【作者】 吴长辉；

【导师】 哀建国； 张海娟；

【作者基本信息】 浙江农林大学 ， 农业推广， 2012， 硕士

【摘要】 里白(Diplopterygium glaucum)里白科里白属多年生大型蕨类植物，具有重要的药用价值和观赏价值。本研究以蕨类植物里白的孢子为试验材料，利用组织培养的方法，研究了孢子萌发诱导过程中外植体灭菌、培养基选择、植物生长调节剂的种类、蔗糖浓度、温度、光照、溶液pH值等因素对孢子繁殖的影响，此研究对于蕨类植物的组培快繁及其配子体的发育将对蕨类植物的开发利用、种质保存及种的鉴定、科属的划分、蕨类植物的起源、演化、生态分布等具有重要作用。主要结果如下：1、消毒灭菌过程中，2%NaClO和70%酒精组合灭菌效果最好，不同的灭菌时间影响到孢子繁殖的污染问题，总体上来看，2%NaClO灭菌30min，而70%酒精灭菌15s，成活率达到85.0%效果最好。2、不同的培养基类型对里白孢子繁殖的效果不同，几个培养基中，1/2MS培养基的孢子萌发率最高，达到81.1%，萌发时间也最短，在第13d便开始萌发，是较好的培养基。3、生长素和细胞分裂素对孢子的繁殖具有一定影响。总体上来说看，各种生长素均能促进里白孢子萌发，2,4-D的效果最佳，形成孢子繁殖的时间最短，为7d，并且孢子繁殖效率最高，达到70.0%。各种细胞分裂素中，TDZ的诱导效果最佳，孢子萌发时间也最短，在浓度为2.0mg·L-1诱导率最高，达到61.1%，仅需要8d左右孢子就开始萌发。所有培养基组合中，最佳的培养基组合为改良MS+2,4-D0.2mg·L-1+NAA0.2mg·L-1+TDZ3.0mg·L-1。4、不同的蔗糖浓度对里白孢子诱导影响不同，孢子诱导率随着蔗糖浓度的不同呈现不同的变化，当蔗糖浓度为30.0g·L-1时，里白孢子诱导率达到最高的91.48%。5、当光照强度为80μmol·m-2·s-1时，孢子萌发率最高，达到92.17%。孢子需要在光照条件下启动萌发，20-50μmol·m-2·s-1的低光强就能诱导其萌发。6、温度的高低也会影响到里白孢子的繁殖情况。试验结果表明，25℃时孢子萌发所需时间最短，为7d左右。7、里白孢子繁殖过程中，培养基溶液pH值5.6时的萌发率最高，为83.2%里白孢子萌发率没有显著性差异，但是当pH值达到7.6时显著降低。更多还原

【Abstract】 This study using the fern spores of Diplopterygium glaucum as the experimental material,the spore germination induction explant sterilization, selection of culture medium, plant growthregulators, sucrose concentration, temperature, type of light, pH value of solution and other factorson the spore propagation were studied in this text, this text is good to the rapid propagation andutilization of gametophytes, it plays an important role in the genera, divided fern origin, evolution,ecological distribution of Diplopterygium glaucum. The results are showed as follows:1．The results showed that2%NaClO and70%alcohol combination sterilizing are best.2%NaClO30min70%alcohol sterilization and sterilization,15s, the survival rate of85is best.2． The results are different when using different medium,1/2MS medium sporegermination rate is highest, reaching81.1%, germination time is the shortest, at the13D began togermination, is a better culture medium.3. Auxin and cytokinin on spore reproduction has certain effect. On the whole, all sorts ofauxin can promote the white spore germination,2,4-D were the best, formation of spore breedingtime is the shortest,7d, and spore propagation efficiency highest, up to70%. All kinds ofcytokinins, TDZ induction effect is best, spore germination time is the shortest, at a concentrationof2mg/L induce the highest rate, reached61.1%, only need about8D spores begin germination.All medium combinations, the best combination of medium was modified MS+2,4-D0.2mg/L+NAA0.2mg/L+TDZ3mg/L.4．Different concentrations of sucrose on white spore inducing effects of different inductionrate, spore as the sucrose concentration had different changes, when the sucrose concentration is30.0g/L, white spore inducing rate reaches a maximum of91.48%.5. When the light intensity of80μmol·m-2·s-1, spore germination rate is the highest,reaching92.17%. Spore is needed in light conditions the initiation ofgermination,20-50/μmol·m-2·s-1low intensity can induce the germination.6. Temperature also affects the white spore reproduction. The test results show that, at25degrees C spore germination in the shortest time required, is about7d.7. White spore propagation process, culture medium solution pH value5.6the highestgermination rate,83.2%white spore adorable The rate of no significant difference, but when pH value reaches7.6significantly reduced.更多还原