Seed fertilization, development, and germination in Hydatellaceae (Nymphaeales): Implications for endosperm evolution in early angiosperms

New data on endosperm development in the early-divergent angiosperm Trithuria (Hydatellaceae) indicate that double fertilization results in formation of cellularized micropylar and unicellular chalazal domains with contrasting ontogenetic trajectories, as in waterlilies. The micropylar domain ultimately forms the cellular endosperm in the dispersed seed. The chalazal domain forms a single-celled haustorium with a large nucleus; this haustorium ultimately degenerates to form a space in the dispersed seed, similar to the chalazal endosperm haustorium of waterlilies. The endosperm condition in Trithuria and waterlilies resembles the helobial condition that characterizes some monocots, but contrasts with Amborella and Illicium, in which most of the mature endosperm is formed from the chalazal domain. The precise location of the primary endosperm nucleus governs the relative sizes of the chalazal and micropylar domains, but not their subsequent developmental trajectories. The unusual tissue layer surrounding the bilobed cotyledonary sheath in seedlings of some species of Trithuria is a belt of persistent endosperm, comparable with that of some other early-divergent angiosperms with a well-developed perisperm, such as Saururaceae and Piperaceae. The endosperm of Trithuria is limited in size and storage capacity but relatively persistent.     

A new type of specialized morphophysiological dormancy and seed storage behaviour in Hydatellaceae, an early-divergent angiosperm family

Background and Aims

Recent phylogenetic analysis has placed the aquatic family Hydatellaceae as an early-divergent angiosperm. Understanding seed dormancy, germination and desiccation tolerance of Hydatellaceae will facilitate ex situ conservation and advance hypotheses regarding angiosperm evolution.

Methods

Seed germination experiments were completed on three species of south-west Australian Hydatellaceae, Trithuria austinensis, T. bibracteata and T. submersa, to test the effects of temperature, light, germination stimulant and storage. Seeds were sectioned to examine embryo growth during germination in T. austinensis and T. submersa.

Key Results

Some embryo growth and cell division in T. austinensis and T. submersa occurred prior to the emergence of an undifferentiated embryo from the seed coat (‘germination’). Embryo differentiation occurred later, following further growth and a 3- to 4-fold increase in the number of cells. The time taken to achieve 50 % of maximum germination for seeds on water agar was 50, 35 and 37 d for T. austinensis, T bibracteata and T. submersa, respectively.

Conclusions

Seeds of Hydatellaceae have a new kind of specialized morphophysiological dormancy in which neither root nor shoot differentiates until after the embryo emerges from the seed coat. Seed biology is discussed in relation to early angiosperm evolution, together with ex situ conservation of this phylogenetically significant group.     

Pollen tube development in two species of Trithuria (Hydatellaceae) with contrasting breeding systems

Trithuria (Hydatellaceae; Nymphaeales) is unique among early-divergent angiosperms in that its species are extremely small and most have exceptionally short, annual life histories. Given the evolution of these extremes of size and development, we sought to understand whether post-pollination processes still varied predictably with breeding system in Trithuria. To address this question, we studied two Western Australian species, Trithuria austinensis (dioecious, obligately outcrossing) and Trithuria submersa (bisexual, highly selfing). To document developmental timing, carpels were hand-pollinated, collected at sequential time points, and examined with light and fluorescence microscopy. In both species, pollen tubes first entered ovules<1 h after pollination, but the pollen tube pathway of outcrossing T. austinensis was almost four times longer and its pollen tube growth rates were up to six times faster (≤2,166 vs. 321 μm/h) than those of T. submersa. T. austinensis also exhibited greater male investment, slower pollen germination, and greater pollen tube attrition. These differences in male gametophyte development are predicted for outcrossers versus selfers in phylogenetically derived angiosperms. These new data for Hydatellaceae reinforce the idea that an acceleration of pollen tube development occurred in the Nymphaeales stem lineage, before the origin of Hydatellaceae. We infer that a recent evolutionary transition to selfing in T. submersa has been accompanied by predictable modifications to reproductive development, which, because of the ancient relationship between Hydatellaceae and all other angiosperms, suggests that traits underlying the lability of flowering plant post-pollination biology were present early in their history.     

