油松种胚的组织培养和植株再生的初步研究
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摘要
油松(Pinus tabulaeformis Carr.)为我国特有树种,是我国华北、西北及东北南部的主要造林树种之一,同时也是世界上重要的木材及松脂生产的资源。目前林业上主要依靠实生苗和优树嫁接的繁殖方法,繁殖系数低,苗木高低不一,林相参差不齐,远远不能满足生产的要求。而通过组织培养的方法不仅能为植树造林提供大量苗木,又能为遗传转化改良树种打下扎实的基础。目前关于油松组培成苗的报道非常少,主要存在增殖技术不成熟,生根难等问题。为此,本文以油松成熟种子为材料,进行了不定芽诱导与生根诱导条件的探索和优化,,试图建立成熟的油松组织培养技术,为其苗木的无性繁殖做出贡献。主要实验结果如下:
1、不定芽的诱导研究结果表明,诱导油松种胚不定芽的最佳条件是以不经萌发的种子为材料,在光照条件下培养,且最佳培养基为DCR+6-BA 2 mg.L~(-1)+NAA 0.1mg.L~(-1),诱导率可以达到41.11%。
2、不定芽增殖研究结果表明,不定芽增殖培养的最适附加物的组合是蔗糖30g.L~(-1)+肌醇500mg.L~(-1)+活性炭2g.L~(-1)。方差分析表明,蔗糖浓度的影响是显著,但肌醇与活性炭浓度的影响不显著。继代培养时间以4周为宜。
3、生根培养的研究结果表明,单芽的取材时间以培养12周后的不定芽为宜,在1/2DCR+NAA 0.1 mg.L~(-1)+IBA 0.1 mg.L~(-1)+KT 0.1 mg.L~(-1)培养基中生根率可以达到31.67%。
Pinus tabulaeformis Carr.,as a peculiar plant of our country,is one of staple trees for forestation in the North,Northwest and South of Northeast in China,as well as a most important raw material for wood and turpentine in the world.Seedlings and Grafted trees,the mainly ways of vegetative propagation at present,have many disadvantages,such as low progenitive rate,different tallness of trees, and so on,at which,requirements can't be meet.However,studying the technology of tissue culture can supply a lot of seedlings for forestation,besides,it can provide substantial foundation for genetic improvement of tree species.Currently,reports on Pinus tabulaeformis Carr breeding are too little and with many problem,such as faulty technology,hard to root,and so on.At this,in this paper,a lot of discovery and optimization about Adventitious Shoot Induction and root formation Induction are done on mature seeds of Pinus tabulaeformis Carr;and try to establish a consummate technology on its breeding,to make a contribution to asexual multiplication of planting stock.Main results are as follows:
1.Studying on adventitious shoot induction,and the result showed that,the best condition of inducing adventitious shoot is in the illumination environment,using the seed that have not germinated as matiarial.The inducement rate was higher in the medium of DCR+6-BA2 mg.L~(-1)+NAA0.1 mg.L~(-1) than in other media.,and the callus induction rate was 44.11%.
2.The study of the multiplication of adventitious buds showed that the most effective subculture was DCR+sucrose30g.L~(-1)+inositol 500mg.L~(-1)+activated carbon2g.L~(-1).And the result showed the effect of sucrose concentration was notable,while others were not.The period for subculture was 4 weeks.
3.The study of the callus induction of root showed that the effective single bud was from the adventitious buds which cultured for 12 weeks,and the media was 1/2DCR+NAA 0.1 mg.L~(-1)+IBA 0.1 mg.L~(-1)+KT 0.1 mg.L~(-1).The radiation rate was 31.67%.
1、不定芽的诱导研究结果表明,诱导油松种胚不定芽的最佳条件是以不经萌发的种子为材料,在光照条件下培养,且最佳培养基为DCR+6-BA 2 mg.L~(-1)+NAA 0.1mg.L~(-1),诱导率可以达到41.11%。
2、不定芽增殖研究结果表明,不定芽增殖培养的最适附加物的组合是蔗糖30g.L~(-1)+肌醇500mg.L~(-1)+活性炭2g.L~(-1)。方差分析表明,蔗糖浓度的影响是显著,但肌醇与活性炭浓度的影响不显著。继代培养时间以4周为宜。
3、生根培养的研究结果表明,单芽的取材时间以培养12周后的不定芽为宜,在1/2DCR+NAA 0.1 mg.L~(-1)+IBA 0.1 mg.L~(-1)+KT 0.1 mg.L~(-1)培养基中生根率可以达到31.67%。
Pinus tabulaeformis Carr.,as a peculiar plant of our country,is one of staple trees for forestation in the North,Northwest and South of Northeast in China,as well as a most important raw material for wood and turpentine in the world.Seedlings and Grafted trees,the mainly ways of vegetative propagation at present,have many disadvantages,such as low progenitive rate,different tallness of trees, and so on,at which,requirements can't be meet.However,studying the technology of tissue culture can supply a lot of seedlings for forestation,besides,it can provide substantial foundation for genetic improvement of tree species.Currently,reports on Pinus tabulaeformis Carr breeding are too little and with many problem,such as faulty technology,hard to root,and so on.At this,in this paper,a lot of discovery and optimization about Adventitious Shoot Induction and root formation Induction are done on mature seeds of Pinus tabulaeformis Carr;and try to establish a consummate technology on its breeding,to make a contribution to asexual multiplication of planting stock.Main results are as follows:
1.Studying on adventitious shoot induction,and the result showed that,the best condition of inducing adventitious shoot is in the illumination environment,using the seed that have not germinated as matiarial.The inducement rate was higher in the medium of DCR+6-BA2 mg.L~(-1)+NAA0.1 mg.L~(-1) than in other media.,and the callus induction rate was 44.11%.
2.The study of the multiplication of adventitious buds showed that the most effective subculture was DCR+sucrose30g.L~(-1)+inositol 500mg.L~(-1)+activated carbon2g.L~(-1).And the result showed the effect of sucrose concentration was notable,while others were not.The period for subculture was 4 weeks.
3.The study of the callus induction of root showed that the effective single bud was from the adventitious buds which cultured for 12 weeks,and the media was 1/2DCR+NAA 0.1 mg.L~(-1)+IBA 0.1 mg.L~(-1)+KT 0.1 mg.L~(-1).The radiation rate was 31.67%.
引文
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