Phenotypic and Genetic Diversity of Local <Emphasis Type="Italic">Perilla</Emphasis> (<Emphasis Type="Italic">Perilla frutescens</Emphasis> (L.) Britt.) from Northern Thailand

Phenotypic and Genetic Diversity of Local Perilla ( Perilla frutescens (L.) Britt.) from Northern Thailand. Perilla frutescens (L.) Britt., an important oil and culinary crop in Asia, is a valuable genetic resource. Despite its nutritional value and historic and cultural importance, research on Perilla has been scarce, particularly as far as its genetic diversity is concerned. The aims of the present study were to assess variability within and between 29 seed samples of P. frutescens collected from farmers in northern Thailand, and evaluation conducted of their genetic, morphological, and agronomic characteristics, and the seed composition, including polyunsaturated fatty acids omega-3, omega-6, and omega-9, and the vitamin E γ-tocopherols. Perilla frutescens (L.) Britt. of northern Thailand is genetically variable, and structured according to origin of collection which was the consequence of local adaptation. The discovery of high levels of polyunsaturated fatty acids, namely α-linolenic acid and γ-tocopherols, in some Perilla samples indicates the potential for utilizing Perilla for its high omega-3 content including as a vitamin E supplement for humans, a prospect that should be taken into account when planning conservation strategies or when Perilla variability is used in breeding programs.     

Synergistic plant–microbe interactions between endophytic bacterial communities and the medicinal plant <Emphasis Type="Italic">Glycyrrhiza uralensis</Emphasis> F.

Little is known about the composition, diversity, and geographical distribution of bacterial communities associated with medicinal plants in arid lands. To address this, a collection of 116 endophytic bacteria were isolated from wild populations of the herb Glycyrrhiza uralensis Fisch (licorice) in Xinyuan, Gongliu, and Tekesi of Xinjiang Province, China, and identified based on their 16S rRNA gene sequences. The endophytes were highly diverse, including 20 genera and 35 species. The number of distinct bacterial genera obtained from root tissues was higher (n?=?14) compared to stem (n?=?9) and leaf (n?=?6) tissue. Geographically, the diversity of culturable endophytic genera was higher at the Tekesi (n?=?14) and Xinyuan (n?=?12) sites than the Gongliu site (n?=?4), reflecting the extremely low organic carbon content, high salinity, and low nutrient status of Gongliu soils. The endophytic bacteria exhibited a number of plant growth-promoting activities ex situ, including diazotrophy, phosphate and potassium solubilization, siderophore production, auxin synthesis, and production of hydrolytic enzymes. Twelve endophytes were selected based on their ex situ plant growth-promoting activities for growth chamber assays to test for their ability to promote growth of G. uralensis F. and Triticum aestivum (wheat) plants. Several strains belonging to the genera Bacillus (n?=?6) and Achromobacter (n?=?1) stimulated total biomass production in both G. uralensis and T. aestivum under low-nutrient conditions. This work is the first report on the isolation and characterization of endophytes associated with G. uralensis F. in arid lands. The results demonstrate the broad diversity of endophytes associated with wild licorice and suggest that some Bacillus strains may be promising candidates for biofertilizers to promote enhanced survival and growth of licorice and other valuable crops in arid environments.     

Effects of using different host plants and long-term fertilization systems on population sizes of infective arbuscular mycorrhizal fungi

