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1.

Background and aim

Saccharothrix algeriensis NRRL B-24137, isolated from a Saharan soil, has been described as a potential biocontrol agent against Botrytis cinerea and other phytopathogens. However, the plant protection mechanisms involved still need to be described. The aim of this study was to determine this protection phenomenon as well as parts of the mechanisms involved, using Arabidopsis thaliana seedlings and B. cinerea.

Methods

The bacterial colonization process was evaluated on A. thaliana seedlings using fluorescence in situ hybridization. Protection of A. thaliana seedlings inoculated with NRRL B-24137 against B. cinerea was then evaluated. Parts of the mechanisms involved in the systemic protection against B. cinerea were evaluated using known mutants of genes involved in jasmonate (JA)/ethylene (ET)/salicylic acid (SA) signaling. Other Arabidopsis mutants, AtrhbohD-3, AtrhbohF-3, and ups1-1 were also screened to determine other parts of the mechanisms involved.

Results

The results showed that the strain NRRL B-24137 colonized, epi- and endophytically, the roots of Arabidopsis seedlings but the strain was not a systemic colonizer during the time of the experiment. The strain NRRL B-24137 also reduced B. cinerea symptoms and the protection was linked to known mechanisms of induced systemic resistance (ISR; JA/ET signaling), as well as to functionality of AtrbohF oxidase and of UPS1. Crosstalk between ET/JA and SA signaling could also be involved.

Conclusions

The isolate NRRL B-24137, after colonizing the root systems of A. thaliana, induces an ISR against B. cinerea, which is JA/ET dependent, but could also require SA crosstalk and protection could also require NAPDH oxidases and UPS1 functionalities.  相似文献   

2.
A study was conducted to examine whether Beauveria bassiana (Balsamo) Vuillemin (Ascomycota: Hypocreales) can colonize grapevine leaf tissues and subsequently confer protection against downy mildew caused by Plasmopara viticola (Berk. and Curt.) Berl. and de Toni. Following the foliar inoculation of plants with conidial suspensions of selected B. bassiana strains, colonization of leaves by the fungus was determined using culture-based and PCR techniques at different time intervals. Seven days following B. bassiana inoculation, grapevine plants were challenged with P. viticola and symptoms were assessed by calculating the disease incidence and severity. Although all tested strains were able to colonize grapevine plants, percent colonization differed significantly among strains. Disease incidence and severity were, on the other hand, significantly reduced in B. bassiana-inoculated plants compared to control plants irrespective of strain. This study is one of very few studies investigating the promising role B. bassiana could play as a plant disease antagonist.  相似文献   

3.

Aim

It is necessary to understand the roles of root exudates involved in plant-microbe interactions to inform practical application of beneficial rhizosphere microbial strains.

Methods

Colonization of Bacillus amyloliquefaciens SQR9 (isolated from cucumber rhizosphere) and Bacillus subtilis N11 (isolated from banana rhizosphere) of their original host was found to be more effective as compared to the colonization of the non-host plant. Organic acids in the root exudates of the two plants were identified by High performance liquid chromatography (HPLC). The chemotactic response and effects on biofilm formation were assessed for SQR9 and N11 in response to cucumber and banana root exudates, as well as their organic acids components.

Results

Citric acid detected exclusively in cucumber exudates could both attract SQR9 and induce its biofilm formation, whereas only chemotactic response but not biofilm formation was induced in N11. Fumaric acid that was only detected in banana root exudates revealed both significant roles on chemotaxis and biofilm formation of N11, while showing only effects on biofilm formation but not chemotaxis of SQR9.

Conclusion

The relationship between PGPR strain and root exudates components of its original host might contribute to preferential colonization. This study advances a clearer understanding of the mechanisms relevant to application of PGPR strains in agricultural production.  相似文献   

4.

Background and Aim

Climate change models are limited by lack of baseline data, in particular carbon (C) allocation to – and dynamics within – soil microbial communities. We quantified seasonal C-assimilation and allocation by plants, and assessed how well this corresponds with intraradical arbuscular mycorrhizal fungal (AMF) storage and structural lipids (16:1ω5 NLFA and PLFA, respectively), as well as microscopic assessments of AMF root colonization.

