Inefficient photosynthesis in the Mediterranean orchid Limodorum abortivum is mirrored by specific association to ectomycorrhizal Russulaceae

Among European Neottieae, Limodorum abortivum is a common Mediterranean orchid. It forms small populations with a patchy distribution in woodlands, and is characterized by much reduced leaves, suggesting a partial mycoheterotrophy. We have investigated both the photosynthetic abilities of L. abortivum adult plants and the diversity of mycorrhizal fungi in Limodorum plants growing in different environments and plant communities (coniferous and broadleaf forests) over a wide geographical and altitudinal range. Despite the presence of photosynthetic pigments, CO2 fixation was found to be insufficient to compensate for respiration in adult plants. Fungal diversity was assessed by morphological and molecular methods in L. abortivum as well as in the related rare species Limodorum trabutianum and Limodorum brulloi. Phylogenetic analyses of the fungal internal transcribed spacer (ITS) sequences, obtained from root samples of about 80 plants, revealed a tendency to associate predominantly with fungal symbionts of the genus Russula. Based on sequence similarities with known species, most root endophytes could be ascribed to the species complex encompassing Russula delica, Russula chloroides, and Russula brevipes. Few sequences clustered in separate groups nested within Russula, a genus of ectomycorrhizal fungi. The morphotypes of ectomycorrhizal root tips of surrounding trees yielded sequences similar or identical to those obtained from L. abortivum. These results demonstrate that Limodorum species with inefficient photosynthesis specifically associate with ectomycorrhizal fungi, and appear to have adopted a nutrition strategy similar to that known from achlorophyllous orchids.     

Germination and growth ofErythrorchis ochobinesis (Orchidaceae) accelerated by monokaryons and dikaryons ofLenzites betulinus andTrametes hirsuta

Four sib-monokaryons and two reconstituted dikaryons of two basidiomycetes,Lenzites betulinus andTrametes hiruta, accelerated the seed germination ofErythrorchis ochobiensis, an achlorophyllous orchid. All isolates ofL. betulinus and three isolates ofT. hirsuta induced the development of plants from germinated seeds. Although three monokaryotic isolates ofT. hirsuta failed to induce the development of plants, the reconstituted dikaryons induced the development.     

Importance of woody debris in seed germination of Tipularia discolor (Orchidaceae)

Concerns about declining populations of terrestrial orchids make it important to identify the environmental factors crucial to seedling recruitment. This study shows that seedlings of Tipularia discolor (cranefly orchid) primarily occur on decomposing wood. Extensive searches of decomposing logs and stumps in mature and successional forests revealed seedlings at 24 sites, of which 15 could be identified as originating from seven different deciduous trees and one conifer. Seeds were planted in natural habitats to test the hypothesis that germination requires decomposing wood. In one experiment, seeds were placed into soil at sites where adult plants were abundant; no germination resulted. In a second experiment, germination of seeds sown in ambient soil was compared with sowings in plots amended with decomposing wood collected from a stump where spontaneous seedlings grew. Germination was much more frequent in plots amended with decomposing wood. We conclude that germination of T. discolor is stimulated in substrates that contain decomposing wood; judging from the occurrence of spontaneous seedlings, wood from at variety of tree species offer a suitable substrate.     

Can orchid mycorrhizal fungi be persistently harbored by the plant host?

The environmental distribution of non-obligate orchid mycorrhizal (OM) symbionts belonging to the ‘rhizoctonia’ complex remains elusive. Some of these fungi, indeed, are undetectable in soil outside the host rhizosphere. A manipulation experiment was performed to assess the importance of neighbouring non-orchid plants and soil as possible reservoirs of OM fungi for Spiranthes spiralis, a widespread photosynthetic European terrestrial orchid species. Fungi of S. spiralis roots were identified by DNA metabarcoding before and 4 months after the removal of the surrounding vegetation and soil. Although such a treatment significantly affected fungal colonization of newly-formed orchid roots, most OM fungi were consistently associated with the host roots. Frequency patterns in differently aged roots suggest that these fungi colonize new orchid roots from either older roots or other parts of the same plant, which may thus represent an environmental source for the subsequent establishment of the OM symbiosis.     

