Differential effects of <Emphasis Type="Italic">Paenibacillus</Emphasis> spp. on cucumber mycorrhizas

The effects of 17 Paenibacillus strains on root colonization by Glomus intraradices or Glomus mosseae and plant growth parameters (shoot and root weight) of mycorrhizal cucumber plants were examined. The Paenibacillus strains were originally isolated from mycorrhizal (G. intraradices) and non-mycorrhizal cucumber rhizosphere and/or hyphosphere, except for strain EJP73, which originated from a Pinus sylvestris-Lactarius rufus ectomycorrhiza. Root colonization of cucumber plants by G. intraradices or G. mosseae was unaffected by all seven strains of Paenibacillus polymyxa, but was decreased or increased by four strains of Paenibacillus macerans and strain EJP73 of Paenibacillus sp. Overall, shoot dry weight of cucumber grown in symbioses with either G intraradices or G. mosseae was unaffected by inoculation with all of the Paenibacillus strains, except for strain MB02-429 of P. macerans, which increased the shoot dry weight in the cucumber-G. mosseae symbiosis. On the other hand, several Paenibacillus strains caused altered root growth. Three strains of P. polymyxa and four strains of P. macerans increased the root fresh weight of the cucumber–G. intraradices symbiosis, whereas three strains of P. polymyxa and one strain of P. macerans as well as Paenibacillus sp. EJP73, decreased the root fresh weight of the cucumber–G. mosseae symbiosis. In conclusion, our results show that bacteria from several species of Paenibacillus differentially affect cucumber mycorrhizas.     

Potential role of D-<Emphasis Type="Italic">myo</Emphasis>-inositol-3-phosphate synthase and 14-3-3 genes in the crosstalk between <Emphasis Type="Italic">Zea mays</Emphasis> and <Emphasis Type="Italic">Rhizophagus intraradices</Emphasis> under drought stress

Arbuscular mycorrhizal (AM) symbiosis is known to stimulate plant drought tolerance. However, the mechanisms underlying the synergistic responses of the symbiotic partners to drought stress are largely unknown. A split-root experiment was designed to investigate the molecular interactions between a host plant and an AM fungus (AMF) under drought stress. In the two-compartment cultivation system, an entire or only a half root system of a maize plant was inoculated with an AMF, Rhizophagus intraradices, in the presence of localized or systemic drought treatment. Plant physiological parameters including growth, water status, and phosphorus concentration, and the expression of drought tolerance-related genes in both roots and R. intraradices were recorded. Although mycorrhizal inoculation in either one or both compartments systemically decreased abscisic acid (ABA) content in the whole root system subjected to systemic or local drought stress, we observed local and/or systemic AM effects on root physiological traits and the expression of functional genes in both roots and R. intraradices. Interestingly, the simultaneous increase in the expression of plant genes encoding D-myo-inositol-3-phosphate synthase (IPS) and 14-3-3-like protein GF14 (14-3GF), which were responsible for ABA signal transduction, was found to be involved in the activation of 14-3-3 protein and aquaporins (GintAQPF1 and GintAQPF2) in R. intraradices. These findings suggest that coexpression of IPS and 14-3GF is responsible for the crosstalk between maize and R. intraradices under drought stress, and potentially induces the synergistic actions of the symbiotic partners in enhancing plant drought tolerance.     

Influence of Salinity on the <Emphasis Type="Italic">In Vitro</Emphasis> Development of <Emphasis Type="Italic">Glomus intraradices</Emphasis> and on the <Emphasis Type="Italic">In Vivo</Emphasis> Physiological and Molecular Responses of Mycorrhizal Lettuce Plants

