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1.

Aims

Rytidosperma species are native Australian grasses which have different growth rates and phosphorus (P) requirements. This study examined the role of root morphology traits in response to P supply.

Methods

Nine Rytidosperma species ranging from slow- to fast-growth were examined along with Lolium perenne and Bromus hordeaceus. Plants were grown in a glasshouse for 47 days in soil supplied with six levels of P between 0 and 60 mg P per pot. Root mass, length and diameter, root hair length and density, and extent of mycorrhizal colonisation were measured.

Results

Across all species there was a positive correlation (P < 0.001) between P uptake and root mass, length and root hair cylinder volume (RHCV; estimated using root diameter, root hair length and root length) at all levels of P supply. An exception was the RHCV of B. hordeaceus, where expected P uptake was not achieved due to a markedly reduced root length at low-P supply. For the Rytidosperma species, morphological plasticity for specific root length, root mass fraction and root hair length ranged from 1.5-fold to 2.7-fold between high- and low-P supply. However, across all species and P levels no single root morphological trait was identified for universally increasing the size of the root system and P uptake.

Conclusions

Fast-growing species took up more P as a result of an overall larger root mass, greater root length and larger RHCV.
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2.

Background

In this report the phytochemical profile of Nitraria. Retusa (N. Retusa) leaf extracts were identified and their ability to induce apoptosis in human chronic myelogenous erythroleukaemia (K562) was evaluated.

Methods

Apoptosis of the human chronic myelogenous erythroleukaemia (K562) was evidenced by investigating DNA fragmentation, PARP cleavage and caspases 3 and 8 inducing activities, in the presence of N. retusa extracts.

Results

Our study revealed that the tested extracts from N. Retusa contain many useful bioactive compounds. They induced in a time-dependent manner the apoptosis the tested cancerous our cell line. This result was confirmed by ladder DNA fragmentation profile and PARP cleavage, as well as a release in caspase-3 and caspase-8 level.

Conclusion

Our results indicate that the tested compounds have a significant antiproliferative effect which may be due to their involvement in the induction of the extrinsic apoptosic pathway.
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3.

Background and aims

In Malawi, strategies are being sought to boost maize production through improvements in soil fertility. This study assessed the impact of intercropping maize (Zea mays) with pigeon pea (Cajanus cajan) in Lixisols of Malawi on yield, biological N fixation, soil aggregation, and P forms within soil aggregates.

Methods

Maize and pigeon pea were grown intercropped in pots, with varying degrees of root interaction in order to understand the relative importance of biochemical versus physical rhizospheric interactions. Following harvest, soils were separated into aggregate fractions using wet-sieving, and the nutrient content of all fractions was assessed.

Results

The proportion of macroaggregates and microaggregates increased by 52 and 111%, respectively, in the intercropping treatment compared to sole maize, which significantly increased organic P storage in the microaggregates of intercropped compared to sole maize (84 versus 29 mg P kg?1, respectively). Biologically fixed N increased from 89% in the sole pigeon pea to 96% in the intercropped system.

Conclusions

Intercropping maize with pigeon pea can have a significant and positive impact on soil structure as well as nutrient storage in these high P-sorbing soils. This is caused primarily by physical root contact and to a lesser degree by biochemical activities.
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4.

Introduction

Botanicals containing iridoid and phenylethanoid/phenylpropanoid glycosides are used worldwide for the treatment of inflammatory musculoskeletal conditions that are primary causes of human years lived with disability, such as arthritis and lower back pain.

Objectives

We report the analysis of candidate anti-inflammatory metabolites of several endemic Scrophularia species and Verbascum thapsus used medicinally by peoples of North America.

Methods

Leaves, stems, and roots were analyzed by ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) and partial least squares-discriminant analysis (PLS-DA) was performed in MetaboAnalyst 3.0 after processing the datasets in Progenesis QI.

Results

Comparison of the datasets revealed significant and differential accumulation of iridoid and phenylethanoid/phenylpropanoid glycosides in the tissues of the endemic Scrophularia species and Verbascum thapsus.

Conclusions

Our investigation identified several species of pharmacological interest as good sources for harpagoside and other important anti-inflammatory metabolites.
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5.

Objective

To improve the diagnosis and treatment of Penicilliosis marneffei without human immunodeficiency virus infection.

Methods

Analyze and review the clinical features, diagnosis and treatment of six cases of P. marneffei without human immunodeficiency virus infection at The First Affiliated Hospital of Fujian Medical University.

