Gametophyte development and reproduction of the Asian fern Polystichum polyblepharum (Roem. ex Kunze) C. Presl (Dryopteridaceae,Polypodiopsida)

Abstract

The gametophytic generation of Polystichum polyblepharum (Dryopteridaceae), including spore germination, morphological development of the gametophytes, major vegetative features and reproduction strategies, was studied. Spore germination was of the Vittaria model and the developmental pattern was of the Aspidium model. Adult gametophytes were cordate and hairy, with unicellular hairs located at the margins and at the prothallus surface. The marginal ones were secretory. The gametophytes produced archegonia located in the central area between the notch and the rhizoids, but antheridia were never detected. Archegonia were of the normal type described for leptosporangiate ferns. Apogamous sporophytes appeared from a cellular outgrowth developed just under the apical notch. Initially, the outgrowth appeared completely surrounded by glandular hairs and scales, which persisted throughout the subsequent stages of leaf development.     

Alternation of generations – unravelling the underlying molecular mechanism of a 165‐year‐old botanical observation

Characteristically, land plants exhibit a life cycle with an ‘alternation of generations’ and thus alternate between a haploid gametophyte and a diploid sporophyte. At meiosis and fertilisation the transitions between these two ontogenies take place in distinct single stem cells. The evolutionary invention of an embryo, and thus an upright multicellular sporophyte, in the ancestor of land plants formed the basis for the evolution of increasingly complex plant morphologies shaping Earth's ecosystems. Recent research employing the moss Physcomitrella patens revealed the homeotic gene BELL1 as a master regulator of the gametophyte‐to‐sporophyte transition. Here, we discuss these findings in the context of classical botanical observations.     

Influence of metabolic stress on the inheritance of cell determination in the moss, Pottia intermedia

Epigenetically-determined apogamy in aposporous regenerants of the moss Pottia intermedia persists during vegetative propagation, the capacity of apogamy being inherited by individual aposporous protonemal cells. To test Bauer-Lazarenko's proposal that stable apogamy in mosses may be due to some self-replicating cytoplasmic factor, the effects of different metabolic stress treatments on the expression of apogamy have been tested. Chronic metabolic stress caused by long-term growth of autotrophic aposporous protonema on mineral medium with 0.25% of casamino acids and on Murashiga-Skoog (MS) medium with sucrose and phytohormones, as well as by transitory action of high kinetin concentration, have a much stronger influence on the expression of apogamy, than short-term stress treatments with RNase and Pb(2+). Apogamy has been found to be lost stably, after prolonged growth on MS medium containing kinetin and ABA. The proposal that the capacity for apogamy is related to the release of aposporous protonemal cells from a putative factor for apogamy is discussed.     

In vitro regeneration patterns of Platycerium bifurcatum leaf cell suspension culture

A simple suspension culture system of Platycerium bifurcatum was developed where sporophytes could be regenerated directly from leaf cells or indirectly through an aposporous gametophyte stage under the same culture conditions. Single cells and aggregates of up to 100 cells developed aposporous gametophytes which later gave rise to sporophytes. Such gametophytes started apogamy when they were mostly less than 0.7 mm in length, bearing only rhizoids. In most cases, only one sporophyte was regenerated from one gametophyte. Aggregates of 500–1000 or more cells, on the other hand, regenerated sporophytes directly. Intercellular interaction was considered to be the physiological cause, and the separation of leaf cells to a certain degree drove the cells to embark on different regeneration paths. It is suggested that the possible existence of a threshold size of cell aggregates separates the two regeneration patterns. Received: 3 March 1997 / Revision received: 11 April 1997 / Accepted: 3 June 1997     

Reproduction and Hybridisation in the Genus Rosa,Section Caninae (Ser.) Rehd.

Crossing experiments were carried out in order to investigate the importance of apogamy in relation to autogamy, geitonogamy and xenogamy in the reproductive biology of pentaploid members of the genus Rosa, section Caninae (Ser.) Rehd. About 10,000 seeds from 900 crossing experiments were examined, 4500 of them for both filling of the grain and viability of the embryo. The results were compared with experimental reports and literature dating from 1753 to 1995. Apomictic reproduction was shown to be possible in all species of the section tested, but viable seed yields were only about 5% of those produced by xenogamy. This contradicts the hypothesis of Fagerlind (1940,1944,1945,1950) and Custafsson and Hakansson (1942), but supports the results of Matsson (1912) and Schwertschlager (1915). The number of viable seeds increased in the order apogamy, autogamy, geitonogamy to xenogamy. Reciprocal crossings showed significant differences in the number of seeds produced in the hip. This contradicts the hypothesis on the genomic constitution of the pentaploid Caninae with unbalanced heterogamy, as advocated by Hurst (1927) and Custafsson and Hakansson (1942).     

