Endophytic System Research Articles

Exploring the Role of Salicylic Acid in Regulating the Colonization Ability of Bacillus subtilis 26D in Potato Plants and Defense against Phytophthora infestans

Plant colonization by endophytic bacteria is mediated by different biomolecules that cause dynamic changes in gene expression of both bacteria and plant. Phytohormones, in particular, salicylic acid, play a key role in the regulation of endophytic colonization and diversity of bacteria in methaphytobiome. For the first time it was found that salicylic acid influenced motility in biofilms and transcription of the surfactin synthetase gene of the endophytic strain Bacillus subtilis 26D in vitro. Treatment of Solanum tuberosum plants with salicylic acid, along with B. subtilis 26D, increased the number of endophytic cells of bacteria in potato internal tissues and level of transcripts of bacterial surfactin synthetase gene and decreased transcription of plant PR genes on the stage of colonisation with endophytes. Thus, the production of surfactin plays an important role in endophytic colonization of plants, and salicylic acid has an ability to influence this mechanism. Here we firstly show that plants treated with salicylic acid and B. subtilis 26D showed enhanced resistance to the late blight pathogen Phytophthora infestans, which was accompanied by increase in transcriptional activity of plant PR-genes and bacterial surfactin synthetase gene after pathogen inoculation. Therefore, it is suggested that salicylic acid can modulate physiological status of the whole plant–endophyte system and improve biocontrol potential of endophytic strains.

Biomechanics of the parasite-host interaction of the European mistletoe.

The European mistletoe (Viscum album) is an epiphytic hemiparasite that attaches to its host by an endophytic system. Two aspects are essential for its survival: the structural integrity of the host-parasite interface must be maintained during host growth and the functional integrity of the interface must be maintained during ontogeny and under mechanical stress. We investigated the mechanical properties of the mistletoe-host interaction. Intact and sliced mistletoe-host samples, with host wood as reference, were subjected to tensile tests up to failure. We quantified the rough fractured surface by digital microscopy and analysed local surface strains by digital image correlation. Tensile strength and deformation energy were independent of mistletoe age but exhibited markedly lower values than host wood samples. Cracks initiated at sites with a major strain of about 30%, especially along the mistletoe-host interface. The risk of sudden failure was counteracted by various sinkers and a lignification gradient that smooths the differences in the mechanical properties between the two species. Our results improve the understanding of the key mechanical characteristics of the host-mistletoe interface and show that the mechanical connection between the mistletoe and its host is age-independent. Thus, functional and structural integrity is ensured over the lifetime of the mistletoe.

Advances on the Visualization of the Internal Structures of the European Mistletoe: 3D Reconstruction Using Microtomography.

The European mistletoe (Viscum album) is a dioecious epiphytic evergreen hemiparasite that develops an extensive endophyte enabling the absorption of water and mineral salts from the host tree, whereas the exophytic leaves are photosynthetically active. The attachment mode and host penetration are well studied, but little information is available about the effects of mistletoe age and sex on haustorium-host interactions. We harvested 130 plants of Viscum album ssp. album growing on host branches of Aesculus flava for morphological and anatomical investigations. Morphometric analyses of the mistletoe and the (hypertrophied) host interaction site were correlated with mistletoe age and sex. We recorded the morphology of the endophytic systems of various ages by using X-ray microtomography scans and corresponding stereomicroscopic images. For detailed anatomical studies, we examined thin stained sections of the mistletoe-host interface by light microscopy. The diameter and length of the branch hypertrophy showed a positive linear correlation with the age of the mistletoe. Correlations with their sex were only found for ratios between host branch and hypertrophy size. A female bias of about 76% was found. In a 4-year-old mistletoe, several small, almost equally sized sinkers and the connected cortical strands extend over more than 5 cm within the host branch. In older mistletoes, one main sinker was predominant and occupied an increasingly large proportion of the stem cross-section. Bands of vessels ran along the axis of the wedge-shaped haustoria and sinkers and bent sideways toward the mistletoe-host interface. At the interface, the vascular elements of the host wood changed their direction and formed vortices near the haustorium.