The effect of cultivation of mycorrhizal and non-mycorrhizal plants and mineral fertilization on the arbuscular mycorrhizal fungal (AMF) community structure of maize (Zea mays L.) plants was studied. Soil samples were collected from two field experiments treated for 5 years with three fertilization systems (Control – no fertilization; Mineral – NPK fertilization; and Organic – Farmyard manure fertilization). Soil samples containing soil and root fragments of rapeseed (Brassica napus L., non-mycorrhizal plant) and wheat (Triticum aestivum L., mycorrhizal plant) collected from the field plots were used as native microbial inoculum sources to maize plants. Maize plants were sown in pots containing these inoculum sources for four months under glasshouse conditions. Colonization of wheat roots by AMF, AMF community structure, AMF diversity (Shannon’s index), AMF dominance (Simpson’s index) and growth of maize were investigated. Sixteen AMF species were identified from rhizosphere soil samples as different species of genera Acaulospora, Claroideoglomus, Dentiscutata, Funneliformis, Gigaspora, Quatunica, Racocetra, and Rhizoglomus. Maize plants grown in manure-fertilized soils had a distinct AMF community structure from plants either fertilized with mineral NPK-fertilizer or non-fertilized. The results also showed that inoculum from non-mycorrhizal plants combined with mineral fertilization decreased AMF diversity (Shannon’s index), AMF dominance (Simpson’s index) and growth of maize. Our findings suggest that non-mycorrhizal plants, such as B. napus, can negatively affect the presence and the effects of soil inoculation on maize growth. Also, our results highlight the importance of considering the long-term effect of rapeseed cultivation system on the reduction of population sizes of infective AMF, and its effect on succeeding annual crops.     

Copper and zinc concentrations of medicinal herbs and soil surrounding ponds on agricultural land

This study measured copper and zinc concentrations in medicinal herbs and soil sampled from three agricultural fields with ponds from mid-June till the end of July 2014. Six herb species were tested: Potentilla anserina L., Mentha arvensis, Achillea millefolium L., Comarum palustre L., Lysimachia vulgaris L., and Lycopus europaeus L. Two of the ponds were in the borough of Jab?onna Lacka and one in the borough of Sabnie. The fields around each pond were divided into transects with three concentric soil moisture sections, each with different soil water contents. Soil and plant samples were taken from each transect to determine the concentrations of copper and zinc by inductively coupled plasma atomic emission spectrometry. Despite the use of fertilizers and pesticides in the fields, copper and zinc had not polluted the soil or the plants growing in the soil depressions. Out of the six plants tested, only C. palustre had an increased concentration of zinc. Different farming methods used on the fields affected the concentrations of copper and zinc in the soil of the different depressions. The highest concentrations of these metals were found in the soil of the periodically wet sections.     

Native and exotic foundation grasses differ in traits and responses to belowground tri-trophic interactions

A plant’s growth and fitness are influenced by species interactions, including those belowground. In primary successional systems, belowground organisms are known to have particularly important control over plant growth. Exotic plant invasions in these and other habitats may in part be explained by altered associations with belowground organisms compared to native plants. We investigated the growth responses of two foundation grasses on Great Lakes sand dunes, the native grass Ammophila breviligulata and the exotic grass Leymus arenarius, to two groups of soil organisms with important roles in dune succession: arbuscular mycorrhizal fungi (AMF) and plant-parasitic nematodes (PPN). We manipulated the presence/absence of two generalist belowground species known to occur in Great Lakes dunes, Rhizophagus intraradices (AMF) and Pratylenchus penetrans (PPN) in a factorial greenhouse experiment and assessed the biomass production and root architectural traits of the plants. There were clear differences in growth and above- and belowground architecture between Ammophila and Leymus, with Leymus plants being bigger, taller, and having longer roots than Ammophila. Inoculation with Rhizophagus increased above- and belowground biomass production by ~32% for both plant species. Inoculation with Pratylenchus decreased aboveground biomass production by ~36% for both plant species. However belowground, the exotic Leymus was significantly more resistant to PPN than the native Ammophila, and gained more benefits from AMF in belowground tri-trophic interactions than Ammophila. Overall, our results indicate that differences in plant architecture coupled with altered belowground interactions with AMF and PPN have the potential to promote exotic plant invasion.     

Characterization of root-associated microbiota in medicinal plants <Emphasis Type="Italic">Astragalus membranaceus</Emphasis> and <Emphasis Type="Italic">Astragalus mongholicus</Emphasis>