Increased salinization of arable land is expected to have devastating global effects in the coming years. Arbuscular mycorrhizal fungi (AMF) have been shown to improve plant tolerance to abiotic environmental factors such as salinity, but they can be themselves negatively affected by salinity. In this study, the first in vitro experiment analyzed the effects of 0, 50, or 100 mM NaCl on the development and sporulation of Glomus intraradices. In the second experiment, the effects of mycorrhization on the expression of key plant genes expected to be affected by salinity was evaluated. Results showed that the assayed isolate G. intraradices DAOM 197198 can be regarded as a moderately salt-tolerant AMF because it did not significantly decrease hyphal development or formation of branching absorbing structures at 50 mM NaCl. Results also showed that plants colonized by G. intraradices grew more than nonmycorrhizal plants. This effect was concomitant with a higher relative water content in AM plants, lower proline content, and expression of Lsp5cs gene (mainly at 50 mM NaCl), lower expression of the stress marker gene Lslea gene, and lower content of abscisic acid in roots of mycorrhizal plants as compared to nonmycorrhizal plants, which suggest that the AM fungus decreased salt stress injury. In addition, under salinity, AM symbiosis enhanced the expression of LsPIP1. Such enhanced gene expression could contribute to regulating root water permeability to better tolerate the osmotic stress generated by salinity.     

<Emphasis Type="Italic">Mycena</Emphasis> sp., a mycorrhizal fungus of the orchid <Emphasis Type="Italic">Dendrobium officinale</Emphasis>

An endophytic fungus, F-23, was isolated from the roots of Dendrobium officinale Kimura et Migo, an endangered Chinese medicinal plant. The sequence of the ITS region indicated that the isolate belongs to the genus Mycena. After 4 months of inoculation, the root systems of D. officinale that were inoculated with F-23 fungus were much larger than the control’s root systems. We also observed that the hyphae of F-23 penetrated the epidermal cells within the host’s roots and spread from cell to cell. A large number of pelotons existed in the root cortical cells of D. officinale inoculated with F-23 fungus. Intracellular hyphae crossing through the host walls were also observed using SEM (scanning electron microscopy). In contrast, light microscopy and SEM showed that the transverse sections of the roots of control plants remained uncolonized. Therefore, the F-23 fungus can form mycorrhizal associations with the roots of its host plant, D. officinale, and enhance the growth of seedlings and roots. In brief, Mycena sp. was identified and shown to be a mycorrhizal fungus of the epiphytic orchid, D. officinale. This might be of potential use to the mass cultivation of D. officinale under artificial conditions.     

The responsiveness of the <Emphasis Type="Italic">IAA2</Emphasis> promoter to IAA and IBA is differentially affected in <Emphasis Type="Italic">Arabidopsis</Emphasis> roots and shoots by flavonoids

The structural features of flavonoids which are involved in the modulation of auxin distribution in Arabidopsis thaliana were evaluated. An auxin-inducible promoter IAA2 fused to a reporter gene (GUS) was used to monitor the tissue responsiveness to auxins. The following aspects were investigated: 1) the influence of flavonoids (quercetin, naringenin, kaempferol, myricetin and isorhamnetin) on the distribution of indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA) in roots and leaves, 2) differences in flavonoid uptake into roots and shoots depending on flavonoid concentration in the medium, and 3) influence of structurally different flavonoids on the gravitropic response and growth of roots. The same flavonoids differently affected IAA and IBA distribution in leaves and roots. There were several structural requirements for the flavonoids which resulted in the changes of auxin response/distribution. Great differences between the ability of shoots and roots to take up quercetin were showed. Also, flavonoids influenced gravitropism and root growth of Arabidopsis seedlings in a structure-dependent manner.     

