【作者】 陈静文；

【导师】 曹同；

【作者基本信息】 上海师范大学 ， 水生生物学， 2006， 硕士

【摘要】 本论文分别选取用于分子生物学实验的模式植物——小立碗藓；在生态系统中起着重要作用的“先锋植物”——真藓；以及有药用价值的小蛇苔为对缘，进行组织培养的实验研究。筛选了这三种苔藓植物的最佳培养条件，诱导了小立碗藓和真藓的愈伤组织，完成了小立碗藓整个生活史的培养，并对真藓的土培进行了研究。同时，对小立碗藓总DNA的提取做了初步探讨。主要实验结果如下： （1） 采用改良的固体Knop为基本培养基，添加一定量的酒石酸铵，当pH6.5，糖浓度为0.5％时，在（25±1）℃，光照周期12h／12h，光照强度3500Lx条件下培养小立碗藓的效果最好。将小立碗藓移至室外培养，散光照射，喷洒营养液，可在低温条件下诱导出孢子体。成熟孢子在诱导孢子萌发的培养基上培养6d后能够萌发，三个月就可完成小立碗藓的生活史。 （2） 当糖浓度为2％，添加KT 0.05mg·L^-1或添加6-BA 0.05 mg·L^-1，诱导小立碗藓愈伤组织的效果最为理想。诱导愈伤组织分化的实验结果表明：未添加任何植物激素的Knop培养基上分化出正常的配子体数量最多。添加2，4-D或IAA的培养基上能长出较多的原丝体。 （3） 真藓在改良的Knop培养基上，（25±1）℃，光照强度3500Lx，光照周期12h／12h条件下培养时生长情况良好。 （4） 当糖浓度为2％，6-BA 0.05 mg·L^-1和KT 0.1 mg·L^-1时，真藓愈伤组织的诱导率为100％，愈伤较大，褐化最少。 （5） 真藓土培的最适条件为：（25±1）℃，泥土+砂+营养土的基质：营养液的种类对其生长的影响并不明显。 （6） 小蛇苔在（18±1）℃，光照强度3000Lx，光照周期12h／12h条件下培养时生长情况良好，与培养基的种类和pH值无明显关系。 本研究首次报道真藓愈伤组织的诱导，申请小立碗藓愈伤组织诱导与分化的发明专利，为苔藓植物组织培养的深入研究提供科学依据和技术方法。更多还原

【Abstract】 Three representative species of bryophytes were selected for study of tissue culture: Physcomitrella patens which as model plant is widely used in molecular biology; Bryum argenteum which as pioneer plant has important ecological function in different ecosystems and Conocephalum supradecompositum which as medicine plants have potential economic value. The experiments of selecting best culture condition of three bryophytes, inducing calli of P. patens and B. argenteum and culturing whole life history of P. patens were carried out. Moreover, extracting DNA from P. patens is tried and tested. Main results of the experiments can be summarized as follows:（1） Physcomitrella patens can be cultured best on the improved medium Knop added tartrate and 0.5% glucose under （25 ± 1） ℃, light intensity 3500Lx, and 12h day / 12h night when pH 6.5. The sporophyte of P. patens can be induced under low temperature. Then, spore can generate on the medium after 6 days culture. Culture of the whole life history of P. patens takes about 3 monthes.（2） Calli of P. patens were induced better on medium Knop plus 6-BA 0.05 mg L^-1 or KT 0.05mg.L^-1 when the concentration of glucose is 2%. In the differentiation experiment, normal gametophyte of P. patens were differentiated in medium Knop. Moreover, the much moss of protonemata were gained in medium Knop added 2,4-D or IAA.（3） Bryum argenteum can be cultured best on the improved medium Knop under light intensity 3500Lx, and 12h day / 12h night when the temperature in the range of（25±1） ℃.（4） Calli of B. argenteum was induced better in medium Knop plus 6-BA 00.5mg L^-1 and KT 0.1 mg L^-1 by using agar medium containing 2% glucose.（5） Bryum argenteum was grown best when the temperature in the range of （ 25 ± 1） ℃ and the medium blent with soil, sand and nurture soil. The types of culture solutions seemed no evident influence on its growth.（ 6） Conocephalum supradecompositum can be cultured best on the improved更多还原