Comparison of photosynthetic performances of marine picocyanobacteria with different configurations of the oxygen-evolving complex

The extrinsic PsbU and PsbV proteins are known to play a critical role in stabilizing the Mn₄CaO₅ cluster of the PSII oxygen-evolving complex (OEC). However, most isolates of the marine cyanobacterium Prochlorococcus naturally miss these proteins, even though they have kept the main OEC protein, PsbO. A structural homology model of the PSII of such a natural deletion mutant strain (P. marinus MED4) did not reveal any obvious compensation mechanism for this lack. To assess the physiological consequences of this unusual OEC, we compared oxygen evolution between Prochlorococcus strains missing psbU and psbV (PCC 9511 and SS120) and two marine strains possessing these genes (Prochlorococcus sp. MIT9313 and Synechococcus sp. WH7803). While the low light-adapted strain SS120 exhibited the lowest maximal O₂ evolution rates (P_max per divinyl-chlorophyll a, per cell or per photosystem II) of all four strains, the high light-adapted strain PCC 9511 displayed even higher P^Chl_max and P^PSII_max at high irradiance than Synechococcus sp. WH7803. Furthermore, thermoluminescence glow curves did not show any alteration in the B-band shape or peak position that could be related to the lack of these extrinsic proteins. This suggests an efficient functional adaptation of the OEC in these natural deletion mutants, in which PsbO alone is seemingly sufficient to ensure proper oxygen evolution. Our study also showed that Prochlorococcus strains exhibit negative net O₂ evolution rates at the low irradiances encountered in minimum oxygen zones, possibly explaining the very low O₂ concentrations measured in these environments, where Prochlorococcus is the dominant oxyphototroph.     

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.     

Phylogeny,antimicrobial, antioxidant and enzyme-producing potential of fungal endophytes found in <Emphasis Type="Italic">Viola odorata</Emphasis>

Viola odorata, a medicinal plant, is traditionally used to treat common cold, congestion and cough. Given its medicinal properties and occurrence in the northwestern Himalayas, we isolated and characterized endophytic fungi from this plant morphologically, microscopically and by internal transcribed spacer-based rDNA sequencing. In total, we isolated 27 morphotypes of endophytes belonging to phyla Ascomycota and Basidiomycota. The roots showed the highest diversity of endophyte as well as fungal dominance, followed by leaves and leaf nodes. The fungal extract of VOR16 (Fusarium oxysporum) displayed potent antimicrobial activity against Salmonella typhimurium, Klebsiella pneumoniae and Escherichia coli, with a minimum inhibitory concentration of 0.78, 0.78 and 1.56 μg/mL, respectively, while fungal extract VOLF4 (Aspergillus sp.) exhibited promising antioxidant activity (IC₅₀ of 17.4 μg/mL). To identify the components responsible for various bioactivities, we analyzed the content of penicillin G in the extract of bioactive endophytes. The results suggested that the expression of penicillin G under the fermentation conditions applied was too low to display antimicrobial effects. Thus, the activity may be contributed by a different, novel secondary metabolite. The antioxidant activity of VOLF4 may be attributed to its high content of flavonoids. Of the endophytic fungi assessed, 27% were found to be enzyme producers. The highest zone of clearance was observed in VOLN5 (Colletotrichum siamense) for protease production. Only VOR5 (Fusarium nematophilum) was found to be a producer of cellulase, glutenase, amylase and protease. In summary, this is the first report of the isolation of endophytes, namely Fusarium nematophilum, Colletotrichum trifolii, C. destructivum, C. siamense and Peniophora sp., from V. odorata and their bioactive and enzyme-producing potential.     

Correction to: Neighbourhood stories: role of neighbour identity,spatial location and order of arrival in legume and non-legume initial interactions

Background and aims

We characterized fungal endophytes of seeds of invasive, non-native Phragmites from three sites in the Great Lakes region to determine if fungal symbiosis could contribute to invasiveness through their effects on seed germination and seedling growth.

Methods

Field-collected seeds were surface sterilized and plated on agar to culture endophytes for ITS sequencing. Prevalence of specific endophytes from germinated and non-germinated seeds, and from seedlings, was compared.

