Arbuscular mycorrhizal colonization of tomato by Gigaspora and Glomus species in the presence of root flavonoids

The effect of flavonoids isolated from arbuscular mycorrhizal (AM) colonized and noncolonized clover roots on the number of entry points and percentage of root colonization of tomato (Lycopersicum esculentum L.) by Gigaspora rosea, Gi margarita, Glomus mosseae and G. intrarradices symbionts was determined. With fungi of both genera, a correlation between the number of entry points and the percentage of root colonization was found in the presence of some of the tested flavonoids. The flavonoids acacetin and rhamnetin, present in AM clover roots, inhibited the formation of AM penetration structures and the AM colonization of tomato roots, whereas the flavonoid 5,6,7,8,9-hydroxy chalcone, which could not be detected in AM clover root, inhibited both parameters. The flavonoid quercetin, which was present in AM clover roots, stimulated the penetration and root colonization of tomato by Gigaspora. However, the flavonoids 5,6,7,8-hydroxy-4'-methoxy flavone and 3,5,6,7,4'-hydroxy flavone, which was not found in AM clover root, increased the number of entry points and the AM colonization of tomato roots by Gigaspora. These results indicated that flavonoids could be imnplicated in the process of regulation of AM colonization in plant root, but its role is highly complex and depend not only on flavonoids, but also on AM fungal genus or even species.     

Flavonoids in roots of white clover: interaction of arbuscular mycorrhizal fungi and a pathogenic fungus

The effects of two arbuscular mycorrhizal fungi (AMF) (Glomus mosseae and G. claroideum) and a pathogenic fungus (Pythium ultimum) on the production of eight flavonoids in roots of two white clover (Trifolium repens L.) cultivars were evaluated. Quantification of AM and pathogenic fungi in the roots showed that the AM symbiosis significantly reduced P. ultimum biomass and in some cases prevented infection. The flavonoid productions in clover roots varied depending on the presence of beneficial and/or pathogenic fungi, fungal isolate or plant cultivar. Only plants colonized with G. claroideum showed detectable concentrations of either coumestrol or kaempferol (cultivar-dependant). In addition, inoculation with G. claroideum resulted in significantly higher concentrations of coumestrol in cv. Sonja and medicarpin in cv. Milo. A low production of coumestrol and kaempferol in mycorrhizal plants may be G. mosseae-specific. Only the concentrations of formononetin and daidzein increased in clover roots in response to infection with P. ultimum. These flavonoids are supposedly stress metabolites, synthesized or produced from glycosides in response to pathogen infection. However, the presence of one or both AMF significantly lowered the formononetin and daidzein concentrations, and overruled the inductive effect of P. ultimum. Therefore the antagonistic action of AM against the pathogen must take place through another mechanism.     

Accumulation of cyclohexenone derivatives in barley, wheat and maize roots in response to inoculation with different arbuscular mycorrhizal fungi

Glomus intraradices , Glomus mosseae, and Gigaspora rosea leads to the accumulation of cyclohexenone derivatives. Mycorrhizal roots of all plants accumulate in response to all three fungi blumenin [9-O-(2′-O-glucuronosyl)-β-glucopyranoside of 6-(3-hydroxybutyl)-1,1,5-trimethyl-4-cyclohexen-3-one], 13-carboxyblumenol C 9-O-gentiobioside, nicoblumin [9-O-(6′-O-β-glucopyranosyl)-β-glucopyranoside of 13-hydroxy-6-(3-hydroxybutyl)-1,1,5-trimethyl-4-cyclohexen-3-one] and another, as yet unidentified, cyclohexenone derivative. The accumulation of all four compounds in three tested mycorrhizal plants colonized by the three arbuscular mycorrhizal fungi indicates no fungus-specific induction of these compounds. Accepted: 6 October 1999     

