Microinjection of heme oxygenase genes rescues phytochrome-chromophore-deficient mutants of the moss Ceratodon purpureus

 In protonemal tip cells of the moss Ceratodon purpureus (Hedw.) Brid., phototropism and chlorophyll accumulation are regulated by the photoreceptor phytochrome. The mutant ptr116 lacks both responses as a result of a defect in the biosynthesis of phytochromobilin, the chromophore of phytochrome, at the point of biliverdin formation. The rescue of the phototropic response and of chlorophyll synthesis were tested by injecting different substances into tip cells of ptr116. Microinjection was first optimised with the use of fluorescent dyes and an expression plasmid containing a green fluorescent protein (GFP) gene. Injected phycocyanobilin, which substitutes for phytochromobilin, rescued both the phototropic response and light-induced chlorophyll accumulation in ptr116. The same results were obtained when expression plasmids with heme oxygenase genes of rat (HO-1) and Arabidopsis thaliana (L.) Heynh. (HY1) were injected. Heme oxygenase catalyses the conversion of heme into biliverdin. Whereas HY1 has a plastid target sequence and is presumably transferred to plastids, HO-1 is proposed to be cytosolic. The data show that ptr116 lacks heme oxygenase enzyme activity and indicate that heme oxygenases of various origin are active in Ceratodon bilin synthesis. In addition, it can be inferred from the data that the intracellular localisation of the expressed heme oxygenase is not important since the plastid enzyme can be replaced by a cytosolic one. Received: 8 March 1999 / Accepted: 30 July 1999     

From axenic spore germination to molecular farming

The first bryophyte tissue culture techniques were established almost a century ago. All of the techniques that have been developed for tissue culture of seed plants have also been adapted for bryophytes, and these range from mere axenic culture to molecular farming. However, specific characteristics of bryophyte biology—for example, a unique regeneration capacity—have also resulted in the development of methodologies and techniques different than those used for seed plants. In this review we provide an overview of the application of in vitro techniques to bryophytes, emphasising the differences as well as the similarities between bryophytes and seed plants. These are discussed within the framework of physiological and developmental processes as well as with respect to potential applications in plant biotechnology.     

Classification and ordination of cryptogamic communities in Wilkes Land,Continental Antarctica

The entirely cryptogamic vegetation of Bailey and Clark Peninsulas, Windmill Islands, Budd Coast, Wilkes Land, Antarctica, is described for the first time. The vegetation of this area is exceptionally well developed and diverse and represents one of the most important botanical sites on the continent. The macroflora comprises three species of moss, one liverwort, three fruticose lichens, four foliose and over 20 crustose lichens; several macroalgae also occur. Seventy stands of relatively homogeneous vegetation were analysed and the percentage cover afforded by every species within 20 quadrats per site was recorded. A subjective classification was developed by visual ordering of the data sets and a hierarchical system erected which incorporates one moss- and one lichen-dominated sub-formation; the former includes two associations and seven sociations, while the latter comprises one association which includes four sociations. The data were then arranged by centroid linkage analysis to produce and objective classification, and subsequently ordinated by principal components analysis to generate groups of stands, the inter-relations of which were interpreted in ecological and environmental terms. The objective classification and ordination strongly support the subjectively derived groupings or sociations. Examples of plant interactions are qualitatively described.     

Osmotic stress effects on the freezing tolerance of the antarctic nematodePanagrolaimus davidi

 The freezing and freezing survival of the Antarctic nematode Panagrolaimus davidi after exposure to solutions of different osmotic concentrations has been examined using a thermoelectric cooling stage and multi-specimen cooling block to see if there is any evidence that freeze-induced desiccation prevents inoculative freezing. The nematodes froze in all the test solutions used (up to 1138 mosmol ⋅ l^-1) and at all cooling rates and nucleation temperatures tested. Freezing survival was at its maximum in 0.1 mol ⋅ l^-1 NaCl in artificial tap water after 1 h exposure to the test solution and in artificial tap water after 24 h exposure. Hyperosmotic and hyposmotic stress adversely affected the nematodes’ ability to survive freezing. In non-frozen controls survival declined with increasing osmolality of the test solution. Measurements of the osmolality of water extracted from a variety of moss samples indicate that the nematodes are exposed to an osmotic concentration of about 9 mosmol ⋅ l^-1 in their natural habitat. This is close to that of artificial tap water. Our experiments, and measurements of freeze concentration effects in the literature, indicate that freeze-induced desiccation is unlikely to prevent inoculative freezing and the survival of nematodes over the winter. Accepted: 5 May 1996     

Filial mistletoes: the functional morphology of moss sporophytes

Background

A moss sporophyte inherits a haploid set of genes from the maternal gametophyte to which it is attached and another haploid set of genes from a paternal gametophyte. Evolutionary conflict is expected between genes of maternal and paternal origin that will be expressed as adaptations of sporophytes to extract additional resources from maternal gametophytes and adaptations of maternal gametophytes to restrain sporophytic demands.