Although the quality of herbal medicine is tightly associated with plant genotype and location, microbial traits of most herbs remain unclear. In this study, bacterial communities residing Astragali Radix, which is derived from Astragalus membranaceus and A. mongholicus roots, have been characterized by automated ribosomal intergenic spacer analysis (ARISA) and pyrosequencing of 16S rRNA amplicons. The samples were collected from four representive locations and differing in genotype and planting pattern. The spatial resolution study by ARISA firstly distinguished between the two anatomically-based parts, the periderm and secondary vascular tissue, demonstrating that microbial communities residing in the former were more diverse and clearly separated by host genotype and location as compared with those residing in the latter. Taxonomic coverage revealed that phyla of Proteobacteria, Planctomycetes and Bacteroidetes dominated the bacterial assemblages across the samples. The community diversity in A. mongholicus was more abundant than it was in A. membranaceus, especially A. mongholicus in Shanxi province of China. In addition, the conventional cultivation exerted consistently negative effects on microbiota complexity when compared with the planting pattern “imitating wild conditions”, which was regardless of the host genotype. With the focus on microbiota members in Shanxi, taxa associated with the genotype, geography and planting pattern were finally discriminated and used as indicators for the screening of endophytic bacteria that contain ACC deaminase. Taken together, microbiota should be an important trait of herbal medicines and the periderm should be a specialized niche for microbiota research in medicinal plants.     

<Emphasis Type="Italic">Epichloë</Emphasis> exudates promote in vitro and in vivo arbuscular mycorrhizal fungi development and plant growth

Background and aims

We studied, through exudates employment, the effect of Epichloë (endophytic fungi), both independently and in association with Bromus auleticus (grass), on arbuscular mycorrhizal fungi (AMF) colonization, host and neighbouring plants biomass production and soil changes.

Methods

Through in vitro and greenhouse experiments, Epichloë endophytes effect on AMF development was evaluated. In vitro studies of exudates effect on Gigaspora rosea and Rhizophagus intraradices were performed using root or endophyte exudates. A 6-month greenhouse experiment was conducted to determine Bromus auleticus endophytic status effect and endophyte exudates role in biomass production, neighbouring plants mycorrhizal colonization and soil properties.

Results

Endophyte exudates and E+ plant root exudates promoted in vitro AMF development in the pre-infective stage of G. rosea and in carrot root culture mycelium of R. intraradices in a dose-response relationship, while control media and E- plants exudates had no effect. R. intraradices colonization and plant growth was clearly increased by endophytes and their exudates.

Conclusions

This is the first work evidencing the direct effect of Epichloë endophytes and infected plants root exudates on AMF extramatrical development. While higher levels of AMF colonization were observed in E+ plants, no clear effect was detected in neighbouring plants colonization, plant biomass or soil properties.

     

Distribution of arbuscular mycorrhizal fungi in four semi-mangrove plant communities

To better understand the diversity and species composition of arbuscular mycorrhizal fungi (AMF) in mangrove ecosystems, the AMF colonization and distribution in four semi-mangrove plant communities were investigated. Typical AMF hyphal, vesicle and arbuscular structures were commonly observed in all the root samples, indicating that AMF are important components on the landward fringe of mangrove habitats. AMF spores were extracted from the rhizospheric soils, and an SSU rDNA fragment from each spore morph-type was amplified and sequenced for species identification. AMF species composition and diversity in the roots of each semi-mangrove species were also analyzed based on an SSU-ITS-LSU fragment, which was amplified, cloned and sequenced from root samples. In total, 11 unique AMF sequences were obtained from spores and 172 from roots. Phylogenetic analyses indicated that the sequences from the soil and roots were grouped into 5 and 14 phylotypes, respectively. AMF from six genera including Acaulospora, Claroideoglomus, Diversispora, Funneliformis, Paraglomus, and Rhizophagus were identified, with a further six phylotypes from the Glomeraceae family that could not be identified to the genus level. The AMF genus composition in the investigated semi-mangrove communities was very similar to that in the intertidal zone of this mangrove ecosystem and other investigated mangrove ecosystems, implying possible fungal adaptation to mangrove conditions.     

