Effect of salinity on mycorrhizal onion and tomato in soil with and without additional phosphate

Summary Vesicular-arbuscular mycorrhizal fungi (VAM) are known to increase plant growth in saline soils. Previous studies, however, have not distinguished whether this growth response is due to enhanced P uptake or a direct mechanism of increased plant salt tolerance by VAM. In a glasshouse experiment onions (Allium cepa L.) were grown in sterilized, low-P sandy loam soil amended with 0, 0.8, 1.6 mmol P kg^–1 soil with and without mycorrhizal inoculum. Pots were irrigated with saline waters having conductivities of 1.0, 2.8, 4.3, and 5.9 dS m^–1. Onion colonized withGlomus deserticola (Trappe, Bloss, and Menge) increased growth from 394% to 100% over non-inoculated control plants when soil P was low ( 0.2 mmol kg^–1 NaHCO₃-extractable P) at soil saturation extract salinities from 1.1 dS m^–1 to 8.8 dS m^–1. When 0.8 and 1.6 mM P was added no dry weight differences due to VAM were observed, however, K and P concentrations were higher in VAM plants in saline treatments.Glomus fasciculatum (Gerdeman and Trappe) andGlomus mosseae (Nicol. and Gerd.) isolates increased growth of VAM tomato 44% to 193% in non-sterilized, saline soil (10 dS m^–1 saturation extract) despite having little effect on growth in less saline conditions when soil P was low. Higher tomato water potentials, along with improved K nutrition by VAM in onion, indicate mechanisms other than increased P nutrition may be important for VAM plants growing under saline stress. These effects appear to be secondary to the effects of VAM on P uptake.     

Contribution of the VA mycorrhizal hyphae in acquisition of phosphorus and zinc by maize grown in a calcareous soil

An investigation was carried out to test whether the mechanism of increased zinc (Zn) uptake by mycorrhizal plants is similar to that of increased phosphorus (P) acquisition. Maize (Zea mays L.) was grown in pots containing sterilised calcareous soil either inoculated with a mycorrhizal fungus Glomus mosseae (Nicol. and Gerd.) Gerdemann and Trappe or with a mixture of mycorrhizal fungi, or remaining non-inoculated as non-mycorrhizal control. The pots had three compartments, a central one for root growth and two outer ones for hyphal growth. The compartmentalization was done using a 30-m nylon net. The root compartment received low or high levels of P (50 or 100 mg kg^–1 soil) in combination with low or high levels of P and micronutrients (2 or 10 mg kg^–1 Fe, Zn and Cu) in the hyphal compartments.Mycorrhizal fungus inoculation did not influence shoot dry weight, but reduced root dry weight when low P levels were supplied to the root compartment. Irrespective of the P levels in the root compartment, shoots and roots of mycorrhizal plants had on average 95 and 115% higher P concentrations, and 164 and 22% higher Zn concentrations, respectively, compared to non-mycorrhizal plants. These higher concentrations could be attributed to a substantial translocation of P and Zn from hyphal compartments to the plant via the mycorrhizal hyphae. Mycorrhizal inoculation also enhanced copper concentration in roots (135%) but not in shoots. In contrast, manganese (Mn) concentrations in shoots and roots of mycorrhizal plants were distinctly lower, especially in plants inoculated with the mixture of mycorrhizal fungi.The results demonstrate that VA mycorrhizal hyphae uptake and translocation to the host is an important component of increased acquisition of P and Zn by mycorrhizal plants. The minimal hyphae contribution (delivery by the hyphae from the outer compartments) to the total plant acquisition ranged from 13 to 20% for P and from 16 to 25% for Zn.     

Influence of the arbuscular mycorrhizal fungus Glomus mosseae on uptake of arsenate by the As hyperaccumulator fern Pteris vittata L.

We report for the first time some effects of colonization by an arbuscular mycorrhizal (AM) fungus (Glomus mosseae) on the biomass and arsenate uptake of an As hyperaccumulator, Pteris vittata. Two arsenic levels (0 and 300 mg As kg^–1) were applied to an already contaminated soil in pots with two compartments for plant and hyphal growth in a glasshouse experiment. Arsenic application had little or no effect on mycorrhizal colonization, which was about 50% of root length. Mycorrhizal colonization increased frond dry matter yield, lowered the root/frond weight ratio, and decreased frond As concentration by 33–38%. Nevertheless, transfer of As to fronds showed a 43% increase with mycorrhizal colonization at the higher soil As level. Frond As concentrations reached about 1.6 g kg^–1 (dry matter basis) in non-mycorrhizal plants in the As-amended soil. Mycorrhizal colonization elevated root P concentration at both soil As levels and mycorrhizal plants had higher P/As ratios in both fronds and roots than did non-mycorrhizal controls.     