Interactions between Lotus japonicus genotypes and arbuscular mycorrhizal fungi

Abstract

Interactions between three genotypes (Ljsym 71-1, Ljsym 71-2 and Ljsym 72) of Lotus japoicus and one isolate from each of four species of arbuscular mycorrhizal fungi (Glomus sp. R-10, Glomus intraradices, Glomus etunicatum, and Gigaspora margarita) were investigated and compared with the wild-type ‘Gifu’ B-129. All the three genotypes showed no or defective internal colonization after inoculation with these AM fungi. In Ljsym72 mutant, the AM fungi produced deformed appressoria on the root surface, but failed to form any internal structures (internal hyphae, arbuscules and vesicles) except only in Glomus intraradices. The Ljsym71-1 and Ljsym71-2 mutants had more deformed appressoria and occasionally formed internal hyphae, arbuscules and vesicles, depending on AM fungi used. Wild-type ‘Gifu’ (nod⁺myc⁺) plants had typical colonization. The colonization of mutants by several fungi varied and provides a basis for studying recognition and compatibility between plants and mycorrhizal fungal species. These mutants also will be useful in studies of the genetics of the symbiosis between plant species and AM fungi.     

In situ analysis of anastomosis in representative genera of arbuscular mycorrhizal fungi

Arbuscular mycorrhizal fungi (AMF) form obligate symbiotic associations with plants. As a result, the role of hyphal interactions in the establishment and maintenance of common mycorrhizal networks is poorly understood because of constraints on methods for in situ analysis. We designed a rhizohyphatron that allows the examination of intact mycelia growing from whole mycorrhizal plants. Plants preinoculated with spores were cultivated in a compartment with a connecting tube from which hyphae extend through a fine nylon mesh onto agar-coated slides. Species selected from each of the five AMF genera were used to assess and characterize the anastomosis behavior in the rhizohyphatron. Hyphal networks of Paraglomus occultum, Ambispora leptoticha, Scutellospora heterogama, and Gigaspora gigantea growing on the agar-coated slides showed no evidence of hyphal fusion. In contrast, anastomosis occurred in the hyphal networks of Glomus clarum and Glomus intraradices at an average frequency of less than 15% for both species. The rhizohyphatron developed in this study will provide knowledge of the biology and genetics of self/non-self recognition in AMF and help to better understand Glomeromycotan life history strategies.     

Weed control and cover crop management affect mycorrhizal colonization of grapevine roots and arbuscular mycorrhizal fungal spore populations in a California vineyard

Arbuscular mycorrhizal (AM) fungi naturally colonize grapevines in California vineyards. Weed control and cover cropping may affect AM fungi directly, through destruction of extraradical hyphae by soil disruption, or indirectly, through effects on populations of mycorrhizal weeds and cover crops. We examined the effects of weed control (cultivation, post-emergence herbicides, pre-emergence herbicides) and cover crops (Secale cereale cv. Merced rye, × Triticosecale cv.Trios 102) on AM fungi in a Central Coast vineyard. Seasonal changes in grapevine mycorrhizal colonization differed among weed control treatments, but did not correspond with seasonal changes in total weed frequency. Differences in grapevine colonization among weed control treatments may be due to differences in mycorrhizal status and/or AM fungal species composition among dominant weed species. Cover crops had no effect on grapevine mycorrhizal colonization, despite higher spring spore populations in cover cropped middles compared to bare middles. Cover crops were mycorrhizal and shared four AM fungal species (Glomus aggregatum, G. etunicatum, G. mosseae, G. scintillans) in common with grapevines. Lack of contact between grapevine roots and cover crop roots may have prevented grapevines from accessing higher spore populations in the middles.     

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.     

Traits related to differences in function among three arbuscular mycorrhizal fungi

