【作者】 郭红娟；

【导师】 王任翔；

【作者基本信息】 广西师范大学 ， 生物化学与分子生物学， 2010， 硕士

【摘要】 蕨类植物是高等植物的三大门类之一,在系统演化上介于苔藓植物与种子植物之间。地球上现存的蕨类植物有12000多种,其中大部分为草本植物。我国的蕨类植物资源十分丰富,约有2 600多种,占全世界种的1/5以上。自古以来民间就有以蕨类植物为治病偏方的历史,许多种类都有药用功效。近些年分析化学用在蕨类植物上,越来越多的药用化学成分被发现。而且蕨类植物许多种类十分优美,具有很高的观赏价值。蕨类植物与人类生活关系越来越紧密,加之在生物学上的重要地位,促使更多的人参与到对其的研究中。本文对几种药用蕨类植物进行了三个方面的研究。全文主体分四个部分。第一部分综述了蕨类植物的繁殖技术、配子体发育和叶表皮结构及发育的研究进展,并阐述了本论文研究的意义和目的。第二部分对蜈蚣草(Pteris vittata Linn.)、镰羽贯众(Cyrtomiu balansae (Christ) C. Chr.)、乌毛蕨(Blechnum orientale L.)、凤尾草(Pteris multifida Poir)、光亮瘤蕨(Phynatodes lucida (Roxb.) Ching)等五种药用蕨类做了孢子繁殖技术的研究,通过对其进行组织培养和常规培养及营养液纸桥培养等方法,探讨适合其繁殖的方式和激素对其生长的影响。研究结果:(1)蜈蚣草是一种繁殖比较容易的蕨类植物,MS基本培养基就能满足其大量繁殖,当然,加入适量浓度的6-BA(1.0~1.5mg/L)其繁殖速度会更快。蜈蚣草原叶体在培养基中增殖成GGB;(2)凤尾草组织培养中,发现实验用的几种激素对其影响都不是很大,而在常规培养中用MS营养液代替普通水培养凤尾草可以是一条很好的繁殖凤尾草的方法,这样既简单方便成苗率又高;(3)镰羽贯众组织培养取得的结果不是很理想,但发现在常规培养中如果对其实施全天日照,镰羽贯众孢子萌发和原叶体生长发育都很快,而且会进行无性生殖;(4)乌毛蕨的原叶体可以被2,4-D诱导出愈伤组织,但诱导出的愈伤组织因激素浓度不同而形态各异,而且都不能正常生长诱导出孢子体;MS培养基中培养孢子体诱导率不是很高;(5) 2,4-D可以诱导光亮瘤蕨原叶体产生很多黄色的假根,但几种激素都没能诱导光亮瘤蕨的原叶体长出孢子体。第三部分对蜈蚣草、镰羽贯众、凤尾草、光亮瘤蕨、乌毛蕨、巢蕨(Neottopteris nidus(L.)J.Sm.)、渐尖毛蕨(Cyclosorus acuminatus (Houtt.) Nakai)七种药用蕨类的配子体发育进行了研究。蜈蚣草孢子萌发为书带蕨型(Vittaria-type),配子体发育为水蕨型(Ceratopteris-type),配子体发育过程中无毛状体;凤尾草孢子萌发为书带蕨型,配子体发育过程为水蕨型,发育过程无毛状体产生,与蜈蚣草相同;光亮瘤蕨孢子萌发类型为密穗蕨型(Anemia-type),配子体发育为水蕨型;巢蕨孢子萌发类型为书带蕨型,配子体发育类型为水蕨型;乌毛蕨孢子萌发为书带蕨型,配子体发育为铁线蕨型(Adiantum-type);渐尖毛蕨孢子萌发类型为书带蕨型,配子体发育应为铁线蕨型;镰羽贯众孢子萌发为书带蕨型,配子体发育为三叉蕨型(Aspidium-type)。第四部分对巢蕨、镰羽贯众、渐尖毛蕨、乌毛蕨、假鞭叶铁线蕨(Adiantum malesianum Ghatak)、蜈蚣草、凤尾草、肾蕨(Nephrolepis cordifolia (L.) Presl)等8种药用蕨类植物进行了组培苗的叶表皮观察,总结了它们上下叶表皮的特征和发育的过程,对气孔器的发育和类型也做了详细叙述。在这8种蕨类中共观察到8种气孔复体类型:极细胞型(Polocytic type)、并极细胞型(Copolocytic type)、腋细胞型(Axillocytic type)、并腋细胞型(Coaxillocytic type)、不等细胞型(Anisocytic type)、不规则四细胞型(Anomotetracytic type)、辐射状细胞型(Aotinocytic type)、横列型(Diacytic type)。这些气孔器类型在每种叶表皮中所占比例都不同,每种中都有1~3种占优势的类型。气孔的发育主要是气孔母细胞经过两次和三次分裂而形成,都具同源的副卫细胞。更多还原