Growth response of Atriplex nummularia to inoculation with arbuscular mycorrhizal fungi at different salinity levels

Chenopods are generally regarded as non-host plants for mycorrhizal fungi and are believed not to benefit from colonization by mycorrhizal fungi. Perennial Atriplex nummularia Lindl., growing under field conditions, showed a relatively high level of colonization by mycorrhizal fungi (10–30% of root length colonized) in spring and summer. Accordingly, two glasshouse experiments were designed to assess the effects of inoculation with mycorrhizal fungi (with a single species or a mixture of different species) on growth, nutrient uptake, and rhizosphere bacterial community composition of A. nummularia at high and low salinity levels (2.2 and 12 dSm^–1). Only low and patchy colonization by mycorrhizal fungi (1–2 of root length colonized) was detected in inoculated plants under glasshouse conditions which was unaffected by salinity. Despite the low colonization, inoculation increased plant growth and affected nutrient uptake at both salinity levels. The effects were higher at an early stage of plant development (6weeks) than at a later stage (9–10 weeks). Salinity affected the bacterial community composition in the rhizosphere as examined by ribosomal intergenic spacer amplification (RISA) of 16S rDNA, digitization of the band patterns and multivariate analysis. The effects of inoculation with mycorrhizal fungi on growth of A. nummularia may be attributed to (i) direct effects of mycorrhizal fungi on plant nutrient uptake and/or (ii) indirect effects via mycorrhizal-induced changes in the bacterial community composition.     

Chemical alteration of the rhizosphere of the mycorrhizal-colonized wheat root

Plexiglass pot growth chamber experiments were conducted to evaluate the chemical alterations in the rhizosphere of mycorrhizal wheat roots after inoculation with Glomus intraradices [arbuscular mycorrhizal fungus (AMF)]. Exchange resins were used as sinks for nutrients to determine whether the inoculated plant can increase the solubility and the uptake of P and micronutrients. Treatments included: (1) soil (bulk soil); (2) AMF inoculation no P addition (I–P); (3) no inoculation with no P addition (NI–P); (4) AMF inoculation with addition of 50 mg P (kg soil)^–1 (I+P), and (5) no inoculation with addition of 50 mg P (kg soil)^–1 (NI+P). The AMF inoculum was added at a rate of four spores of G. intraradices (g soil)^–1. The exchange resin membranes were inserted vertically 5 cm apart in the middle of Plexiglass pots. Spring wheat (Triticum aestivum cv. Len) was planted in each Plexiglass pot and grown for 2 weeks in a growth chamber where water was maintained at field capacity. Rhizosphere pH and redox potential (Eh), nutrient bioavailability indices and mycorrhizal colonization were determined. Mycorrhizal inoculation increased the colonization more when P was not added, but did not increase the shoot dry weight at either P level. The rhizosphere pH was lower in the inoculated plants compared to the noninoculated plants in the absence of added P, while the Eh did not change. The decrease in pH in the rhizosphere of inoculated plants could be responsible for the increased P and Zn uptake observed with inoculation. In contrast, Mn uptake was decreased by inoculation. The resin-adsorbed P was increased by inoculation, which, along with the bioavailability index data, may indicate that mycorrhizal roots were able to increase the solubility of soil P.     

The effect of in vitro pretreatments on subsequent shoot organogenesis from excised Rubus and Malus leaves

Shoot regeneration from Rubus leaves was obtained on a medium containing MS salts, vitamins and sugars, Staba vitamins, casein hydrolysate (100 mg l^–1) and 10 M thidiazuron. Shoot regeneration from Malus leaves was obtained on N⁶ rice anther medium with 5 M thidiazuron. In vitro pretreatment of source shoots with either colchicine or thidiazuron enhanced the organogenic potential of detached leaves of two Rubus hybrids. The response to colchicine was quadratic and occurred at non-mutagenic concentrations (75–250 M). The response to thidiazuron was exponential between 0 and 5 M. When applied as a pretreatment, the effectiveness of several different cytokinins (benzyladenine, thidiazuron, zeatin) at enhancing Malus and Rubus organogenesis was related to the shoot proliferation activity of the cytokinin and to treatment-induced variation in leaf and petiole size.Abbreviations BA benzyladenine - 2,4-D 2,4-dichlorophenoxyacetic acid - IBA indolebutyric acid - MS Murashige & Skoog basal medium devoid of plant growth regulators - OI organogenesis-initiating subculture - PTI colchicine pretreatment subculture - PTII cytokinin pretreatment subculture - NAA naphthaleneacetic acid - TDZ thidiazuron - zeatin trans-zeatin     