Nonflowers near the base of extant angiosperms? Spatiotemporal arrangement of organs in reproductive units of Hydatellaceae and its bearing on the origin of the flower

Reproductive units (RUs) of Trithuria, the sole genus of the early-divergent angiosperm family Hydatellaceae, are compared with flowers of their close relatives in Cabombaceae (Nymphaeales). Trithuria RUs combine features of flowers and inflorescences. They differ from typical flowers in possessing an "inside-out" morphology, with carpels surrounding stamens; furthermore, carpels develop centrifugally, in contrast to centripetal or simultaneous development in typical flowers. Trithuria RUs could be interpreted as pseudanthia of two or more cymose partial inflorescences enclosed within an involucre, but the bractlike involucral phyllomes do not subtend partial inflorescences and hence collectively resemble a typical perianth. Teratological forms of T. submersa indicate a tendency to fasciation and demonstrate that the inside-out structure-the primary feature that separates RUs of Hydatellaceae from more orthodox angiosperm flowers-can be at least partially modified, thus producing a morphology that is closer to an orthodox flower. The Trithuria RU could be described as a "nonflower", i.e., a structure that contains typical angiosperm carpels and stamens but does not allow recognition of a typical angiosperm flower. The term nonflower could combine cases of secondary loss of flower identity and cases of a prefloral condition, similar to those that gave rise to the angiosperm flower. Nonhomology among some angiosperm flowers could be due to iterative shifts between nonfloral construction and flower/inflorescence organization of reproductive organs. Potential testing of these hypotheses using evolutionary-developmental genetics is explored using preliminary data from immunolocalization of the floral meristem identity gene LEAFY in T. submersa, which indicated protein expression at different hierarchical levels.     

Restionaceae seedlings: Morphology,anatomy and systematic implications

Restionaceae differ from most monocot families in having both epigeal and hypogeal germination. The green cotyledons associated with epigeal germination have a central vascular strand as found in most epigeal monocotyledons. In some genera the cotyledon may have a hairpin‐like structure, also described for Anthericaceae. The cotyledon of the hypogeal seedlings is short, without green pigment and largely remains embedded in the seed coat. Hypogeal germination is correlated with large, woody, indehiscent, frequently myrmecochorous nuts, while epigeal germination is found in species with smaller indehiscent nutlets or seeds, dispersed in a variety of ways. The primitive condition is most likely epigeal germination. In hypogeal seedlings of some African and Australian taxa an epicotyledonary rhizome is found between the primary root and the first leaves. Seedlings of African Restionaceae frequently have elongated culm internodes, whereas in the Australian species studied, internodes are very short, resulting in a cluster of seedling leaves. The leaf blades, which in most species are only found on the seedlings, are very simple anatomically. However, they appear to be unifacial, similar to the leaf blades of Anarthria (Anarthriaceae). The anatomical specialisations in the blades mirror those recorded for the culm anatomy. These observations are consistent with the hypothesis that Centrolepidaceae may be neotonous Restionaceae. They also corroborate the morphology of the African Restionaceae, and the presently accepted phylogeny of the African genera of Restionaceae.     