Symbiotic germination and development of myco-heterotrophic plants in nature: ontogeny of Corallorhiza trifida and characterization of its mycorrhizal fungi

The processes of symbiotic germination and seedling development were analysed in the myco-heterotrophic orchid Corallorhiza trifida , seeds of which were buried in 'packets' either adjacent to or at varying distances from adult plants in defined communities of ectomycorrhizal tree species. Germination occurred within eight months of burial under Betula – Alnus and within seven months under Salix repens . It was always associated with penetration of the suspensor by a clamp-forming mycorrhizal fungus. Four distinct developmental stages were defined and the rates of transition through these stages were plotted. There was no evidence of a relationship between extent of germination or rate of development and the presence of naturally distributed plants of C. trifida at the spatial scale of 1 m. The best germination and the most rapid rate of development of C. trifida seedlings occurred in a Salix repens community located at a considerable distance from any extant C. trifida population. Determination of internal transcribed spacer (ITS) RFLPs and of gene sequences of the fungi involved in symbiotic germination and growth of C. trifida , revealed them to belong exclusively to the Thelephora – Tomentella complex of the Thelephoraceae. These fungi are known also to be ectomycorrhizal associates of trees. It is hypothesized that the rate of growth of the C. trifida seedlings is determined by the ability of the fungal symbionts to transfer carbon from their ectomycorrhizal co-associates.     

Sulla Struttura del Frutto della “Stubborn” L

Novel methods are required to break the seed dormancy of temperate-zone orchids and aid the conservation of rare species. Zeatin is produced in increasing concentrations during the development of pea (Pisum sativum) seeds. We hypothesised that hot water extracts of pea seeds stimulate germination of the orchid Ophrys apifera in vitro, particularly for extractions made during later pea development. Pea seeds, exposed to 0, 3, 6, 9, 12 or 15 days of periodic wetting, were extracted in hot water and the extracts were added to Malmgren's growth medium. Germination of O. apifera on this medium was quantified after 7 months, stimulated by a range of pure hormones. Pea seed extract collected later in pea development (at 6–15 days) inhibited germination of O. apifera. However, extracts taken at 0 and 3 days significantly increased germination from 3.8 ± 0.32% in the control to 9.1 ± 1.84% and 7.6 ± 0.79%, respectively: increases comparable to the most effective of the pure hormones. Dried peas therefore provide an economical alternative source of germination stimulants for orchids. We briefly report how sufficient mature plants of O. apifera were produced to allow a population to be multiplied to 15 times its original size during a habitat restoration project.     

菌根真菌提高植物抗旱性机制的研究回顾与展望

菌根真菌与全世界约97％的维管植物具有广泛的共生关系.大量研究结果显示菌根植物相比于非菌根植物对于干旱胁迫具有更高的耐受性,说明菌根真菌在植物抗旱过程中发挥着重要作用.本文对近年来国内外在菌根真菌协助植物抵御干旱作用机制方面的研究进行了归纳和总结,主要包括在干旱胁迫下菌根真菌对植物生理学特性的影响机制、菌根真菌提高植物...     

