Phosphorus effect on phosphatase activity in endomycorrhizal maize

Success of a mycorrhizal symbiosis is influenced by the availability of phosphorus (P) in the soil. Maize ( Zea mays L. cv. Great Lakes 586) plants were grown under five different levels of soil P, either in the presence or absence of formononetin or the vesicular‐arbuscular mycorrhizal (VAM) fungus Glomus intraradices Schenck and Smith. We detected physiological differences in mycorrhizal roots very early in the development of symbiosis, before the onset of nutrient‐dependent responses. Under low P levels, VAM roots accumulated a greater shoot dry weight (13%), root P concentration (15%) and protein concentration (30%) than non-VAM roots, although root growth was not statistically significantly different. At higher P levels, mycorrhizal roots weighed less than non-VAM roots (10%) without a concomitant host alteration of growth or root P concentration. Mycorrhizal colonization decreased as soil P increased. Formononetin-treatment enhanced colonization of the root by G. intraradices and partially overcame inhibition of VAM colonization by high soil P concentrations. This is the first report that formononetin improves root colonization under high levels of soil P. Acid phosphatase (ACP) and alkaline phosphatase (ALP) activities were closely related to the level of fungal colonization in corn roots. ACP activity in corn roots responded more to soil P availability than did ALP activity (38% more). These results suggest that ACP was involved in the increased uptake of P from the soil, while ALP may be linked to active phosphate assimilation or transport in mycorrhizal roots. Thus, soil P directly affected a number of enzymes essential in host-endophyte interplay, while formononetin enhanced fungal colonization.     

Initial and residual toxicity of soil-applied thiram on the vesicular-arbuscular mycorrhizal symbiosis

The effect of the non-systemic fungicide thiram on the vesicular-arbuscular mycorrhizal (VAM) symbiosis and on Leucaena leucocephala was evaluated in a greenhouse experiment. In the uninoculated soil treated with P at a level optimal for mycorrhizal activity, mycorrhizal colonization of roots was low, and did not change as the concentration of thiram in the soil increased with the from 0 to 1000 mg/kg. When this soil was inoculated VAM fungus Glomus aggregatum, with VAM colonization was enhanced significantly, but decreased increase in thiram concentration until it coincided with the level observed in the uninoculated soil. Similarly, symbiotic effectiveness was reduced, its expression delayed or completely eliminated with increase in the concentration of thiram. Amending soil to a P level sufficient for non-mycorrhizal host growth fully compensated for thiram-induced loss of VAM activity if the thiram levels did not exceed 125 mg/kg. In soil treated with 50 mg thiram/kg, the toxicity of the fungicide dissipated within 66 days of application. At higher concentrations, the toxicity of the chemical on the mycorrhizal symbiosis appeared to be enhanced.Contribution from the Hawaii Institute of Tropical Agriculture and Human Resources Journal Series No. 3716     

Response of citronella Java (Cymbopogon winterianus Jowitt) to VA mycorrhizal fungi and soil compaction in relation to P supply

Nutrient acquisition and growth of citronella Java (Cymbopogon winterianus Jowitt) was studied in a P-deficient sandy soil to determine the effects of mycorrhizal symbiosis and soil compaction. A pasteurized sandy loam soil was inoculated either with rhizosphere microorganisms excluding VAM fungi (non-mycorrhizal) or with the VAM fungus, Glomus intraradices Schenck and Smith (mycorrhizal) and supplied with 0, 50 or 100 mg P kg^-1 soil. The soil was compacted to a bulk density of 1.2 and 1.4 Mg m^-3 (dry soil basis). G. intraradices substantially increased root and shoot biomass, root length, nutrient (P, Zn and Cu) uptake per unit root length and nutrient concentrations in the plant, compared to inoculation with rhizosphere microorganisms when the soil was at the low bulk density and not amended with P. Little or no plant response to the VAM fungus was observed when the soil was supplied with 50 or 100 mg P kg^-1 soil and/or compacted to the highest bulk density. At higher soil compaction and P supply the VAM fungus significantly reduced root length. Non-mycorrhizal plants at higher soil compaction produced relatively thinner roots and had higher concentrations and uptake of P, Zn and Cu than at lower soil compaction, particularly under conditions of P deficiency. The quality of citronella Java oil measured in terms citronellal and d-citronellol concentration did not vary appreciably due to various soil treatments.     

