An introduced ant enhances pollination in the non-photosynthetic plant Balanophora fungosa.

In a striking contrast, matK is one of the most rapidly evolving plastid genes and also one of the few plastid genes to be retained in all nonphotosynthetic plants examined to date. DNA sequences of this region were obtained from photosynthetic and nonphotosynthetic plants of Orobanchaceae and their relatives. The resulting plastid DNA phylogeny was congruent with that recently obtained from analyses of rps2 and provided much better resolution. This phylogeny was then used to examine the relative degrees of evolutionary constraint of both the matK gene and the non-protein-coding regions that flank it inside the trnK intron. The method of subtree contrasts was introduced to compare levels of constraint. matK has evolved with a low but significant level of constraint on its amino acid sequence in both photosynthetic and nonphotosynthetic plants. Constraint is greater in photosynthetic than in nonphotosynthetic plants of this group. Domain X, thought to contain the active site of the protein, is not significantly more constrained than the rest of the protein. The portions of the flanking regions that are thought to form paired stem structures also show constraint, but in this case, there is no significant difference in degree of constraint between photosynthetic and nonphotosynthetic plants.

Genomic DNA was isolated using three DNA extraction commercial kits and three CTAB-based methods for two non-photosynthetic plants, Balanophora japonica and Mitrastemon kanehirai. The quality of the isolated DNA was evaluated and subjected to following restriction enzyme digestions. All six procedures yielded DNA of sufficient quality for PCR, and the method described by Barnwell et al. (1998) performed well in isolating DNA from both species for restriction enzyme digestion. In addition, we succeeded to enrich plastid DNA content by using the methods depending on a high salt buffer to deplete nuclear material. The 'high salt' methods based on protocol presented by Milligan (1989) were able to increase plastid DNA effectively and significantly reduce nuclear DNA from M. kanehirai. The plastid DNA enrichment protocols are inexpensive and not time-consuming, and may be applicable to other non-photosynthetic plants.

Mycoheterotrophic plants are non-photosynthetic plants that obtain nutrients from fungi. Even though most of these plants are associated with the mycorrhizal partners of surrounding photosynthetic plants, recent studies have suggested that some mycoheterotrophic orchids indirectly obtain carbon from decaying organic matter through associations with saprotrophic fungi. However, such suggestions have been based primarily on indirect evidence, such as the 13C and 15N abundances of fungi and plants. It was recently reported that some mycoheterotrophs yield elevated Δ 14C values, owing to the indirect acquisition of 14C-enriched bomb carbon from dead wood. The approach was based on the anthropogenic change of Δ 14C values; atmospheric CO2 were globally elevated by nuclear weapons testing in the 1950s and early 1960s, but have steadily declined since its peak after the atmospheric nuclear test ban treaty of 1963. The study has provided novel evidence that mycoheterotrophic plants can exploit both mycorrhizal and saprotrophic fungi. We suggest that the radiocarbon analysis is also useful for investigating the nutritional modes of mixotrophic plants as well as for investigating whether the recruitment of wood-decaying fungi into novel mycorrhizal partnerships preceded the evolution of full mycoheterotrophy.

The forfeiting of photosynthetic capabilities has occurred independently many times throughout eukaryotic evolution. But almost all non-photosynthetic plants and algae still retain a colorless plastid and an associated genome, which performs fundamental processes apart from photosynthesis. Unfortunately, little is known about the forces leading to photosynthetic loss; this is largely because there is a lack of data from transitional species. Here, we compare the plastid genomes of two “transitional” green algae: the photosynthetic, mixotrophic Auxenochlorella protothecoides and the non-photosynthetic, obligate heterotroph Prototheca wickerhamii. Remarkably, the plastid genome of A. protothecoides is only slightly larger than that of P. wickerhamii, making it among the smallest plastid genomes yet observed from photosynthetic green algae. Even more surprising, both algae have almost identical plastid genomic architectures and gene compositions (with the exception of genes involved in photosynthesis), implying that they are closely related. This close relationship was further supported by phylogenetic and substitution rate analyses, which suggest that the lineages giving rise to A. protothecoides and P. wickerhamii diverged from one another around six million years ago.

Chemical composition and effect on intestinal Caco-2 cell viability and lipid profile of fixed oil from Cynomorium coccineum L.