Spatial repartition and genetic relationship of green and albino individuals in mixed populations of Cephalanthera orchids

Several green orchids of the Neottieae tribe acquire organic carbon both from their mycorrhizal fungi and from photosynthesis. This strategy may represent an intermediate evolutionary step towards mycoheterotrophy of some non‐photosynthetic (albino) orchids. Mixed populations of green and albino individuals possibly represent a transient evolutionary stage offering opportunities to understand the evolution of mycoheterotrophy. In order to understand the emergence of albinos, we investigated patterns of spatial and genetic relationships among green and albino individuals in three mixed populations of Cephalanthera damasonium and one of C. longifolia using spatial repartition and Amplified fragment length polymorphism (AFLP) markers. Two of these populations were monitored over two consecutive flowering seasons. In spatial repartition analyses, albino individuals did not aggregate more than green individuals. Genetic analyses revealed that, in all sampled populations, albino individuals did not represent a unique lineage, and that albinos were often closer related to green individuals than to other albinos from the same population. Genetic and spatial comparison of genets from the 2‐year monitoring revealed that: (i) albinos had lower survival than green individuals; (ii) accordingly, albinos detected in the first year did not correspond to the those sampled in the second year; and (iii) with one possible exception, all examined albinos did not belong to any green genet from the same and/or from the previous year, and vice versa. Our results support a scenario of repeated insurgence of the albino phenotypes within the populations, but unsuccessful transition between the two contrasting phenotypes. Future studies should try to unravel the genetic and ecological basis of the two phenotypes.     

Cytoskeleton in mycorrhizal symbiosis

An understanding of the role played by the cytoskeleton in formation and function of mycorrhizas has been hampered by the technical difficulty of working with mycorrhizal material. Recently, however, improved labelling techniques suitable for both plant and fungal symbionts in combination with either epifluorescence microscopy or laser scanning confocal microscopy have resulted in new information. As well, molecular methods have made it possible to monitor changes of cytoskeletal elements during mycorrhiza development. Currently we know that the cytoskeletal systems of both plant and fungal partners undergo changes during both ecto- and endomycorrhizal symbiosis. However, little information is available concerning the regulatory factors or the cause and effect relationship of cytoskeletal changes and cellular events. In this article, research involving the cytoskeleton of mycorrhizas is reviewed in detail, whereas basic information of the cytoskeleton of plant and fungal cells is only briefly discussed as background. A brief comparison is also made between the information on mycorrhizas with that of biotrophic pathogenic fungi and the Rhizobium–legume symbiosis.     

Coelonin,A 9,10-dihydrophenanthrene from the orchids Coelogyne ochracea and Coelogyne elata

2,7-Dihydroxy-4-methoxy-9,10-dihydrophenanthrene was isolated and identified from the whole plant of Coelogyne ochracea and C. elata.     

Identity and specificity of the fungi forming mycorrhizas with the rare mycoheterotrophic orchid Rhizanthella gardneri

Fully subterranean Rhizanthella gardneri (Orchidaceae) is obligately mycoheterotrophic meaning it is nutritionally dependent on the fungus it forms mycorrhizas with. Furthermore, R. gardneri purportedly participates in a nutrient sharing tripartite relationship where its mycorrhizal fungus simultaneously forms ectomycorrhizas with species of Melaleuca uncinata s.l. Although the mycorrhizal fungus of R. gardneri has been morphologically identified as Thanatephorus gardneri (from a single isolate), this identification has been recently questioned. We sought to clarify the identification of the mycorrhizal fungus of R. gardneri, using molecular methods, and to identify how specific its mycorrhizal relationship is. Fungal isolates taken from all sites where R. gardneri is known to occur shared almost identical ribosomal DNA (rDNA) sequences. The fungal isolate rDNA most closely matched that of other Ceratobasidiales species, particularly within the Ceratobasidium genus. However, interpretation of results was difficult as we found two distinct ITS sequences within all mycorrhizal fungal isolates of R. gardneri that we assessed. All mycorrhizal fungal isolates of R. gardneri readily formed ectomycorrhizas with a range of M. uncinata s.l. species. Consequently, it is likely that R. gardneri can form a nutrient sharing tripartite relationship where R. gardneri is connected to autotrophic M. uncinata s.l. by a common mycorrhizal fungus. These findings have implications for better understanding R. gardneri distribution, evolution and the ecological significance of its mycorrhizal fungus, particularly in relation to nutrient acquisition.