Development of white spruce somatic embryos: II. Continual shoot meristem development during germination

Summary This study compares the development of shoot apical meristems of white spruce somatic and zygotic embryos during germination. In mature somatic embryos, the functional part of the shoot apical meristem was bi-layered. After partial drying, a normal shoot meristem was formed from these two cell layers during germination. Other cells within the meristem were vacuolated and separated by intercellular air spaces. In the absence of the partial drying treatment, somatic embryos enlarged in size primarily due to vacuolation of cells and the formation of large intercellular air spaces. A majority of these somatic embryos failed to form a functional shoot apical meristem. Compared with somatic embryos, the shoot apical meristem of a mature zygotic embryo was well organized with a densely cytoplasmic apical layer. The cells within the meristem were tightly packed. Judging from the cell profiles during germination, all cells within the meristem of the zygotic embryo took part in the formation of the vegetative shoot apical meristem.     

Study of aberrant morphologies and lack of conversion of somatic embryos of chickpea (Cicer arietinum L.)

Summary Somatic embryos which originated from mature embryo axes of the chickpea (Cicer arietinum L.) showed varied morphologies. Embryos were classified based on shape of the embryo and number of cotyledons. “Normal” (zygotic-like) embryos were bipolar structures with two cotyledons and a well-developed shoot and root apical meristem, whereas “aberrant” embryos were horn-shaped, had single and multiple cotyledons, and were fasciated. Histological examination revealed the absence of a shoot apical meristem in horn-shaped embryos. Fasciated embryos showed diaxial fusion of two embryos. Secondary embryogenesis was also observed, in which the embryos emerged from the hypocotyl and cotyledonary region of the primary somatic embryo. This report documents the absence of an apical meristem as a vital factor in the lack of conversion of aberrant somatic embryos.     

Seed and early plantlet structure of the Mediterranean seagrass Posidonia oceanica

Seeds from mature fruits of the Mediterranean seagrass Posidonia oceanica deposited in the intertidal zone by sea surface currents revealed an advanced state of embryo development. The fruit dehisces by three longitudinal openings, which originate from the base or point of fruit attachment. Within the fruit the seed is positioned with its radical end at the fruit base, and the apical or plumular end protected until the seed is completely released. Structural observations of the collected mature seeds suggest the possible onset of germination. The mature seed is characterized by an enlarged hypocotyl with abundant starch reserves, a well-defined vascular system with a predominant central vascular strand to mobilize those reserves, a well-developed plumule, and root system initials, which will assure anchorage to the sea floor. Thus, within the dispersal unit, the future plant organs and growing points are well established and the carbohydrate-rich endosperm will assure the availability of sufficient nutrient supplies for short-term development of the seedling.     

Reserve mobilization in the Arabidopsis endosperm fuels hypocotyl elongation in the dark, is independent of abscisic acid, and requires PHOSPHOENOLPYRUVATE CARBOXYKINASE1

Arabidopsis thaliana is used as a model system to study triacylglycerol (TAG) accumulation and seed germination in oilseeds. Here, we consider the partitioning of these lipid reserves between embryo and endosperm tissues in the mature seed. The Arabidopsis endosperm accumulates significant quantities of storage lipid, and this is effectively catabolized upon germination. This lipid differs in composition from that in the embryo and has a specific function during germination. Removing the endosperm from the wild-type seeds resulted in a reduction in hypocotyl elongation in the dark, demonstrating a role for endospermic TAG reserves in fueling skotomorphogenesis. Seedlings of two allelic gluconeogenically compromised phosphoenolpyruvate carboxykinase1 (pck1) mutants show a reduction in hypocotyl length in the dark compared with the wild type, but this is not further reduced by removing the endosperm. The short hypocotyl phenotypes were completely reversed by the provision of an exogenous supply of sucrose. The PCK1 gene is expressed in both embryo and endosperm, and the induction of PCK1:beta-glucuronidase at radicle emergence occurs in a robust, wave-like manner around the embryo suggestive of the action of a diffusing signal. Strikingly, the induction of PCK1 promoter reporter constructs and measurements of lipid breakdown demonstrate that whereas lipid mobilization in the embryo is inhibited by abscisic acid (ABA), no effect is seen in the endosperm. This insensitivity of endosperm tissues is not specific to lipid breakdown because hydrolysis of the seed coat cell walls also proceeded in the presence of concentrations of ABA that effectively inhibit radicle emergence. Both processes still required gibberellins, however. These results suggest a model whereby the breakdown of seed carbon reserves is regulated in a tissue-specific manner and shed new light on phytohormonal regulation of the germination process.     