The Beltsville method for soilless production of vesicular-arbuscular mycorrhizal fungi

A low-cost, low-maintenance system for soilless production of vesicular-arbuscular mycorrhizal (VAM) fungus spores and inoculum was developed and adapted for production of acidophilic and basophilic isolates. Corn (Zea mays) plants were grown with Glomus etunicatum, G. mosseae or Gigaspora margarita in sand automatically irrigated with modified Hoagland's solution. Sand particle size, irrigation frequency, P concentration, and buffer constituents were adjusted to maximize spore production. Modified half-strength Hoagland's solution buffered with 4-morpholine ethane-sulfonic acid (MES) automatically applied 5 times/day resulted in production of 235 G. etunicatum spores/g dry wt. of medium (341000 spores/pot) and 44 G. margarita spores/g dry wt. of medium (64800 spores/pot). For six basophilic isolates of G. mosseae, CaCO₃ was incorporated into the sand and pots were supplied with the same nutrient solution as for acidophilic isolates. The increased pH from 6.1±0.2 to 7.2±0.2 resulted in spore production ranging from 70 to 145 spores/g dry wt. (102000–210000 spores/pot). Spore production by all isolates grown in the soilless sand system at Beltsville has exceeded that of traditional soil mixtures by 32–362% in 8–12 weeks.     

Performance of vesicular-arbuscular mycorrhizae in saline-alkali soil in relation to various amendments

The present study shows that vesicular-arbuscular mycorrhizal colonization in plants grown in saline-alkali soil is host dependent and it is significantly affected by various amendments given to reclaim such soils. Maximum reduction in vesicular-arbuscular mycorrhizal colonization was observed in the plants raised in saline-alkali soil amended with gypsum and maximum colonization was observed after plant-growth promoting rhizobacterium or farm yard manure amendments. Plants possessing high level of vesicular-arbuscular mycorrhizal colonization (50–70%) included Acacia nilotica, Albizia lebbeck and Dalbergia sissoo. Casuarina equisetifolia possessed moderate level of vesicular-arbuscular mycorrhizae colonization (30–50%) and Azadirachta indica possessed low level of colonization (20–40%).     

Mycorrhization alleviates benzo[a]pyrene-induced oxidative stress in an in vitro chicory root model

Among chemicals that are widely spread both in terrestrial and aquatic ecosystems, benzo[a]pyrene is a major source of concern. However, little is known about its adverse effects on plants, as well as about the role of mycorrhization in protection of plant grown in benzo[a]pyrene-polluted conditions. Hence, to contribute to a better understanding of the adverse effects of polycyclic aromatic hydrocarbons on the partners of mycorrhizal symbiotic association, benzo[a]pyrene-induced oxidative stress was studied in transformed Cichorium intybus roots grown in vitro and colonized or not by Glomus intraradices. The arbuscular mycorrhizal fungus development (colonization, extraradical hyphae length, and spore formation) was significantly reduced in response to increasing concentrations of benzo[a]pyrene (35–280 μM). The higher length of arbuscular mycorrhizal roots, compared to non-arbuscular mycorrhizal roots following benzo[a]pyrene exposure, pointed out a lower toxicity of benzo[a]pyrene in arbuscular mycorrhizal roots, thereby suggesting protection of the roots by mycorrhization. Accordingly, in benzo[a]pyrene-exposed arbuscular mycorrhizal roots, statistically significant decreases were observed in malondialdehyde concentration and 8-hydroxy-2′-desoxyguanosine formation. The higher superoxide dismutase activity detected in mycorrhizal chicory roots could explain the benzo[a]pyrene tolerance of the colonized roots. Taken together, these results support an essential role of mycorrhizal fungi in protecting plants submitted to polycyclic aromatic hydrocarbon, notably by reducing polycyclic aromatic hydrocarbon-induced oxidative stress damage.     