Diversity in phosphorus (P) acquisition strategies was assessed among three species of arbuscular mycorrhizal fungi (AMF) isolated from a single field in Switzerland. Medicago truncatula was used as a test plant. It was grown in a compartmented system with root and root-free zones separated by a fine mesh. Dual radioisotope labeling (³²P and ³³P) was employed in the root-free zone as follows: ³³P labeling determined hyphal P uptake from different distances from roots over the entire growth period, whereas ³²P labeling investigated hyphal P uptake close to the roots over the 48 hours immediately prior to harvest. Glomus intraradices, Glomus claroideum and Gigaspora margarita were able to take up and deliver P to the plants from maximal distances of 10, 6 and 1 cm from the roots, respectively. Glomus intraradices most rapidly colonized the available substrate and transported significant amounts of P towards the roots, but provided the same growth benefit as compared to Glomus claroideum, whose mycelium was less efficient in soil exploration and in P uptake and delivery to the roots. These differences are probably related to different carbon requirements by these different Glomus species. Gigaspora margarita provided low P benefits to the plants and formed dense mycelium networks close to the roots where P was probably transiently immobilized. Numerical modeling identified possible mechanisms underlying the observed differences in patterns of mycelium growth. High external hyphal production at the root-fungus interface together with rapid hyphal turnover were pointed out as important factors governing hyphal network development by Gigaspora, whereas nonlinearity in apical branching and hyphal anastomoses were key features for G. intraradices and G. claroideum, respectively.     

Separated components of root exudate and cytosol stimulate different morphologically identifiable types of branching responses by arbuscular mycorrhizal fungi

Two morphologically distinct hyphal branching responses by the AM fungus, Glomus intraradices, were stimulated by separated components of carrot root exudate. Complex branching up to the sixth order was induced by compounds most soluble in 35 % methanol, whereas the formation of more lateral branches (second order) was stimulated by compounds most soluble in 70 % methanol. This same 70 % alcohol soluble fraction also stimulated a completely different type of branching pattern in another fungus, Gigaspora gigantea. This pattern consisted of a very periodic distribution of dense clusters of hyphal branches that had a very high degree of complexity. In contrast to exudate components, separated cytosolic components of carrot roots did not stimulate any of the observed hyphal branching patterns. Alcohol-soluble fractions actually inhibited hyphal tip growth of G. gigantea and induced the formation of “recovery” branches that were identical to those induced by an inhibitor found in the exudate of Chard (Beta vulgaris ssp. cicla), a non-host plant.     

Overlapping expression patterns and differential transcript levels of phosphate transporter genes in arbuscular mycorrhizal,P<Subscript>i</Subscript>-fertilised and phytohormone-treated <Emphasis Type="Italic">Medicago truncatula</Emphasis> roots

A microarray carrying 5,648 probes of Medicago truncatula root-expressed genes was screened in order to identify those that are specifically regulated by the arbuscular mycorrhizal (AM) fungus Gigaspora rosea, by P_i fertilisation or by the phytohormones abscisic acid and jasmonic acid. Amongst the identified genes, 21% showed a common induction and 31% a common repression between roots fertilised with P_i or inoculated with the AM fungus G. rosea, while there was no obvious overlap in the expression patterns between mycorrhizal and phytohormone-treated roots. Expression patterns were further studied by comparing the results with published data obtained from roots colonised by the AM fungi Glomus mosseae and Glomus intraradices, but only very few genes were identified as being commonly regulated by all three AM fungi. Analysis of P_i concentrations in plants colonised by either of the three AM fungi revealed that this could be due to the higher P_i levels in plants inoculated by G. rosea compared with the other two fungi, explaining that numerous genes are commonly regulated by the interaction with G. rosea and by phosphate. Differential gene expression in roots inoculated with the three AM fungi was further studied by expression analyses of six genes from the phosphate transporter gene family in M. truncatula. While MtPT4 was induced by all three fungi, the other five genes showed different degrees of repression mirroring the functional differences in phosphate nutrition by G. rosea, G. mosseae and G. intraradices. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Growth stimulatory substances for vesicular-arbuscular mycorrhizal fungi in Bahia grass (Paspalum notatum Flügge.) roots

Three kinds of the growth stimulatory substances for vesicular-arbuscular mycorrhizal (VAM) fungi were isolated from the 25% methyl alcohol (MeOH) eluates of Bahia grass roots fractionated by flash chromatography. One of them was identified as eupalitin (3,5-dihydroxy-6,7-dimethoxy-4-hydroxy flavone). At 10 ppm, this compound significantly stimulated the in vitro hyphal growth of a VAM fungus, Gigaspora ramisporophora Spain. The others were not identified. These results suggest that the flavonoids such as eupalitin, and the unidentified compounds may act as signal molecules in VAM symbiosis.     