RUSTY-LEAF MISTLETOE (SCURRULA FERRUGINEA), A PARASITIC PLANT ON COCOA AND ITS MANAGEMENT

The mistletoe ( Scurrula ferruginea ), a parasitic plant, is quite widespread in Malaysia. A severe infestation of S. ferruginea was reported on mature hybrid cocoa (9-12-year-old plantings) in a cocoa estate near Merlimau, Malacca in the 1980s. The effect on crop yield was not known, but the vigour, yield and photosynthetic capability had been greatly reduced. The control measures included: (i) removal of infected trees or their infected branches by pruning or poisoning, and (ii) chemical treatments designed to kill the endophytic system. A total of 588 hectares were pruned manually to remove the mistletoes in the cocoa estate near Merlimau, in May/June 1989. The average cost of pruning was estimated at RM 34.50 per hectare. In a poisoning trial using six herbicides (fluroxypyr, triclopyr, dicamba, paraquat, glufosinate-ammonium and 2,4,5-T) the costs was as high as pruning. Manual pruning was therefore the recommended method of controlling cocoa mistletoe. Integrated Weed Management is perhaps the next step towards more effective and more efficient control of mistletoes over very large areas inclusive of cocoa estates, orchards and other agricultural/non-agricultural areas. Keywords: Scurrula ferruginea, mistletoe, parasitic plant, cocoa, management

A novel experimental system using the liverwort Marchantia polymorpha and its fungal endophytes reveals diverse and context-dependent effects.

Fungal symbioses are ubiquitous in plants, but their effects have mostly been studied in seed plants. This study aimed to assess the diversity of fungal endophyte effects in a bryophyte and identify factors contributing to the variability of outcomes in these interactions. Fungal endophyte cultures and axenic liverwort clones were isolated from wild populations of the liverwort, Marchantia polymorpha. These collections were combined in a gnotobiotic system to test the effects of fungal isolates on the growth rates of hosts under laboratory conditions. Under the experimental conditions, fungi isolated from M.polymorpha ranged from aggressively pathogenic to strongly growth-promoting, but the majority of isolates caused no detectable change in host growth. Growth promotion by selected fungi depended on nutrient concentrations and was inhibited by coinoculation with multiple fungi. The M.polymorpha endophyte system expands the resources for this model liverwort. The experiments presented here demonstrate a wealth of diversity in fungal interactions even in a host reported to lack standard mycorrhizal symbiosis. In addition, they show that some known pathogens of vascular plants live in M.polymorpha and can confer benefits to this nonvascular host. This highlights the importance of studying endophyte effects across the plant tree of life.

Plant Growth, Antibiotic Uptake, and Prevalence of Antibiotic Resistance in an Endophytic System of Pakchoi under Antibiotic Exposure.

Antibiotic contamination in agroecosystems may cause serious problems, such as the proliferation of various antibiotic resistant bacteria and the spreading of antibiotic resistance genes (ARGs) in the environment or even to human beings. However, it is unclear whether environmental antibiotics, antibiotic resistant bacteria, and ARGs can directly enter into, or occur in, the endophytic systems of plants exposed to pollutants. In this study, a hydroponic experiment exposing pakchoi (Brassica chinensis L.) to tetracycline, cephalexin, and sulfamethoxazole at 50% minimum inhibitory concentration (MIC) levels and MIC levels, respectively, was conducted to explore plant growth, antibiotic uptake, and the development of antibiotic resistance in endophytic systems. The three antibiotics promoted pakchoi growth at 50% MIC values. Target antibiotics at concentrations ranging from 6.9 to 48.1 µg·kg−1 were detected in the treated vegetables. Additionally, the rates of antibiotic-resistant endophytic bacteria to total cultivable endophytic bacteria significantly increased as the antibiotics accumulated in the plants. The detection and quantification of ARGs indicated that four types, tetX, blaCTX-M, and sul1 and sul2, which correspond to tetracycline, cephalexin, and sulfamethoxazole resistance, respectively, were present in the pakchoi endophytic system and increased with the antibiotic concentrations. The results highlight a potential risk of the development and spread of antibiotic resistance in vegetable endophytic systems.