Green plants that feed on fungi: facts and questions about mixotrophy

Nutrient mediation of sink strength in the Orobanche minor – Red clover association

The goal of the present chapter is to give the reader a clearer sense of some of the many processes that involve mineral nutrients and ranges of rates at which those processes occur. Mineral nutrition of plants involves the acquisition of elements from the environment, and the organization and functioning of essential plant nutrients are a consequence of the interaction of deoxyribonucleic acid (DNA) of the plant with the environment. Because of the great complexity of plants, processes involving essential nutrient elements vary from relatively slow to relatively fast. Plants require 17 essential nutrients, and several other elements, such as cobalt, sodium, and silicon, have been found to stimulate the growth of some plants (Epstein and Bloom, 2005). Green plants obtain carbon from the air, whereas nonphotosynthetic plants such as fungi obtain carbon as saprophytes, breaking down organic materials of living or dead organisms. Unicellular and multicellular, photosynthetic, nonvascular plants such as algae obtain carbon and other essential elements with little transport from the environment to the site of photosynthesis, and the products of photosynthesis are transported relatively short distances to become new living tissue or to be expelled from the plant to the environment. Vascular plants, on the other hand, have evolved to move essential plant nutrients within the xylem and the phloem over distance far greater than the dimensions of the uncharged atoms, molecules, or ions containing the essential plant nutrients. The anatomy of vascular plants as well as nonvascular plants can be understood to include sources and sinks, between which mineral nutrients move. In addition, the plant itself can be viewed as a sink, which, as a result of the organizing force of DNA, is the recipient of essential nutrients of the plant that originate from the environment, which can be considered a source. Several means of measuring rates of transport of mineral nutrients in plants, other than velocity, include volume transfer (cm3 h−1), mass transfer (g h−1), and specific mass transfer (g cm−2 h−1) (Canny, 1960). CONTENTS

How well is photosensory system conserved in non-photosynthetic plants? : Analysis of phytochrome genes(Abstracts of The 37th Annual Meeting of the Japanese Society for Chemical Regulation of Plants)

Due to their reduced morphology, non-photosynthetic plants have been one of the most challenging groups to delimit to species level. The mycoheterotrophic genus Monotropastrum, with the monotypic species M. humile, has been a particularly taxonomically challenging group, owing to its highly reduced vegetative and root morphology. Using integrative species delimitation, we have focused on Japanese Monotropastrum, with a special focus on an unknown taxon with rosy pink petals and sepals. We investigated its flowering phenology, morphology, molecular identity, and associated fungi. Detailed morphological investigation has indicated that it can be distinguished from M. humile by its rosy pink tepals and sepals that are generally more numerous, elliptic, and constantly appressed to the petals throughout its flowering period, and by its obscure root balls that are unified with the surrounding soil, with root tips that hardly protrude. Based on genome-wide single-nucleotide polymorphisms, molecular data has provided clear genetic differentiation between this unknown taxon and M. humile. Monotropastrum humile and this taxon are associated with different Russula lineages, even when they are sympatric. Based on this multifaceted evidence, we describe this unknown taxon as the new species M. kirishimense. Assortative mating resulting from phenological differences has likely contributed to the persistent sympatry between these two species, with distinct mycorrhizal specificity.

Holoparasites are nonphotosynthetic plants that acquire all resources from hosts. The holoparasite Cuscuta gronovii is native to much of the US with a broad host range including Verbesina alternifolia, an understory perennial. Both species grow in moderate to moist soils and occur in habitats that may experience prolonged or episodic drought. We applied the Wise-Abrahamson Limiting Resource Model (LRM) developed for plant-herbivore relations to examine the effects of pattern of drought stress on tolerance of V. alternifolia to parasitism by C. gronovii. Individual plants were assigned one of six treatments that were combinations of parasite (none or addition of parasite) and drought stress (well-watered, continuously-stressed, or pulse-stressed). After pulse-stressed plants had experienced two wet-dry cycles all plants were harvested. Parasitism strongly reduced both shoot and root mass and well-watered hosts exhibited the greatest decline, indicating reduced tolerance to parasitism when water was readily available. This is consistent with the LRM if parasitism limits photosynthates available to the host. However, parasitism increased allocation to shoot and this effect did not differ between well-watered and drought-stressed plants, indicating equal tolerance. This outcome is in accord with an alternative prediction of the LRM if hosts are not carbon limited. Total pot productivity was reduced by parasitism and drought stress, and this effect was greater for pulse-stressed than for continuously-stressed hosts. We discuss the applicability of the LRM for understanding the effects of drought on tolerance to parasitism.