Toxicity of diesel fuel to germination,growth and colonization of <Emphasis Type="Italic">Glomus intraradices</Emphasis> in soil and <Emphasis Type="Italic">in vitro</Emphasis> transformed carrot root cultures

Phytoremediation is the use of selected plants to decontaminate polluted environments. Arbuscular mycorrhizal fungi (AMF) may potentially be useful for phytoremediation, but it is not known how petroleum hydrocarbons influence AMF spore germination and hyphal growth. To address this question, germination of spores and germ tube growth of Glomus intraradices Schenck and Smith and Glomus aggregatum Schenck and Smith were assessed in soil contaminated with up to 3% (w/v) of F2 diesel oil or HAGO reference oil. Hyphal growth, colonization and progeny spore production were assessed in vitro using transformed root cultures of Daucus carota and G. intraradices spores in a F2 diesel contaminated medium. In addition, extraradical hyphal growth of G. intraradices colonizing Daucus carota in the presence of F2 diesel was studied. Neither F2 diesel nor HAGO reference oil affected spore germination or germ tube growth in soil. However, in the presence of plant roots, germ tube growth of G. intraradices was reduced and delayed in the presence of F2 diesel and root colonization was not detected. Hyphal growth of pre-colonized carrot roots by G. intraradices was reduced and delayed in F2 contaminated medium compared to controls. F2 diesel did not inhibit spore germination of these AMF species but did reduce colonization, germ tube and hyphal growth. These results suggest that AMF inoculum can be established in petroleum-contaminated sites. However, it may prove beneficial to plant pre-colonized plants to increase the probability of sufficient AMF colonization and growth. The likely mechanism(s) of petroleum toxicity in this plant-microbe system was discussed.     

Plant Responses to Drought Stress and Exogenous ABA Application are Modulated Differently by Mycorrhization in Tomato and an ABA-deficient Mutant (<Emphasis Type="Italic">Sitiens</Emphasis>)

The aims of the present study are to find out whether the effects of arbuscular mycorrhizal (AM) symbiosis on plant resistance to water deficit are mediated by the endogenous abscisic acid (ABA) content of the host plant and whether the exogenous ABA application modifies such effects. The ABA-deficient tomato mutant sitiens and its near-isogenic wild-type parental line were used. Plant development, physiology, and expression of plant genes expected to be modulated by AM symbiosis, drought, and ABA were studied. Results showed that only wild-type tomato plants responded positively to mycorrhizal inoculation, while AM symbiosis was not observed to have any effect on plant development in sitiens plants grown under well-watered conditions. The application of ABA to sitiens plants enhanced plant growth both under well-watered and drought stress conditions. In respect to sitiens plants subjected to drought stress, the addition of ABA had a cumulative effect in relation to that of inoculation with G. intraradices. Most of the genes analyzed in this study showed different regulation patterns in wild-type and sitiens plants, suggesting that their gene expression is modulated by the plant ABA phenotype. In the same way, the colonization of roots with the AM fungus G. intraradices differently regulated the expression of these genes in wild-type and in sitiens plants, which could explain the distinctive effect of the symbiosis on each plant ABA phenotype. This also suggests that the effects of the AM symbiosis on plant responses and resistance to water deficit are mediated by the plant ABA phenotype.     

Clustered root distribution in mature stands of <Emphasis Type="Italic">Fagus sylvatica</Emphasis> and <Emphasis Type="Italic">Picea abies</Emphasis>

Distribution of small roots (diameter between 2 mm and 5 mm) was studied in 19 pits with a total of 72 m² trench profile walls in pure stands of Fagus sylvatica and Picea abies. Root positions within the walls were marked and transformed into x-coordinates and y-coordinates. In a GIS-based evaluation, zones of potential influence around each root were calculated. The total potential influence produced isoline maps of relative root influence zones, thus indicating small root clustering. The questions studied were (1) whether there were marked clusters of small roots in the soil and (2) whether trees surrounding the pit (defined as tree density) correlate with the root abundance and distribution on the trench profile walls. Small roots of both species showed maximum abundance in the top 20 cm of the soil, where pronounced root clusters occurred next to areas with only low root accumulation. The area of root clusters did not differ significantly between the two stands. Weighted clumping, WC, calculated as a product of root class, and its area was used as an index of root clustering, which again did not differ between beech and spruce stands. However, evaluations on a single root level showed that beech achieved the same degree of clustering with lower number of roots. Regardless of soil properties related to root clusters, a significantly higher clustering acquired per root for beech than for spruce suggests beech to be more efficient in belowground acquisition of space. Because none of the parameters describing root clustering were correlated with tree density around the investigated soil profiles, clusters of small roots are inherently present within the tree stands.     