Molecular inter-kingdom interactions of endophytes isolated from Lychnophora ericoides

The importance of microbial natural products has been widely demonstrated in the search for new antibiotics. However, the functional role of microbial metabolites in nature remains to be deciphered. Several natural products are known to mediate microbial interactions through metabolic exchange. One approach to investigate metabolic exchange in the laboratory is through microbial interactions. Here, we describe the chemical study of selected endophytes isolated from the Brazilian medicinal plant Lychnophora ericoides by pairwise inter-kingdom interactions in order to correlate the impact of co-cultivation to their metabolic profiles. Combining mass spectrometry tools and NMR analyses, a total of 29 compounds were identified. These compounds are members of polyene macrocycles, pyrroloindole alkaloids, angucyclines, and leupeptins chemical families. Two of the identified compounds correspond to a new fungal metabolite (29) and a new actinobacterial angucycline-derivative (23). Our results revealed a substantial arsenal of small molecules induced by microbial interactions, as we begin to unravel the complexity of microbial interactions associated with endophytic systems.

Bioremediation of phenol in soil through using a mobile plant–endophyte system

Plant–endophyte remediation of volatile pollutants in soil is an emerging technology. For more efficient application, plant–endophyte systems were formed through stimulation of transfer of degradative plasmids in plant tissue by co-inoculation of corn, wheat or tomato seedlings with Pseudomonas fluorescens TP13 carrying a self-transmissible degradative plasmid, and P. fluorescens streptomycin-resistant P13 strain. The corn–TP13–P13 (CTP) system had higher degradation activity than other plant–endophyte systems. Transplanting the CTP, from loam to sandy clay loam soil, from greenhouse to field trials, almost completely removed phenol from contaminated soils in 15 d. Intact transplantation of the CTP to contaminated soils was more efficient than co-transplanting of phenol-degrading bacteria and plant in detoxification of phenol. After the experiments the harvested CPT still exhibited remarkable bioremediation activity. The number of degradative plasmid-carrying endophytic bacteria in the CTP system was just slightly more than in the corn seedlings inoculated with TP13 alone, but the former substantially surpassed the latter in phenol-degrading activity, probably due to stimulation of transfer of the degradative plasmids among endophytic bacteria in plant tissues. More degradative plasmid-carrying bacteria colonized bioremediating soil and plant tissues, and higher plasmid transfer frequency and C23O activity of transconjugant were found in soils for the CTP system compared with other treatments. These results showed that the CTP system is a valuable tool to degrade volatile organic pollutants and transfer of degradative plasmids in plant tissues is important for construction of a mobile plant–endophyte system applied in bioremediation of volatile pollutants.

Aligning Microtomography Analysis with Traditional Anatomy for a 3D Understanding of the Host-Parasite Interface - Phoradendron spp. Case Study.

The complex endophytic structure formed by parasitic plant species often represents a challenge in the study of the host-parasite interface. Even with the large amounts of anatomical slides, a three-dimensional comprehension of the structure may still be difficult to obtain. In the present study we applied the High Resolution X-ray Computed Tomography (HRXCT) analysis along with usual plant anatomy techniques in order to compare the infestation pattern of two mistletoe species of the genus Phoradendron. Additionally, we tested the use of contrasting solutions in order to improve the detection of the parasite’s endophytic tissue. To our knowledge, this is the first study to show the three-dimensional structure of host-mistletoe interface by using HRXCT technique. Results showed that Phoradendron perrottetii growing on the host Tapirira guianensis forms small woody galls with a restricted endophytic system. The sinkers were short and eventually grouped creating a continuous interface with the host wood. On the other hand, the long sinkers of P. bathyoryctum penetrate deeply into the wood of Cedrela fissilis branching in all directions throughout the woody gall area, forming a spread-out infestation pattern. The results indicate that the HRXCT is indeed a powerful approach to understand the endophytic system of parasitic plants. The combination of three-dimensional models of the infestation with anatomical analysis provided a broader understanding of the host-parasite connection. Unique anatomic features are reported for the sinkes of P. perrottetii, while the endophytic tissue of P. bathyoryctum conformed to general anatomy observed for other species of this genus. These differences are hypothesized to be related to the three-dimensional structure of each endophytic system and the communication stablished with the host.

