接触式反应器培养,在3L反应器中接种15g根状茎进行完全浸没培养45 d后,根状茎鲜重可达145.5 g,增殖系数为8.7,显著好于10g和20 g接种量处理；活性炭浓度为0.3 g·L-1时,其根状茎鲜重和增殖系数均显著优于其它处理,分别为151.2 g和9.08;0.10vvm是促进墨兰根状茎增殖的最适宜通气量,过高或过低都不利于根状茎的增殖生长；光照强度在1600 Lux时最适合于墨兰根状茎增殖生长；在3 L反应器(工作体积2 L)内进行墨兰根状茎增殖培养的研究,测定培养基中的酸碱度(pH)和电导率(EC)的结果,发现pH和EC值随培养时间增加先呈下降的趋势,分别在40 d和45 d时出现最低值,以后则出现上升的趋势；培养基中总糖浓度在培养45 d时基本耗尽,根状茎的鲜重、干重和增殖系数在45d时均达到最高,因此反应器根状茎增殖培养周期以45 d为佳。 【英文摘要】Cymbidium Sinense Willd. (C. sinense Willd.) is one of the advanced floweres which have great ornamental value. However, large scale production is impossible due to low propagation rate. Tissue culture is the important way for solving the problem of low propagation. In this study, in order to establish the system of tissue culture and rapid propagation for C.sinense Willd., rhizomes. were used as explants to investigate the factors affecting proliferation of rhizome, proliferation of shoots and rooting in vitro; In addition, the effects of several physical and chemical parameters on rhizome proliferation and growth during air-lift bioreactor culture were examined, which provide the theoretical basis reference for large scale production of C.sinense Willd. The several factors affecting proliferation of rhizome were investigated, The results showed that it was not suitable wh