摘要
为了在短期内大量扩繁栀子优良种苗,满足工厂化需要,本文从栀子的组培苗增殖、生根以及移栽等方面进行了研究。同时,为了降低组培苗的生产成本,提出利用生物反应器进行栀子增殖和瓶外生根培养的可行性,并对影响瓶外生根的几种因素进行了研究。
通过研究植物生长调节剂对栀子组培苗增殖和生根的影响,发现:在MS培养基中添加BA 1.0mg/L和NAA 0.1mg/L有利于促进芽的分化和生长:对栀子组培苗,NAA效果好于IBA和IAA,且NAA浓度为0.1mg/L时,生根率可达100%,组培苗长势良好;MS培养基浓度、MS培养基中总氮量及NO_3~-/NH_4~+影响栀子的增殖,MS培养基基本浓度(1.0MS),总氮量为60mM和90mM和NO_3~-/NH_4~+为40mM/20mM时,利于栀子的增殖培养;通过糖浓度对栀子增殖影响的探明,糖浓度MS培养基中加入30g/L蔗糖时,对栀子组培增殖的效果好于本试验的其它糖浓度;改变培养瓶封口的透气性可调节组培微环境条件,本研究结果,培养瓶口密闭时,植株有徒长趋势,通气口直径为0.5cm条件下,增殖率高,植株生长良好,但通气口直径继续增加时,培养基出现干裂现象,增殖数减少。
在栀子增殖阶段,利用生物反应器比较了完全浸没式培养、接触式培养和间歇浸没式培养对增殖效果影响,结果表明,利用生物反应器进行栀子苗扩繁有可行性,在完全浸没培养条件下,增殖苗数可达7.1个,比固体培养高或相当,但叶片卷曲;而间歇浸没式培养虽然增殖率低于完全浸没培养,但卷叶也比完全浸没培养少。接触式培养增殖率最低,卷曲叶也最少。
在生根阶段采用瓶外生根技术是可行的,以泡沫板做为支持物的生根率及根的生长好于珍珠岩;营养液中pH和EC影响栀子生根,发现在pH为5时,不论是地上部,还是地下部根的生长均良好,低于或高于pH 5都不利于栀子的生根培养,尤其是pH为3时,根系短小肥大,不利于幼苗驯化移栽。当营养液中EC为1.0mS·cm~(-1)时,根系生长良好,且形成的侧根多,适合栀子瓶外生根培养。通过对瓶外生根苗(7天和14天后)与组培苗及温室植株叶片气孔的观察,发
This study was carried out to investigate the effect of proliferation, rooting, and acclimatization of Gardenia jasminoides Eills for large-scale culture. In order to reduce cost of plantlets in vitro, the feasibility of bioreactor culture during proliferation stage and several factors affecting ex vitro rooting were also investigated.
Plant growth regulators effected proliferation and rooting of Gardenia jasminoides, the result showed: MS medium with BA of 1.0mg/L and NAA of 0.1mg/L was favorable for differentiation and growth of shoots. NAA was more effective than IBA and IAA for rooting, optimal plantlet growth and 100% of rooting were found in 0.1 mg/L NAA treatment. In addition, standard MS strength (1.0MS), 60mM or 90mM nitrogen, 40 mM NO_3~- mixed 20 mM NH_4~+ treatments, and 30g/L sucrose were suitable for proliferation of Gardenia jasminoides in vitro. Proliferation and shoot growth were also promoted by ventilation of vessels, a higher proliferation rate and favorable growth were obtained by treatment of 0.5 cm ventilation diameter, however succulent growth of shoot was found in non-ventilation treatment (0 cm). Otherwise less adventitious shoots were obtained in large ventilation treatments (over 0.5 cm).
To investigate the feasibility of bioreactor culture during proliferation stage, three culture methods, namely immersion, raft and ebb & flood culture were compared. Immersion culture resulted in highest adventitious shoots numbers (7.1) among the three methods, but showed a large of curled leaves. In ebb & flood culture , although the proliferation rate was lower than immersion culture, but fewer curled leaves was. observed. Raft culture was not suitable for proliferation of Gardenia jasminoides.
The feasibility of rooting ex vitro was found during rooting stage. Root formation and growth were batter in foam than perlite treatment; pH and EC in
引文
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