Results

Two cases were diagnosed in the ENT Department, three cases in the respiratory department and one case in the dermatological department. Penicillium marneffei infection was confirmed by sputum culture, blood culture and tissue biopsy. After definite diagnosis, one refused further treatment, and others showed significant improvement.

Conclusion

Penicilliosis marneffei is insidious onset and easy to be escaped and misdiagnosed. To achieve early diagnosis and appropriate treatment, doubtful cases should be alerted for the diagnoses as P. marneffei.
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6.

Objective

To generate Candida antarctica lipase A (CAL-A) mutants with modified fatty acid selectivities and improved lipolytic activities using error-prone PCR (epPCR).

Results

A Candida antarctica lipase A mutant was obtained in three rounds of epPCR. This mutant showed a 14 times higher ability to hydrolyze triacylglycerols containing conjugated linoleic acids, and was 12 and 14 times more selective towards cis-9, trans-11 and trans-10, cis-12 isomers respectively, compared to native lipase. Lipolytic activities towards fatty acid esters were markedly improved, in particular towards butyric, lauric, stearic and palmitic esters.

Conclusion

Directed molecular evolution is an efficient method to generate lipases with desirable selectivity towards CLA isomers and improved lipolytic activities towards esters of fatty acids.
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7.

Aims

During the first days after harvest of Lolium perenne L., N remobilized from roots and stubble forms the main N source for regrowth. Low N uptake from the soil during this period may lead to N loss if N fertilizer is applied too soon. Furthermore, temporary N deprivation has been found to stimulate root growth. We therefore hypothesized that a strategic delay in N application after harvest may improve N-use efficiency of L. perenne grassland by increasing root biomass and reducing N loss.

Methods

In a laboratory and field experiment with L. perenne, we delayed N fertilizer application after harvest for 0, 3, 6, 9 and 12 days, repeated this for up to six harvest cycles, and determined effects on herbage yield, herbage N uptake and root biomass.

Results

In both experiments, delaying N application for up to 12 days had no significant effect on root biomass or total herbage N uptake, but it significantly reduced total herbage yield in the laboratory experiment. Total yield tended to be highest when N application was delayed for 3 days. Two growth periods in the field experiment showed significantly higher N uptake when N application was delayed, possibly due to rainfall-induced N losses in the treatments with shorter delay.

Conclusions

Our results do not provide evidence that delaying N application improves N-use efficiency of L. perenne grassland by increasing root biomass. However, strategic timing of N fertilizer application based on rainfall forecasts could contribute to improve N-use efficiency by reducing N losses from leaching and denitrification.
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8.

Introduction

Mass spectrometry imaging (MSI) experiments result in complex multi-dimensional datasets, which require specialist data analysis tools.

Objectives

We have developed massPix—an R package for analysing and interpreting data from MSI of lipids in tissue.

Methods

massPix produces single ion images, performs multivariate statistics and provides putative lipid annotations based on accurate mass matching against generated lipid libraries.

Results

Classification of tissue regions with high spectral similarly can be carried out by principal components analysis (PCA) or k-means clustering.

Conclusion

massPix is an open-source tool for the analysis and statistical interpretation of MSI data, and is particularly useful for lipidomics applications.
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9.

Introduction

Swine dysentery caused by Brachyspira hyodysenteriae is a production limiting disease in pig farming. Currently antimicrobial therapy is the only treatment and control method available.

Objective

The aim of this study was to characterize the metabolic response of porcine colon explants to infection by B. hyodysenteriae.

Methods

Porcine colon explants exposed to B. hyodysenteriae were analyzed for histopathological, metabolic and pro-inflammatory gene expression changes.

Results

Significant epithelial necrosis, increased levels of l-citrulline and IL-1α were observed on explants infected with B. hyodysenteriae.

Conclusions

The spirochete induces necrosis in vitro likely through an inflammatory process mediated by IL-1α and NO.
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10.

Background and aims

Identification of organic P species is important to understand their origin, turnover in soils and their effects on soil fertility. Attention has been recently devoted to microbial inocula, referred to as Bioeffectors, that are capable to increase P bioavailability and plant uptake. Nevertheless, little is known on the effect of Bioeffectors on soil P forms and their dynamics in agricultural soils upon different P fertilization.