Antioxidant Enzyme Activities in Arbuscular Mycorrhizal (<Emphasis Type="Italic">Glomus intraradices</Emphasis>) Fungus Inoculated and Non-inoculated Maize Plants Under Zinc Deficiency

A greenhouse experiment was conducted to examine the changes in antioxidant enzyme activities of arbuscular mycorrhizal (AM) fungus Glomus intraradices Schenck and Smith inoculated (M+) and non-inoculated (M−) maize (Zea mays L.) plants (variety COHM5) under varying levels of zinc (0, 1.25, 2.5, 3.75 and 5.0 mg kg⁻¹). Roots and shoots sampled at 45 days after sowing (DAS) were estimated for its antioxidant enzymes (superoxide dismutase, peroxidase) IAA oxidase, polyphenol oxidase, acid phosphatase and nutritional status especially P and Zn concentrations. Mycorrhizal inoculation significantly (P ≤ 0.01) increased all the four antioxidant enzymes in both roots and shoots at 45 DAS regardless of Zn levels. All enzyme activities except SOD increased progressively with increasing levels of Zn under M+ and M− conditions. The SOD activity got decreased in roots and shoots at 2.5 and 3.75 mg Zn kg⁻¹. Acid phosphatase activity in M+ roots and shoots were higher in all levels of Zn but the values decreased with increasing levels of Zn particularly in roots. Mycorrhizal fungus inoculated plants had higher P and Zn concentrations in both stages in comparison to non-inoculated plants. Our overall data suggest that mycorrhizal symbiosis plays a vital role in enhancing activities of antioxidant enzymes and nutritional status that enables the host plant to sustain zinc deficient conditions.     

Effect of <Emphasis Type="Italic">Glomus</Emphasis> species on physiology and biochemistry of <Emphasis Type="Italic">Catharanthus roseus</Emphasis>

The present study on efficacy of different Glomus species, an arbuscular mycorrhizal (AM) fungus (G. aggregatum, G. fasciculatum, G. mosseae, G. intraradices) on various growth parameters such as biomass, macro and micronutrients, chlorophyll, protein, cytokinin and alkaloid content and phosphatase activity of pink flowered Catharanthus roseus plants showed that all Glomus species except G. intraradices enhanced the chlorophyll, protein, crude alkaloid, phosphorus, sulphur, manganese and copper contents of C. roseus plants along with phosphatase activity significantly over uninoculated plants. However only G. mosseae and G. fasciculatum exhibited superior symbiotic relationship with the plant. G. mosseae was found to be the best for increasing the crude alkaloid content (8.19%) in leaf and also in increasing the quantity of important alkaloids vincristine and vinblastine.     

Survival and growth of parasitic <Emphasis Type="Italic">Maculinea alcon</Emphasis> caterpillars (Lepidoptera,Lycaenidae) in laboratory nests of three <Emphasis Type="Italic">Myrmica</Emphasis> ant species

The Alcon blue butterfly (Maculinea alcon) parasitizes the nests of several Myrmica ant species. In Denmark, it uses M. rubra and M. ruginodis, but never M. scabrinodis. To further examine the basis of this specificity and local co-adaptation between host and parasite, the pattern of growth and survival of newly-adopted caterpillars of M. alcon in Myrmica subcolonies was examined in the laboratory. M. alcon caterpillars were collected from three populations differing in their host use, and reared in laboratory nests of all three ant species collected from each M. alcon population. While there were differences in the pattern of growth of caterpillars from different populations during the first few months after adoption, which depended on host ant species and the site from which the ants were collected, there was no evidence of major differences in final size achieved. Survival was, however, much higher in nests of M. rubra than in nests of M. ruginodis and M. scabrinodis, even for caterpillars from a population that is never known to use M. rubra as a host in the field. The caterpillars of M. alcon thus do not show local adaptation in their pattern of growth and survival, but instead show a pattern that may reflect different nestmate recognition abilities of the host ants, related to their sociogenetic organisation. The pattern of observed host ant use in the field seems to result from a combination of differences in local host availability and locally adapted infectivity, modulated by smaller differences in survivorship in the nests of the different host ants.     