Methods

Coastal Hypochoeris radicata plants were labeled with 13CO2 in February, July and October, and 13C-allocation to fine roots and NLFA 16:1ω5, as well as overall lipid contents and AM colonization were quantified.

Results

C-allocation to fine roots and AMF storage lipids differed seasonally and mirrored plant C-assimilation, whereas AMF structural lipids and AM colonization showed no seasonal variation, and root colonization exceeded 80 % throughout the year. Molecular analyzes of the large subunit rDNA gene indicated no seasonal AMF community shifts.

Conclusions

Plants allocated C to AMF even at temperatures close to freezing, and fungal structures persisted in roots during times of low C-allocation. The lack of seasonal differences in PLFA and AM colonization indicates that NLFA analyses should be used to estimate fungal C-status. The implication of our findings for AM function is discussed.  相似文献   

5.

Background and aims

The root surface of a plant usually exceeds the leaf area and is constantly exposed to a variety of soil-borne microorganisms. Root pathogens and pests, as well as belowground interactions with beneficial microbes, can significantly influence a plants' performance. Unfortunately, the analysis of these interactions is often limited because of the arduous task of accessing roots growing in soil. Here, we present a soil-free root observation system (SF-ROBS) designed to grow maize (Zea mays) plants and to study root interactions with either beneficial or pathogenic microbes.

Methods

The SF-ROBS consists of pouches lined with wet filter paper supplying nutrient solution.

Results

The aspect of maize grown in the SF-ROBS was similar to soil-grown maize; the plant growth was similar for the shoot but different for the roots (biomass and length increased in the SF-ROBS). SF-ROBS-grown roots were successfully inoculated with the hemi-biotrophic maize fungal pathogen Colletotrichum graminicola and the beneficial rhizobacteria Pseudomonas putida KT2440. Thus, the SF-ROBS is a system suitable to study two major belowground phenomena, namely root fungal defense reactions and interactions of roots with beneficial soil-borne bacteria.

Conclusions

This system contributes to a better understanding of belowground plant microbe interactions in maize and most likely also in other crops.  相似文献   

6.

Aims

This study aimed to determine the effect of arbuscular mycorrhizal (AM) fungi and phosphorus (P) supply levels on β-carotene concentrations in sweet potato (Ipomoea batatas L.) tubers.

Methods

Two commercial AM fungal isolates of Glomus intraradices (IFP Glintra) and Glomus mosseae (IFP Glm) which differ in their life cycles were used. Sweet potato plants were grown in a horizontal split-root system that consisted of two root compartments. A root-free fungal compartment that allowed the quantification of mycelial development was inserted into each root compartment. The two root compartments were inoculated either with the same or with different AM isolates, or remained free of mycorrhizal propagules. Each fungal treatment was carried out in two P supply levels.

Results

In the low P supply level, mycorrhizal colonization significantly increased β-carotene concentrations in sweet potato tubers compared with the non-mycorrhizal plants. Glomus intraradices appeared to be more efficient in increasing β-carotene concentrations than G. mosseae. Dual inoculation of the root system with the two mycorrhizal fungi did not result in a higher increase in tuber β-carotene concentrations than inoculation with the single isolates. Improved P nutrition led to higher plant tuber biomass but was not associated with increased β-carotene concentrations.

Conclusions

The results indicate a remarkable potential of mycorrhizal fungi to improve β-carotene concentrations in sweet potato tubers in low P fertilized soils. These results also suggest that β-carotene metabolism in sweet potato tubers might be specifically activated by root mycorrhizal colonization.  相似文献   

7.

Background and aims

Bacterial endophytes can colonize various plants and organs. However, endophytic bacteria (other than rhizobia) colonizing root nodules in legumes have been rarely analyzed. The present study aimed to examine the colonization and spread of gfp-tagged Paenibacillus polymyxa in soybean plants under gnobiotic conditions.