Benzyladenine-induced adventitious shoot regeneration from hypocotyls of adzukibean (Vigna angularis {Willd.} Ohwi & Ohashi)

A complete in vitro plant regeneration systemvia adventitious shoot-bud formation from seedlinghypocotyls of adzukibean was developed. Cotyledonarynode (CN) and root explants from 4-d-old invitro-germinated seedlings of 4 cultivars werecultured on agar-solidified R medium consisting of MSsalts, B5 vitamins, 3.0% (w/v) sucrose and 4.4 µM6-benzyladenine (BA). Shoot buds arose adventitiouslyat the basipetal cut of the hypocotyl in 40 to 85%of type 1 CN (8–10 mm) explants 6 weeks later with cvsBansei aki azuki and Tanbadainagon azuki exhibitingsignificantly higher response. Shorter type 2 (4–5 mm) CN explants responded poorly (0–6%) in culturewhile intact roots were regenerative but at lowerfrequencies (5 to 31%). In cv Bansei aki azuki,optimum shoot initiation (78.7%) was achieved whentype 1 CN explants from BA-preconditioned seedlingswere re-cultured in R medium (SC0). BA supplied at8.8 µM during SC0 significantly reduced the shootinitiation response to 46.7%. In the same manner,substituting equimolar concentration (4.4 µM) ofkinetin (KN), zeatin (Z) or thidiazuron (TDZ) to BAduring SC0 yielded no or significantly lowerresponses. Mean shoot production ranged from 6.2 to9.8 shoots per explant 6 weeks after subculture (SC1)and varied significantly among the 4 cultivars used. Efficient rooting (93–96%) was achieved on excisedshoots transferred to MS salts + B5 vitamins basalmedium alone. The tissue culture system covered only16 weeks to completion. Regenerated plantlets weretransferred to soil with 86–100% survival rate andall surviving plants were morphologically normal andseed-fertile.     

Some isolates of the nematophagous fungus Pochonia chlamydosporia promote root growth and reduce flowering time of tomato

The fungal parasite of nematode eggs Pochonia chlamydosporia is also a root endophyte known to promote growth of some plants. In this study, we analysed the effect of nine P. chlamydosporia isolates from worldwide origin on tomato growth. Experiments were performed at different scales (Petri dish, growth chamber and greenhouse conditions) and developmental stages (seedlings, plantlets and plants). Seven P. chlamydosporia isolates significantly (P < 0.05) increased the number of secondary roots and six of those increased total weight of tomato seedlings. Six P. chlamydosporia isolates also increased root weight of tomato plantlets. Root colonisation varied between different isolates of this fungus. Again P. chlamydosporia significantly increased root growth of tomato plants under greenhouse conditions and reduced flowering and fruiting times (up to 5 and 12 days, respectively) versus uninoculated tomato plants. P. chlamydosporia increased mature fruit weight in tomato plants. The basis of the mechanisms for growth, flowering and yield promotion in tomato by the fungus are unknown. However, we found that P. chlamydosporia can produce Indole‐3‐acetic acid and solubilise mineral phosphate. These results suggest that plant hormones or nutrient ability could play an important role. Our results put forward the agronomic importance of P. chlamydosporia as biocontrol agent of plant parasitic nematodes with tomato growth promoting capabilities.     

Regulation of resistance and susceptibility in wheat–powdery mildew pathosystem with exogenous cytokinins

Dose–response relationship between resistance of wheat seedlings (Triticum aestivum, cultivar Zarya) to Erysiphe graminis f. sp. tritici Marchal. (Syn. Blumeria graminis), a causal organism of wheat powdery mildew and exogenous zeatin has been investigated. Two-week-old seedlings were inoculated with the pathogen. Zeatin or zeatinriboside were added to the nutrient solution immediately after inoculation. The dose–response curve of cytokinin in the most cases was multiphasic, with peaks of increased susceptibility occurring at 0.25–1.5 and 1.5–9 μM cytokinin, separated by a region of increased resistance at 0.5–3 μM cytokinin. The change in mineral nutrition or simultaneous treatment with thidiazuron revealed alterations of the dose–response curve ranging from a curve with maximum of resistance to a curve with maximum of susceptibility. Both multiphase nature of dose–response and its variability were proposed as possible explanations for earlier observed discrepancies in experimental data on modification of disease resistance by cytokinins. A mathematical model for two metabolic processes with substrate inhibition connected in-series was suggested to explain the multiphase dose–response. In this model, the product of the first reaction was used as substrate for the second reaction. Numerical experiments showed the changes in the shape of dose–response curve with changes in parameters dependent of cytokinin metabolism.     