Methods

We investigated the effects of the application of different commercial inocula strains (Trichoderma harzianum T 22, Pseudomonas sp., and Bacillus amyloliquefaciens) alone or in combination with different P fertilizers (triple superphosphate, rock phosphate, and both composted cow- and horse-manure) on soil organic P forms. P forms were characterized by liquid-state 31P–NMR spectroscopy, while plant P uptake from P-treated soil was followed in a greenhouse pot experiment under maize cultivation.

Results

NMR spectra showed that the type of P fertilizer and bioeffectors inoculation, affected the abundance and the composition of organic P forms. The specific capacity of all bioeffectors, and especially Pseudomonas, was related to an increased content of diesters P forms. Pseudomonas, and, to a lesser extent, B. amyloliquefaciens showed the largest increase in combination with organic P amendments, which also provided the largest plant P uptake. This suggests a key role of Diester-P forms in determining P availability in agroecosystems.

Conclusions

Microbial inoculation plays an important role in the dynamics of soil P, inducing a rapid P cycling that prevents P fixation and losses from soils, thus enhancing the P fertilizer use efficiency in agricultural soils.
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11.
12.

Background and aims

Intraspecific aggregation of plant individuals can promote species coexistence by delaying competitive exclusions. However, such impacts may differ among species with contrasting spatial architecture and rely on the spatial distribution of resources.

Methods

We grew a phalanx clonal plant Carex neurocarpa (with aggregated ramets) and a guerilla one Bolboschoenus planiculmis (with diffused ramets) in monocultures or in 1:1 mixtures with an even or a clustered distribution pattern of the two species in homogeneous or heterogeneous soils.

Results

After 16 months, shoot biomass and ramet number were greater in mixtures than in monocultures in C. neurocarpa, but smaller in B. planiculmis. However, the growth of neither C. neurocarpa nor B. planiculmis differed between even and clustered mixtures. Soil nutrient heterogeneity did not significantly affect the growth of either species, but increased relative yield of B. planiculmis and decreased that of C. neurocarpa.

Conclusions

The relative importance of intra- vs. interspecific competition depends on the spatial architecture of plants, and soil nutrient heterogeneity slows down competitive exclusion by decreasing differences in competitive ability between plants. However, our results do not support the idea that intraspecific aggregation of individuals alters competitive interactions between species.
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13.

Introduction

Acylsugar specialized metabolites function as defenses against insect herbivores, and are the most abundant specialized metabolites produced in Solanaceous trichomes. Metabolite profiling provides the foundation for determining the genetic basis of specialized metabolism and its evolution.

Objectives

To profile and identify acylsugar specialized metabolites in three Petunia species: P. axillaris, P. integrifolia and P. exserta.

Methods

Metabolites were profiled using ultra-high performance liquid chromatography/time-of-flight mass spectrometry (UHPLC/TOF MS). Metabolites were purified using solid phase extraction and HPLC, and structures were established using NMR spectroscopy.

Results

Twenty-eight distinct acylsucrose formulas, representing a sampling of more than 100 different detected chemical forms, were purified from three Petunia species and structures have been proposed based on one- and two-dimensional NMR data. 15 of the 28 purified acylsugars were sucrose pentaesters that possess a malonyl group on the fructose ring. These malonate esters can be readily distinguished from other acylsugars based on distinct masses of pseudomolecular ions and fragment ions generated using multiplexed collision-induced dissociation. Chemical diversity of acylsugars was observed between Petunia species, particularly with respect to the lengths of acyl chains and specific acylation positions.

Conclusions

These findings suggest substrate selectivity of various acyltransferases in Petunia species.
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14.

Background

The fungus Colletotrichum is a plant pathogen that causes the anthracnose disease, resulting in huge losses in various crops including the rose-scented geranium (Pelargonium graveolens). Although the bacterial community associated with plants has an important role in the establishment of plant diseases, little is known about what happens in P. graveolens.

Aims

To increase the knowledge about the bacterial community associated with P. graveolens and its relationship with anthracnose disease symptoms.

Methods

Quantitative PCR and high-throughput sequencing were combined to determine the presence of the fungus Colletotrichum and to reveal the bacterial communities associated with different plant parts – root, stem and leaf – and in the rhizosphere and bulk soil, and also to determine the respective bacterial communities associated with P. graveolens leaves symptomatic and asymptomatic for anthracnose disease.

Results

The fungus Colletotrichum was detected in all plant parts and in the surrounding soil. Bacterial communities varied spatially in plants, and the disease symptoms also influenced the composition of the bacterial community. Abundances of operational taxonomic units (OTUs) assigned to the phylum Actinobacteria and to the genus Streptococcus were greatly increased in asymptomatic leaves.

