Effect of inoculation method and plant growth medium on endophytic colonization of sorghum by the entomopathogenic fungus <Emphasis Type="Italic">Beauveria bassiana</Emphasis>

A study was conducted to determine the effect of inoculation method and plant growth medium on colonization of sorghum by an endophytic Beauveria bassiana. Colonization of leaves, stems, and roots by B. bassiana was assessed 20-days after application of the fungus. Although B. bassiana established as an endophyte in sorghum leaves, stems, and roots regardless of inoculation method (leaf, seed, or soil inoculation), plant growth medium (sterile soil, non-sterile soil, or vermiculite) apparently influenced colonization rates. Seed inoculation with conidia caused no stem or leaf colonization by the fungus in non-sterile soil but did result in substantial endophytic colonization in vermiculite and sterile soil. Leaf inoculation did not result in root colonization, regardless of plant growth medium. Endophytic colonization was greater in leaves and stems than roots. Endophytic colonization by B. bassiana had no adverse effects on the growth of sorghum plants. Leaf inoculation with a conidial suspension proved to be the best method to introduce B. bassiana into sorghum leaves for plants growing in either sterile or non-sterile soil. Further research should focus on the virulence of endophytic B. bassiana against sorghum stem borers.     

Colonization of barley roots by Fusarium culmorum and influence of Pseudomonas fluorescens on the process

The stages of barley root colonization by Fusarium culmorum were studied in sterile vermiculite by the method of fluorescent antibodies. The influence of the antagonistic bacterium Pseudomonas fluorescens on the process of root colonization by F. culmorum was demonstrated. In vermiculite inoculated with F. culmorum, the fungus density on the roots increased gradually. In the case of joint inoculation of vermiculite with the fungus and the bacterium, the F. culmorum density on the roots changed abruptly. It was shown that the site of primary colonization of the roots by the fungus was mainly the zone of root hairs. When Pseudomonas fluorescens was present on the roots, F. culmorum colonized not only root hairs, but also the elongation zone, during the first two days. Introduction of Pseudomonas fluorescens into vermiculite resulted in lower intensity of barley root rot.     

An ecological view of the formation of VA mycorrhizas

In spite of the major advances in understanding the functioning of symbioses between plants and arbuscular mycorrhizal fungi, details of the ecology of mycorrhizal fungi are not well documented. The benefits of the association are related to the timing and extent of colonization of roots, and fungi differ in their contribution to plant growth and presumably to soil aggregation. Knowledge of the processes that lead to successful colonization of roots by beneficial fungi at appropriate times for the host plants will form the basis of guidelines for soil management to maximize the benefits from the symbiosis. Fungi differ in the manner and extent to which they colonize roots. They also differ in their capacity to form propagules. The importance of hyphae, spores and propagules within living or dead mycorrhizal roots also differs among species and for the same species in different habitats. The relationships between colonization of roots and propagule formation, and between propagule distribution and abundance and subsequent mycorrhiza formation, for different fungi in field environments, are not well understood. Methods for quantifying mycorrhizal fungi are not especially suitable for distinguishing among different fungi within roots. Consequenctly, the dynamics of colonization of roots by different fungi, within and between seasons, have been little studied. Research is required that focuses on the dynamics of fungi within roots as well as on changes in the abundance of propagules of different fungi within soil. Interactions between fungi during the colonization of roots, the colonization of soil by hyphae and sporulation are all poorly understood. Without knowledge of these processes, it will by difficult to predict the likely success of inoculation with introduced fungi. Such knowledge is also required for selecting soil management procedures to enhance growth and survival of key species within the population. The relative tolerance of various fungi to perturbations in their surroundings will provide a basis for identifying those fungi that are likely to persist in specific environments. The processes that influence mycorrhizal fungi in field soils can be identified in controlled studies. However, greater emphasis is required on studying these processes with mixed populations of fungi. The role played by diversity within populations of mycorrhizal fungi is virtually unexplored.     

Studies on fungi in the root region

Summary A study of the colonization of barley root surfaces by sterile dark fungi is described in which the root washing-segment plating technique and direct observation were employed to study the pattern of root colonization. Data are presented showing sterile dark fungi to be rapid colonizers of young roots and to persist on the roots with increasing root age.     

