大型绿藻浒苔藻段及组织块的生长和发育特征

通过将浒苔叶状体分为基部、中部和顶端三部分分别进行切段和切碎处理,在实验室条件下,用液体浅层培养的方法,系统地研究了其组织和细胞的生长和发育特性。显微观察的结果显示：切段培养条件下,基部和中部的藻段均可在其形态学下端形成假根,在形态学上端产生类似叶状体的突起。藻段的发育具极性,但是其极性并不绝对的,在1.0 mm的基部藻段两端都观察到了假根的形成。虽然顶端的藻段和组织块全都形成和释放了孢子,未见明显的营养生长,但是在培养早期,其下端仍然具有形成假根的能力。浒苔各部位藻段和组织块释放的和滞留于孢子囊内的孢子都可以立即萌发成苗。快速生长的中基部藻段形成了气囊,致使其漂浮于培养基上。有很多藻段和组织块形成和释放了生殖细胞,释放到外界以及滞留于孢子囊内的孢子均可立即附着萌发。数据分析表明：藻段的生长具有极性,不同部位相同长度的藻段生长率差异明显,基部藻段的生长率高于中部藻段,顶部藻段无明显的营养生长。藻段的生长与其原始长度和在藻体中所处的位置有密切关系;藻段和组织块的再生与藻体的完整性及其在藻体中所处的位置有关。     

浒苔(Enteromorpha prolifera)藻体发育的显微观察

通过显微镜观察以及石蜡切片法观察了浒苔属浒苔藻体的发育过程,阐述了浒苔从孢子释放到形成成熟藻体这一发育过程的显微特征。结果表明:浒苔孢子由成熟体细胞分化发育而成,每个成熟体细胞可形成10～30个孢子;孢子首先分裂为2细胞结构,2细胞具有明显的极性,基细胞发育成为假根,顶端细胞不断进行横分裂形成丝状体;丝状体顶端以下细胞通过纵分裂形成管状叶状体,顶端细胞始终保持横分裂;纵分裂分为切向分裂与径向分裂,切向分裂增加管状叶状体管周细胞数从而使管体增大,径向分裂形成管外壁突起细胞,最终发育为分枝;管状叶状体管腔内部有绒毛状的突起及糖蛋白成分的网架结构;当管状叶状体管周细胞达到30～50h,管腔内部突起消失,网架结构收缩拉紧,管壁细胞贴近形成片状叶状体;整个浒苔藻体始终保持极性发育。     

紫外辐射和低盐胁迫对浒苔生长的影响

探究了不同紫外曝光量及低盐胁迫对浒苔生长的影响。结果显示,紫外辐射对浒苔生长有抑制作用,甚至使浒苔呈负增长,而且在盐度为24时抑制作用比盐度为12时大,藻体湿重下降明显。2.3×10⁴KJ·m^-2～4.7×10⁴KJ·m^-2的紫外曝光量使藻体变细、变软、变散,在显微镜下观察发现部分藻体受到损伤,死亡后内容物外溢;1.9×10⁴KJ·m^-2紫外曝光量对浒苔生长率影响较小。实验设置了0、6、12、24四个盐度梯度,培养42d,在盐度为0的情况下,前21 d浒苔藻体颜色没有明显变化,但藻体变软,变散,随后,藻体颜色慢慢变黄、变白直至死亡;其它各组生长均正常,显示出浒苔的盐度适应范围广,对低盐具有一定的耐受性,但不能长时间耐受0盐度。     

竹叶蕨配子体发育的培养观察

首次在光学显微镜下观察竹叶蕨孢子及其萌发、丝状体发育、片状体和生长点的形成及分化、原叶体细胞形态、假根及性器的发育等方面所表现出的显微特征。初步讨论竹叶蕨科从鳞始蕨科中分立出来的合理性,以及原叶体边缘细胞的形态、叶绿体对光的敏感性、假根的形态和精子器的形成及分化的系统学意义。     