Interpretation

The seta and stomata of peristomate mosses are interpreted as sporophytic devices for increasing nutrient transfer. The seta connects the foot, where nutrients are absorbed, to the developing capsule, where nutrients are needed for sporogenesis. Its elongation lifts stomata of the apophysis above the boundary layer, into the zone of turbulent air, thereby increasing the transpirational pull that draws nutrients across the haustorial foot. The calyptra is interpreted as a gametophytic device to reduce sporophytic demands. The calyptra fits tightly over the intercalary meristem of the sporophytic apex and prevents lateral expansion of the meristem. While intact, the calyptra delays the onset of transpiration.

Predictions

Nutrient transfer across the foot, stomatal number and stomatal aperture are predicted to be particular arenas of conflict between sporophytes and maternal gametophytes, and between maternal and paternal genomes of sporophytes.     

Chlorophyll concentrations and a/b ratios in mosses collected from exposed and shaded habitats in Kansas

Abstract

Chlorophyll concentrations and a/b ratios of 31 moss species in 16 families collected from various habitats in Kansas were lower than those of most higher vascular plants. Most of the mosses examined had higher chlorophyll concentrations and lower chlorophyll a/b ratios when collected from habitats with low solar irradiance, relative to mosses collected from habitats characterized by high irradiance. This was true for mosses collected from under a forest canopy compared with those collected from exposed sandstone outcrops, and for the same populations of mosses collected before and after forest canopy closure. Such changes in the characteristics of the light-harvesting pigment system appear to represent adaptations enhancing the efficiency of light capture.     

potamolejeunea Holtii (Spruce) nov.comb.

Abstract

Bryophytes usually have anti-feeding properties to defend against microbial and herbivore attack; however, the consumption of Haplocladium microphyllum (Hedw.) Broth. capsules by Agrotis sp. larvae is rather common in Shanghai in the spring. To test whether H. microphyllum is the only moss eaten, and why the gametophytes of H. microphyllum are not eaten, a series of quantitative experiments were carried out to understand the feeding habits of Agrotis larvae on the given moss materials at three growth stages of larval life. The results show that the larvae can feed on the capsules of six moss species to different degrees: Funaria hygrometrica Hedw., H. microphyllum, Physcomitrium sphaericum (C.F.Ludw. ex Schkuhr) Brid., Trematodon longicollis Michx., Ditrichum pallidum (Hedw.) Hampe, and Pogonatum inflexum (Lindb.) Sande Lac. The capsules of the first four species were grazed heavily by the larvae, compared with limited consumption of the latter two (D. pallidum and P. inflexum), which even induced a high mortality rate among the larvae. With the growth of the larval instar, the daily demand for moss capsules by the larvae increases gradually. The lipid content of the capsules may play an important role when the larva selects its feeding target.     

Seasonal impact of waterfowl on communities of macrophytes in a shallow lake

From 2005 to 2007, we established bird-proof enclosures in a small, shallow and semi-permanent lake, lacking fish, at Brown Moss, Shropshire, UK, to investigate the effects of aquatic birds on seasonal growth of submerged and emergent macrophytes. The highest density of birds on the lake was in winter (110 individuals ha⁻¹) and the lowest in summer 2005 (6 ha⁻¹). Plant growth varied with season but there were significantly different (F = 8.03, p < 0.05, df = 1) standing crops of macrophytes between bird-proof enclosures (proportion of volume occupied, 0.47 ± 0.04) and control treatments (0.36 ± 0.11). Different densities of birds occurred in different areas and this was reflected in their effects. Ducks, mainly mallard (Anas platyrhynchos, Linnaeus), and teal (Anas crecca, Linnaeus), damaged plants by direct consumption, uprooting and trampling, whereas larger birds, such as mute swan (Cygnus olor, Gmelin), were able to remove Typha latifolia (Linnaeus). In summer, grazing pressure was reduced as the population of birds declined. Waterfowl caused seasonal impacts on the re-development of the water plant community. However, waterfowl herbivory had low potential to shift a macrophyte-dominated state into a phytoplankton-dominated state because aquatic plants could recover, during the growing season, when bird populations declined.     

Clues about the ancestral roles of plant MADS-box genes from a functional analysis of moss homologues

Classic MIKC-type MADS-box genes (MIKC ^c genes) are indispensable elements in the genetic programming of pattern formation, including the segmental organisation of angiosperm flowers, in seed plants. Since little is known about the functions of MIKC ^c genes in non-seed plants, a functional analysis of moss MIKC ^c homologues was performed using the genetically amenable, simple model plant, Physcomitrella patens. Expression of moss homologues was knocked down using an antisense RNA approach or abolished by generating transformants with gene knockouts. The knocked down (“antisense”) transformants displayed a multifaceted mutant phenotype comprising delayed gametangia formation, diminished sporophyte yield and, in the most extremely affected cases, abnormal sporophyte development and altered leaf morphogenesis. Knocked out transformants were phenotypically normal. Analysis of in situ MIKC ^c gene expression using transgenic strains containing MIKC ^c promoter–GUS fusions showed that these genes are generally expressed ubiquitously in vegetative and reproductive tissues. We conclude that MIKC ^c genes play significant roles in morphogenetic programming of the moss. Functional redundancy characterises some members of the gene group. Our findings provide clues concerning the ancestral roles of some MIKC ^c genes that may be represented in the genomes of diverse extant plant taxa. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.