Results

One-third of 740 seeds yielded endophyte isolates. Fifteen taxa were identified with Alternaria sp. representing 54% of all isolates followed by Phoma sp. (21%) and Penicillium corylophilum (12%). Overall germination of seeds producing an isolate (36%) was significantly higher than seeds not producing an isolate (20%). Penicillium in particular was strongly associated with increased germination of seeds from one site. Sixty-three isolates and 11 taxa were also obtained from 30 seedlings where Phoma, Penicillium and Alternaria respectively were most prevalent. There was a significant effect of isolating an endophyte from the seed on seedling growth.

Conclusions

These results suggest that many endophyte taxa are transmitted in seeds and can increase seed germination and seedling growth of invasive Phragmites. The role of fungal endophytes in host establishment, growth and invasiveness in nature requires further research.

     

Study of the oxidation of alkanes in their mixtures with oxygen and air under the action of a pulsed volume nanosecond discharge

The slow oxidation of alkanes (from methane to hexane) in their stoichiometric mixtures with oxygen or air under the action of nanosecond pulsed discharges was investigated. The discharges were excited in a tube of diameter 5 cm and length of 20 cm by 25-ns voltage pulses with an amplitude of 10 kV and a repetition rate of 40 Hz. The initial pressure in the mixture was varied in the range 0.76–10.1 torr. The current, the electric field strength, and the power deposited in a discharge were measured with a nanosecond time resolution. In time-resolved and time-integrated measurements, the intensities of the following bands were determined: CO ₂ ⁺ (B²Σ → X²Π, δv=0), CH(A²Δ, v′=0 → X²Π, v″=0), OH(A²Σ, v′=0 → X²Π, v″=0), CO(B¹Σ, v′=0 → A¹Π, v″=2), NO(A²Σ → X²Π, δv=3), N₂(C³Π, v′=1 → B³Π, v″=7), N₂(B³Π, v′=6 → A³Σ, v″=3), and N ₂ ⁺ (B²Σ, v′=0 → X²Σ, v″=2). The methane concentration was measured from the absorption of He-Ne laser radiation. Based on the results of optical measurements, the times of the complete oxidation of hydrocarbons were determined.     

Nitrogen fixing bacterial diversity in a tropical estuarine sediments

Microorganisms play a significant role in biogeochemical cycles, especially in the benthic and pelagic ecosystems. Role of environmental parameters in regulating the diversity, distribution and physiology of these microorganisms in tropical marine environment is not well understood. In this study, we have identified dinitrogen (N₂) fixing bacterial communities in the sediments by constructing clone libraries of nitrogenase (nifH) gene from four different stations in the Cochin estuary, along the southeastern Arabian Sea. N₂ fixing bacterial clones revealed that over 20 putative diazotrophs belong to alpha-, beta-, gamma-, delta- and epsilon- proteobacteria and firmicutes. Predominant genera among these were Bradyrhizobium sp. (α-proteobacteria), Dechloromonas sp. (β-proteobacteria); Azotobactor sp., Teredinibacter sp., Methylobacter sp., Rheinheimera sp. and Marinobacterium sp. (γ-proteobacteria); Desulfobacter sp., Desulfobulbus sp. and Desulfovibrio sp. (δ -proteobacteria); Arcobacter sp. and Sulfurospirillum sp. (ε-proteobacteria). Nostoc sp. was solely identified among the cyanobacterial phylotype. Nitrogen fixing Sulfate reducing bacteria (SRBs) such as Desulfobulbus sp., Desulfovibrio sp., Desulfuromonas sp., Desulfosporosinus sp., Desulfobacter sp., were also observed in the study. Most of the bacterial nifH sequences revealed that the identities of N₂ fixing bacteria were less than 95% similar to that available in the GenBank database, which suggested that the sequences were of novel N₂ fixing microorganisms. Shannon-Weiner diversity index of nifH gene ranged from 2.95 to 3.61, indicating an inflated diversity of N₂ fixing bacteria. Canonical correspondence analysis (CCA) implied positive correlation among nifH diversity, N₂ fixation rate and other environmental variables.     