Methods

Inoculation with gfp-tagged Paenibacillus. polymyxa HKA ?15 alone and in combination with Bradyrhizobium japonicum were done on soybean seedlings. In situ localization was detected through confocal microscopy and PCR.

Results

Inoculation with P. polymyxa-gfp strain alone and in combination with B. japonicum DS-1 had a stimulatory effect on the plant growth. There was an increase in shoot (7.2 %) and root dry weights (14.5 %) when the two strains were co - inoculated over that of B. japonicum inoculation alone. In vivo simultaneous visualization using Confocal Laser Scanning Microscopy (CLSM) showed the localization of the gfp-tagged P. polymyxa cells in the root nodules and its spread in the root tissue, both tap as well as lateral roots. Systemic spread into aerial tissue did not occur as indicated by the absence of bacteria. CLSM observations of the presence of gfp-tagged P. polymyxa in the nodule and roots tissues was corroborated with PCR amplification of the gfp-gene from these tissues.

Conclusions

CLSM and PCR methods confirmed that P. polymyxa invades roots and root nodules of soybean, but the spread is restricted to root tissue only. The strain improves plant growth when inoculated singly or in combination with B. japonicum.  相似文献   

8.

Aim

Few studies have analyzed life-history strategies of arbuscular mycorrhizal fungi (AMF), in terms of the different propagule types they produce, and their ability to colonize new seedlings. The aim was to assess whether life-history strategies influence AMF successional dynamics and assemblages.

Methods

Rosemary (Rosmarinus officinalis L.) seedlings, grown in a mesocosm system, were colonized by either the AMF hyphae coming from a living rosemary plant, or from spores germinating in soil. The AMF community established in the plantlets was monitored every 3 months during 2 years, using terminal restriction fragment length polymorphism of genes coding for rDNA.

Results

The two different sources of AMF propagules resulted in a different initial community colonizing rosemary roots. AMF propagating from hyphae attached to living mycorrhizal-roots seemed to colonize faster and were season-dependent. AMF taxa originating from soil-borne propagules were most frequent over time and exhibit the dominant colonization strategy in this system. The evolution of the AMF community also revealed different strategies in succession.

Conclusions

AMF associated with rosemary evidenced contrasting life-history strategies in terms of source of inoculum for new colonization and hence survival. The observed successional dynamics of AMF have implications for understanding the ecological processes in Mediterranean environments and seasonality of colonization processes.  相似文献   

9.

Background and aims

Accurate predictions of nutrient acquisition by plant roots and mycorrhizas are critical in modelling plant responses to climate change.

Methods

We conducted a field experiment with the aim to investigate root nutrient uptake in a future climate and studied root production by ingrowth cores, mycorrhizal colonization, and fine root N and P uptake by root assay of Deschampsia flexuosa and Calluna vulgaris.

Results

Net root growth increased under elevated CO2, warming and drought, with additive effects among the factors. Arbuscular mycorrhizal colonization increased in response to elevated CO2, while ericoid mycorrhizal colonization was unchanged. The uptake of N and P was not increased proportionally with root growth after 5 years of treatment.

Conclusions

While aboveground biomass was unchanged, the root growth was increased under elevated CO2. The results suggest that plant production may be limited by N (but not P) when exposed to elevated CO2. The species-specific response to the treatments suggests different sensitivity to global change factors, which could result in changed plant competitive interactions and belowground nutrient pool sizes in response to future climate change.  相似文献   

10.

Background and aims

Seeds are inhabited by diverse bacterial and fungal taxa whose colonization patterns are little understood. We hypothesized, however, that specific niches within seeds host microbes.

Methods

In this study, the putative presence of bacteria, inhabiting the seed endosphere of an angiosperm, the melon Cucumis melo reticulatus group cv. ‘Dulce’, was examined by scanning electron microscopy (SEM) and confocal laser-scanning microscopy coupled with double labeling of oligonucleotide probes for fluorescence in situ hybridization (DOPE-FISH).