VA-mycorrhiza mediated P effect on growth and yield of sunflower (Helianthus annuus L.) at different P levels

A field trial was conducted to study the response of sunflower (Helianthus annuus L.) to different phosphorus levels (16, 24 or 32 kg P ha^–1) and inoculation with vesicular-arbuscular mycorrhizal fungus, Glomus fasciculatum on vertisol during summer 1993. At the vegetative stage of sunflower, percent mycorrhizal root colonization, spore count, dry biomass and P uptake did not differ significantly between inoculated and uninoculated control plants. However, at later stages (flowering and maturity) percent root colonization, spore count, total dry biomass and total P uptake were significantly higher in inoculated plants than in uninoculated control plants. The total dry biomass, P content and seed yield increased with increasing P level in uninoculated plants, whereas no significant difference was observed between 16 and 32 kg P ha^–1 in inoculated plants. The positive effect of mycorrhizal inoculation decreased with increasing P level above 16 kg P ha^–1, due to decreased percent root colonization and spore count at higher P levels.     

The effect of Mg on Ca,K and P content of date palm seedlings under mycorrhizal and non-mycorrhizal conditions

The effects of the presence or absence of Mg in the nutrient solution and of vesicular-arbuscular mycorrhizal fungi (VAMF) on the content and partitioning of Ca, K and P between root and shoot of date palm (Phoenix dactylifera) seedlings were examined under greenhouse conditions using soil as basal medium. Mg content of the soil was 14.95 µmol/g dry soil. The infection percentages after inoculation of VAMF were 66.0% and 55.5%, respectively, on application of –Mg and +Mg nutrient solution. Ca content of both roots and shoots did not change by these treatments; but a highly significant decrease in shoots was recorded on –Mg and +VAMF treatment. K content of root was significantly elevated by –Mg and +VAMF treatment but no changes were observed in shoots. P content of both roots and shoots increased significantly with +VAMF regardless of the presence or absence of Mg.     

Plant regeneration in Kentucky bluegrass (Poa pratensis L.) via coleoptile tissue cultures

Summary Plant regeneration in Kentucky bluegrass (Poa pratensis L. cv. Touchdown) via culture of seedling tissues was investigated. When coleoptile, leaf, and stem sections of dark-germinated seedlings were cultured on Murashige and Skoog (MS) medium, different types of callus were produced, depending on the expiant source and growth regulator combinations. Only compact-friable callus (type 3) and moderately compact, friable callus (type 2) produced shoots upon subculture. The nonstructured watery callus (type 4) produced roots without shoots. Shoot differentiation from callus tissues was highest when the culture medium contained 0.2 mgL^–1 picloram + 0.01 mgL^–1 -naphthaleneacetic acid (NAA). Calli grown from coleoptiles had higher shoot regeneration frequency (32%) than that obtained from either stem sections (12%) or young leaf tissues (2%) of the same seedlings. Some organogenic callus lines produced exclusively green plants, while others produced albino shoots or a mixture of green and albino shoots. The green plants were multiplied in a medium containing 0.1 mgL^–1 BAP plus either 0.2 mgL^–1 picloram or 0.1 mgL^–1 indole-3-acetic acid (IAA). Over 90% of the cultures in the shoot proliferation medium produced roots in 4 weeks. The rooted plants were successfully established in soil medium and grown in the greenhouse.Abbreviations BAP 6-benzylaminopurine - 2,4-D 2,4-dichlorophenoxyacetic acid - IAA indole-3-acetic acid - MS Murashige and Skoog (1962) medium - NAA -naphthaleneacetic acid - picloram 4-amino-3,5,6-trichloropicolinic acid - TDZ thidiazuron     

An efficient mass propagation system for cassava (Manihot esculenta Crantz) based on nodal explants and axillary bud-derived meristems