莲幼苗的胚根发育及其筛分子研究

报道了莲无初生根型幼苗的胚根发育过程,其胚根由分生组织转化成贮藏组织,致使胚根不突破种皮;莲幼苗初生维管系统中的筛分子为筛管分子。莲的子叶和胚根连为一体的特殊结构与其生活环境和系统演化密切相关。     

DELLA-mediated cotyledon expansion breaks coat-imposed seed dormancy

Seed dormancy is a key adaptive trait in plants responsible for the soil seed bank. The long established hormone-balance theory describes the antagonistic roles of the dormancy promoting plant hormone abscisic acid (ABA), and the germination promoting hormone gibberellin (GA) in dormancy control. Light, temperature, and other dormancy-breaking signals function to modulate the synthesis and perception of these hormones in the seed. However, the way in which these hormones control dormancy in the imbibed seed remains unknown. Here, we show that the DELLA protein regulators of the GA response are required for dormancy and describe a model through which hormone signal integration and dormancy regulation is achieved. We demonstrate that cotyledon expansion precedes radicle emergence during Arabidopsis seed germination and that a striking correlation exists between final seedling cotyledon size and seed dormancy in the DELLA mutants. Furthermore, twelve previously characterized seed-dormancy mutants are also defective in the control of cotyledon size in a manner consistent with their effect on germination potential. We propose that DELLA-mediated, light-, temperature-, and hormone-responsive cotyledon expansion prior to radicle emergence overcomes dormancy imposed by the seed coat and underlies seed-dormancy control in Arabidopsis.     

Starch-accumulating (S-type) sieve-element plastids in Hydatellaceae: implications for plastid evolution in flowering plants

TEM investigation of sieve-element plastids in three species of Trithuria, the sole genus of the small aquatic family Hydatellaceae, show that P-type plastids are absent from this genus and only starch-accumulating (S-type) sieve-element plastids are present. This discovery is consistent with the recent transfer of Hydatellaceae from the highly derived monocot order Poales (grasses and their allies) to the early-divergent angiosperm order Nymphaeales (waterlilies) based on molecular phylogenetic data. Species of Poales consistently possess P2-subtype plastids, in common with other monocots, but only S-type plastids are present in Nymphaeales. The results confirm that Hydatellaceae do not belong in monocots. Optimisation of the two major types of sieve-element plastid onto a recent phylogeny of early-divergent angiosperms confirms that S-type is the primitive form and indicates that P-type sieve-element plastids have evolved more than once in angiosperms.     

Morphology of Hydatellaceae, an anomalous aquatic family recently recognized as an early-divergent angiosperm lineage

The family Hydatellaceae was recently reassigned to the early-divergent angiosperm order Nymphaeales rather than the monocot order Poales. This dramatic taxonomic adjustment allows comparison with other early-divergent angiosperms, both extant and extinct. Hydatellaceae possess some monocot-like features that could represent adaptations to an aquatic habit. Ecophysiological parallels can also be drawn from fossil taxa that are known from small achene-like diaspores, as in Hydatellaceae. Reproductive units of Hydatellaceae consist of perianthlike bracts enclosing several pistils and/or stamens. In species with bisexual reproductive units, a single unit resembles an "inside-out" flower, in which stamens are surrounded by carpels that are initiated centrifugally. Furthermore, involucre development in Trithuria submersa, with delayed growth of second whorl bracts, resembles similar delayed development of the second perianth whorl in Cabomba. Several hypotheses on the homologies of reproductive units in Hydatellaceae are explored. Currently, the most plausible interpretation is that each reproductive unit represents an aggregation of reduced unisexual apetalous flowers, which are thus very different from flowers of Nymphaeales. Each pistil in Hydatellaceae is morphologically and developmentally consistent with a solitary ascidiate carpel. However, ascidiate carpel development, consistent with placement in Nymphaeales, is closely similar to pseudomonomerous pistil development as in Poaes.     