波叶仙鹤藓的孢子培养及发育生物学研究

首次成功地进行了人工条件下波叶仙鹤藓(Atrichum undulatum(Hedw.)P.Beauv)单细胞孢子的培养,并详细研究、拍摄了其植物体(配子体)形态发育的全过程。孢子无后熟或体眠期,3-4天即可萌发,萌发率可达95％以上;培养10天左右绿丝体系统、轴丝体系统及假根开始分化,约40天开始出现芽体原基,50天芽体陆续形成,80天左右植物体完成营养生长,开始性器发育。初生假根发育不良,基原细胞不分裂,由绿丝体产生轴丝体的方式复杂,轴丝体细胞分裂面有球形和斜向、横向三种,芽原基分化时,首先在轴丝体上产生光合能力很强的细胞群,其基部产生芽原基,芽原基的分化成功率仅有50％左右,芽体形成后,原丝体系统多枯萎或特化为假根。并就波叶仙鹤藓的形态发育、生理生态、生殖和进化等方面也进行了理论探讨。     

植物离体茎段嫁接

植物体茎段嫁接系统是在无菌条件下将茎切段嫁接后放入培养基中、使接穗和砧木分别与含不同成分的培养基接触,再署光下培养的一个模拟植物正常生理过程、环境条件可控的实验系统,离体茎段嫁接体的发育与整体类拟,包括接穗与砧木粘连、愈伤组织产生、次生甩间连丝形成和维管束分化等几个步骤,发育进程受植物激素如生长素和细胞分裂素调节。该系统的建立为阐明嫁接体发机理及嫁接亲和性机制提供了重要的依据。     

植物的微体繁殖

植物微体繁殖是植物在离体的条件下,以其组织、器官或细胞作为外植体,在一定的培养基中和培养条件下,经过形态发生的不同时期与细胞的生长、发育和分化,形成再生植株的过程。1902年哈布兰特(Haberlandt)提出植物细胞全能性理论,1958年斯蒂沃德(Steward)从胡萝卜根的悬浮细胞诱导分化,形成完整的再生小植株,首次证实了细胞全能性,至今,植     

粗枝软骨藻Chondria crassicaulis组织培养初探

在固体培养基上研究了粗枝软骨藻(Chondria crassicaulis)再生的极性,并利用三种不同的培养液和不同的温度对粗枝软骨藻进行切段培养,探索了部分外界因素对粗枝软骨藻生长的影响.发现软骨藻在人工培养条件下的再生出芽过程并无明显极性.在10℃、15℃和22℃三个温度梯度和PES、改良的ASP1和改良的ES三种培养液中,15℃改良的ES培养液中较有利于再生芽的出现.同时我们还观察了粗枝软骨藻四分孢子的发育过程,发现软骨藻具有十字型和四面锥型两种不同的四分孢子囊,软骨藻在四分孢子萌发过程中可形成四种不同的孢子苗.     

风信子试管苗芽体培养中的全息现象初探

风信子试管苗,取其芽体及切块在离体培养时存在全息现象,主要表现为,在风信子芽体的不同位置的切段上。不定芽发生的数量和频率呈梯度变化;在风信子芽体的不同大小的切段上,循着切段越小不定芽的分化数越少这一规律变化,这些现象都遵循了生物全息律。适当的激素浓度对全息胚的表现具有一定的促进作用。     

枸杞悬浮培养条件下的胚状体发生

在MS培养基上诱导宁夏枸杞下胚轴切段形成愈伤组织,并进行细胞悬浮培养。观察了悬浮培养条件下胚状体的发生过程。<1>观察发现,细胞经悬浮培养几天以后,先形成胚性细胞团,再有胚性细胞团块形成一个或几个胚状体。较大的胚状体也可以形成新的次生胚状体。<1>影响胚状体形成的关键因素是激素的种类及其含量。实验采用的基本培养基为MS,比较适宜胚状体发生的激素是0.2mg/L 2,4-D。同样浓度的2,4-D,会抑制胚状体的进一步发育,用0.2mg/L6-BA代替2,4-D, 则胚状体可以进一步发育并形成小植株。     