Results

SEM images showed microbial-like structures in different tissues and FISH revealed endophytic bacteria colonizing the outer and inner seed parts, on perisperm/endosperm envelope, inside the cotyledons as parts of the embryo, and, to a lesser extent, inside embryonic hypocotyl-root axis tissues. Alphaproteobacteria were shown to inhabit the seed coat and the envelope surrounding the embryonic hypocotyl-root tissues, but could not be seen in the cotyledons, whereas Betaproteobacteria were only detected in the outer seed coat. Some Gammaproteobacteria were also seen in the outer seed coat, but were mainly visualized in the cotyledons with a few inside the seed’s embryonic hypocotyl-root tissues, among other bacteria. Firmicutes were visualized inside the seed coat, but mostly inside the cotyledon tissues, on the perisperm/endosperm envelope and inside the embryonic hypocotyl-root axis tissues. Microscopy revealed Actinobacteria inside the inner and outer seed coat and inside the embryonic parts such as cotyledons, with a few inside the hypocotyl-root axis.

Conclusions

This is the first demonstration of niches for the most active groups of bacteria inhabiting different seed tissues of an angiosperm.
  相似文献   

11.

Aims

Nutrients play a key role in arbuscular mycorrhizal (AM) symbiosis. We quantified the response of AM symbiosis of seedlings and adult plants of Plantago lanceolata to fertilization under field conditions in managed grasslands differing in nutrient availability and soil moisture.

Methods

The AM symbiosis was measured as the total extent of AM fungal colonization and frequency of arbuscules or vesicles, and as the relative proportions of morphotypes. We further examined the effects of the surrounding vegetation upon AM symbiosis.

Results

Fertilization decreased total AM colonization and relative arbuscular frequency of the whole mycorrhizal community and of Acaulospora and “fine endophyte” morphotypes in seedling roots, but it had no effect upon the mycorrhiza in adult plants. The decline in arbuscular frequency in seedling roots due to fertilization was greater at the sites with higher nutrient availability and lower N:P ratio. Seedlings surrounded by more forbs had a greater total AM colonization and higher vesicular frequency.

Conclusions

Increased nutrient availability in the initial stages of seedling development has a prominent effect upon AM symbiosis development, but these effects seem to diminish over the long term, as evidenced by the results obtained for adult plants and from the limited effects of parameters characterizing long-term nutrient availability.  相似文献   

12.
13.

Key message

This study established an efficient method of regenerating plants of Ficus lyrata and producing purple-leaved F. lyrata plants through genetic transformation using a VvMybA1 gene of grapevine.

Abstract

Ficus lyrata, a species with unique violin- or guitar-shaped leaves, was regenerated from leaf-derived calli cultured on Murashige and Skoog (MS) basal medium supplemented with 4.5 μM N-phenyl-N’-1, 2, 3-thiadiazol-5-yl urea (TDZ) and 0.5 μM α-naphthalene acetic acid (NAA). Leaf discs were inoculated with Agrobacterium tumefaciens strain EHA 105 harboring a binary vector DEAT that contains the VvMybA1 gene and neomycin phosphotransferase (npt II) gene and subsequently cultured on the established regeneration medium supplemented with 100 mg l?1 kanamycin. Results showed that 87.5 % of the leaf discs produced kanamycin-resistant callus, and 68.8 % of them produced adventitious shoots. Transgenic plants with three leaf colors including green, green-purple, and purple were produced. Regular and quantitative real-time PCR analyses confirmed the integration of transgenes into the host genome. Semi-quantitative RT-PCR analysis indicated that the VvMybA1 gene was responsible for the purple-colored phenotype. Purple-leaved plants with strong color stability grew vigorously in a greenhouse. This study illustrated the feasibility of using a genetically engineered VvMybA1 gene for drastic modification of leaf color of an important woody ornamental plant.  相似文献   

14.
15.

Background and aims

Year of release of a cultivar reflects the agricultural and breeding practices of its time; we hypothesize that there are differences in mycorrhizal responsiveness of new high yielding and old crop plants and landraces. We evaluated the importance of the year of release on mycorrhizal responsiveness, arbuscular mycorrhizal (AM) fungal root colonization and P efficiency. We also analyzed the effect of experimental treatments, P efficiency (P acquisition and P utilization efficiency) and AM fungal root colonization on a potential mycorrhizal responsiveness trend for year of release.