Diversity of Culturable Plant Growth-Promoting Bacterial Endophytes Associated with Sugarcane Roots and Their Effect of Growth by Co-Inoculation of Diazotrophs and Actinomycetes

Application of environmentally friendly agents to reduce the use of chemicals and to enhance growth of plants is an ultimate goal of sustainable agriculture. The use of plant growth-promoting endophytes has become of great interest as a way to enhance plant growth and additionally protect plants from phytopathogens. In this study, 135 isolates of endophytic bacteria including actinomycetes were isolated from roots of commercial sugarcane plants cultivated in Thailand and were characterized for plant growth-promoting (PGP) traits. Based on morphological and 16S rRNA sequence analysis, the endophytes were distributed into 14 genera of which the most dominant species belong to Bacillus, Enterobacter, Microbispora, and Streptomyces. Two strains of endophytic diazotrophs, Bacillus sp. EN-24 and Enterobacter sp. EN-21; and two strains of actinomycetes, Microbispora sp. GKU 823 and Streptomyces sp. GKU 895, were selected based on their PGP traits including 1-aminocyclopropane-1-decarboxylate deaminase, indole-3-acetic acid, nitrogen fixation, phosphate solubilization, and siderophore production for evaluation of sugarcane growth enhancement by individual and co-inoculation. Sixty days after co-inoculation by endophytic diazotrophs and actinomycetes, the growth parameters of sugarcane plants were significantly greater than that of individual and un-inoculated plants. The results indicated that these endophytes have high potential as PGP agents that could be applied to promote sugarcane growth and could be developed as active added value biofertilizers in the future.     

Mixotrophic cultivation of <Emphasis Type="Italic">Chlorococcum</Emphasis> sp. under non-controlled conditions using a digestate from pig manure within a biorefinery

A number of business opportunities may arise from microalgae and wastewater treatment becoming an integrated system, as biofuels and high-added value products could be obtained simultaneously. This study, performed under controlled and non-controlled conditions, aimed at cultivating Chlorococcum sp. using a digestate from pig manure as culture medium and assessing its growth and biochemical composition for further applications. Under controlled conditions, cultures of Chlorococcum sp. were established testing various digestate dilutions (v/v). It was found that all tested dilutions (up to 8% v/v) promoted a higher biomass density, compared to the control culture in modified Bold’s Basal Medium (modified BBM). Under non-controlled conditions, it was found that the biomass productivity using the digestate diluted 5.6% v/v (23.4 mg L^?1 day^?1) was statistically similar to the one obtained using modified BBM (26.4 mg L^?1 day^?1). The volatile fatty acids contained in the digestate might have allowed mixotrophic growth for Chlorococcum sp. The intracellular lipid content in Chlorococcum sp. remained constant throughout the experiments in both, treatment and control cultures, while carbohydrates increased from 20 to 45% of the cell dry weight in the treatment and from 20 to 42% in the control one. It was concluded that conditions of nitrogen starvation and fluctuating irradiance and temperature benefit carbohydrate accumulation in this strain, since intracellular carbohydrate content increased nearly two-fold during this period. Additionally, the obtained biomass has the potential to be used as feedstock for bioethanol production. This system can meet the concept of a microalgae-based biorefinery, in which biofuels and high-added value products are produced from microalgae and wastewater.     

Amino acid composition of leaf,grain and bracts of japonica rice (<Emphasis Type="Italic">Oryza Sativa</Emphasis> ssp. <Emphasis Type="Italic">japonica</Emphasis>) and its response to nitrogen fertilization

Heat stress is one of the main abiotic stresses that limit plant growth. The effects of high temperature on oxidative damage, PSII activity and D1 protein turnover were studied in three wheat varieties with different heat susceptibility (CS, YN949 and AK58). The results showed that heat stress induced lower lipid peroxidation in AK58 and YN949 than CS, which was related to different changes of SOD, CAT, POD and H₂O₂. Similarly, AK58 and YN949 performed better PSII photochemical efficiency (F_v/F_m, ΦPSII and ETR) under high temperature, which was attributed to rapid synthesis and degradation of D1 protein. Moreover, higher expression of D1 protein turnover-related genes (PsbA, STN8, PBCP, Deg1, Deg2, Deg5, Deg8, FtsH1/5 and FtsH2/8) and SOD activity in AK58 and YN949 under normal conditions also established a basis for acclimatizing high temperatures, thereby alleviating PSII photoinhibition and reducing oxidative damage when exposed to heat stress.     