Influence of Chicken Manure Fertilization on Antibiotic-Resistant Bacteria in Soil and the Endophytic Bacteria of Pakchoi.

Animal manure is commonly used as fertilizer for agricultural crops worldwide, even though it is believed to contribute to the spread of antibiotic resistance from animal intestines to the soil environment. However, it is unclear whether and how there is any impact of manure fertilization on populations and community structure of antibiotic-resistant endophytic bacteria (AREB) in plant tissues. To investigate the effect of manure and organic fertilizer on endophytic bacterial communities, pot experiments were performed with pakchoi grown with the following treatments: (1) non-treated; (2) chicken manure-treated and (3) organic fertilizer-treated. Manure or organic fertilizer significantly increased the abundances of total cultivable endophytic bacteria (TCEB) and AREB in pakchoi, and the effect of chicken manure was greater than that of organic fertilizer. Further, 16S rDNA sequencing and the phylogenetic analysis indicated that chicken manure or organic fertilizer application increased the populations of multiple antibiotic-resistant bacteria (MARB) in soil and multiple antibiotic-resistant endophytic bacteria (MAREB) in pakchoi. The identical multiple antibiotic-resistant bacterial populations detected in chicken manure, manure- or organic fertilizer-amended soil and the vegetable endophytic system were Brevundimonas diminuta, Brachybacterium sp. and Bordetella sp., suggesting that MARB from manure could enter and colonize the vegetable tissues through manure fertilization. The fact that some human pathogens with multiple antibiotic resistance were detected in harvested vegetables after growing in manure-amended soil demonstrated a potential threat to human health.

Transformation of methane in peatland environments

Abstract Wetlands and particularly peatlands are the main natural source of methane. Data indicate that 10-45% of methane emission comes from these sources. Methane emission from wetlands is the result of the balance between methanogenesis and methanotrophic processes and is actively affected by the wetland plant community composition. There are many factors affecting the balance of CH4: for instance, vegetation has a strong effect on CH4 emissions from wetland ecosystems by influencing methane production, consumption and transport in the soil. The effects of plants on methane fluxes may be mediated by: molecular diffusion, internal transport through plant aerenchyma tissues and ebullition. Methane is formed in the process of methanogenesis under anaerobic conditions. It may then be emitted into the atmosphere directly from the soil or by internal transport through the plant. Alternatively, it may undergo methane oxidation by methanotrophic bacteria, both free-living in the root zone and associated with the host plant in symbiosis. Sphagnum moss is of particular importance for this processes as it contains methanotrophic bacteria in its endophytic system. Methanotrophic bacteria live inside the dead hyaline cells or on the surface of Sphagnum leaves and are able to oxidise methane produced in the soil during methanogenesis, making peatlands a natural biofilter for methane, one of the main greenhouse gases

Prospects for an integrated control of Loranthaceae species parasitizing <em>Vitellaria paradoxa</em> C. F. Gaertn in Burkina Faso

This study investigated the potential components of an integrated management of Loranthaceae species (mistletoes) which constitute a major constraint to the survival and fruit production of many trees such as Vitellaria paradoxa in West Africa. Three types of pruning, five doses of each of two herbicides and host resistance were evaluated in mistletoe-infested parklands. Natural enemies were surveyed with a view to biological control. Pruning of the host branch along with the parasite’s haustorium (endophytic system) led to the total elimination of the parasite. When the endophytic system was spared, the time of pruning and species of parasite significantly influenced the number of sprout stalks per tuff. Their interaction was also significant. Death of at least 80% of treated Loranthaceae tufts was recorded for doses of 15 g l -1 of glyphosate and 20 g l -1 of 2,4-D. The artificial infestation of stands with parasite seeds revealed the likely existence of genetic resistance to mistletoe in V. paradoxa . The agroforestry parkland survey showed that birds, chiefly Pogoniulus chrysonocus , and insects are potential natural enemies of Loranthaceae. Results showed that Loranthaceae control could be achieved with techniques already used to successfully manage weeds. However, there is need to improve control components and their integration to successfully manage mistletoes in Burkina Faso. Keywords : Vitellaria paradoxa , Loranthaceae, mechanical, herbicides, host resistance, bio-control