EMBRYOGENESIS AND SEEDLING DEVELOPMENT IN CASSIPOUREA ELLIPTICA (SW.) POIR. (RHIZOPHORACEAE)

Embryogenesis in Cassipourea elliptica (Sw.) Poir, begins with a first division of the zygote which may be oriented transversely, obliquely, or rarely longitudinally. The orientation of the second division is also variable. Though the differentiation of suspensor and embryo proper occurs early, some derivatives of the terminal cell sometimes contribute to the suspensor. Provascular tissue “differentiates” after the initiation of the cotyledons. The radicle apical meristem originates subterminally, 5–10 cell layers from the juncture of the embryo proper and the suspensor. After germination, during early seedling establishment, radicle apical organization is of an unspecialized, columellate type. Vascular differentiation occurs before germination, and there are two loci of initial xylem differentiation: one in the hypocotyl and another in the median trace of the cotyledons. After germination, additional xylem differentiates de novo (without lateral or longitudinal continuity with already-mature vessels) inside the arcs of phloem in the hypocotyl, a pattern reported in few angiosperms. The cotyledonary node is one-trace, unilacunar.     

Organ identity of the thalloid plant body of Griffithella hookeriana and Polypleurum stylosum – Podostemoideae (Podostemaceae)

The morphological nature of the thalloid plant body of podostemads has remained controversial for long. The present investigation was carried out on two members of the Podostemoideae i.e. Griffithella hookeriana and Polypleurum stylosum to understand their organ identity. The origin of the plant body was traced from the embryo by germinating the seeds under aseptic conditions. Mature embryo of both species does not show an identifiable shoot apical meristem (SAM) and root apical meristem (RAM). Upon germination, the radicular pole does not form a primary root but differentiates adhesive hairs. At the cotyledonary junction, SAM is initiated that differentiates 6–9 leaves apically (primary axis) and a primordium laterally. This primordium subsequently emerges from the hypocotyl and develops into a thalloid plant. The latter has been interpreted as a flattened stem because it not only shows tunica-corpus like organization at the tip but also originates endogenously from the same SAM that forms the `primary axis'.     

Succinyl CoA Synthetase of Tobacco

The isouition and partial purification of succinyl CoA synthetase from Nicotiana tabacum is described. Enzynio activity had a pH optimum of pH 7.0 and apparent Km values for succinic acid. ATF and MgCl(2) . are 3.5 × 10(-2) 5.7 × 10(-3) and 4.0 × 10(-3) M, respectively. The euzyme is specific in requirement for ATP. Enzyme activity was low in seed, decreased during germination and seedling development, then increased to maximum in physiologically mature leaf tissue. Plauts with the apical apical meristem removed had less euzyme activity in leaves but more enzyme activity in the roots than plants with apical meristem intact. Most enzyme activity observed was associated with the soluble fractions of the cell.     

Anatomical Studies of After-Ripening of Coptis chinensis Seed

The seeds of Coptis chinensis Franeh took 9–10 months to germinate after they were collected and buried under ground.. This is due to the fact that the embryo of Coptis chinensis was not fully developed after shedding but remained in the stage of globular or early heart-shaped embryo, and the latter of which has to pass through a rather long period before it developed into torpedo-embryo stage. Embryo completed its growth about January of the following year. The delay in seed germination is because of the embryo not being fully developed after shedding. However, the primordium of the first foliage leaf initiated early in the apical meristem of the young embryo. Perceiving the whole process of the young embryo growth, it seems possible that, if a suitable condition is envisaged to shorten the heart-shaped embryo stage, the whole germinating process of seed might be somewhat curtailed.     