Comparative proteomic analysis of leaf tissue from tomato plants colonized with <Emphasis Type="Italic">Rhizophagus irregularis</Emphasis>

A comparative proteomic approach was performed to analyze the differential accumulation of leaf proteins in response to the symbiosis between Solanum lycopersicum and the arbuscular mycorrhizal fungus (AMF) Rhizophagus irregularis. Protein profiling was examined in leaves from tomato plants colonized with AMF (M), as well as non-colonized plants fertilized with low phosphate (20 μM P; NM-LP) and non-colonized plants fertilized with regular phosphate Hoagland’s solution (200 μM P; NM-RP). Comparisons were made between these groups, and 2D-SDS-PAGE revealed that 27 spots were differentially accumulated in M vs. NM-LP. Twenty-three out of the 27 spots were successfully identified by mass spectrometry. Two of these proteins, 2-methylene-furan-3-one reductase and auxin-binding protein ABP19a, were up-accumulated in M plants. The down-accumulated proteins in M plants were associated mainly with photosynthesis, redox, and other molecular functions. Superoxide dismutase, harpin binding protein, and thioredoxin peroxidase were down-accumulated in leaves of M tomato plants when compared to NM-LP and NM-RP, indicating that these proteins are responsive to AMF colonization independently of the phosphate regime under which they were grown. 14-3-3 protein was up-accumulated in NM-RP vs. NM-LP plants, whereas it was down-accumulated in M vs. NM-LP and M vs. NM-RP, regardless of their phosphate nutrition. This suggests a possible regulation by P nutrition and AMF colonization. Our results demonstrate AMF-induced systemic changes in the expression of tomato leaf proteins, including the down-accumulation of proteins related to photosynthesis and redox function.     

Silicon nutrition alleviates the lipid peroxidation and ion imbalance of <Emphasis Type="Italic">Glycyrrhiza uralensis</Emphasis> seedlings under salt stress

Soil salinity reduces growth of Glycyrrhiza uralensis in arid and semi-arid areas of north-west in China. Silicon (Si) nutrition may alleviate salt stress in many crops including grain crop, fruit crop, and vegetable crop. In this study, the alleviating effects of Si on growth characteristics, antioxidant enzyme activity (SOD and POD) and MDA concentration, and K⁺ and Na⁺ concentrations in G. uralensis seedlings subjected to 50 mM NaCl stress were investigated. The results showed that NaCl stress imposed significant reduction in root length, secondary root number, leaf number, and stem and total dry weight of G. uralensis. NaCl stress also significantly reduced the activities of SOD and POD, and ration of K⁺/Na⁺, but significantly increased MDA concentration in leaves of G. uralensis seedling. The addition of Si increased SOD and POD activities, and reduced MDA concentration, which resulting in greater reactive oxygen species detoxification and lower lipid peroxidation. Si also significantly increased the ratio of K⁺/Na⁺ in stem and leaves of G. uralensis. In conclusion, Si could alleviate adverse effects of salt stress probably by decreasing Na⁺ concentration and improving antioxidant enzyme activity of G. uralensis, and these alleviating effects were dependent on Si concentration and on Si processing time.     

Micropropagation of Ajuga species: a mini review

The genus Ajuga L., belonging to Lamiaceae family, is widespread. The demand for Ajuga species has risen sharply because of their medicinal, ornamental, and pharmacological properties. These wide-ranging plants are being rapidly depleted due to over-collection for ornamental and medicinal purposes, as well as by habitat destruction and deforestation. Ajuga boninsimae, A. bracteosa, A. ciliate, A. genevensis, A. incisa, A. makinoi, A. multiflora, A. pyramidalis, A. shikotanensis, A. reptans, and A. vestita are categorized and protected as endangered plants. In vitro plant culture has therefore emerged for the conservation and mass clonal propagation of rare plants. This mini-review covers the current in vitro scenario in the propagation of Ajuga species. Adventitious or axillary shoots are initiated on the leaf, petiole and internodes, as well as roots, nodes, and shoot tip explants. Shoot induction is predominantly dependent on plant growth regulators added to the culture medium. Full- or half-strength Murashige and Skoog medium with or without auxin is used for in vitro rooting. Rooted shoots need to be acclimatized in the greenhouse with an estimated 82–100% survival rate.     