Natural (non-pathogenic) death of the cortex of wheat and barley seminal roots,as evidenced by nuclear staining with acridine orange

Summary Nuclear staining with acridine orange was used to assess cell viability in the cortex of wheat and barley seminal roots from glasshouse and field experiments. Results from this method correlated well with nuclear assessments made in unstained or Feulgen-stained roots, and other evidence is presented to support the validity of the method. The pattern of root cortex death (RCD) was similar in wheat and barley and consistent over a wide range of conditions. Behind the extending root tip and zone of nucleate root hairs, nuclei disappeared progressively from the outer five (of six) cortical cell layers of the root axes, starting in the epidermis. Stainable nuclei remained in the sixth cell layer, next to the endodermis, and in most cell layers around the bases of root laterals and in a small region immediately below the grain. The onset of cell death was apparently related more to the age of a root region than to its distance behind the root tip, and it was not closely correlated with endodermal or stelar development assessed by staining with phloroglucinol/HCl. The rate of RCD was much faster in wheat than barley in both glasshouse and field conditions, and faster in some spring wheat cultivars than in others in the glasshouse. RCD occurred in sterile vermiculite and perlite and was not enhanced by the presence of soil microorganisms; nor was it enhanced in soil by the addition of the non-pathogenic fungal parasitesPhialophora radicicola var..graminicola orMicrodochium bolleyi. RCD is suggested to be endogenously controlled by the amount of photosynthate reaching the cortex. Its implications for growth of soil microorganisms and especially for growth and biological control of root-infecting fungi are discussed.     

Influence of liming,inoculum level and inoculum placement on root colonization of subterranean clover

Arbuscular mycorrhizal (AM) fungi differ in their response to soil pH. Thus, change in soil pH may influence the relative abundance of mycorrhizal fungi inside roots. Root colonization by two AM fungi was studied in relation to addition of lime (CaCO3), quantity of inoculum and inoculum placement. Addition of CaCO3 to an acid soil decreased the colonization of roots by Acaulospora laevis but increased colonization by Glomus invermaium when both fungi were present. In acid soil (pH 4.7), almost all roots were colonized by A. laevis, while G. invermaium was dominant when soil pH was increased to pH 7.3. This occurred regardless of whether the inoculum was banded or mixed throughout the soil. There was no effect of CaCO3 on the relative abundance of fungi inside roots at intermediate rates of CaCO3 application (pH 5.3-6.3) when both fungi were inoculated together. In this experiment, both fungi colonized roots at all levels of CaCO3 when inoculated alone, except for A. laevis at the highest level of CaCO3. We conclude that soil pH affects the competitive ability of these two AM fungi during mycorrhiza formation primarily by affecting hyphae growth in soil and thus the relative abundance of hyphae at the root surface and subsequently inside the root.     

Patterns of arbuscular mycorrhiza down the profile of a heavy textured soil do not reflect associated colonization potential

Colonization of roots by arbuscular mycorrhizal (AM) fungi in several annual crops in two consecutive seasons was compared with, in the second season, the density of fungal propagules in the soil with the use of a bioassay. Root density decreased down the soil profile in both years in all crops, and a high proportion of roots were mycorrhizal throughout the profile. AM colonization decreased down the profile in cotton and lablab in the second season only. The bioassay indicated that most propagules of AM fungi in soils under cotton were located near the surface, with virtually no propagules at 1 m. The absence of propagules at depth indicates a lack of mycelium deep in the soil, and suggests that mycorrhizas are primarily initiated in the surface soil and that the fungi colonize the root system mostly through secondary spread down the profile. The use of AM colonization in the field as an indicator of propagule density and symbiotic function should be qualified by an understanding of the depth in the soil from which roots were extracted.     

Studies on fungi in the root region

Summary Data are given on the fungi isolated from 30-day-old adventitious roots and seedling roots of severalAllium species by means of the root-washing technique. These fungi were typical root-surface forms includingCylindrocarpon, Fusarium, and sterile forms. The amount of root colonization observed, however, was not so great as with other higher plant roots.The nature and role of bulb surface contaminants is discussed in relation to the colonization of young roots, and is compared with the role of seed-coat contaminants in such colonization.     