Salinity and flooding stress effects on mycorrhizal and non-mycorrhizal citrus rootstock seedlings

Seedlings of the rootstocks Pineapple sweet orange (SwO), Carrizo citrange (CC), and sour orange (SO) were grown in low phosphorus (P) sandy soil and either inoculated with the vesicular-arbuscular mycorrhizal (VAM) fungus,Glomus intraradices, or were non-mycorrhizal (NM) and fertilized with P. VAM and NM seedings of similar shoot size and adequate P-status were selected for study of salinity and flooding stress. One-third of each of the VAM and NM plants were given 150 mM NaCl for a period of 24 days. One-third of the plants were placed into plastic bags and flooded for 21 days while the remaining third were non-stressed controls. In general, neither stress treatment affected mycorrhizal colonization. Salinity stress reduced the hydraulic conductivity of roots, leaf water potential, stomatal conductance and net assimilation of CO₂ (ACO₂) of mycorrhizal and non-mycorrhizal seedlings to a similar extent. VAM plants of CC and SO accumulated more Cl in leaves than NM plants. Cl was higher in non-mycorrhizal roots of SwO and CC than in mycorrhizal roots. Flooding the root zone for 3 weeks did not produce visible symptoms in the shoot but did influence plant water relations and reduce ACO₂ of all 3 rootstocks. VAM and NM plants of each rootstock were affected similarly by flooding. Comparable reduction in nitrogen and P content of both mycorrhizal and non-mycorrhizal plants suggested that flooding stress was primarily affecting root rather than hyphal nutrient uptake. Florida Agricultural Experimental Station Journal Series No. 7773.     

Genotype dependent variation in mycorrhizal colonization and response to inoculation of pearl millet

Summary Genotypes of pearl millet (Pennisetum americanum L. Leeke) were examined for differences in vesicular-arbuscular mycorrhizal (VAM) colonization and response to inoculation. For thirty genotypes tested across three field locations there was a range of mycorrhizal colonization intensity between 25 and 56%. In another experiment with two male-sterile lines, restorer lines and their derived crosses, grown in pots filled with non-sterilized soil there were significant differences between genotypes for colonization by mycorrhiza. This showed hostgenotype dependence for mycorrhizal colonization.Root growth rates, mycorrhizal root length, percentage root colonization and plant growth and P uptake were studied in ten genotypes. A set of 3 genotypes with similar root lengths varied significantly with regard to mycorrhizal root length and the percentage colonization. This supports the suggestion that VAM colonization and spread is dependent on the host genotype. The growth responses differed significantly between the genotypes and they also differed in their responses to P uptake and VAM inoculation. The utility of host-genotype dependent differences in VAM symbiosis in plant breeding is discussed.Journal Article No. 453     

Host dependent differential spread of <Emphasis Type="Italic">Glomus intraradices</Emphasis> on various Ri T-DNA transformed roots <Emphasis Type="Italic">in vitro</Emphasis>

Carrots (Daucus carota) are used extensively for culturing arbuscular mycorrhizal fungi under the root organ culture system (ROC) in vitro. Four different cultivars of carrot and one of clover (Trifolium subterraneum) root cultures were used to investigate symbiotic events with Glomus intraradices when introduced to different host-transformed roots. The investigation was directed to study the state of mycorrhization, its pattern and variation, if any and, the differences in nutritional uptake of each AM-host symbiosis. The findings suggest the role of the host on the differential expression of G. intraradices with respect to colonization, spore production, intraradical and extramatrical spread of the fungus, nutritional kinetics of each host-fungus symbiosis, pattern of growth and differences in anatomical and morphological features of host roots.     

Growth of Glomus intraradices and its effect on linseed (Linum usitatissimum L.) in hydroponic culture

This paper presents a hydroponic system for culturing and maintaining the VAM fungus Glomus intraradices in symbiosis with linseed (Linum usitatissimum L.) under greenhouse conditions in pure nutrient solution. It was possible to obtain large quantities of mycorrhizal host plant roots as well as extramatrical mycelium and chlamydospores free of impeding residues of solid substrate components. Starting from linseed donor plants inoculated in sand and transferred to the nutrient solution, new infections arose within the fast growing root system, hyphae spread out into the liquid and infected mycorrhiza-free receptor plants. Data for infection rates and plant growth parameters are presented. In comparsion to other culture systems for VAM fungi, the advantages of this hydroponic system are discussed and potential uses suggested.     