Genomic Analysis of the Hydrocarbon-Producing, Cellulolytic, Endophytic Fungus Ascocoryne sarcoides

The microbial conversion of solid cellulosic biomass to liquid biofuels may provide a renewable energy source for transportation fuels. Endophytes represent a promising group of organisms, as they are a mostly untapped reservoir of metabolic diversity. They are often able to degrade cellulose, and they can produce an extraordinary diversity of metabolites. The filamentous fungal endophyte Ascocoryne sarcoides was shown to produce potential-biofuel metabolites when grown on a cellulose-based medium; however, the genetic pathways needed for this production are unknown and the lack of genetic tools makes traditional reverse genetics difficult. We present the genomic characterization of A. sarcoides and use transcriptomic and metabolomic data to describe the genes involved in cellulose degradation and to provide hypotheses for the biofuel production pathways. In total, almost 80 biosynthetic clusters were identified, including several previously found only in plants. Additionally, many transcriptionally active regions outside of genes showed condition-specific expression, offering more evidence for the role of long non-coding RNA in gene regulation. This is one of the highest quality fungal genomes and, to our knowledge, the only thoroughly annotated and transcriptionally profiled fungal endophyte genome currently available. The analyses and datasets contribute to the study of cellulose degradation and biofuel production and provide the genomic foundation for the study of a model endophyte system.

Histopathology of the endophytic system and aerial shoots of Arceuthobium americanum infected by Colletotrichum gloeosporioides

Infection of the endophytic system of Arceuthobium americanum parasitizing Pinus contorta var. latifolia by the fungus Colletotrichum gloeosporioides was assessed on naturally infected and inoculated localized dwarf mistletoe plants. Two different techniques were utilized to attempt to inoculate C. gloeosporioides into the endophytic system: the fungus was applied as Stabileze formulated product onto cut A. americanum shoots and as a mycelial plug to wounded A. americanum infected host branches. The fungus was cultured from infected shoots and the basal cup region of naturally infected A. americanum, but not pine tissues that included the endophytic system. Following inoculation of cut shoots, the shoots were abscised from the dwarf mistletoe plant, and the fungus was not isolated 1 year after inoculation. The fungus was recovered from the wounded region of infections that were inoculated with a mycelial plug, but not outside of the wounded area. Hyphae with morphology that matched that of C. gloeosporioides were observed microscopically in the shoots and basal cup region of naturally infected A. americanum, but not in the samples from the inoculated treatments. Thus, we were unable to confirm that C. gloeosporioides infected the endophytic system of A. americanum parasitizing P. contorta var. latifolia, although xylem continuity between the aerial and endophytic systems was observed.

Endosymbiosis and population differentiation in wild and cultivated Lolium perenne (Poaceae)

Endophytic fungi are common inhabitants of cool-season grasses that can affect host growth and reproduction. One prerequisite for the expected coevolution between symbionts is that there exist variable effects of the endosymbiont on its host in different environments. Here, the impact of endophytes ( Neotyphodium lolii ) on survival, growth, and reproduction of wild populations of Lolium perenne from its native range (Italy, Morocco, Turkey, Tunisia) and two United States cultivars was explored. Two unmanaged, common gardens 30 m apart in New Jersey, USA, were planted with endophyte-infected and uninfected plants of the six populations in August 2008. Plants were scored for tiller number and lengths, and survival in October and December 2008 and May and September 2009; fl owering tillers (spikes) were counted in spring 2009. A greenhouse experiment was designed to assess tiller production and growth of four populations. There were significant effects of population and garden on tiller number and lengths but no detectable effects of endophytes. Populations from Italy and Morocco and the two US cultivars grew well in one garden, but Tunisian plants grew the least in either garden. Overwinter survival and postfl owering survival were unaffected by endophytes. Spike production was strongly influenced by population. Although populations varied, there were no differences in growth between infected and uninfected plants in the greenhouse. Due to the absence of unambiguous endophyte-mediated effects on growth and reproduction of these differentiated populations, putative coevolutionary relations between the endosymbiont and its native host species remain obscure in this grass – endophyte system.