Citrus replant problem in Iraq I. Possible role of soil fungi and nematodes

The role of soil fungi and nematodes in citrus replant problems was investigated. Several pathogenic and non-pathogenic fungi and the nematodeTylenchulus semipenetrans were isolated in about equal abundance from soils which displayed or did not display replant problems. However, in almost all trees showing evidence of decline, infection by the fungusPhytophthora citrophthora had advanced through the roots until it had reached the tree crown. Fumigation of old citrus soil with methyl bromide greatly increased the growth of sour organe. The application of Nemacure, Benomyl and Ridomil 5G confirmed the important role of pathogenic and non-pathogenic fungi and nematodes in the citrus replant problem. Inoculation of methyl bromide-treated soil with the pathogenic and non-pathogenic fungi separately or in combination further confirmed the previous experiments and also revealed an important role ofP. citrophthora andFusarium solani when replanting citrus in old citrus orchards.     

Effects of soil extracts from repeated plantation woodland of Chinese-fir on microbial activities and soil nitrogen mineralization dynamics

Effects of soil extracts from repeated plantation woodlands of Chinese-fir on soil fungi growth, the activities of microbial communities, and rates of net soil nitrogen mineralization were investigated. Soil extracts from replanted woodlands significantly inhibited soil non-pathogenic fungi growth, reduced soil respiration activities, and net soil nitrogen mineralization rates. However, soil extracts from replanted woodland increased the growth of pathogenic fungi. The combination of soil extracts and pathogenic fungi did not significantly reduce the growth of Chinese-fir seedlings when compared to the soil extracts alone. The combination of soil extracts with pathogenic and non-pathogenic fungi significantly increased the growth of Chinese-fir seedlings when compared to the combination of soil extracts and pathogenic fungi. The results suggest that the allelochemicals from soil extracts, rather than pathogenic fungi, are the key factor regulating the productivity and nitrogen cycling in repeated plantation woodlands.     

Arbuscular mycorrhizal colonization of giant sequoia (Sequoiadendron giganteum) in response to restoration practices

Interactions with soil microbiota determine the success of restoring plants to their native habitats. The goal of our study was to understand the effects of restoration practices on interactions of giant sequoia Sequoiadendron giganteum with arbuscular mycorrhizal (AM) fungi (Glomeromycota). Natural regeneration of Sequoiadendron is threatened by the absence of severe fires that create forest canopy gaps. Generating artificial canopy gaps offers an alternative tool for giant sequoia restoration. We investigated the effect of regeneration practices, including (i) sapling location within gaps, (ii) gap size and (iii) soil substrate, on AM fungal colonization of giant sequoia sapling roots in a native giant sequoia grove of the Sierra Nevada, California. We found that the extent of AM fungal root colonization was positively correlated with sapling height and light availability, which were related to the location of the sapling within the gap and the gap size. While colonization frequency by arbuscules in saplings on ash substrate was higher relative to saplings in mineral soil, the total AM fungal root colonization was similar between the substrates. A negative correlation between root colonization by Glomeromycota and non-AM fungal species indicated antagonistic interactions between different classes of root-associated fungi. Using DNA genotyping, we identified six AM fungal taxa representing genera Glomus and Ambispora present in Sequoiadendron roots. Overall, we found that AM fungal colonization of giant sequoia roots was associated with availability of plant-assimilated carbon to the fungus rather than with the AM fungal supply of mineral nutrients to the roots. We conclude that restoration practices affecting light availability and carbon assimilation alter feedbacks between sapling growth and activity of AM fungi in the roots.     

Impact of soil stockpiling on ericoid mycorrhizal colonization and growth of velvetleaf blueberry (Vaccinium myrtilloides) and Labrador tea (Ledum groenlandicum)