Novel Cryphonectriaceae from La Réunion and South Africa,and their pathogenicity on <Emphasis Type="Italic">Eucalyptus</Emphasis>

Fungi in the Cryphonectriaceae are important canker pathogens of plants in the Melastomataceae and Myrtaceae (Myrtales). These fungi are known to undergo host jumps or shifts. In this study, fruiting structures resembling those of Cryphonectriaceae were collected and isolated from dying branches of Syzygium cordatum and root collars of Heteropyxis natalensis in South Africa, and from cankers on the bark of Tibouchina grandifolia in La Réunion. A phylogenetic species concept was used to identify the fungi using partial sequences of the large subunit and internal transcribed spacer regions of the nuclear ribosomal DNA, and two regions of the β-tubulin gene. The results revealed a new genus and species in the Cryphonectriaceae from South Africa that is provided with the name Myrtonectria myrtacearum gen. et sp. nov. Two new species of Celoporthe (Cel.) were recognised from La Réunion and these are described as Cel. borbonica sp. nov. and Cel. tibouchinae sp. nov. The new taxa were mildly pathogenic in pathogenicity tests on a clone of Eucalyptus grandis. Similar to other related taxa in the Cryphonectriaceae, they appear to be endophytes and latent pathogens that could threaten Eucalyptus forestry in the future.     

Effect of vermicompost fertilizer on photosynthetic characteristics of chickpea (Cicer arietinum L.) under drought stress

Water availability is an important factor for plant growth in arid environments. In recent decades, vermicompost (VC) fertilizer has been used in agriculture as a safe and effective fertilizer with high water-holding capacity. The aim of the present study was to characterize effects of VC fertilizer on photosynthetic activity of chickpea (Cicer arietinum L. cv. Karaj) under drought conditions at three different growth stages. Tests were carried out with four volumetric ratios of VC to soil, i.e., 0:100, 10:90, 20:80, and 30:70, and three levels of drought stress, i.e., no stress (NS), moderate drought (MS), and severe drought (SS) (100, 75, and 25% of field capacity, respectively). Evaluations were performed at the seedling, flowering, and podding stage. We found that the VC treatment under NS conditions significantly increased total chlorophyll content [Chl (a+b)], intercellular CO₂ concentration (C_i), net photosynthetic rate (P_N), transpiration rate (E), and maximal quantum yield of PSII photochemistry (F_v/F_m) at all three stages. The VC addition of 10 and 20% significantly enhanced the Chl content and F_v/F_m under MS and F_v/F_m, C_i, and P_N under SS at the flowering stage. In conclusion, our results proved a positive effect of the VC fertilizer on photosynthesis of chickpea under NS conditions, but it was not found under MS and SS.     

Intrapopulation heterogeneity of the fluorescence parameters of the marine plankton alga <Emphasis Type="Italic">Thalassiosira weissflogii</Emphasis> at various nitrogen levels

Fluorescence of the marine alga Thalassiosira weissflogii (Grunow) Fryxell et Hasle with open (F _o ) and closed (F _m ) reaction centers of photosystem 2 (PS 2) and its relative variable fluorescence (F _v/F _m ) were measured at various levels of inorganic nitrogen. A significant heterogeneity of the population in terms of these parameters was revealed. Some cells within the population were more sensitive to nitrogen deficiency, and their photosynthetic apparatus was disrupted to a greater extent. The cells within a population also differed in terms of their ability to recover after incubation at low nitrogen levels. Enhancement of nitrogen deficiency resulted in an increase in the variability of the F _o and F _v/F _m values of the cells. Fluorescence variability decreased at a less pronounced deficiency. Fluorescence variability should be taken into consideration in the studies concerning responses of algae to changes in nutrient contents.     

The eurythermal adaptivity and temperature tolerance of a newly isolated psychrotolerant Arctic <Emphasis Type="Italic">Chlorella</Emphasis> sp.

A new strain of Chlorella sp. (Chlorella-Arc), isolated from Arctic glacier melt water, was found to have high specific growth rates (μ) between 3 and 27 °C, with a maximum specific growth rate of 0.85 day^?1 at 15 °C, indicating that this strain was a eurythermal strain with a broad temperature tolerance range. To understand its acclimation strategies to low and high temperatures, the physiological and biochemical responses of the Chlorella-Arc to temperature were studied and compared with those of a temperate Chlorella pyrenoidosa strain (Chlorella-Temp). As indicated by declining F _v/F _m, photoinhibition occurred in Chlorella-Arc at low temperature. However, Chlorella-Arc reduced the size of the light-harvesting complex (LHC) to alleviate photoinhibition, as indicated by an increasing Chl a/b ratio with decreasing temperatures. Interestingly, Chlorella-Arc tended to secrete soluble sugar into the culture medium with increasing temperature, while its intracellular soluble sugar content did not vary with temperature changes, indicating that the algal cells might suffer from osmotic stress at high temperature, which could be adjusted by excretion of soluble sugar. Chlorella-Arc accumulated protein and lipids under lower temperatures (<15 °C), and its metabolism switched to synthesis of soluble sugar as temperatures rose. This reflects a flexible ability of Chlorella-Arc to regulate carbon and energy distribution when exposed to wide temperature shifts. More saturated fatty acids (SFA) in Chlorella-Arc than Chlorella-Temp also might serve as the energy source for growth in the cold and contribute to its cold tolerance.     