Nodes from 3- to 5-week-old in vitro plants of different cassava cultivars were cultured for 2–3 days on solid Murashige and Skoog basal medium supplemented with cytokinin to induce the enlargement of axillary buds. Subculture of these buds on the same medium resulted in multiple shoot formation within 4–6 weeks. Of the four cytokinins tested (6-benzylaminopurine (BAP), thidiazuron (TDZ), zeatin, and kinetin), BAP induced shoot development most efficiently. The best results were obtained with cultivar TMS 30555, in which 63% of the explants each produced at least 25 shoots on medium with 10 mg/l BAP. In cultivars that did not produce shoots, the addition of the surfactant Pluronic F-68 (2% wt/vol) raised the percentage of explants forming at least 5 shoots from 0 to 20–60%. Axillary buds were also used to dissect meristems and test their ability to regenerate into shoots. Shoot formation from meristems of six different cultivars was observed after preculture on medium with 5 mg/l BAP followed by transfer to 10 mg/l BAP.Abbreviations MS Murashige and Skoog - BAP 6-Benzylaminopurine - TDZ Thidiazuron     

Arbuscular mycorrhizal fungi affect both penetration and further life stage development of root-knot nematodes in tomato

The root-knot nematode Meloidogyne incognita poses a worldwide threat to agriculture, with an increasing demand for alternative control options since most common nematicides are being withdrawn due to environmental concerns. The biocontrol potential of arbuscular mycorrhizal fungi (AMF) against plant-parasitic nematodes has been demonstrated, but the modes of action remain to be unraveled. In this study, M. incognita penetration of second-stage juveniles at 4, 8 and 12 days after inoculation was compared in tomato roots (Solanum lycopersicum cv. Marmande) pre-colonized or not by the AMF Glomus mosseae. Further life stage development of the juveniles was also observed in both control and mycorrhizal roots at 12 days, 3 weeks and 4 weeks after inoculation by means of acid fuchsin staining. Penetration was significantly lower in mycorrhizal roots, with a reduction up to 32%. Significantly lower numbers of third- and fourth-stage juveniles and females accumulated in mycorrhizal roots, at a slower rate than in control roots. The results show for the first time that G. mosseae continuously suppresses root-knot nematodes throughout their entire early infection phase of root penetration and subsequent life stage development.     

Effects of a vesicular-arbuscular mycorrhizal fungus and other soil microorganisms on growth,mineral nutrient acquisition and root exudation of soil-grown maize plants

Maize (Zea mays L. cv. Alize) plants were grown in a calcareous soil in pots divided by 30-m nylon nets into three compartments, the central one for root growth and the outer ones for hyphal growth. Sterle soil was inoculated with either (1) rhizosphere microorganisms other than vesicular-arbuscular mycorrhizal (VAM) fungi, (2) rhizosphere microorganisms together with a VAM fungus [Glomus mosseae (Nicol. and Gerd.) Gerdemann and Trappel], or (3) with a gamma-irradiated inoculum as control. Plants were grown under controlled-climate conditions and harvested after 3 or 6 weeks. VAM plants had higher shootroot ratios than non-VAM plants. After 6 weeks, the concentrations of P, Zn and Cu in roots and shoots had significantly increased with VAM colonization, whereas Mn concentrations had significantly decreased. Root exudates were collected on agar sheets placed on the interface between root and hyphal compartments. Six-week-old VAM and non-VAM plants had similar root exudate compositions of 72–73% reducing sugars, 17–18% phenolics, 7% organic acids and 3% amino acids. In another experiment in which root exudates were collected on agar sheets with or without antibiotics, the amounts of amino acids and carbohydrates recovered were similar in VAM and non-VAM plants. However, threeto sixfold higher amounts of carbohydrates, amino acids and phenolics were recovered when antibiotics were added to the agar sheets. Thus, the high microbial activity in the rhizosphere and on the rhizoplane limits the exudates recovered from roots.     

Instructions for Authors

Efficient shoot regeneration of Vanda coerulea was achieved using thin shoot tip sections and thidiazuron. Protocorm-like bodies or proliferating shoot buds was observed when thin shoot tip sections were cultured on Vacin and Went's (VW) (1949) basal medium supplemented with 11.35 µM thidiazuron. The highest percentage of protocorm-like bodies (95%) survived and ultimately produced healthy shoots with 2 – 3 leaves when subjected to a 4 week thidiazuron treatment. A culture period longer than 8 weeks with thidiazuron resulted in the formation of fasciated or distorted shoots. Shoots produced roots when cultured on half strength VW basal medium supplemented with 11.42 µM IAA. The well rooted shoots were transferred to pots containing charcoal chips, coconut husk and broken tiles (2:2:1) and a 98% survival rate was achieved.     

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.