Mycorrhizas in agroforestry: spread and sharing of arbuscular mycorrhizal fungi between trees and crops: complementary use of molecular and microscopic approaches

The spread of arbuscular mycorrhizal (AM) fungi from tree to crop roots was examined by molecular and microscopic methods in a glasshouse study. Growth of Calliandra calothyrsus Meissner trees inoculated with isolates of the AM fungi Glomus etunicatum Becker and Gerdemann and Gigaspora albida Schenck and Smith was monitored over an 18-month period. Three successive ‘intercrops’ of beans or maize were sown at 25, 50 and 75 cm distances from the tree and harvested during this period. At each crop harvest, the distribution of tree and crop roots and the spread of the inoculant fungi were determined using traditional microscopic methods and fungal specific primers. Both inoculants greatly improved the growth of the trees and colonization spread to the crops once the trees were 6 months old. However, benefits of inoculation to crop growth were not observed due to increased competition from the larger inoculated trees growing in a restricted soil volume. Of the two inoculant fungi, Glomus etunicatum appeared to be more mobile as it spread more rapidly, formed higher levels of colonization at increasing distances from the tree and was responsible for most of the mycorrhizal cross-contamination. In contrast, colonization of tree and crop roots by Gigaspora albida was higher nearest the tree. This work demonstrated the benefits of mycorrhizal fungus inoculation for tree growth and confirmed that trees and crops share the same AM fungi. Trees may therefore act as reservoirs of mycorrhizal fungi, either inoculant or indigenous, for surrounding crops or other annual vegetation. It was also shown that tree pruning, the normal practice in agroforestry systems, did not reduce mycorrhizal colonization or prevent spread to crops. However, the slow rates of inoculant spread found here suggest that it may take years before inoculants benefit the growth of crops sown several metres from the tree. The work also demonstrated that microscopic quantification of mycorrhizal colonization and the use of molecular probes to identify specific fungi within roots can complement each other effectively. Molecular probes were more sensitive at detecting mycorrhizal fungi than microscopic methods, but did not discriminate between full mycorrhizal structures and traces of hyphae.     

Cooccurring plants forming distinct arbuscular mycorrhizal morphologies harbor similar AM fungal species

Arbuscular mycorrhizal (AM) fungal spores were isolated from field transplants and rhizosphere soil of Hedera rhombea (Miq) Bean and Rubus parvifolius L., which form Paris-type and Arum-type AM, respectively. DNA from the spore isolates was used to generate molecular markers based on partial large subunit (LSU) ribosomal RNA (rDNA) sequences to determine AM fungi colonizing field-collected roots of the two plant species. Species that were isolated as spores and identified morphologically and molecularly were Gigaspora rosea and Scutellospora erythropa from H. rhombea, Acaulospora longula and Glomus etunicatum from R. parvifolius, and Glomus claroideum from both plants. The composition of the AM fungal communities with respect to plant trap cultures was highly divergent between plant species. Analysis of partial LSU rDNA sequences amplified from field-collected roots of the two plant species with PCR primers designed for the AM fungi indicated that both plants were colonized by G. claroideum, G. etunicatum, A. longula, and S. erythropa. G. rosea was not detected in the field-collected roots of either plant species. Four other AM fungal genotypes, which were not isolated as spores in trap cultures from the two plant species, were also found in the roots of both plant species; two were closely related to Glomus intraradices and Glomus clarum. One genotype, which was most closely related to Glomus microaggregatum, was confined to R. parvifolius, whereas an uncultured Glomeromycotan fungus occurred only in roots of H. rhombea. S. erythropa was the most dominant fungus found in the roots of H. rhombea. The detection of the same AM fungal species in field-collected roots of H. rhombea and R. parvifolius, which form Paris- and Arum-type AM, respectively, shows that AM morphology in these plants is strongly influenced by the host plant genotypes as appears to be the case in many plant species in natural ecosystems, although there are preferential associations between the hosts and colonizing AM fungi in this study.     