Nutritional regulation of <Emphasis Type="Italic">ANR1</Emphasis> and other root-expressed MADS-box genes in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>

The ANR1 MADS-box gene in Arabidopsis thaliana (L.) Heynh. has previously been identified as a key regulator of lateral root growth in response to signals from external nitrate (NO₃⁻). We have used quantitative real-time PCR to investigate the responsiveness of ANR1 and 11 other root-expressed MADS-box genes to fluctuations in the supply of N, P and S. ANR1 expression in roots of hydroponically grown Arabidopsis plants was specifically regulated by changes in the N supply, being induced by N deprivation and rapidly repressed by N re-supply. This pattern of N responsiveness differs from the NO₃⁻ -inducibility of ANR1 previously observed in Arabidopsis root cultures [H.M. Zhang and B.G. Forde (1998) Science 279:407–409]. Seven of the other MADS-box genes responded to N in a manner similar to ANR1, but less strongly, while four (AGL12, AGL17, AGL18 and AGL79) were unaffected. Six of the N-regulated genes (ANR1, AGL14, AGL16, AGL19, SOC1 and AGL21) belong to just two clades within the type II MADS-box lineage, while the other two (AGL26 and AGL56) belong to the poorly characterized type I lineage. Only SOC1 was additionally found to respond to changes in the P and S supply, suggesting a possible role in a general response to nutrient stress. Studies with an ANR1 transposon-insertion mutant provided no evidence for regulatory interactions between ANR1 and the other root-expressed MADS-box genes. The implications of the current data for our understanding of the role of ANR1 and other MADS box genes in the nutritional regulation of lateral root growth are discussed.     

Expression analysis of <Emphasis Type="Italic">Clavata1</Emphasis>-like and <Emphasis Type="Italic">Nodulin21</Emphasis>-like genes from <Emphasis Type="Italic">Pinus sylvestris</Emphasis> during ectomycorrhiza formation

The ecology and physiology of ectomycorrhizal (EcM) symbiosis with conifer trees are well documented. In comparison, however, very little is known about the molecular regulation of these associations. In an earlier study, we identified three EcM-regulated Pinus expressed sequence tags (EST), two of which were identified as homologous to the Medicago truncatula nodulin MtN21. The third EST was a homologue to the receptor-like kinase Clavata1. We have characterized the expression patterns of these genes and of auxin- and mycorrhiza-regulated genes after induction with indole-3-butyric acid in Pinus sylvestris and in a time course experiment during ectomycorrhizal initiation with the co-inoculation of 2,3,5-triiodobenzoic acid, an auxin transport inhibitor. Our results suggest that different P. sylvestris nodulin homologues are associated with diverse processes in the root. The results also suggest a potential role of the Clv1-like gene in lateral root initiation by the ectomycorrhizal fungus.     