The question of cotyledon homology in angiosperms

The flowering plants (Magnoliophyta) are separated into two large classes distinguished by the morphology of their embryos. The embryos of monocots (class Liliopsida) have a single terminal cotyledon, while the embryos of dicots (class Magnoliopsida) usually have two lateral cotyledons. The cotyledons of monocots and dicots also differ in form, and there are no true intermediates. In addition, the third leaf of Nymphaealean seedlings appears to be identical to the single cotyledon of monocots. From this it is concluded that the cotyledons of monocots and dicots are not homologous. In addition, dissimilarity of cotyledons and succeeding leaves in dicots, together with recent genetic studies, suggests that the two cotyledons of dicots are not homologous with the succeeding leaves of the same plant. This interpretation is consistent with the view that the Nymphaealean embryo’s third leaf is homologous to the first leaf (cotyledon) of monocots. Because dicotyledonous embryos are common among seed plants and are present in the Gnetopsids, the most likely scenario is that the dicots share a widespread seed plant symplesiomorphy and that the monocots have lost this character state. A less parsimonious hypothesis of monocotyledonous embryos as plesiomorphic for angiosperms is also discussed. Genetic analysis of early embryo development in a variety of vascular plants may be the only way to conclusively determine the evolutionary origin of the distinctive difference between monocot and dicot embryos.     

Ontogeny of the mycoheterotrophic species Afrothismia hydra (Burmanniaceae)

The complete ontogeny of the mycoheterotrophic Afrothismia hydra (Burmanniaceae) from seed to seed dispersal is presented. The oblong–ovoidal seeds are up to 0.7 mm long. They germinate with root tissue only, disrupting the seed coat and developing a primary ovoid root tubercle. At the proximal end of the tubercle, a second tubercle arises and further root initials indicate the sequential growth of more root tubercles with filiform extensions resulting in a small root aggregate. The seed coat often remains attached to this structure. When the root aggregate enlarges, a central axis to which all roots are connected becomes visible. This axis has a growth pole where new root tubercles arise. The same growth pole will later develop into a stem with scale leaves finally terminating in a flower. Flowers develop sympodially when the mature plant is only several centimetres long. After anthesis, the corolla tube disintegrates, leaving a pyxidium which opens by means of a peculiar elongating placenta, here called 'placentophore'. The placentophore elevates the placenta with attached seeds above the flowering level and is interpreted as an adaptation to ombrohydrochory. The reduction of hypocotyl, cotyledon and primary shoot is discussed with regard to the classical germination concepts of monocotyledons and with mycoheterotrophic dicotyledons.  © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society , 2008, 157 , 31–36.     

Embryology in Trithuria submersa (Hydatellaceae) and relationships between embryo, endosperm, and perisperm in early-diverging flowering plants

? Premise of the study: Despite their highly reduced morphology, Hydatellaceae bear the unmistakable embryological signature of Nymphaeales, including a starch-rich maternal perisperm and a minute biparental endosperm and embryo. The co-occurrence of perisperm and endosperm in Nymphaeales and other lineages of flowering plants, and their respective functions during the course of seed development and embryo germination, remain enigmatic. ? Methods: Development of the embryo, endosperm, and perisperm was examined histologically from fertilization through germination in flowers and fruits of Trithuria submersa. ? Key results: The embryo of T. submersa initiates two cotyledons prior to seed maturity/dormancy, and their tips remain in contact with the endosperm throughout germination. The endosperm persists as a single layer of cells and serves as the interface between the embryo and the perisperm. The perisperm contains carbohydrates and proteins, and functions as the main storage tissue. The endosperm accumulates proteins and aleurone grains and functions as a transfer cell layer. ? Conclusions: In Nymphaeales, the multiple roles of a more typical endosperm have been separated into two different tissues and genetic entities: a maternal perisperm (nutrient acquisition, storage, mobilization) and a minute biparental endosperm (nutrient transfer to the embryo). The presence of perisperms among several other ancient lineages of angiosperms suggests a modest degree of developmental and functional lability for the nutrient storage tissue (perisperm or endosperm) within seeds during the early evolution of flowering plants. Finally, we examine the evolutionary developmental hypothesis that, contrary to longstanding assumptions, an embryo-nourishing perisperm along with a minute endosperm may represent the plesiomorphic condition for flowering plants.     