【Abstract】 Ferns is one of the three catagories of higher plants,which is between bryophyte and spermatophyte from the perspect of phylogeny evolution. There exists about 12,000 kinds of Ferns in the earth,most of them are herbage plants. Our country has quite a lot of ferns,with an about of 2,600 kinds,taking above 1/5 of the world. From the ancient to the current day, there has history of using ferns for curing disease as folk prescription,and many kinds of ferns have medicinal. In the recent years, more and more chemical constituents in the ferns are discovered using analytical chemistry. And a lot of ferns look beautiful, with a high value of ornamental value. Adding to its importance in biology, ferns is getting closer and closer to humans’lives, which makes more people taking part in researching.The article is mainly doing research on some kinds of medicinal ferns from three aspects, which could be divided into four parts.The first part summarizes something about ferns, including Propagation technology, gametophyte development and development of the research on the structure and growth of leaf epidermis.With the methods of using tissue culture, routine culture and solution culture, the second part researches on spores propagation technique through five medicinal ferns, which are Pteris vittata Linn, Cyrtomiu balansa (Christ) C.Chr.,Blechnum orientale L., Pteris multifida Poir, Phynatosorus cuspidatus (D. Don) Pic. Serm , trying to find out its proper way of propagation and the hormone’s effect. It turns out that :(1)Pteris vittata Linn is easier to propagate and it could largely breed in MS routine culture, apparently it will grow quicker if adding a density of 6-BA(1.0~1.5mg/L);Prothallium grows to GGB in the culture medium;(2) It is also discovered in the experiment that the hormones does not have much effect on Pteris multifida Poir ,but in the routine culture ,using MS nutrient solution instead of using regular water may be a good method for Pteris multifida Poir‘s propagation. It is simple and with a high seedling rate. (3)It is not very ideal result of tissue culture of Cyrtomiu balansa (Christ) C. Chr, but it’s found that in the routine culture Cyrtomiu balansa (Christ) C. Chr.’s spores and prothallus not only grow fast but also could be vegetative propagation if they’re lightened all the day and night. (4)Prothallium of Blechnum orientale L. could be induced callus by 2,4-D,but the induced callus performs different properties depending on the density of hormone, and they could not normally grow and induce sporophyte. It performs low-efficiency of inducing cultured sporophyte in MS culture medium. (5)2,4-D could induce prothallium of Phynatosorus cuspidatus (D. Don) Pic. Serm quite a lot of rhizoid, but all used hormones could not induce sporophyte of prothallium of Phynatosorus cuspidatus (D. Don) Pic. Serm..The third part does research on gametophyte development of seven medicinal ferns belowed as: Pteris vittata Linn., Cyrtomiu balansa (Christ) C. Chr., Pteris multifida Poir, Phynatosorus cuspidatus (D. Don) Pic. Serm., Blechnum orientale L., Neottopteris nidus (L.) J.Sm., Cyclosorus acuminatus. spore-germination type of Pteris vittata Linn is Vittaria-type, gametophyte development type is Ceratopteris-type, in the course of gametophyte development does not have trichomatas. Spore-germination of Pteris multifida Poir is Vittaria-type, gametophyte development type is Ceratopteris-type, in the course of gametophyte development does not have trichomatas, performing the same as Pteris vittata Linn. Spore-germination type of Phynatosorus cuspidatus (D. Don) Pic. Serm is Anemia-type, gametophyte development type is Ceratopteris-type;spore-germination of Neottopteris nidus(L.) J.Sm. is Vittaria-type, gametophyte development type is Ceratopteris-type; Spore germination type of Blechnum orientale L. is Vittaria-type, gametophyte development type is Adiantum-type; Spore-germination type of Cyclosorus acuminatus is Vittaria-type, gametophyte development is Adiantum-type; Spore-germination type of Cyrtomiu balansa (Christ) C. Chr. is Vittaria-type, gametophyte development type is Aspidium-type.The fourth part does some observation on leaf epidermis of eight medicinal ferns belowed as: Neottopteris nidus(L.) J.Sm., Cyrtomiu balansa (Christ) C. Chr., Cyclosorus acuminatus, Blechnum orientale L.,Adiantum malesianum Ghatak, Pteris vittata Linn., Pteris multifida Poir,Nephrolepis auriculata(L.)Trimen., and sums up characteristics and growing course of up and lower leaf epidermis and also carefully describes the development and type of stomatal apparatus. Eight types of stomatal apparatus are observed in these eight ferns:Polocytic type,Copolocytic type,Axillocytic type,Coaxillocytic type,Anisocytic type,Anomotetracytic type,Aotinocytic type,Diacytic type.These stomatal apparatus takes up different percentage in every kind of leaf epiderm, and every stomatal apparatus has one to three advantage kind. Development of stomatal mainly means stomatal’s mother cells are formed through two and three splittings, and they all have Subs diary cells.更多还原