<Emphasis Type="Italic">In vitro</Emphasis> culture of <Emphasis Type="Italic">Chrysanthemum</Emphasis> plantlets using light-emitting diodes

Effects of illumination spectrum on the morphogenesis of chrysanthemum plantlets (Chrysanthemum morifolium Ramat. ‘Ellen’) grown in vitro were studied using an illumination system consisting of four groups of light-emitting diodes (LEDs) in the following spectral regions: blue (450nm), red (640nm), red (660nm), and far-red (735nm). Taking into account all differences in shoot height, root length, and fresh and dry weight (FW and DW, respectively), observed while changing the total photon flux density (PFD), the optimal total PFD for growth of chrysanthemum plantlets in vitro was estimated. For 16 h photoperiod and typical fractions of the spectral components (14%, 50%, 28%, and 8%, respectively), the optimal total PFD was found to be 40 μmol m⁻² s⁻¹. Our study shows that the blue component in the illumination spectrum inhibits the plantlet extension and formation of roots and simultaneously increases the DW to FW ratio and content of photosynthetic pigments. We demonstrate photomorphogenetic effects in the blue region and its interaction with the fractional PFD of the far-red spectral component. Under constant fractional PFD of the blue component, the root number, length of roots and stems, and fresh weight of the plantlets have a correlated nonmonotonous dependence on the fractional PFD of the far-red component.     

Contribution of <Emphasis Type="Italic">Glomus intraradices</Emphasis> inoculation to nutrient acquisition and mitigation of ionic imbalance in NaCl-stressed <Emphasis Type="Italic">Trigonella foenum-graecum</Emphasis>

The study aimed to investigate the effects of an AM fungus (Glomus intraradices Schenck and Smith) on mineral acquisition in fenugreek (Trigonella foenum-graecum) plants under different levels of salinity. Mycorrhizal (M) and non-mycorrhizal (NM) fenugreek plants were subjected to four levels of NaCl salinity (0, 50, 100, and 200 mM NaCl). Plant tissues were analyzed for different mineral nutrients. Leaf senescence (chlorophyll concentration and membrane permeability) and lipid peroxidation were also assessed. Under salt stress, M plants showed better growth, lower leaf senescence, and decreased lipid peroxidation as compared to NM plants. Salt stress adversely affected root nodulation and uptake of NPK. This effect was attenuated in mycorrhizal plants. Presence of the AM fungus prevented excess uptake of Na⁺ with increase in NaCl in the soil. It also imparted a regulatory effect on the translocation of Na⁺ ions to shoots thereby maintaining lower Na⁺ shoot:root ratios as compared to NM plants. Mycorrhizal colonization helped the host plant to overcome Na⁺-induced Ca²⁺ and K⁺ deficiencies. M plants maintained favorable K⁺:Na⁺, Ca²⁺:Na⁺, and Ca²⁺:Mg²⁺ ratios in their tissues. Concentrations of Cu, Fe, and Zn²⁺ decreased with increase in intensity of salinity stress. However, at each NaCl level, M plants had higher concentration of Cu, Fe, Mn²⁺, and Zn²⁺ as compared to NM plants. M plants showed reduced electrolyte leakage in leaves as compared to NM plants. The study suggests that AM fungi contribute to alleviation of salt stress by mitigation of NaCl-induced ionic imbalance thus maintaining a favorable nutrient profile and integrity of the plasma membrane.     

Host-related variability in arbuscular mycorrhizal fungal structures in roots of <Emphasis Type="Italic">Hedera rhombea</Emphasis>, <Emphasis Type="Italic">Rubus parvifolius</Emphasis>, and <Emphasis Type="Italic">Rosa multiflora</Emphasis> under controlled conditions