Different levels of hyphal self-incompatibility modulate interconnectedness of mycorrhizal networks in three arbuscular mycorrhizal fungi within the Glomeraceae

Arbuscular mycorrhizal fungi (AMF) live in symbiosis with most plant species and produce underground extraradical hyphal networks functional in the uptake and translocation of mineral nutrients from the soil to host plants. This work investigated whether fungal genotype can affect patterns of interconnections and structural traits of extraradical mycelium (ERM), by comparing three Glomeraceae species growing in symbiosis with five plant hosts. An isolate of Funneliformis coronatus consistently showed low ability to form interconnected ERM and self-incompatibility that represented up to 21 % of hyphal contacts. The frequency of post-fusion self-incompatible interactions, never detected before in AMF extraradical networks, was 8.9 %. In F. coronatus ERM, the percentage of hyphal contacts leading to perfect hyphal fusions was 1.2–7.7, while it ranged from 25.8–48 to 35.6–53.6 in Rhizophagus intraradices and Funneliformis mosseae, respectively. Low interconnectedness of F. coronatus ERM resulted also from a very high number of non-interacting contacts (83.2 %). Such findings show that AMF genotypes in Glomeraceae can differ significantly in anastomosis behaviour and that ERM interconnectedness is modulated by the fungal symbiont, as F. coronatus consistently formed poorly interconnected networks when growing in symbiosis with five different host plants and in the asymbiotic stage. Structural traits, such as extent, density and hyphal self-compatibility/incompatibility, may represent key factors for the differential performance of AMF, by affecting fungal absorbing surface and foraging ability and thus nutrient flow from soil to host roots.     

Illumina-based analysis of the rhizosphere microbial communities associated with healthy and wilted Lanzhou lily (<Emphasis Type="Italic">Lilium davidii</Emphasis> var. <Emphasis Type="Italic">unicolor</Emphasis>) plants grown in the field

Lanzhou lily (Liliumdavidii var. unicolor) is the best edible lily as well as a traditional medicinal plant in China. The microbes associated with plant roots play crucial roles in plant growth and health. However, little is known about the differences of rhizosphere microbes between healthy and wilted Lanzhou lily (Lilium davidii var. unicolor) plants. The objective of this study was to compare the rhizosphere microbial community and functional diversity of healthy and wilted plants, and to identify potential biocontrol agents with significant effect. Paired end Illumina Mi-Seq sequencing of 16S rRNA and ITS gene amplicons was employed to study the bacterial and fungal communities in the rhizosphere soil of Lanzhou lily plants. BIOLOG technology was adopted to investigate the microbial functional diversity. Our results indicated that there were major differences in the rhizosphere microbial composition and functional diversity of wilted samples compared with healthy samples. Healthy Lanzhou lily plants exhibited lower rhizosphere-associated bacterial diversity than diseased plants, whereas fungi exhibited the opposite trend. The dominant phyla in both the healthy and wilted samples were Proteobacteria and Ascomycota, i.e., 34.45 and 64.01 %, respectively. The microbial functional diversity was suppressed in wilted soil samples. Besides Fusarium, the higher relative abundances of Rhizoctonia, Verticillium, Penicillium, and Ilyonectria (Neonectria) in the wilted samples suggest they may pathogenetic root rot fungi. The high relative abundances of Bacillus in Firmicutes in healthy samples may have significant roles as biological control agents against soilborne pathogens. This is the first study to find evidence of major differences between the microbial communities in the rhizospheric soil of healthy and wilted Lanzhou lily, which may be linked to the health status of plants.     

Nutrient allocation and photochemical responses of <Emphasis Type="Italic">Populus</Emphasis> × <Emphasis Type="Italic">canadensis</Emphasis> ‘Neva’ to nitrogen fertilization and exogenous <Emphasis Type="Italic">Rhizophagus irregularis</Emphasis> inoculation

Arbuscular mycorrhizal fungi (AMF) can promote plant growth performance, but their effectiveness varies depending on soil nitrogen (N) availability. To clarify the effectiveness of exogenous AMF along an N-fertilization gradient (0, 2, 10, 20, and 30 mM), the impacts of exogenous Rhizophagus irregularis and N on the growth, photochemical activity, and nutritional status of Populus?×?canadensis ‘Neva’ in natural soil were evaluated in a pot experiment. The results showed that the 10 mM N level was the optimal fertilization regime with the highest promotion effect on plant growth and the maximum quantum yield of photosystem II (PSII) (F_v/F_m). Excess N (20 and 30 mM) fertilization reduced the actual quantum yield of PSII (ФPSII) and the F_v/F_m of the plants. Regardless of the N availability, inoculated plants exhibited greater F_v/F_m values than did non-inoculated plants. The biomass of inoculated plants was significantly higher compared with the control under low N levels (0 and 2 mM). Under high N levels, inoculated plants showed significant increases in ФPSII. Moreover, the nutrient imbalance of plants inoculated with exogenous R. irregularis was eased by increasing P, Fe, Mn and Cu uptake in roots and higher P, Ca, Mg, Fe, Mn and Zn concentrations in leaves. Moreover, the F_v/F_m and ФPSII exhibited positive correlations with P, Ca, Mg and Zn concentrations in leaves. In conclusion, inoculation with exogenous R. irregularis can benefit plant fitness by improving the photochemical capacity and nutrient composition of poplar under different N levels.     