Response of vesicular-arbuscular mycorrhizas of maize to various rates of P addition to different rooting zones

Colonization of plant roots by vesicular-arbuscular mycorrhizal fungi is known to be reduced as the phosphorus nutrition of the plant is increased. It is generally accepted that the concentration of P in the plant rather than the soil regulates VAM colonization. Whether it is the shoot P concentration, the mean P concentration in the root system or the P concentration in the specific root being colonized is not known, but is of agronomic significance because fertilizer P is frequently applied in concentrated zones which would be expected to result in higher P concentration in roots growing in the fertilized zone than in the remainder of the root system. Growth chamber and field experiments were conducted to determine the effect on colonization of supplying varying amounts of P to different portions of the rooting zone. In growth chamber studies using a split-pot technique, the proportion of maize (Zea mays L.) root length containing arbuscules in a high-P zone was lower than that of roots of the same plant growing in a low- or medium-P zone. Root P concentration was higher in the high-P zone. In a field experiment conducted over a two-year period, VAM colonization of roots of young maize plants growing in fertilized soil was affected differently than that of roots growing outside the fertilized zone. A small addition of fertilizer P increased colonization of roots in the fertilized soil, but further additions resulted in an abrupt decline followed by a slower further decline, although colonization was not eliminated even by rates of 1600 g P g^-1 soil. Colonization of roots growing outside the fertilized zone declined gradually with increasing P addition but the overall decline was less than for roots in the fertilized zone. The data support the hypothesis that it is P concentration in the portion of the root system being colonized rather than the general P status of the plant which regulates VAM colonization. The agronomic implication of this is that, although a fertilizer band may reduce VAM colonization of roots in the band volume, roots growing outside this volume may be well colonized so the mycorrhizal symbiosis may be an important contributor to P nutrition.     

Residual toxicity of soil-applied chlorothalonil on mycorrhizal symbiosis in Leucaena leucocephala

The residual effect of the fungicide chlorothalonil on the vesicular-arbuscular mycorrhizal (VAM) symbiosis was evaluated in a greenhouse experiment. The soil used was an oxisol (Tropeptic Eutrustox) treated with P to obtain target levels near-optimal for VAM activity or sufficient for nonmycorrhizal host growth. In the uninoculated soil treated with the former P level, the fungicide reduced VAM colonization of roots and completely suppressed symbiotic effectiveness measured in terms of pinnule P content. When this soil was inoculated with Glomus aggregatum, symbiotic effectiveness was significantly reduced but not eliminated by 50 mg of the fungicide kg⁻¹. At higher chlorothalonil levels, VAM effectiveness but not VAM colonization was completely suppressed in the inoculated soil. The pattern with which chlorothalonil influenced tissue P content and dry matter yield at the time of harvest closely paralleled its effect on VAM effectiveness. In the soil treated with P level sufficient for nonmycorrhizal host growth, the adverse effect of the fungicide on the above variables was appreciably milder than when the host relied on VAM fungi for its P supply. The toxic effect of the fungicide, therefore, was partly offset by P fertilization, suggesting that VAM fungi were more sensitive to chlorothalonil than the host. Our results demonstrate that although the toxic effect of chlorothalonil declined as a function of time, a significant level of toxicity persisted 12.5 weeks after the chemical was applied to soil. Contribution from Hawaii Institute of Tropical Agriculture and Human Resources Journal Series No. 3625. Contribution from Hawaii Institute of Tropical Agriculture and Human Resources Journal Series No. 3625.     

Influence of three vesicular-arbuscular mycorrhizal fungi (Glomaceae) on the activity of specific enzymes in the root system of Cucumis sativus L.

The influence of three vesicular-arbuscular mycorrhizal (VAM) Glomus species on the activity of enzymes in the roots of Cucumis sativus was tested. Cucumber plants were grown in a split-root system, in which colonized and uncolonized roots of a single plant could be separated. The activity of the host root malate dehydrogenase (MDH), glucose 6-phosphate dehydrogenase (Gd), glutamate oxaloacetate transaminase (GOT) and glutamate dehydrogenase (GDH) was measured on a densitometer after separation of the host and fungal enzymes on polyacrylamide gels.The results showed that only minor changes in the activity of the host root enzymes occurred after VAM inoculation. Gd was stimulated by VAM and phosphorus, and one of the fungi decreased the activity of GDH in the host plant when both parts of the root system were colonized.     