Accumulation of reactive oxygen species in arbuscular mycorrhizal roots

We investigated the accumulation of reactive oxygen species (ROS) in arbuscular mycorrhizal (AM) roots from Medicago truncatula, Zea mays and Nicotiana tabacum using three independent staining techniques. Colonized root cortical cells and the symbiotic fungal partner were observed to be involved in the production of ROS. Extraradical hyphae and spores from Glomus intraradices accumulated small levels of ROS within their cell wall and produced ROS within the cytoplasm in response to stress. Within AM roots, we observed a certain correlation of arbuscular senescence and H₂O₂ accumulation after staining by diaminobenzidine (DAB) and a more general accumulation of ROS close to fungal structures when using dihydrorhodamine 123 (DHR 123) for staining. According to electron microscopical analysis of AM roots from Z. mays after staining by CeCl₃, intracellular accumulation of H₂O₂ was observed in the plant cytoplasm close to intact and collapsing fungal structures, whereas intercellular H₂O₂ was located on the surface of fungal hyphae. These characteristics of ROS accumulation in AM roots suggest similarities to ROS accumulation during the senescence of legume root nodules.     

Time-course study and partial characterization of a protein on hyphae of arbuscular mycorrhizal fungi during active colonization of roots

Material on the surface of hyphal walls of arbuscular mycorrhizal fungi (AMF) during active colonization of plant roots was detected by a monoclonal antibody. Pot-cultured isolates of Glomus, Acaulospora, Gigaspora, Scutellospora, and Entrophospora had immunofluorescent material (IM) on younger, thinner, intact hyphae, but IM was scant to absent on thicker, melanized or lysing hyphae. Colonization of corn (Zea mays L.), Sudangrass (Sorghum sudanense (Piper) Staph.) or red clover (Trifolium pratense L.) was examined during 5 months of plant growth by removing cores and performing an indirect immunoassay on roots with attached hyphae. Fresh spores of some Glomus spp. had IM on the outer layer of the spore wall. Abundant IM was seen on root hairs of plants colonized by some isolates, and some IM was detected on root surfaces of all plants examined even during early colonization. After cultures were dried, hyphae, roots and spores had little to no IM. Uninoculated control roots had very rare, small patches of IM. An immunoreactive protein was extracted from hyphae of Gigaspora and Glomus isolates by using 20mM citrate (pH 7.0) at 121°C for 90 min. Gel electrophoresis profiles indicated that all isolates tested had the same banding patterns. Lectin-binding of extracted protein is suggestive of a glycoprotein. The immunofluorescence assay can be used to examine root sections for active colonization by AMF, and the potential use of the protein to quantify AMF activity in soil is discussed.     

AM 真菌影响三叶草根系抗氧化酶活性的系统效应

本文对三叶草接种AM 真菌根内球囊霉, 用盆栽试验和分根试验测定根系的菌根侵染率和抗氧化酶活性, 研究AM 真菌对根系抗氧化酶活性的影响以及该影响的系统性。结果表明, 盆栽试验中接种根内球囊霉显著提高了根系中SOD、POD、CAT 的活性, 表明AM 真菌可以促进根系的抗氧化酶活性; 分根试验中一半根系接种了根内球囊霉的植株, 其另一半未接种的根系SOD、POD 活性也增加, 表明AM 真菌对根系抗氧化酶系统的促进具有系统效应。由于抗氧化酶系统是植物产生抗逆性的生理生化基础, 可以推测, AM 真菌对根系抗氧化酶活性的系统性提高有助于保护根系整体, 而非仅仅保护受侵染根段。     

Plasma membrane ATPase and H+ transport activities in microsomal membranes from mycorrhizal tomato roots