DNA synthesis and mRNA accumulation during germination of embryos of the orchidSpathoglottis plicata

Summary Germination of embryos of the orchid,Spathoglottis plicata Blume involves the formation of a protocorm in which DNA synthesis and cell divisions are confined to the proximal end whereas cells at the distal end undergo enlargement. Although³H-thymidine was incorporated into the distal cells of the embryo during the early period of germination, DNA synthesis was not followed by mitosis and cytokinesis. Poly(A)-RNA detected by in situ hybridization of sections to³H-poly-(U) was present uniformly in all cells of the embryo of the dry seed. However, coincident with the formation of the apical meristem, there was an increase in the density of auto-radiographic silver grains in the cells of the embryo, with a concentration of silver grains in the proximal part. The results indicate that regulatory events in the embryo prior to seed maturity determine the fate of its proximal and distal parts during germination.Dedicated to the memory of Professor John G. Torrey     

Developmental morphology of the Asian one-leaf plant, Monophyllaea glabra (Gesneriaceae) with emphasis on inflorescence morphology

We examined the developmental morphology of the tropical Asian one-leaf plant Monophyllaea glabra, which is believed to have diverged first in the phylogenetic tree of the genus. The embryo within the seed consists of two cotyledons and a hypocotyl with no shoot or root apical meristems. The endogenous root meristem is formed nearer the hypocotyl end than in other examined Monophyllaea species. One of the cotyledons grows to form the macrocotyledon by means of the basal meristem. The groove meristem arises between the anisocotyledons, shifts toward the macrocotyledon, and is transformed to the inflorescence apex, which produces inflorescence axes in the axils of all ventral bracts of two rows, and secondary inflorescences in the axils of the lower dorsal bracts of the other two rows. The macrocotyledon may act as a ventral bract for the first inflorescence axis at the reproductive stage. This organization suggests that a common ancestor of Monophyllaea and Whytockia with decussate inflorescences diverged in one direction to become Monophyllaea and in another to become Whytockia.     

Structure of the zygotic embryos and seedlings of Butia capitata (Arecaceae)

Butia capitata, an endemic palm of the Brazilian savanna threatened by deforestation, demonstrates low germinability due to seed dormancy. The present study characterizes the structure of the zygotic embryo and describes germination and seedling development. Pyrenes were sown into sandy soil substrates to germinate, and their embryos were also cultivated in vitro in MS medium; structural evaluations were made during their development. Seedling growth through the endocarp germ pore culminates in the protrusion of the cotyledonary petiole, with the root and leaf sheaths subsequently being emitted laterally from its extremity. The embryos are composed of the cotyledon (whose proximal third has a haustorial function) and a diminutive embryo axis that is contained within the cotyledonary petiole. The protoderm, ground meristem, and procambium can be observed in their typical positions in the embryo axis and cotyledon. The development of the vegetative axis could be observed on the second day of in vitro cultivation, with elongation of the embryo axis and the beginning of the differentiation of the first eophyll. Elongation of the cotyledonary petiole and the differentiation of the parenchyma and tracheary elements were observed during the second to fifth day. Although the hypocotyl–radicle axis is less differentiated than the plumule, root protrusion occurs on the eighth day, and the leaf sheaths are only emitted between the twelfth and the sixteenth days; the haustorium atrophied during this stage. The embryonic structure of B. capitata does not impose limitations on seed germination as dormancy is of the non-profound physiological type, and the 50 % elongation of the cotyledonary petiole serves as a morphological indicator of germination.     

Cotton somatic embryo morphology affects its conversion to plant

Somatic embryos differentiated from hypocotyl explant in cotton (Gossypium hirsutum L.) exhibited very divergent morphologies. Six different types of somatic embryos based on cotyledon development were observed. The growth hormones (2,4-dichlorophenoxyacetic acid and kinetin) used in induction and maintenance media did not affect embryo rooting and germination. The 95 % conversion of normal embryos (with two cotyledons) was achieved, while an overall conversion was only 38 %. Horn shaped embryos failed to exhibit shoot growth. Poorly developed apical meristems were responsible for lower conversion percentages in some of embryo classes. However, regenerated plants phenotypically resembled to seed grown control plants regardless of somatic embryo morphology.     