Development and organization of the central vacuole of Acetabularia acetabulum

* Here we analyzed the shape of the central vacuole of Acetabularia acetabulum by visualizing its development during diplophase (from juvenility through reproduction) and haplophase (from meiosis through mating). * Light microscopy and whole-organism applications of a pH-sensitive dye, neutral red, were used to visualize the anatomy of the central vacuole. We studied connectivity within the thallus by locally applying dye to morphologically distinct regions (rhizoid, stalk, apex, hairs) and observing dye movements. * In vegetative thalli most of the rhizoid, stalk and young hairs stained with dye. In reproductive structures (caps, gametangia) dye also stained the majority of the interiors. When applied to small areas, dye moved at different rates through each region of the thallus (e.g. within the stalk). Dye moved from younger hairs, but not from older hairs, into the stalk. Errors in incorporation of central vacuole into gametangia occurred at <10(-5). * These data indicate that the central vacuole of A. acetabulum is a ramified polar organelle with, potentially, a gel-like sap that actively remodels its morphology during development.     

Involvement of auxin and a homeodomain-leucine zipper I gene in rhizoid development of the moss Physcomitrella patens

Differentiation of epidermal cells is important for plants because they are in direct contact with the environment. Rhizoids are multicellular filaments that develop from the epidermis in a wide range of plants, including pteridophytes, bryophytes, and green algae; they have similar functions to root hairs in vascular plants in that they support the plant body and are involved in water and nutrient absorption. In this study, we examined mechanisms underlying rhizoid development in the moss, Physcomitrella patens, which is the only land plant in which high-frequency gene targeting is possible. We found that rhizoid development can be split into two processes: determination and differentiation. Two types of rhizoids with distinct developmental patterns (basal and mid-stem rhizoids) were recognized. The development of basal rhizoids from epidermal cells was induced by exogenous auxin, while that of mid-stem rhizoids required an unknown factor in addition to exogenous auxin. Once an epidermal cell had acquired a rhizoid initial cell fate, expression of the homeodomain-leucine zipper I gene Pphb7 was induced. Analysis of Pphb7 disruptant lines showed that Pphb7 affects the induction of pigmentation and the increase in the number and size of chloroplasts, but not the position or number of rhizoids. This is the first report on the involvement of a homeodomain-leucine zipper I gene in epidermal cell differentiation.     

Spore Germination and Rhizoid Differentiation in Onoclea sensibilis: A Two-Dimensional Electrophoretic Analysis of the Extant Soluble Proteins

Following a geometrically asymmetrical cell division during germination of spores of the fern Onoclea sensibilis L., the small cell differentiates into a rhizoid and the large cell divides to form the protonema. Using silver-staining of two-dimensional gels, we have examined the soluble proteins of spores during germination and of separated rhizoid protoplasts and protonemal cells. Of over 500 polypeptides followed, nearly 25% increased or decreased in prominence during spore germination and the initial phases of rhizoid elongation. Soluble proteins from purified protoplasts of young rhizoids were quantitatively different from those of protonemal cells and germinated spores. Nine polypeptides which appeared after cell division were substantially more prominent in rhizoid protoplasts than in whole germinated spores and have been putatively designated rhizoid-specific polypeptides. The differences in the soluble protein composition of young rhizoids and protonemal cells probably reflect the differential organelle distribution between the two cells as well as differential net protein synthesis in the cytoplasms of the two cells.     

Negative phototropic response of rhizoid cells in the fern Adiantum capillus-veneris

In general, phototropic responses in land plants are induced by blue light and mediated by blue light receptor phototropins. In many cryptogam plants including the fern Adiantum capillus-veneris, however, red as well as blue light effectively induces a positive phototropic response in protonemal cells. In A. capillus-veneris, the red light effect on the tropistic response is mediated by phytochrome 3 (phy3), a chimeric photoreceptor of phytochrome and full-length phototropin. Here, we report red and blue light-induced negative phototropism in A. capillus-veneris rhizoid cells. Mutants deficient for phy3 lacked red light-induced negative phototropism, indicating that under red light, phy3 mediates negative phototropism in rhizoid cells, contrasting with its role in regulating positive phototropism in protonemal cells. Mutants for phy3 were also partially deficient in rhizoid blue light-induced negative phototropism, suggesting that phy3, in conjunction with phototropins, redundantly mediates the blue light response.     