Conclusions

The bacterial community associated to geranium leaves responds to anthracnose symptoms.
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15.

Objectives

To improve the stability and sweetness of the sweet-tasting protein, monellin, by using site-directed mutagenesis and a Pichia pastoris expression system with a GAPDH constitutive promoter.

Results

Both wild-type and E2 N mutant of single-chain monellin gene were cloned into the PGAPZαA vector and expressed in Pichia pastoris. The majority of the secreted recombinant protein, at 0.15 g/l supernatant, was monellin. This was purified by Sephadex G50 chromatography. The sweetness threshold of wild-type and E2 N were 30 μg/ml and 20 μg/ml, respectively. Compared with the proteins expressed in Escherichia coli, the thermostability of both proteins was improved. The N-terminal sequence is determinative for the sweetness of the proteins expressed in yeast strains.

Conclusions

Site-directed mutagenesis, modification of the N-terminus of monellin, and without the need of methanol induction in P. pastoris expression system, indicate the possibility for large-scale production of this sweet-tasting protein.
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16.

Objectives

To achieve biosynthesis of caffeoylmalic acid from glucose in engineered Escherichia coli.

Results

We constructed the biosynthetic pathway of caffeoylmalic acid in E. coli by co-expression of heterologous genes RgTAL, HpaBC, At4CL2 and HCT2. To enhance the production of caffeoylmalic acid, we optimized the tyrosine metabolic pathway of E. coli to increase the supply of the substrate caffeic acid. Consequently, an E. coliE. coli co-culture system was used for the efficient production of caffeoylmalic acid. The final titer of caffeoylmalic acid reached 570.1 mg/L.

Conclusions

Microbial production of caffeoylmalic acid using glucose has application potential. In addition, microbial co-culture is an efficient tool for producing caffeic acid esters.
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17.

Background and aims

Layered profiles of designed soils may provide long-term benefits for green roofs, provided the vegetation can exploit resources in the different layers. We aimed to quantify Sedum root foraging for water and nutrients in designed soils of different texture and layering.

Methods

In a controlled pot experiment we quantified the root foraging ability of the species Sedum album (L.) and S. rupestre (L.) in response to substrate structure (fine, coarse, layered or mixed), vertical fertiliser placement (top or bottom half of pot) and watering (5, 10 or 20 mm week?1).

Results

Water availability was the main driver of plant growth, followed by substrate structure, while fertiliser placement only had marginal effects on plant growth. Root foraging ability was low to moderate, as also reflected in the low proportion of biomass allocated to roots (5–13%). Increased watering reduced the proportion of root length and root biomass in deeper layers.

Conclusions

Both S. album and S. rupestre had a low ability to exploit water and nutrients by precise root foraging in substrates of different texture and layering. Allocation of biomass to roots was low and showed limited flexibility even under water-deficient conditions.
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18.

Objectives

Lycopene biosynthetic genes from Deinococcus radiodurans were co-expressed in Lactococcus lactis to produce lycopene and improve its tolerance to stress.

Results

Lycopene-related genes from D. radiodurans, DR1395 (crtE), DR0862 (crtB), and DR0861 (crtI), were fused in line with S hine-Dalgarno (SD) sequences and co-expressed in L. lactis. The recombinant strain produced 0.36 mg lycopene g-1 dry cell wt after 48 h fermentation. The survival rate to UV irradiation of the recombinant strain was higher than that of the non-transformed strain.

Conclusion

The L. lactis with co-expressed genes responsible for lycopene biosynthesis from D. radiodurans produced lycopene and exhibited increased resistance to UV stress, suggesting that the recombinant strain has important application potential in food industry.
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19.

Objective

To construct a promoter probe vector, pBE-bgaB, to screen strong promoters from Bacillus amyloliquefaciens.

Results

266 colonies containing active promoter elements from the genomic DNA of B. amyloliquefaciens were identified. Among these, promoter P41 exhibited the strongest β-Gal activity in Escherichia coli and B. amyloliquefaciens. Sequence analysis showed that promoter P41 contained P ykuN , a ykuN gene encoding flavodoxin. Optimization of the ribosome-binding site from P41 to P382 improved β-Gal activity by ~ 200%.

Conclusion

A new strong promoter for protein expression and genetic engineering of Bacillus species.
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20.
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