Assessment of Colletotrichum gloeosporioides as a biological control agent for management of hemlock dwarf mistletoe (Arceuthobium tsugense)

The ascomycete Colletotrichum gloeosporioides was tested for biological control of Arceuthobium tsugense on western hemlock (Tsuga heterophylla) in British Columbia (BC), Canada. A field trial was conducted near Nanaimo, BC, using C. gloeosporioides isolate PFC 2415 in three treatments applied in late August 2002. The treatments consisted of C. gloeosporioides formulated with ‘Stabileze’ (incorporation of fungi in a water-absorbent starch matrix with oil and sucrose, then granulating the matrix with hydrated silica) which were sprayed on (1) intact mistletoe swellings, (2) swellings on which all mistletoe shoots had been cut at 0.5 cm from the base of the shoot and (3) C. gloeosporioides formulated in a sucrose and gelatin preparation sprayed on intact mistletoe swellings. The ultimate goal of this research was to substantially reduce the ability of the treated dwarf mistletoe plants to produce seed inoculum. The ‘Stabileze’ and sucrose–gelatin treatments formulated with C. gloeosporioides reduced the current berry crop by 36.8 and 40.5%, respectively (p < 0.05). While the results for shoots appeared promising, heavy background infection and/or secondary infection, especially on controls, limited the ability to detect clear treatment effects. Careful culturing from various live and dead host tissues showed that C. gloeosporioides was unable to invade and kill the mistletoe endophytic system within the living xylem and phloem of the host.

Sinker Structure of Phoradendron californicum (Viscaceae) Confounds its Presumed Close Relationship to Other Acataphyllous Species

Phoradendron is the largest genus of New World mistletoes, with about 250 species in two subgenera, Boreales and Aequatoriales, corresponding, respectively, to northern acataphyllous and southern cataphyllous groups. The typically acataphyllous P. californicum of western North America is controversial because recent phylogenetic work has nested it in the southern cataphyllous clade. Seedling establishment, stem anatomy, and endophytic system structure of this species were studied. Seedling haustorial holdfasts have gland cavities, structures considered absent in the Viscaceae clade of Santalales. The stem epidermis has a thick cuticle, deeply sunken stomata, and branched multicellular trichomes. The stem has an outer cortex of palisade chlorenchyma and an inner cortex of large isodiametric parenchyma cells. The boundary area between the outer and inner cortex contains druses and an unusual ring of small xylic bundles lacking protoxylem fibers and phloem. Sinkers are of two types: uniseriate, with only parenchyma that often has thick-walled transfer cells at its interface with vessels of the host; and multiseriate, with parenchyma and vessel elements that often are in direct contact and share simple perforation plates with vessels of the host. Sinker morphology is also dimorphic in the cataphyllous P. fragile but only unimorphic (multiseriate) in the acataphyllous P. juniperinum and P. serotinum. The dimorphic sinker system of P. californicum may be functionally partitioned, whereas these functions are combined in the unimorphic sinkers of other acataphyllous species. Differences in sinker morphology may reflect evolutionary trends in Phoradendron. This study also supports the hypothesis that P. californicum is more closely aligned with the mainly tropical cataphyllous species of the genus.