Effects of caffeine on germination and differentiation in spores of the fern, Onoclea sensibilis

During spore germination in the fern, Onoclea sensibilis L., the nucleus moves from a central position to one end, and an asymmetrical cell division partitions the spore into two cells of greatly unequal size. The smaller cell differentiates directly into a rhizoid, whereas the larger cell and its derivatives give rise to the prothallus. In the presence of 5 mM caffeine, the nuclei of most of the spores undergo mitotic replication, whereas cell wall formation is blocked. Multinucleate single cells are produced, which are capable of growth, but no rhizoid differentiation occurs. In some cases a partial cell wall is produced, but the nucleus moves through the discontinuity back to the center of the spore, and the enucleate, incompletely partitioned small “cell” fails to differentiate into a rhizoid. In less than 1% of the spores a broad protuberance, whose wall is yellow-brown, is formed in a multinucleate single cell. The color, staining reaction to ruthenium red, and ultrastructural appearance of the protuberance resemble that of the rhizoid wall. It appears that infrequently in the caffeine-treated spores, a feature which is characteristic of rhizoids is expressed, in the absence of asymmetric cell division, in a cell which otherwise is unable to produce a rhizoid. The results are interpreted to mean that the spore has a highly localized, persistent differentiated region. For rhizoid differentiation to occur, a nucleus must be confined in that region – a confinement which normally is accomplished by the geometrically asymmetric first cell division of germination.     

ELECTRICAL CONTROL OF RHIZOID FORMATION IN THE RED ALGA,GRIFFITHSIA BORNETIANA

1. Direct galvanic current of 10 to 40 microamperes per square millimeter of cross-section of medium results in anodal determination of rhizoid origin in the differentiated cells of the red alga Griffithsia bornetiana. The current is most effective near the upper end of the range. 2. Within the range used there is an increase in the number of rhizoids produced with increase in current intensity and a decrease in size of rhizoids. 3. Currents of lower intensity require a longer time to produce these effects than comparatively high currents. 4. The orientation of the plants in the electrical field seems to affect the number of rhizoids produced, in that plants with apexes toward the anode produce more rhizoids. 5. Together with anodal rhizoid determination there is migration of chromatophores toward the anodal side of each cell. 6. Displacement of chromatophores (and other cytoplasmic bodies) by the centrifuge does not affect the point of rhizoid origin, but does affect the shoots. 7. Together with anodal determination of rhizoids the algal filaments become graded in color, from bright pink toward the cathode to pale tan toward the anode. 8. Evidence is presented to show that this is not due to a pH change, but to a loss of pigment by chromatophores toward the anode and electrophoresis of the pigment toward the cathode. 9. In conclusion the probability is pointed out that the current acts in morphogenesis by moving particles of different charge.     

Observations on mechanisms of attachment in the green algaUlva mutabilis Føyn

The development of the rhizoid cells of the green alga Ulva mutabilis was investigated at the ultrastructural level paying special attention to the mechanism of attachment of the plant. Cytochemical data concerning the initial settling of the early zygote are also given. On the basis of histochemical staining and enzyme treatment it is concluded that the adhesive material secreted by the rhizoid cells is chemically different from that secreted by the zygote during the initial settling of the alga.     

Early development in fern gametophytes: interpreting the transition to prothallial architecture in terms of coordinated photosynthate production and osmotic ion uptake

Gametophytes of Onoclea sensiblis L. were grown under various light and media-ion conditions to gain a better understanding of the source/sink relationships between photosynthetic and ion-absorbing cells. There was a clear interdependency between green cell and rhizoid functions, such that the growth and development of the rhizoids was completely dependent on the internal delivery of photosynthates from green cells, and conversion of the one-dimensional filament into the two-dimensional prothallus required monovalent cations that could only be provided by rhizoid uptake. The need for monovalent cations was related to osmotic demands of dividing and expanding cells; prothallial development was blocked by monovalent cation deficiency, and the system resorted to Na+ uptake to support cell expansion when K+ was absent. Surgical excisions of filament cells further demonstrated the high degree of coordinated growth between the light-absorbing and ion-absorbing regions. It was also learned that excised sub-apical cells of the protonemata, like the intensively studied apical cell, were capable of remodelling remnants of the filament into a normal prothallus.