Methods

We conducted a meta-analysis on 39 publications working on 320 different crop plant genotypes.

Results

New cultivars were less intensely colonized but were more mycorrhiza-responsive (and possibly dependent) compared to ancestral genotypes. This trend was potentially influenced by the moderator variables density, pre-germination, plant, plant type and AMF species. AM root colonization was also important for the mycorrhizal responsiveness trend for year of release, but P efficiency was not.

Conclusions

With the data available we could find no evidence that new crop plant genotypes lost their ability to respond to mycorrhiza due to agricultural and breeding practices.  相似文献   

16.

Background & Aims

Searching for root traits underpinning efficient nutrient acquisition has received increased attention in modern breeding programs aimed at improved crop productivity. Root models provide an opportunity to investigate root-soil interactions through representing the relationships between rooting traits and the non-uniform supply of soil resources. This study used simulation modelling to predict and identify phenotypic plasticity, root growth responses and phosphorus (P) use efficiency of contrasting Lupinus angustifolius genotypes to localised soil P in a glasshouse.

Methods

Two L. angustifolius genotypes with contrasting root systems were grown in cylindrical columns containing uniform soil with three P treatments (nil and 20 mg P kg?1 either top-dressed or banded) in the glasshouse. Computer simulations were carried out with root architecture model ROOTMAP which was parameterized with root architectural data from an earlier published hydroponic phenotyping study.

Results

The experimental and simulated results showed that plants supplied with banded P had the largest root system and the greatest P-uptake efficiency. The P addition significantly stimulated root branching in the topsoil, whereas plants with nil P had relatively deeper roots. Genotype-dependent root growth plasticity in response to P supply was shown, with the greatest response to banded P.

Conclusions

Both experimental and simulation outcomes demonstrated that 1) root hairs and root proliferation increased plant P acquisition and were more beneficial in the localised P fertilisation scenario, 2) placing P deeper in the soil might be a more effective fertilisation method with greater P uptake than top dressing, and 3) the combination of P foraging strategies (including root architecture, root hairs and root growth plasticity) is important for efficient P acquisition from a localised source of fertiliser P.  相似文献   

17.

Key message

Embryogenic suspension cultures are suitable for EMS mutagenesis in grapevine, and HRM prescreening of EMS-treated somatic embryo clusters allows rapid detection of point mutations before plant regeneration.

Abstract

Somatic embryogenesis is an excellent system for induced mutagenesis and clonal propagation in woody plants. Our work was focused on establishing a procedure for inducing ethyl methanesulfonate (EMS) mutagenesis in grapevine. Embryogenic cell aggregates (ECAs) growing in liquid medium were treated with increasing concentrations of EMS. We found that EMS dramatically affects the viability of ECAs at concentrations above 20 mM (25.5 ± 2.9 % survival), whereas concentrations above 10 mM affect embryogenic potential (22.1 ± 1.7 % of ECAs gave rise to embryos). Embryo masses generated from EMS-treated embryogenic cell aggregates were prescreened by quantitative PCR-High Resolution Melting (qPCR-HRM) to detect single nucleotide polymorphisms (SNPs) in a 1,000-bp VvNCED1-encoding DNA fragment, which served as the target gene. Detected mutations were verified in regenerated plants by PCR and sequencing. qPCR-HRM analysis of the difference plots for the fluorescence signals allowed detection of a mutation in a sample from an embryogenic aggregate treated with 10 mM EMS. To confirm the nature of the mutation, embryos from this aggregate were recovered and germinated, and leaves were collected for PCR and sequencing analysis. The alignment of sequences from regenerated plants with the wild-type sequence revealed a transitional mutation (G/C to A/T) in the 1,000-bp VvNCED1-encoding region. To our knowledge, this is the first time that EMS mutagenesis has been performed using an embryogenic cell suspension of grapevine.  相似文献   

18.