Cloning and expression of <Emphasis Type="Italic">Perilla frutescens FAD2</Emphasis> gene and polymorphism analysis among cultivars

?12 fatty acid desaturase (FAD2) is a key enzyme for linoleic acid and linolenic acid biosynthesis. Perilla frutescens is a special oil plant species with highest linolenic acid content. In this study, based on RACE, two alleles for one FAD2 gene were isolated from P. frutescens cultivar C2: the 3956 bp PfFAD2a and the 3959 bp PfFAD2b, both with a full-length cDNA of 1526 bp, and both encoding a 382aa basic protein. The alleles have identities of over 98%, and their encoded proteins differ only by substitution of a strongly similar residue. Saccharomyces cerevisiae heterologous expression suggested that PfFAD2a/b both encode a bio-functional FAD2 enzyme. Phylogenetic analyses indicated that PfFAD2 shows the highest homologies to FAD2 genes from dicots such as Boraginaceae and Burseraceae. PfFAD2a/b expressions are mainly restricted to developing seeds. PfFAD2a/b expression in the seedling leaf is upregulated by cold (4 °C) and repressed by heat (42 °C). Each of the eight cultivars contains two alleles for one PfFAD2 and 40 SNP sites are found. One allelic gene in cultivars C1 and P1 is pseudogene because of premature stop codon mutation in 5′ coding region. All other normal PfFAD2 genes/allelic genes encode identical or very similar proteins. PfFAD2a/b expression level in developing seeds also varies among the eight cultivars. This study provides systemic molecular and functional features of PfFAD2 and enables its application in the study of plant fatty acids traits.     

Changes in the expression of key genes involved in the biosynthesis of menthol and menthofuran in <Emphasis Type="Italic">Mentha piperita</Emphasis> L. under drought stress

Peppermint (Mentha piperita) is known as an important medicinal plant throughout the world. In the present study, after exposing peppermint plants under drought stress, the qRT-PCR was use to analyze the expression of genes involved in menthol biosynthesis pathway and encoding: limonene synthase (lS), limon-3-hydroxylase (l3oh), trans-isopiperitenol dehydrogenase (ipd), isopiperitenone reductase (ipr), pulegone reductase (pr), menthol dehydrogenase (mdeh), and menthofuran synthase (mfs), which also evaluated the morphological and physiological traits. The results revealed that due to water stress, the gene expression levels of ipd, ipr, and mfs were increased, whereas the gene expression level of pr and mdeh was decreased under water stress conditions. The most of essential oil components (menthol, menthofuran, and plugene), which were analyzed by gas chromatography–mass spectrometry (GC–MS), was positively correlated with genes expression. Drought stress decreased morphological and induces increasing contents of pulegone and menthofuran and reduction in menthol percentages. Results from this study suggest that up-regulation of mfs might contribute to the altered of menthofuran as well as down-regulation of mdeh might cause the reduction of menthol. Furthermore, increasing ls gene expression levels might induce more essential oil yield, while reduction of mfs gene expression levels causes an improvement of essential oil quality.     