The arbuscular mycorrhizal (AM) morphology of three host plant species inoculated with single and mixed fungal culture and the distribution of AM fungal species in roots of the hosts treated with a mixed culture of AM fungi were determined. The aim was to investigate the effect of host plants and AM fungi on AM morphology of coexisting plant species. Noncolonized rooted cuttings of Hedera rhombea (Miq) Bean, Rubus parvifolius L., and Rosa multiflora Thunb. were inoculated with five fungal species as single and mixed culture inocula. The fungal species used were Gigaspora rosea and Scutellospora erythropa, previously isolated from H. rhombea; Acaulospora longula and Glomus etunicatum from R. parvifolius; and Glomus claroideum from both plant species. A few hyphal and arbusculate coils were seen in the mixed culture-inoculated roots of R. parvifolius; all fungal treatments produced this Paris-type AM in H. rhombea and Arum-type AM in R. parvifolius, and R. multiflora indicates that AM morphology is strongly controlled by the identity of the host plants used in this study. AM fungal rDNA was extracted separately from roots of each replicate plant species inoculated with the mixed fungal culture, amplified, cloned, sequenced, and analyzed to determine the AM fungal species and their respective proportions in roots of each plant species. Glomus etunicatum and G. claroideum of the family Glomaceae generally occurred more frequently in R. parvifolius and R. multiflora, which form Arum-types, whereas S. erythropa, of the family Gigasporaceae, was the most frequently detected species in H. rhombea, which produced Paris-type AM. Although the genotype of the plant species used appears to determine the AM morphologies formed, there was preferential association between the hosts and AM fungal inoculants.     

Polymorphism of lectin genes in <Emphasis Type="Italic">Lathyrus</Emphasis> plants

The carbohydrate-binding sequences of the lectin genes from spring vetchling Lathyrus vernus (L.) Bernh., marsh vetchling L. palustris (L.), and Gmelin’s vetchling L. gmelinii (Fitsch) (Fabaceae) were determined. Computer-aided analysis revealed substantial differences between nucleotide and predicted amino acid sequences of the lectin gene regions examined in each of the three vetchling species tested. In the phylogenetic trees based on sequence similarity of carbohydrate-biding regions of legume lectins, the sequences examined formed a compact cluster with the lectin genes of the plants belonging to the tribe Fabeae. In each plant, L. vernus, L. palustris, and L. gmelinii, three different lectin-encoding genes were detected. Most of the substitutions were identified within the gene sequence responsible for coding the carbohydrate-binding protein regions. This finding may explain different affinity of these lectins to different carbohydrates, and as a consequence, can affect the plant host specificity upon development of symbiosis with rhizobium bacteria.     

Arbuscular mycorrhizal colonization and nodulation improve flooding tolerance in <Emphasis Type="Italic">Pterocarpus officinalis</Emphasis> Jacq. seedlings

Pterocarpus officinalis (Jacq.) seedlings inoculated with the arbuscular mycorrhizal fungus, Glomus intraradices, and the strain of Bradyrhizobium sp. (UAG 11A) were grown under stem-flooded or nonflooded conditions for 13 weeks after 4 weeks of nonflooded pretreatment under greenhouse conditions. Flooding of P. officinalis seedlings induced several morphological and physiological adaptive mechanisms, including formation of hypertrophied lenticels and aerenchyma tissue and production of adventitious roots on submerged portions of the stem. Flooding also resulted in an increase in collar diameter and leaf, stem, root, and total dry weights, regardless of inoculation. Under flooding, arbuscular mycorrhizas were well developed on root systems and adventitious roots compared with inoculated root systems under nonflooding condition. Arbuscular mycorrhizas made noteworthy contributions to the flood tolerance of P. officinalis seedlings by improving plant growth and P acquisition in leaves. We report in this study the novel occurrence of nodules connected vascularly to the stem and nodule and arbuscular mycorrhizas on adventitious roots of P. officinalis seedlings. Root nodules appeared more efficient fixing N₂ than stem nodules were. Beneficial effect of nodulation in terms of total dry weight and N acquisition in leaves was particularly noted in seedlings growing under flooding conditions. There was no additive effect of arbuscular mycorrhizas and nodulation on plant growth and nutrition in either flooding treatment. The results suggest that the development of adventitious roots, aerenchyma tissue, and hypertrophied lenticels may play a major role in flooded tolerance of P. officinalis symbiosis by increasing oxygen diffusion to the submerged part of the stem and root zone, and therefore contribute to plant growth and nutrition.     