ATPase activity, ATP-dependent H⁺ transport and the amount of antigenic tomato plasma membrane H⁺-APTase have been analysed in membrane vesicles isolated from Glomus mosseae- or Glomus intraradices-colonized roots and from non-mycorrhizal tomato roots. Microsomal protein content was higher in mycorrhizal than in control roots. The specific activity of the plasma membrane H⁺-ATPase was not affected by mycorrhizal colonization, although this activity increased in membranes isolated from mycorrhizal roots when expressed on a fresh weight basis. Western blot analysis of microsomal proteins using antibodies raised against the Arabidopsis thaliana plasma membrane H⁺ - ATPase showed that mycorrhizal colonization did not change the relative amount of tomato plasma membrane ATPase in the microsomes. However, on a fresh weight basis, there was a greater amount of this protein in roots of mycorrhizal plants. In addition, mycorrhizal membranes showed a higher specific activity of the vanadate-sensitive ATP-dependant H⁺ transport than membranes isolated from control roots. These results suggest that mycorrhiza might regulate the plasma membrane ATPase by increasing the coupling efficiency between H⁺ transport and ATP hydrolysis. The observed effects of mycorrhizal colonization on plasma membrane H⁺-ATPase were independent of the AM fungal species colonizing the root system.     

Cereal phosphate transporters associated with the mycorrhizal pathway of phosphate uptake into roots

A very large number of plant species are capable of forming symbiotic associations with arbuscular mycorrhizal (AM) fungi. The roots of these plants are potentially capable of absorbing P from the soil solution both directly through root epidermis and root hairs, and via the AM fungal pathway that delivers P to the root cortex. A large number of phosphate (P) transporters have been identified in plants; tissue expression patterns and kinetic information supports the roles of some of these in the direct root uptake pathways. Recent work has identified additional P transporters in several unrelated species that are strongly induced, sometimes specifically, in AM roots. The primary aim of the work described in this paper was to determine how mycorrhizal colonisation by different species of AM fungi influenced the expression of members of the Pht1 gene families in the cereals Hordeum vulgare (barley), Triticum aestivum (wheat) and Zea mays (maize). RT-PCR and in-situ hybridisation, showed that the transporters HORvu;Pht1;8 (AY187023), TRIae;Pht1;myc (AJ830009) and ZEAma;Pht1;6 (AJ830010), had increased expression in roots colonised by the AM fungi Glomus intraradices,Glomus sp. WFVAM23 and Scutellospora calospora. These findings add to the increasing body of evidence indicating that plants that form AM associations with members of the Glomeromycota have evolved phosphate transporters that are either specifically or preferentially involved in scavenging phosphate from the apoplast between intracellular AM structures and root cortical cells. Operation of mycorrhiza-inducible P transporters in the AM P uptake pathway appears, at least partially, to replace uptake via different P transporters located in root epidermis and root hairs. Electronic Supplementary Material Supplementary material is available for this article at     

Sucrose synthase levels do not limit or regulate carbon transfer in the arbuscular mycorrhizal symbiosis

Abstract

Sucrose synthase (SuSy) is the main sucrose breakdown enzyme in plant sink tissues, including nodules, and is a possible candidate for the diversion of plant carbon to arbuscular mycorrhizal (AM) fungi in roots. We tested the involvement of SuSy in AM symbiosis of Glomus intraradices and Pisum sativum (pea). We observed that peas deficient in the predominant root isoform of SuSy were colonized successfully by AM fungi similar to wild-type roots. SuSy protein levels did not increase in roots as AM symbiosis developed, although SuSy protein levels did increase in nodules as the rhizobium symbiosis developed. Our results lead us to conclude that, unlike nodule symbiosis, SuSy protein does not limit or regulate carbon transfer in the AM symbiosis.     

Polygalacturonase activity and location in arbuscular mycorrhizal roots of Allium porrum L.

Polygalacturonase activity and location were analysed in leek roots (Allium porrum L.) colonized by Glomus versiforme (Karst.) Berch, an arbuscular mycorrhizal (AM) fungus. Polygalacturonase activity in mycorrhizal roots did not differ quantitatively from that found in nonmycorrhizal roots on all of the four harvesting dates. Fractionation of mycorrhizal root extracts by ion-exchange chromatography showed that expression of polygalacturonase was specific to the mutualistic association. Immunofluorescence and immunogold experiments were carried out to locate the polygalacturonase in mycorrhizal roots using a polyclonal antibody raised against a Fusarium moniliforme endopolygalacturonase. Immunolabelling was observed all over the arbuscules (intracellular fungal structures) but particularly at the interface between the arbuscule and the plant membrane. Since pectins are located in this area, we suggest that polygalacturonase produced during the symbiosis could play a role in plant pectin degradation.