Biodiversity of arbuscular mycorrhizal fungi in the drawdown zone of the Three Gorges Reservoir under different fertilization histories

Impoundment of the Three Gorges Reservoir (TGR) has dramatically influenced the riparian environment and shaped a new drawdown zone, which has experienced long-term winter conditions and short periods of summer flooding. The community structure and diversity of arbuscular mycorrhizal (AM) fungi (AMF) were investigated in three areas with different fertilization histories [Area A (5 years of fertilization), Area B (3 years of fertilization) and Area C (no fertilization)] in the drawdown zone of the TGR. Altogether, 50 AMF species were identified; the genera Acaulospora, Funneliformis and Glomus were predominant. The AM fungal community differed among areas A, B and C. A higher isolation frequency and relative abundance of Acaulospora, Ambispora, Entrophospora and Paraglomus were observed in areas A and B; however, Claroideoglomus, Diversispora, Sclerocystis and Septoglomus were more abundant in Area C. The highest spore density occurred in Area C, and was slightly lower in Area A and lowest in Area B. Conversely, species richness and diversity indices (Shannon–Wiener and evenness indices) were the highest in Area A, followed by areas C and B. Based on nonmetric multidimensional scaling analyses, the distribution of AMF was influenced by plant host, fertilization practice and environmental factors. Among them, the soil physicochemical properties were the main drivers affecting AMF, in which three edaphic attributes (carbon/nitrogen ratio, available phosphorus and potassium content) were significantly correlated (P < 0.001) with the AM fungal community composition in the three areas of the drawdown zone of the TGR.     

<Emphasis Type="Italic">Rhizophagus irregularis</Emphasis> MUCL 41833 can colonize and improve P uptake of <Emphasis Type="Italic">Plantago lanceolata</Emphasis> after exposure to ionizing gamma radiation in root organ culture

Long-lived radionuclides such as ⁹⁰Sr and ¹³⁷Cs can be naturally or accidentally deposited in the upper soil layers where they emit β/γ radiation. Previous studies have shown that arbuscular mycorrhizal fungi (AMF) can accumulate and transfer radionuclides from soil to plant, but there have been no studies on the direct impact of ionizing radiation on AMF. In this study, root organ cultures of the AMF Rhizophagus irregularis MUCL 41833 were exposed to 15.37, 30.35, and 113.03 Gy gamma radiation from a ¹³⁷Cs source. Exposed spores were subsequently inoculated to Plantago lanceolata seedlings in pots, and root colonization and P uptake evaluated. P. lanceolata seedlings inoculated with non-irradiated AMF spores or with spores irradiated with up to 30.35 Gy gamma radiation had similar levels of root colonization. Spores irradiated with 113.03 Gy gamma radiation failed to colonize P. lanceolata roots. P content of plants inoculated with non-irradiated spores or of plants inoculated with spores irradiated with up to 30.35 Gy gamma radiation was higher than in non-mycorrhizal plants or plants inoculated with spores irradiated with 113.03 Gy gamma radiation. These results demonstrate that spores of R. irregularis MUCL 41833 are tolerant to chronic ionizing radiation at high doses.     

Communities of arbuscular mycorrhizal fungi under <Emphasis Type="Italic">Picconia azorica</Emphasis> in native forests of Azores

Arbuscular mycorrhizal fungi (AMF) from the rhizosphere of the endemic Laurisilva tree, Picconia azorica, were characterised at two sites in each of two Azorean islands (Terceira and São Miguel). Forty-six spore morphotypes were found, and DNA extraction was attempted from individual spores of each of these. DNA was obtained from 18 of the morphotypes, from which a 1.5 kb long fragment of the nuclear ribosomal RNA gene (SSU-ITS-LSU) was sequenced. A total of 125 AMF sequences were obtained and assigned to 18 phylotypes. Phylogenetic analysis revealed sequences belonging to the families, Acaulosporaceae, Archaeosporaceae, Claroideoglomeraceae, Gigasporaceae and Glomeraceae. Phylotype richness changed between islands and between sampling sites at both islands suggesting that geographical and historical factors are determinant in shaping AMF communities in native forest of Azores. Ecological analysis of the molecular data revealed differences in AMF community composition between islands. In Terceira, the rhizosphere of P. azorica was dominated by species belonging to Acaulosporaceae and Glomeraceae, while São Miguel was dominated by members of Glomeraceae and Gigasporaceae. This is the first molecular study of AMF associated with P. azorica in native forest of the Azores. These symbiont fungi are key components of the ecosystem. Further research is needed to develop their use as promoters of plant establishment in conservation and restoration of such sites.