Ontogenetic Anatomy of Streptocarpus grandis(Gesneriaceae) with Implications for Evolution of Monophylly

The monophylly of Streptocarpus grandis was examined ontogeneticallyand anatomically. When the seed is shed, the embryo is composedof a hypocotyl and two equal-sized cotyledons, lacking rootand shoot apices. During germination, cell division and subsequentcell enlargement occur in the hypocotyl and cotyledons. Thehypocotyl soon produces a primary root from its distal tip;this involves surface and subsurface cells at the point of attachmentof the suspensor remnant. In the cotyledons, cell enlargementand differentiation occur basipetally, leaving small meristematiccells at the bases. These small cells give rise to the basalmeristem in one of the two cotyledons, which contributes toan accrescent cotyledon. The groove meristem, which later differentiatesinto an inflorescence, arises in place of shoot apices whenthe cotyledons become visibly unequal in size. It later exhibitsa tunica-corpus like configuration and differentiates directlyinto an inflorescence meristem. The evolution of this uniquegrowth of one-leaved Streptocarpus is discussed with regardto morphogenetic data.Copyright 2000 Annals of Botany Company Anisocotyly, developmental anatomy, evolution, Gesneriaceae, one-leaf plant, ontogeny, Streptocarpus grandis     

THE DEVELOPMENT OF THE ACHENE OF POLYGONUM PENSYLVANICUM: EMBRYO,ENDOSPERM, AND PERICARP

A study was made of the ontogeny of the achene of Polygonum pensylvanicum L. from fertilization to maturity. The proembryo is classified as the Polygonum Variation, Asterad Type. Cotyledons are initiated three days after anthesis, and by the fifth day procambium is present in the embryo axis. At approximately seven days after anthesis, the embryo begins to curve and occupy a marginal position in the ovary. By ten days the first foliage leaf primordium is initiated at the stem apex of the embryo. At maturity the embryo consists of two cotyledons, a plumule composed of the stem apex and one leaf primordium, and a hypocotyl with a well-developed radicle. Endosperm nuclei begin to divide before the first division of the zygote. Cell wall formation begins in the endosperm at the micropylar end of the embryo sac and proceeds toward the chalazal region. By the fifth day the endosperm is completely cellular, except for a basal projection; and a peripheral meristem has been established. At approximately ten days the peripheral meristem ceases periclinal cell division and becomes the aleurone. At the time of fertilization the ovary wall has its full complement of cell layers. The walls of the outermost cells elongate and become convoluted. Subsequent thickening and lignification of these cell walls produce the hard epicarp of the mature achene.     

苏铁(Cycas revoluta Thunb.)种子的胚与胚乳解剖结构研究

对苏铁(Cycas revoluta Thunb.)种子的胚和胚乳组织进行了解剖研究。结果表明:苏铁种子为有胚乳种子,兼有胚乳和外胚乳,成熟时具直立型胚。胚乳的表层细胞含有角蜡质,胞核大,不含淀粉粒;中层细胞胞核明显;内层细胞胞核不明显,富含淀粉颗粒,淀粉粒单脐点明显。胚孔端的胚乳内陷成一凹槽,似贮藏窖。成熟的子叶胚为倒生胚胎,位于胚乳细胞解体后形成的囊腔中,子叶胚长度在胚乳中占到种子的1/3至2/3,已达到生理成熟阶段。双子叶直立,半合生。胚状体基部呈喙状突起,喙状突起下端连着一根肠叠着的丝状吸器,吸器基部连着一个小气囊。胚芽由顶端分生组织和数枚真叶组成,此时真叶已具羽状叶原基和绒毛原始体。在胚状体中发现有长管细胞及螺纹加厚的导管,在子叶中脉有数条并列的螺环纹导管。     

黄精种子萌发过程发育解剖学研究

采用石蜡切片技术对成熟黄精种子形态及萌发过程中的形态学变化及解剖结构特征进行了研究,以阐明黄精种子繁殖的生物学机制。结果显示:(1)成熟的黄精种子由外而内依次为种皮、胚乳和胚等3部分组成。其中种皮由一层木质化的细胞组成;胚乳占据种子的大部分结构,胚乳细胞含有大量淀粉,细胞壁增厚;胚处于棒型胚阶段。(2)黄精种子在萌发过程中棒型胚靠近种脐端分化为吸器、子叶联结和子叶鞘,靠近种孔的部位分化出胚根、胚轴和胚芽。(3)黄精种子萌发首先由子叶联结伸长将胚芽和胚根原基推出种孔,紧接着下胚轴膨大形成初生小根茎,吸器留在种子中分解吸收胚乳中的营养物质。(4)通过子叶联结连通吸器和初生小根茎,将胚乳中的营养物质由吸器-子叶联结这个通路转移到初生小根茎中,为初生根茎上胚芽和胚根的进一步分化提供物质保障。(5)黄精种子自然条件下萌发率较低,而且当年不出土。研究表明,黄精种子的繁殖生物学特性是其生态适应的一种重要机制。     