Background and Aims

Several strains of rhizobacteria may be found in the rhizospheric soil, on the root surface or in association with rice plants. These bacteria are able to colonize plant root systems and promote plant growth and crop yield through a variety of mechanisms. The objectives of this study were to isolate, identify, and characterize putative plant growth-promoting rhizobacteria (PGPR) associated with rice cropped in different areas of southern Brazil.

Methods

Bacterial strains were selectively isolated based on their growth on three selective semi-solid nitrogen-free media. Bacteria were identified at the genus level by PCR-RFLP 16S rRNA gene analysis and partial sequencing methodologies. Bacterial isolates were evaluated for their ability to produce indolic compounds and siderophores and to solubilize phosphate. In vitro biological nitrogen fixation and the ability to produce 1-aminocyclopropane-1-carboxylate deaminase were evaluated for each bacterial isolate used in the inoculation experiments.

Results

In total, 336 bacterial strains were isolated representing 31 different bacterial genera. Strains belonging to the genera Agrobacterium, Burkholderia, Enterobacter, and Pseudomonas were the most prominent isolates. Siderophore and indolic compounds producers were widely found among isolates, but 101 isolates were able to solubilize phosphate. Under gnotobiotic conditions, eight isolates were able to stimulate the growth of rice plants. Five of these eight isolates were also field tested in rice plants subjected to different nitrogen fertilization rates.

Conclusions

The results showed that the condition of half-fertilization plus separate inoculation with the isolates AC32 (Herbaspirillum sp.), AG15 (Burkholderia sp.), CA21 (Pseudacidovorax sp.), and UR51 (Azospirillum sp.) achieved rice growth similar to those achieved by full-fertilization without inoculation, thus highlighting the potential of these strains for formulating new bioinoculants for rice crops.  相似文献   

19.

Aims

Non-native shrubs are important invaders of the Eastern Deciduous Forest, dramatically altering forest structure and functioning. Study of invasion mechanisms in this system has emphasized aboveground processes, and plant-soil feedbacks are relatively unexplored as a mechanism of shrub dominance. We tested whether plant-soil feedback in this habitat is affected by competition and whether arbuscular mycorrhizal fungi (AMF) are involved in plant-soil feedback.

Methods

We used a standard two-phase plant-soil feedback experiment run concurrently for each of three invasive shrub species, measuring feedback effects on AMF colonization, aboveground biomass, and the responses of native plant species in greenhouse mesocosms.

Results

Lonicera maackii and Ligustrum vulgare reduced AMF colonization of native roots, both with legacy effects (prior growth in soil) and direct effects (current growth in soil). Elaeagnus umbellata grown with natives left a legacy of increased AMF colonization of native communities.

Conclusions

Our results suggest that woody invasive species can alter the AMF associations of native plants even after the invasive is no longer present. Such consequences merit study with other native species and where environmental factors, such as light availability, might be expected to compound the effects of changes in AMF.  相似文献   

20.

Background and aims

We determined the relationship between site N supply and decomposition rates with respect to controls exerted by environment, litter chemistry, and fungal colonization.

Methods

Two reciprocal transplant decomposition experiments were established, one in each of two long-term experiments in oak woodlands in Minnesota, USA: a fire frequency/vegetation gradient, along which soil N availability varies markedly, and a long-term N fertilization experiment. Both experiments used native Quercus ellipsoidalis E.J. Hill and Andropogon gerardii Vitman leaf litter and either root litter or wooden dowels.

Results

Leaf litter decay rates generally increased with soil N availability in both experiments while belowground litter decayed more slowly with increasing soil N. Litter chemistry differed among litter types, and these differences had significant effects on belowground (but not aboveground) decay rates and on aboveground litter N dynamics during decomposition. Fungal colonization of detritus was positively correlated with soil fertility and decay rates.

Conclusions

Higher soil fertility associated with low fire frequency was associated with greater leaf litter production, higher rates of fungal colonization of detritus, more rapid leaf litter decomposition rates, and greater N release in the root litter, all of which likely enhance soil fertility. During decomposition, both greater mass loss and litter N release provide mechanisms through which the plant and decomposer communities provide positive feedbacks to soil fertility as ultimately driven by decreasing fire frequency in N-limited soils and vice versa.  相似文献   

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