伊贝母种子萌发和籽苗建立

在子叶出土萌发的植物类型中,伊贝母种子萌发过程是罕见的。萌发初期,子叶优先生长。首先伸长而突破种皮,接着长出地面,此后,胚根才开始生长并产生不定根。种子萌发后形成特殊籽苗。在地上部分,子叶变绿,成为第一生长季唯一的同化叶;在地下部分,上胚轴扁平化,芽鳞肉质化,于是形成小鳞茎。籽苗形态属于最简化的类型。伊贝母的籽苗与其营养更新苗相比,是很弱小的。     

蚕豆籽粒内的~(14)C同化物的运转和卸出动态

通过向蚕豆叶片饲喂~(14)CO_2,应用液闪和显微放射性自显影技术表明标记同化物经叶脉和果荚韧皮部筛管快速运输至蚕豆种皮。种皮吸收营养、生长,后期逐步降解、供养子叶。种皮内的两类维管束系统同时输送营养并卸出到种皮内侧的质外体空间里。种皮里的反向维管束韧皮部卸出以共质体方式为主。并提供养分供种皮生长,而大部分的同化物由正向完整维管束韧皮部的筛分子一传递细胞进行质外体方式卸出。膨大中的子叶在早期即已成为生理上十分活跃的库。它对标记同化物的摄入随时间进程而急剧上升。     

Morphogenesis in cucumber seedlings is negatively controlled by gravity

 Seedlings of most cucurbitaceous plants develop a peg (protuberance caused by cell outgrowth) on the transition zone between the hypocotyl and root. The peg is necessary for removing the seed coat after germination. In our spaceflight experiments on the STS-95 space shuttle, Discovery, we found that cucumber (Cucumis sativus L.) seedlings grown under microgravity conditions developed two pegs symmetrically at the transition zone. Thus, cucumber seedlings potentially develop two pegs and do not require gravity for peg formation itself, but on the ground the development of one peg is suppressed in response to gravity. This may be considered as negative control of morphogenesis by gravity. Received: 17 August 1999 / Accepted: 4 October 1999     

Seed germination time course and seedling development of Clintonia udensis Trautv. et Mey. (Uvulariaceae): a typical cool temperate woodland perennial in Japan

Seed germination time course and seedling development mechanisms of Clintonia udensis Trautv. et Mey. (Uvulariaceae) were investigated under experimental condition. Seed germination tests were carried out under four thermal regimes, i. e. 10, 15, 20, and 25°C, after seeds were harvested, and stored at 5°C in wet conditions for 6 months under light‐exposed or shaded conditions. Approximately 63% of all seeds produced had the potential to germinate beyond 4 years and 6 months. The developmental process after germination continued for over 2 years. Phase I: the radicle first breaks through the seed coat 2 years after fructification. Phase II: the radicle becomes much larger with a hypocotyle. Phase III: part of the cotyledon elongates over 20 mm. Phase IV: the plumule further develops in two steps, i. e. the plumule is first formed, while cotyledon is disappearing, and then the plumule appears with second and third radicles, growing with cotyledon.     

Seedling morphology of three mangrove species and its taxonomic implications

Seedling establishment is an important part of the life cycle of mangroves. Very early in the juvenile stage, they face adverse situations such as a variable and unstable substrate as well as a high-saline environment Therefore, the propagules should have some ability to float in saline water. Seedlings differ morphologically from the adult plants. Here, I examined the seedling growth stages in three species of mangrove:Aegialitis rotundifolia Roxb.,Aegiceras corniculatum (L) Blanco, andAvicennia marina (Forsk.) Vierh. that grow in the Sundarbans swamps of West Bengal. In all cases, fruits were the propagating units; seeds were inseparable from those fruits. Germination was rapid and required no dormancy period. The hypocotyi pierced the seed coat rather than emerging from the pericarp — a variation on the traditional viviparous type of germination. Seedlings then became exposed prior to abscission from parent plants. Based on germination patterns and differences in seedling morphology observed here, these species should be classified in a more relevant taxonomic group than was done previously.