Utilization of Seed Reserves and Currently Produced Photosynthates by Embryonic Tissues of Pine Seedlings

The importance of seed reserves for growth of Pinus resinosaAit. during and shortly after seed germination was studied undercontrolled conditions. Tissues in the resting embryo were notcompletely differentiated. Many small, presumably reserve particleswere present in the embryo in addition to reserves in the megagametophyte.During seed germination, procambia in the embryo first differentiatedprotophloem 2 days after seeds were sown. The radicle beganto emerge from the seed coat at 5 days, at which time initialxylem formation was observed. Also, at approximately the sametime, primordia of primary needles were forming in the peripheralzone of the apex. Elements of the photosynthetic apparatus,including stomata and mesophyll with chloroplasts, were differentiatedfirst in the hypocotyl and then in cotyledons between 5 and8 days after seeds were sown. Photosynthetic rates of youngseedlings were correlated with rates of cotyledon expansion.During early developmental stages, reserve particles in megagametophytecells and embryo cells gradually disappeared. Surgical removalof megagametophytes at various stages of seed germination resultedin subsequent growth inhibition of the hypocotyl-radicle axis,with early removal of cotyledons suppressing most growth. Growthof primary needles appeared to be influenced indirectly by megagametophytereserves, probably by changes in amount of photosynthetic tissue.The embryo alone possessed capacity to differentiate such tissuesas primary needle primordia, stomata, and primary and secondaryvascular systems. Megagametophyte reserves appeared to contributeto growth of embryonic tissues only after the embryo itselfinitiated growth. Both current photosynthesis of seedlings andseed reserves contributed importantly to seedling development.     

Immunolocalization of FsPK1 correlates this abscisic acid-induced protein kinase with germination arrest in Fagus sylvatica L. seeds

An enzymatically active recombinant protein kinase, previously isolated and characterized in Fagus sylvatica L. dormant seeds (FsPK1), was used to obtain a specific polyclonal antibody against this protein. Immunoblotting and immunohistochemical analysis of FsPK1 protein in beech seeds showed a strong immunostaining in the nucleus of the cells located in the vascular tissue of the embryonic axis corresponding to the future apical meristem of the root. This protein kinase was found to accumulate in the seeds only when embryo growth was arrested by application of ABA, while the protein amount decreased during stratification, previously proved to alleviate dormancy, and no protein was detected at all when seed germination was induced by addition of GA(3). These results indicate that FsPK1 may be involved in the control of the embryo growth mediated by ABA and GAs during the transition from dormancy to germination in Fagus sylvatica seeds.     

桃儿七种子解剖结构及其萌发生长期形态特征

为探明种皮和胚乳是否是限制桃儿七种子萌发的主要因素,利用组织切片和显微技术,对桃儿七种子及其不同萌发期（1、7、14、21、28 d）解剖结构和播种后一定时期内（7~210 d）的植株生长形态进行观察。桃儿七种子由种皮、胚乳和胚构成。种皮包括外种皮和内种皮,外种皮致密规整,由外至内分别为栅状石细胞和表皮层细胞,内种皮由5~6层海绵细胞组成。胚乳占种子体积的绝大部分,包括珠孔胚乳和外胚乳。胚由胚根、胚轴和子叶组成,被致密种皮、多层珠孔胚乳和外胚乳包围。萌发期1~7 d胚根和胚轴开始伸长,7~14 d两片子叶分离,14~21 d胚根突破珠孔胚乳和种皮,21~28 d胚根、胚轴和子叶继续扩张伸长。种子播种210 d后可平均形成3片功能真叶和5条不定根。致密种皮（物理休眠）和多层胚乳（机械休眠）是限制桃儿七种子萌发的两个主要因素。