Colonization of the <Emphasis Type="Italic">Arabidopsis</Emphasis> rhizosphere by fluorescent <Emphasis Type="Italic">Pseudomonas</Emphasis> spp. activates a root-specific,ethylene-responsive <Emphasis Type="Italic">PR-5</Emphasis> gene in the vascular bundle

Plants of which the roots are colonized by selected strains of non-pathogenic, fluorescent Pseudomonas spp. develop an enhanced defensive capacity against a broad spectrum of foliar pathogens. In Arabidopsis thaliana, this rhizobacteria-induced systemic resistance (ISR) functions independently of salicylic acid but requires responsiveness to jasmonic acid and ethylene. In contrast to pathogen-induced systemic acquired resistance (SAR), ISR is not associated with systemic changes in the expression of genes encoding pathogenesis-related (PR) proteins. To identify genes that are specifically expressed in response to colonization of the roots by ISR-inducing Pseudomonas fluorescens WCS417r bacteria, we screened a collection of Arabidopsis enhancer trap and gene trap lines containing a transposable element of the Ac/Ds system and the GUS reporter gene. We identified an enhancer trap line (WET121) that specifically showed GUS activity in the root vascular bundle upon colonization of the roots by WCS417r. Fluorescent Pseudomonas spp. strains P. fluorescens WCS374r and P. putida WCS358r triggered a similar expression pattern, whereas ISR-non-inducing Escherichia coli bacteria did not. Exogenous application of the ethylene precursor 1-aminocyclopropane-1-carboxylate (ACC) mimicked the rhizobacteria-induced GUS expression pattern in the root vascular bundle, whereas methyl jasmonic acid and salicylic acid did not, indicating that the Ds element in WET121 is inserted in the vicinity of an ethylene-responsive gene. Analysis of the expression of the genes in the close vicinity of the Ds element revealed AtTLP1 as the gene responsible for the in cis activation of the GUS reporter gene in the root vascular bundle. AtTLP1 encodes a thaumatin-like protein that belongs to the PR-5 family of PR proteins, some of which possess antimicrobial properties. AtTLP1 knockout mutant plants showed normal levels of WCS417r-mediated ISR against the bacterial leaf pathogen Pseudomonas syringae pv. tomato DC3000, suggesting that expression of AtTLP1 in the roots is not required for systemic expression of ISR in the leaves. Together, these results indicate that induction of AtTLP1 is a local response of Arabidopsis roots to colonization by non-pathogenic fluorescent Pseudomonas spp. and is unlikely to play a role in systemic resistance.     

Carbon nutrition of mature green orchid <Emphasis Type="Italic">Serapias strictiflora</Emphasis> and its mycorrhizal fungus <Emphasis Type="Italic">Epulorhiza</Emphasis> sp.

We studied the nutritional modes of the orchid Serapias strictiflora and its mycorrhizal fungus Epulorhiza sp. using the differences in carbon isotopic composition (δ¹³C) of C₃ orchid and C₄ maize tissues. We found that if cultivated in substrate lacking any organic compounds, the mycorrhizal extraradical mycelia (δ¹³C = −26.3 ± 0.2 ‰) developed well, despite being fully dependent on nutrition from orchid roots (δ¹³C = −28.6 ± 0.1 ‰). If the mycorrhizal fungus had additional access to and colonized decaying maize roots (δ¹³C = −14.6 ± 0.1 ‰), its isotopic composition (δ¹³C = −21.6 ± 0.4 ‰) reflected a mixture of biotrophy and saprotrophy. No statistically significant differences in δ¹³C of new storage tubers were found between Epulorhiza-associated orchids with (δ¹³C = -28.2 ± 0.1 ‰) and without access to maize roots (δ¹³C = −28.6 ± 0.2 ‰). We conclude that autotrophy is the predominant nutritional mode of mature S. strictiflora plants and that they supply their mycorrhizal fungus with substantial amount of carbon (69 ± 3 % of the fungus demand), even if the fungus feeds saprotrophically.