<Emphasis Type="Italic">PAD4</Emphasis>, <Emphasis Type="Italic">LSD1</Emphasis> and <Emphasis Type="Italic">EDS1</Emphasis> regulate drought tolerance,plant biomass production,and cell wall properties

Key message

Arabidopsis and poplar with modified PAD4, LSD1 and EDS1 genes exhibit successful growth under drought stress. The acclimatory strategies depend on cell division/cell death control and altered cell wall composition.

Abstract

The increase of plant tolerance towards environmental stresses would open much opportunity for successful plant cultivation in these areas that were previously considered as ineligible, e.g. in areas with poor irrigation. In this study, we performed functional analysis of proteins encoded by PHYTOALEXIN DEFICIENT 4 (PAD4), LESION SIMULATING DISEASE 1 (LSD1) and ENHANCED DISEASE SUSCEPTIBILITY 1 (EDS1) genes to explain their role in drought tolerance and biomass production in two different species: Arabidopsis thaliana and Populus tremula × tremuloides. Arabidopsis mutants pad4-5, lsd1-1, eds1-1 and transgenic poplar lines PAD4-RNAi, LSD1-RNAi and ESD1-RNAi were examined in terms of different morphological and physiological parameters. Our experiments proved that Arabidopsis PAD4, LSD1 and EDS1 play an important role in survival under drought stress and regulate plant vegetative and generative growth. Biomass production and acclimatory strategies in poplar were also orchestrated via a genetic system of PAD4 and LSD1 which balanced the cell division and cell death processes. Furthermore, improved rate of cell division/cell differentiation and altered physical properties of poplar wood were the outcome of PAD4- and LSD1-dependent changes in cell wall structure and composition. Our results demonstrate that PAD4, LSD1 and EDS1 constitute a molecular hub, which integrates plant responses to water stress, vegetative biomass production and generative development. The applicable goal of our research was to generate transgenic plants with regulatory mechanism that perceives stress signals to optimize plant growth and biomass production in semi-stress field conditions.

     

Antimicrobial activity,cytotoxicity and chemical analysis of lemongrass essential oil (<Emphasis Type="Italic">Cymbopogon flexuosus</Emphasis>) and pure citral

The aim of this study was to determine the antimicrobial effects of lemongrass essential oil (C. flexuosus) and to determine cytotoxic effects of both test compounds on human dermal fibroblasts. Antimicrobial susceptibility screening was carried out using the disk diffusion method. Antimicrobial resistance was observed in four of five Acinetobacter baumannii strains with two strains confirmed as multi-drug-resistant (MDR). All the strains tested were susceptible to both lemongrass and citral with zones of inhibition varying between 17 to 80 mm. The mean minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of citral (mic—0.14 % and mbc—0.3 % v/v) was lower than that of Lemongrass (mic—0.65 % and mbc—1.1 % v/v) determined using the microtitre plate method. Cell viability using human dermal fibroblasts (HDF; 106-05a) was determined following exposure to both compounds and a control (Grapeseed oil) using the XTT assay and the IC₅₀ determined at 0.095 % (v/v) for citral and 0.126 % (v/v) for lemongrass. Grapeseed oil had no effect on cell viability. Live cell imaging was performed using the LumaScope 500 imaging equipment and changes in HDF cell morphology such as necrotic features and shrinkage were observed. The ability of lemongrass essential oil (EO) and citral to inhibit and kill MDR A. baumannii highlights its potential for use in the management of drug-resistant infections; however, in vitro cytotoxicity does suggest further tests are needed before in vivo or ex vivo human exposure.     