Thismia (commonly known as fairy lanterns) is a genus of strange-looking, elusive, and non-photosynthetic plants. Thismia kobensis was first discovered in Kobe City, Hyogo Prefecture, Japan in 1992, but it was believed to be extinct, given that its type locality was destroyed during the construction of an industrial complex. Here, we have reported the rediscovery of T. kobensis in Sanda City, Hyogo Prefecture, which is located approximately 30 km from the type locality. The new locality of T. kobensis is the northernmost distributional limit of Asian Thismiaceae species. As the original description of the species was based on a single museum specimen that lacked two of the three inner perianth lobes, we have provided an amended description of T. kobensis, highlighting its differences from the morphologically similar species T. huangii. Specifically, our morphological re-examination has revealed that T. kobensis is distinguishable from T. huangii by its short but expanded annulus and many short hairs on each stigma lobe. We have also demonstrated that the genetic distance between these taxa is comparable to that between other closely related species pairs. Finally, we have provided brief notes on the taxonomy, biogeography, evolutionary history, and conservation of T. kobensis and its closely related species, including an enigmatic species T. americana.

Cynomorium coccineum is a non-photosynthetic plant that grows in Mediterranean countries and that is amply used in the traditional medicine. The aim of this study was to extend previous studies on the chemical and biological properties of C. coccineum, evaluating the potential antiviral and antiproliferative activity of the methanolic extract. The MTT assay was used for the in vitro cytotoxic studies against human cancer-derived cell lines, while both MTT and plaque reduction (PRT) methods were used to evaluate the potential inhibitory effect of the extract against a panel of mammal viruses. The results obtained showed no selective activity against any DNA and RNA virus but revealed an interesting antiproliferative activity against human leukaemia-derived cell lines.

Rate variation in genes from all three genomes has been observed frequently in plant lineages with a parasitic and mycoheterotrophic mode of life. While the loss of photosynthetic ability leads to a relaxation of evolutionary constraints in genes involved in the photosynthetic apparatus, it remains to be determined how prevalent increased substitution rates are in nuclear DNA of non-photosynthetic angiosperms. In this study we infer rates of molecular evolution of 18S rDNA of all parasitic and mycoheterotorphic plant families (except Lauraceae and Polygalaceae) using relative rate tests. In several holoparasitic and mycoheterotrophic plant lineages extremely high substitution rates are observed compared to other photosynthetic angiosperms. The position and frequency of these substitutions have been identified to understand the mutation dynamics of 18S rRNA in achlorophyllous plants. Despite the presence of significantly elevated substitution rates, very few mutations occur in major functional and structural regions of the small ribosomal molecule, providing evidence that the efficiency of the translational apparatus in non-photosynthetic plants has not been affected.

BackgroundThe analysis of synonymous and nonsynonymous rates of DNA change can help in the choice among competing explanations for rate variation, such as differences in constraint, mutation rate, or the strength of genetic drift. Nonphotosynthetic plants of the Orobanchaceae have increased rates of DNA change. In this study 38 taxa of Orobanchaceae and relatives were used and 3 plastid genes were sequenced for each taxon.ResultsPhylogenetic reconstructions of relative rates of sequence evolution for three plastid genes (rbcL, matK and rps2) show significant rate heterogeneity among lineages and among genes. Many of the non-photosynthetic plants have increases in both synonymous and nonsynonymous rates, indicating that both (1) selection is relaxed, and (2) there has been a change in the rate at which mutations are entering the population in these species. However, rate increases are not always immediate upon loss of photosynthesis. Overall there is a poor correlation of synonymous and nonsynonymous rates. There is, however, a strong correlation of synonymous rates across the 3 genes studied and the lineage-speccific pattern for each gene is strikingly similar. This indicates that the causes of synonymous rate variation are affecting the whole plastid genome in a similar way. There is a weaker correlation across genes for nonsynonymous rates. Here the picture is more complex, as could be expected if there are many causes of variation, differing from taxon to taxon and gene to gene.ConclusionsThe distinctive pattern of rate increases in Orobanchaceae has at least two causes. It is clear that there is a relaxation of constraint in many (though not all) non-photosynthetic lineages. However, there is also some force affecting synonymous sites as well. At this point, it is not possible to tell whether it is generation time, speciation rate, mutation rate, DNA repair efficiency or some combination of these factors.