Host genotype and the formation and function of VA mycorrhizae

VA mycorrhizae, the most ancient type of mycorrhizal symbiosis, are present in the most phytogenetically advanced groups. Few plants have evolved mechanisms to completely prevent infection by VAM fungi. Yet, plant species that are less dependent on VA mycorrhizae for nutrient acquisition (e.g., grasses) generally have less root colonization in the field than more dependent species (e.g., Citrus). Among closely related Citrus genotypes, there is a greater tendency for less dependent species to limit the rate but not the extent of colonization, even in high-P soils. We hypothesize that colonization represents a significant carbon cost that may be regulated by the host genotype. Carbon expenditure on the fungus at high P may result in mycorrhizal-induced growth depression. The potential value of breeding plants for greater susceptibility to colonization will depend on the cost/benefit of VA mycorrhizae for the specific crop, soil and environmental conditions. Although the genetics and physiology of host control over VAM colonization are barely known, recently discovered mycorrhizal colonization mutants (myc^-) of pea offer great promise for the study of host-fungus compatibility. Florida Agricultural Experiment Station Journal Series No. R-02765. Florida Agricultural Experiment Station Journal Series No. R-02765.     

Resistance Responses of Potato to Vesicular-Arbuscular Mycorrhizal Fungi under Varying Abiotic Phosphorus Levels

In mycorrhizal symbioses, susceptibility of a host plant to infection by fungi is influenced by environmental factors, especially the availability of soil phosphorus. This study describes morphological and biochemical details of interactions between a vesicular-arbuscular mycorrhizal (VAM) fungus and potato (Solanum tuberosum L. cv Russet Burbank) plants, with a particular focus on the physiological basis for P-induced resistance of roots to infection. Root infection by the VAM fungus Glomus fasciculatum ([Thaxt. sensu Gerdemann] Gerdemann and Trappe) was extensive for plants grown with low abiotic P supply, and plant biomass accumulation was enhanced by the symbiosis. The capacity of excised roots from P-deficient plants to produce ethylene in the presence or absence of exogenous 1-amino cyclopropane-1-carboxylic acid (ACC) was markedly reduced by VAM infection. This apparent inhibition of ACC oxidase (ACC_ox) activity was localized to areas containing infected roots, as demonstrated in split-root studies. Furthermore, leachate from VAM roots contained a potent water-soluble inhibitor of ethylene generation from exogenous ACC by nonmycorrhizal (NM) roots. The leachate from VAM-infected roots had a higher concentration of phenolics, relative to that from NM roots. Moreover, the rates of ethylene formation and phenolic concentration in leachates from VAM roots were inversely correlated, suggesting that this inhibitor may be of a phenolic nature. The specific activity of extracellular peroxidase recovered in root leachates was not stimulated by VAM infection, although activity on a fresh weight basis was significantly enhanced, reflecting the fact that VAM roots had higher protein content than NM roots. Polyphenol oxidase activity of roots did not differ between NM and VAM roots. These results characterize the low resistance response of P-deficient plants to VAM infection. When plants were grown with higher abiotic P supply, the relative benefit of the VAM symbiosis to plant growth decreased and root infection was lower. The in vivo ACC_ox activity was also greater in roots of plants grown on high levels of P compared with those grown on low levels, although the influence of VAM infection was partially to counteract the nutritional effect of P on ACC_ox activity. Similar to ACC_ox activity, extracellular peroxidase activity of roots increased linearly with increasing abiotic P supply, thus indicating a greater potential for resistance to VAM infection. These findings suggest that VAM fungi may alter phenolic metabolism of roots so as to hinder ethylene production and the root's ability to invoke a defense response. Raising the abiotic P supply to plants at least partially restores the capacity of roots to produce ethylene and may, in this way, increase the root's resistance to VAM infection.     