Epiparasitism in Phoradendron durangense and P. falcatum (Viscaceae)

Phoradendron, the largest mistletoe genus in the New World, extends from temperate North America to temperate South America. Most species are parasitic on terrestrial hosts, but a few occur only, or primarily, on other species of Phoradendron. We examined relationships among two obligate epiparasites, P. durangense and P. falcatum, and their parasitic hosts. Fruit and seed of both epiparasites were small compared to those of their parasitic hosts. Seed of epiparasites was established on parasitic-host stems, leaves, and inflorescences. Shoots developed from the plumular region or from buds on the holdfast or subjacent tissue. The developing endophytic system initially consisted of multiple separate strands that widened, merged, and often entirely displaced its parasitic host from the cambial cylinder. During establishment the epiparasite invaded the cortex, vascular cylinder, and pith of its parasitic host and spread aggressively within host tissues, extending between and into individual host parenchyma cells, eventually isolating host cells or cell groups. The parasitic host showed little visual response to the epiparasite. Endophytic system growth of the epiparasite within its parasitic host was compared to that of a parasitic Phoradendron within its terrestrial host. The results indicate growth dynamics similar to those of parasitic species on terrestrial hosts. We conclude that the epiparasite/parasitic-host union should not be regarded as a graft union. The harmonious appearance of the union is a result of the growth of the epiparasite replacing entirely tissues of its parasitic host, with little or no hypertrophy of parasitic-host branches.

Contrasting research approaches to managing mistletoes in commercial forests and wooded pasturesThis minireview is one of a collection of papers based on a presentation from the Stem and Shoot Fungal Pathogens and Parasitic Plants: the Values of Biological Diversity session of the XXII International Union of Forestry Research Organization World Congress meeting held in Brisbane, Queensland, Australia, in 2005.

Many mistletoe species are pests in agricultural and forest ecosystems throughout the world. Mistletoes are unusual “weeds” as they are generally endemic to areas where they achieve pest status and, therefore, classical biological control and broad-scale herbicidal control are usually impractical. In North American coniferous forests, dwarf mistletoe ( Arceuthobium spp.) infection results in major commercial losses and poses a public liability in recreation settings. Hyperparasitic fungi have potential as biological control agents of dwarf mistletoe, including species which attack shoots, berries, and the endophytic systems of dwarf mistletoe. Development of an inundative biological control strategy will be useful in situations where traditional silvicultural control is impractical or undesirable. In southern Australia, farm eucalypts are often attacked and killed by mistletoes ( Amyema spp.) in grazed landscapes where tree decline and biodiversity loss are major forms of land degradation. Although long-term strategies to achieve a balance between mistletoe and host abundance are promoted, many graziers want short-term options to treat severely infected trees. Recent research has revisited the efficiency and efficacy of silvicultural treatments and selective herbicides in appropriate situations. The results of recent research on these diverse management strategies in North America and Australia are summarized.

The Endophytic System of Mediterranean Cytinus (Cytinaceae) Developing on Five Host Cistaceae Species

One of the most extreme manifestations of parasitism is found in the genus Cytinus, a holoparasite whose vegetative body is reduced to an endophytic system living within its host root. There are two species of Cytinus in the Mediterranean, C. hypocistis and C. ruber, which parasitize various genera of Cistaceae, one of the most characteristic families of the Mediterranean scrublands. The aim of this work is to describe the endophytic systems of C. hypocistis and C. ruber, and their tissue relationships with their host. Roots from five different hosts infected with C. hypocistis and C. ruber were harvested, and examined by anatomical techniques under light microscopy to elucidate the characteristics of the endophytic system of Cytinus, and to determine if differences in endophytic systems occur between the two species and in response to different hosts. The endophyte structure is similar in both Cytinus species irrespective of the host species. In the initial stages of the endophyte, rows of parenchymal cells spread through the host pericyclic derivatives and phloem, and begin to generate small nodules in the outermost region of the host xylem. Later the nodules anastomose, and bands of parasitic tissue are formed. The host cambium continues to develop xylem tissue, and consequently the endophyte becomes enclosed within the xylem. The bands of parasitic tissue fuse to form a continuous sheath. This mature endophyte has well-developed vascular system with xylem and phloem, and forms sinkers with transfer cells that grow through the host xylem. The endophytic system of Cytinus develops in all host root tissues and reaches its most mature stages in the host xylem. It is more complex than previously reported, showing parenchyma, xylem and phloem tissues. This is the first report of well-developed phloem in a holoparasitic endophytic species.