【作者】 郭万里；

【导师】 李彦舫；

【作者基本信息】 中国人民解放军军需大学 ， 作物遗传育种， 2003， 硕士

【摘要】 本研究采用植物组织培养技术中的器官培养，探索规模化生产樱花(Prunus serrulata)试管苗的途径；还涉及了规模化生产林木试管苗过程中的部分问题：外植体的幼嫩程度和采集季节对建立组培系的影响；香花槐(Robinia)和樱花规模化生产中出现的玻璃化和试管苗弱化(GA引起的徒长)现象，及生产中的对策。 _在探索规模化生产4个樱花品种试管苗的过程中，我们主要采用了腋芽(侧芽)诱导方法建立了腋芽诱导培养系。试管苗初代培养的建立，继代培养和生根培养基的优化，樱花试管苗特殊状态——莲座状问题的解决和应用、继代周期、继代时间和移栽等问题作了系统的试验，并提出了腋芽诱导规模化生产樱花试管苗的可行性程序。4个樱花品种的生长状态在继代培养基中差异不明显，因此可采用一套培养基。但在生根时，它们体现出自身的遗传差异性。经过试验，我们确定了这几种樱花品种的生根培养基最佳激素配比范围。这为规模化生产多品种樱花试管苗提供了生根培养基的激素配比原则。樱花的移栽比较容易，几种樱花在珍珠岩：草炭土为1：1基质上的移栽成活率都达到了90％以上。 由于大部分林木本身的特性，其外植体的采集季节和幼嫩程度都能影响组培系的建立。通过对鸡爪枫(Acer palmatum)幼嫩程度不同的外植体，杜鹃(Rhododendron dauricum)嫩枝顶芽、较老顶芽和芽苞在培养基上的反应的观察。我们认为，林木外植体应采集春天新发的半木质化的外植体为易。 香花槐和樱花试管苗生产中，都出现了玻璃化现象，经研究发现，蔗糖和琼脂对玻璃化影响的方式类似，都是通过调节培养基渗透势和衬质势来防止试管苗玻璃化的发生和抑制玻璃化试管苗进一步恶化。而高强度光照也能抑制玻璃化的进一步恶化。封口方式中则以封口膜对玻璃化试管苗抑制最为显著。这说明培养瓶内微环境是造成玻璃化的诱因，而改善培养瓶内微环境则抑制了玻璃化，同时苗在这种环境中生长状态较好。根据实验和其他学者的发现，推论出诱导试管苗玻璃化的可能途径。同时发现樱花玻璃化现象随品种的不同而存在差异，杨贵妃最不易玻璃化，东樱玻璃化现象严重，冈目和大岛樱处于二者之间。 在香花槐组织培养中，长期使用GA造成了试管苗纤细弱化，通过试验，证明了PP333对GA引起的香花槐试管苗弱化有针对性的抑制作用。同时，适当的PP333浓度还能促进腋芽的发生。在工业化生产试管苗中，PP333与GA类似，用量不能过高，不能长期使用。更多还原

【Abstract】 Shoot tissue culture now be consided of one of the best methods in micropropagation. which can assure plant genetic types. That’s why some vegetables produced in industry by this way, and it is also more economic than other methods.We builded shoot tissue culture systems of four kinds of P. serrulata. Results obtained from these studies show that, it’s the best time for getting new budes of P. serrulata from March to April. The medium Jl( 1/2MS + 6BA0.50 + NAA0.025) is suitable for explant induction, initial culture and subculture, but the explants obtained after May can be cultured in the medium C4(1/2MS+6BA1.0+NAA0.02). The tube-shoots will appear rosellar phenomina after two subcultures, which can effect the growth of tube-shoots seriously, but the optimal medium J11 (1/2MS +6BA0.50 + NAA0.025+GA0.025) can promote elongation of the axillery buds, so the rosella will disappear after one subculture, but GA can not be used too long or in high concentration, because of its resulting in thin stem and excessive growth of tube-shoots. There is another problem, the tube-shoots in J1 and J11 mediums are too tender to be rooted, but the medium J14 (1/2MS +6BA(0.5mg/L)) can resolve the problem better. After several subcultures, the tube-shoots grow slowly little by little. Some investigations discoved that the disorder or higher concentration of hormenes or regulations in plants rleading to the phenomena. We found that the tube-shoots recovered growth after being cultured in the medium J18 (1/2MS) about 20 days. We also found that the growth of four kinds of P. serrulata have no- significant discrepancies in those mediums. That is to say, It’s valuable for mass production of many kinds of P. serrulata by simplifmg the mediums and solving some problems in the course.There are some problems affecting the application of forest trees tissue culture inindustry. Vitrification is around in plant tissue culture, we concluded that suger, agar, light, pH, ionic concentration and contamination proof closure modes effected tube-shoot vitrification from the experiments, suger and agar can inhibit vitrification when their concentration raised to some extent, that will also be happen when air circulation between inner and outer of culture vessels become better and higher ionic concentration and light intensity. On the other side, vitrification are different among four kinds of P. serrulata.GA promotes tube-shoots elongation, but using it longer(about several subcultures) can lead to attenuation of the tube-shoots, such as stem slightness, leaf etiolation, reducing tube-shoots survive rate, and so on.. PP333 can restrain those affections by GA, and it will promote Robinia axillery buds high-frequently occuring if the medium added PP333 (0.05mg/L).A. Palmatum and R. dauricum were used in the studies on the optimal explants. We consider that the best time of collecting explants is spring and it is the seem with others’opinions. On the other side, it is not aways right geting too young explants, especially in forest trees tissue culture. But the half-lignification explants is better than orthers.更多还原