Effects of liming and mycorrhizal colonization on soil phosphate depletion and phosphate uptake by maize (Zea mays L.) and soybean (Glycine max L.) grown in two tropical acid soils

The effects of liming and inoculation with the arbuscular mycorrhizal fungus, Glomus intraradices Schenck and Smith on the uptake of phosphate (P) by maize (Zea mays L.) and soybean (Glycine max [L.] Merr.) and on depletion of inorganic phosphate fractions in rhizosphere soil (Al-P, Fe-P, and Ca-P) were studied in flat plastic containers using two acid soils, an Oxisol and an Ultisol, from Indonesia. The bulk soil pH was adjusted in both soils to 4.7, 5.6, and 6.4 by liming with different amounts of CaCO₃.In both soils, liming increased shoot dry weight, total root length, and mycorrhizal colonization of roots in the two plant species. Mycorrhizal inoculation significantly increased root dry weight in some cases, but much more markedly increased shoot dry weight and P concentration in shoot and roots, and also the calculated P uptake per unit root length. In the rhizosphere soil of mycorrhizal and non-mycorrhizal plants, the depletion of Al-P, Fe-P, and Ca-P depended in some cases on the soil pH. At all pH levels, the extent of P depletion in the rhizosphere soil was greater in mycorrhizal than in non-mycorrhizal plants. Despite these quantitative differences in exploitation of soil P, mycorrhizal roots used the same inorganic P sources as non-mycorrhizal roots. These results do not suggest that mycorrhizal roots have specific properties for P solubilization. Rather, the efficient P uptake from soil solution by the roots determines the effectiveness of the use of the different soil P sources. The results indicate also that both liming and mycorrhizal colonization are important for enhancing P uptake and plant growth in tropical acid soils.     

A tripartite culture system for endomycorrhizal inoculation of micropropagated strawberry plantlets in vitro

The objective of the current investigation was to develop a reliable method to obtain vesicular arbuscular mycorrhizae (VAM) in micropropagated plantlets and to determine their influence on growth. An in vitro system for culturing the VA mycorrhizal fungus Glomus intraradices with Ri T-DNA-transformed carrot roots or nontransformed tomato roots was used in this study as a potential active source of inoculum for the colonization of micropropagated plantlets. After root induction, micropropagated plantlets grown on cellulose plugs (sorbarod) were placed in contact with the primary mycorrhizae in growth chambers enriched with 5000 ppm CO₂ and fed with a minimal medium. After 20 days of tripartite culture, all plantlets placed in contact with the primary symbiosis were colonized by the VAM fungus. As inoculum source, 30-day-old VA mycorrhizal transformed carrot roots had a substantially higher infection potential than 5-, 10-or 20-day-old VAM. Colonized plantlets had more extensive root systems and better shoot growth than control plants. The VAM symbiosis reduced the plantlet osmotic potential. This response may be a useful pre-adaptation for plantlets during transfer to the acclimatization stage.     

Increased Sporulation of Vesicular-Arbuscular Mycorrhizal Fungi by Manipulation of Nutrient Regimens

Adjustment of pot culture nutrient solutions increased root colonization and sporulation of vesicular-arbuscular mycorrhizal (VAM) fungi. Paspalum notatum Flugge and VAM fungi were grown in a sandy soil low in N and available P. Hoagland nutrient solution without P enhanced sporulation in soil and root colonization of Acaulospora longula, Scutellospora heterogama, Gigaspora margarita, and a wide range of other VAM fungi over levels produced by a tap water control or nutrient solutions containing P. However, Glomus intraradices produced significantly more spores in plant roots in the tap water control treatment. The effect of the nutrient solutions was not due solely to N nutrition, because the addition of NH₄NO₃ decreased both colonization and sporulation by G. margarita relative to levels produced by Hoagland solution without P.     

Transformed soybean (Glycine max) roots as a tool for the study of the arbuscular mycorrhizal symbiosis

Ri T-DNA transformed roots have been used effectively in studying the interaction between various plant hosts and arbuscular mycorrhizal (AM) fungi. We investigated the in vitro monoxenic symbiosis between the AM fungus Glomus intraradices and transformed soybean roots (TSRs). Comparisons were made between TSR system and plants of the same genotype. The extraradical fungal structures generated in vitro culture showed normal development. Straight runner hyphae branched into short simple branched absorbing structures and spores were initiated. AM symbiosis was confirmed by the presence of arbuscules and vesicles in cortical cells of the TSRs. The frequency of intraradical colonization in TSRs was higher than in plants grown in soil, whereas the intensity values of intraradical colonization in TSR cultures were similar to those in whole plants. These results show that TSR cultures were able to support the growth and characteristic development of G. intraradices.     