Ultrastructural and cytochemical aspects of induced apogamy following abscisic acid pre-treatment of secondary moss protonema

Abscisic acid (ABA) treatment of secondary protonema of Physcomitrium pyriforme Brid in the presence of sucrose does not prevent cell division but results in shorter cells with vesicular cytoplasm and an accumulation of lipid. When transferred to sucrose medium without ABA and with low irradiance isodiametric intercalary cells are cut off which give rise to apogamous sporophytes either directly or after the formation of a small amount of callus. The organization of the cells leading up to the apogamous sporophyte is described. The cells initiating the sporophyte develop dense cytoplasm and the walls become labyrinthine and callosed, but they do not form any recognizable placenta. It is proposed that labyrinthine walls are a consequence of a perturbation of cell wall metabolism as growth changes from gametophytic to sporophytic. The use of the term transfer cell for this kind of cell is questioned and the need for a causal approach to the investigation of labyrinthine walls is stressed.     

Exogenous and endogenous growth regulators on apogamy in Dryopteris affinis (Lowe) Fraser-Jenkins sp. affinis

This work showed for the first time the relationship between the effect of exogenous auxins and gibberellins on apogamy in Dryopteris affinis (Lowe) Fraser-Jenkins sp. affinis and its endogenous contents during early apogamic events. The addition of NAA (0.53 and 5.37 μM) or GA₃ 2.8 μM to an MS solid medium significantly increased apogamous sporophyte formation. BA induced brown callus that regenerated sporophytes in a hormone-free medium. The endogenous contents of GA₁, GA₃, GA₄, GA₇, GA₉ and IAA were determined by GC–MS in gametophytes cultured on MS solid medium, before and during early stages of apogamous embryo development. The accumulation of both GA₉ and IAA before embryo development was evident as high levels of GA₄ in the earliest analysed stage of embryo development and high levels of GA₃ in elongating shoots were found. The role of gibberellins on apogamy was also supported by data showing a decrease in the percentage of gametophytes developing embryos because of the addition of flurprimidol to the culture medium.     

Electrophoretic evidence for homoeologous chromosome pairing in the apogamous fern species<Emphasis Type="Italic"> Dryopteris nipponensis</Emphasis> (Dryopteridaceae)

Segregation of genotypes through homoeologous chromosome pairing in the apogamous species Dryopteris nipponensis was tested by electrophoretic analysis. Of 284 progeny examined (250 gametophytes and 34 sporophytes), from the parental sporophyte with the Pgi-2 genotype abc, five showed different genotypes from that of the parent (three aac, one bbc and one bcc). This is the first evidence for genetic segregation in the progeny of apogamous fern species. Electronic Publication     

Cytological and Reproductive Studies on Japanese Diplazium (Woodsiaceae; Pteridophyta): Apomictic Reproduction in Diplazium with Evergreen Bi- to Tripinnate Leaves

Diplazium , including polymorphic terrestrial species with evergreen bi- to tripinnate leaves. Diplazium hachijoense, D. virescens var. virescens, var. conterminum, var. okinawaense, and two other unnamed varieties, D. kawakamii var. kawakamii, D. dilatatum var. heterolepia, D. taiwanense, D. × kawabatae (=D. dilatatum × taiwanense), D. × takii (=D. hachijoense × virescens var. virescens), and D.× nakamurae (= D. hachijoense × virescens var. conterminum) are apomictic triploids (2n=n=123). Diplazium amamianum and D. esculentum are sexual diploids (2n=82, n=41) and D. subtripinnatum is a sexual tetraploid (2n= 164, n=82). Diplazium dilatatum var. dilatatum includes both sexual diploid and apomictic triploid populations. Cultivated gametophytes of six triploid taxa produced sporophytes apogamously, confirming their apomictic reproduction. All three putative hybrids, D. × kawabatae, D. × takii, and D. × nakamurae, are triploid, apomictic, and fertile taxa, therefore they are not the result of hybridization between known pairs of Japanese Diplazium plants. Received 16 March 1999/ Accepted in revised form 30 September 1999