ITS2 sequence–structure phylogeny reveals diverse endophytic <Emphasis Type="Italic">Pseudocercospora</Emphasis> fungi on poplars

For matching the new fungal nomenclature to abolish pleomorphic names for a fungus, a genus Pseudocercospora s. str. was suggested to host holomorphic Pseudocercosproa fungi. But the Pseudocercosproa fungi need extra phylogenetic loci to clarify their taxonomy and diversity for their existing and coming species. Internal transcribed spacer 2 (ITS2) secondary structures have been promising in charactering species phylogeny in plants, animals and fungi. In present study, a conserved model of ITS2 secondary structures was confirmed on fungi in Pseudocercospora s. str. genus using RNAshape program. The model has a typical eukaryotic four-helix ITS2 secondary structure. But a single U base occurred in conserved motif of U-U mismatch in Helix 2, and a UG emerged in UGGU motif in Helix 3 to Pseudocercospora fungi. The phylogeny analyses based on the ITS2 sequence–secondary structures with compensatory base change characterizations are able to delimit more species for Pseudocercospora s. str. than phylogenic inferences of traditional multi-loci alignments do. The model was employed to explore the diversity of endophytic Pseudocercospora fungi in poplar trees. The analysis results also showed that endophytic Pseudocercospora fungi were diverse in species and evolved a specific lineage in poplar trees. This work suggested that ITS2 sequence–structures could become as additionally significant loci for species phylogenetic and taxonomic studies on Pseudocerospora fungi, and that Pseudocercospora endophytes could be important roles to Pseudocercospora fungi’s evolution and function in ecology.     

Isolation and molecular characterization of the fungal endophytic microbiome from conventionally and organically grown avocado trees in South Florida

Fungal endophytes are the most ubiquitous and highly diverse microorganisms that inhabit the interior of healthy plants. They are important in plant ecology and offer untapped potential to improve plant health and productivity in agroecosystems. The endophytic assemblage of avocado is poorly understood; therefore, surveys of fungal endophytes of Persea americana Mill. (Avocado) in South Florida organic and conventional orchards were conducted. A total of 17 endophytic fungal species were recovered from healthy avocado terminal branches. Endophytic fungal species were identified by rDNA sequencing of the internal transcribed spacer (ITS) region, using UNITE Species Hypotheses to reliably assign a taxon name, and determined as belonging to the genera Alternaria, Cladosporium, Colletotrichum, Corynespora, Diaporthe, Lasiodiplodia, Neofusicoccum, Neopestalotiopsis, Phyllosticta, and Strelitziana. Endophyte community assemblage differed between organic and conventional agroecosystems. This is the first report of Alternaria eichhorniae, Cladosporium tenuissimum, Corynespora cassiicola, Colletotrichum alatae, Diaporthe fraxini-angustifoliae, Lasiodiplodia gonubiensis, Neofusicoccum algeriense, Neofusicoccum andinum, Neopestalotiopsis foedans, Phyllosticta capitalensis, and Strelitziana africana as endophytes of avocado. Evaluation using pathogenicity tests on avocado leaves and terminal branches showed that endophytic fungal isolates did not cause disease symptoms.     

Turfgrasses as model assay systems for high-throughput <Emphasis Type="Italic">in planta</Emphasis> screening of beneficial endophytes isolated from cereal crops

Cereal crops including maize (Zea mays L.) are inhabited by non-disease causing microbes known as endophytes that can promote plant growth, aid in host nutrient acquisition and promote host pathogen resistance. Screening endophytes for beneficial traits in planta using large, slow-growing cereals is challenging, thus a rapid but relevant in planta system is needed. Here, we propose that turfgrasses can be used as high-throughput assay systems for screening cereal microbes for beneficial nutrient traits. Turfgrasses are genetic relatives of cereals, but small with fast growth rates; they can be grown in test tubes under sterile conditions on defined media. Five turfgrass genotypes were evaluated for traits ideal for assaying endophytes with nutrient acquisition traits. Based on these criteria, annual ryegrass (Lolium multiflorum) was selected as a high-throughput assay system. Annual ryegrass was then used to test a collection of maize endophytes for their ability to promote plant biomass in the absence of nitrogen. Out of 75 bacterial endophytes tested, one strain (an Enterobacter sp) consistently promoted root and shoot biomass. We discuss the potential of annual ryegrass as a model assay system to test cereal endophytes for acquisition of various nutrients, changes in root/shoot architecture as well as anti-pathogen traits.