Effect of liming on spore germination,germ tube growth and root colonization by vesicular-arbuscular mycorrhizal fungi

Summary The effect of soil acidity on spore germination, germ tube growth and root colonization of vesicular-arbuscular mycorrhizal (VAM) fungi was examined using a Florida Ultisol. Soil samples were treated with 0, 4, 8 and 12 meq Ca/MgCO₃/100 g soil and each lime level received 0, 240, and 720 ppm P as superphosphate. Corn (Zea mays L.) was planted in the soil treatments, inoculated with eitherGlomus mosseae orGigaspora margarita spores and grown for 31 days. Acid soil inhibits mycorrhizal formation byG. mosseae through its strong fungistatic effect against the spores. The dolomitic lime increased mycorrhizal formation by both fungal species.G. margarita is much less sensitive to acidic conditions thanG. mosseae. Al ions are a very important component of the fungistatic property against the VAM symbiosis. VAM fungus adaptation may be important for plants growing on infertile acid soils if soil inoculation with these fungi is to contribute significantly to low-input technology for tropical agricultural systems.     

Ultraviolet radiation reduces lichen growth rates

The symbiosis of Leguminosae with arbuscular mycorrhizal fungi (AMF) and N₂-fixing nodulating bacteria (NFNB) can occur simultaneously, forming a tripartite symbiosis. In particular, AMF can colonize root nodules, although this interaction is not yet well elucidated, especially with regard to nodule activity and to the influence of external factors, such as biostimulants. In this study, we hypothesized that the application of the flavonoid formononetin, used to stimulate root colonization by native AMF, increases the AMF colonization of soybean (Glycine max) root nodules, especially under low availability of phosphorus (P). To test this hypothesis, we performed a field experiment in randomized blocks in a 4 × 3 factorial design, with 4 treatments of formononetin (0, 0.46, 0.92 and 1.84 g per kg seed) and 3 of P (0, 60 and 120 kg ha^?1) with 5 replicates. Nodules and roots were collected during the R2 stage (full flowering) and evaluated with respect to AMF colonization. Formononetin stimulated mycorrhizal fungi colonization of active nodules, especially when no P was applied, as also observed for AMF root colonization; however, it had no effect with 60 and 120 kg P ha^?1. Thus, the application of formononetin increases surface AMF colonization of active nodules and roots, but its effect disappears with an increase in P and the inactivity of the nodule.     

The differential behavior of arbuscular mycorrhizal fungi in interaction with Astragalus sinicus L. under salt stress

Three arbuscular mycorrhizal (AM) fungi (Glomus mosseae, Glomus claroideum, and Glomus intraradices) were compared for their root colonizing ability and activity in the root of Astragalus sinicus L. under salt-stressed soil conditions. Mycorrhizal formation, activity of fungal succinate dehydrogenase, and alkaline phosphatase, as well as plant biomass, were evaluated after 7 weeks of plant growth. Increasing the concentration of NaCl in soil generally decreased the dry weight of shoots and roots. Inoculation with AM fungi significantly alleviated inhibitory effect of salt stress. G. intraradices was the most efficient AM fungus compared with the other two fungi in terms of root colonization and enzyme activity. Nested PCR revealed that in root system of plants inoculated with a mix of the three AM fungi and grown under salt stress, the majority of mycorrhizal root fragments were colonized by one or two AM fungi, and some roots were colonized by all the three. Compared to inoculation alone, the frequency of G. mosseae in roots increased in the presence of the other two fungal species and highest level of NaCl, suggesting a synergistic interaction between these fungi under salt stress.     

Transpiration of detached leaves from mycorrhizal and nonmycorrhizal cowpea and rose plants given varying abscisic acid, pH, calcium, and phosphorus

 Vesicular-arbuscular mycorrhizal (VAM) colonization can alter transpiration of host leaves, but scientists remain unclear about the mechanisms involved. We tested whether intact root systems were required to observe effects of root colonization by Glomus intraradices on leaf transpiration, or whether some VAM influence resided in leaves even after they were detached from root systems. We measured the transpiration of detached leaves of VAM and nonmycorrhizal plants exposed to different levels of several substances known to influence stomata locally or act in whole-plant regulation of transpiration: abscisic acid, calcium, phosphorus, and hydrogen ions. In rose, some VAM influence on transpiration resided in leaves, even after they had been separated from their root systems. However, removing leaves from their root systems eliminated the VAM influence on stomatal behavior of cowpeas. Accepted: 22 June 1998