Soil stockpiling is a common practice prior to the reclamation of surface mines. In this study, velvetleaf blueberry and Labrador tea plants were grown from seed in fresh soil, stockpiled soil (1 year), and autoclaved stockpiled soil (1 year) obtained from the Canadian boreal forest. After 7 months of growth, the root colonization intensity with ericoid mycorrhizal (ERM) fungi in both plants growing in stockpiled soil was lower compared to plants growing in the fresh soil. The diversity of ERM fungal species in roots also decreased due to soil stockpiling and Pezoloma ericae was absent from the plants growing in stockpiled soil. Changes in the ERM root colonization in plants growing in stockpiled soil were accompanied by decreases in root and shoot dry weights. Leaf chlorophyll, nitrogen, and phosphorus concentrations of velvetleaf blueberry were higher in fresh soil compared to 1‐year stockpiled soil. Plants grown in the autoclaved stockpiled soil became colonized by the thermotolerant ERM fungus Leohumicola verrucosa and showed higher root and shoot biomass compared to the nonautoclaved stockpiled soil. The results point to the importance of ERM fungi for growth of ericaceous plants, even under favorable environmental conditions and adequate fertilization, and suggest that reduced ERM colonization intensity and ERM fungal diversity in roots likely contributed to the negative effects of soil stockpiling on growth of velvetleaf blueberry and Labrador tea.     

Exotic cheatgrass and loss of soil biota decrease the performance of a native grass

Soil disturbances can alter microbial communities including arbuscular mycorrhizal (AM) fungi, which may in turn, affect plant community structure and the abundance of exotic species. We hypothesized that altered soil microbial populations owing to disturbance would contribute to invasion by cheatgrass (Bromus tectorum), an exotic annual grass, at the expense of the native perennial grass, squirreltail (Elymus elymoides). Using a greenhouse experiment, we compared the responses of conspecific and heterospecific pairs of cheatgrass and squirreltail inoculated with soil (including live AM spores and other organisms) collected from fuel treatments with high, intermediate and no disturbance (pile burns, mastication, and intact woodlands) and a sterile control. Cheatgrass growth was unaffected by type of soil inoculum, whereas squirreltail growth, reproduction and nutrient uptake were higher in plants inoculated with soil from mastication and undisturbed treatments compared to pile burns and sterile controls. Squirreltail shoot biomass was positively correlated with AM colonization when inoculated with mastication and undisturbed soils, but not when inoculated with pile burn soils. In contrast, cheatgrass shoot biomass was negatively correlated with AM colonization, but this effect was less pronounced with pile burn inoculum. Cheatgrass had higher foliar N and P when grown with squirreltail compared to a conspecific, while squirreltail had lower foliar P, AM colonization and flower production when grown with cheatgrass. These results indicate that changes in AM communities resulting from high disturbance may favor exotic plant species that do not depend on mycorrhizal fungi, over native species that depend on particular taxa of AM fungi for growth and reproduction.     

不同基质中番茄菌根苗的培育及其抗南方根结线虫的效果

根结线虫病是番茄主要的土传病害,特别是保护地长期种植会加重病害的发生。接种丛枝菌根真菌（arbuscular mycorrhizal fungi,AMF）能提高植物抗根结线虫的能力,促进植株生长发育,减轻病害。本试验旨在评价常用基质配合施用一种生物有机肥对番茄苗生长和菌根定殖的影响,以获得菌根发育良好且生长健壮的菌根苗,并通过盆栽实验验证菌根苗移栽于大棚连作土壤中对南方根结线虫的防治效果。结果表明,栽培于商业基质的番茄苗的株高、茎粗、地上部及根系干重和壮苗指数均显著高于草炭蛭石混合基质的番茄苗;草炭蛭石混合基质中施加生物有机肥能促进番茄苗的生长,但是显著抑制根内根孢囊霉Rhizophagus intraradices的侵染。草炭蛭石混合基质在培养过程中施加适量Hoagland营养液使番茄苗生长发育较好且根内根孢囊霉R. intraradices侵染率达到71%;商业基质培养的番茄苗壮苗指数更高,但侵染率较低（44%）。在草炭蛭石混合基质中接种变形球囊霉Glomus versiforme和根内根孢囊霉R. intraradices及其混合菌种培育的番茄苗对南方根结线虫Meloidogyne incognita的侵染和繁殖均表现出一定的抑制作用。按照单位重量根系进行统计,混合菌种显著降低了根结线虫的根结和卵块数量（分别降低了22.8%和23.5%）。本研究结果表明番茄菌根苗在草炭蛭石混合基质中AMF发育状况优于常用商业基质,在菌根应用中可优先选择;相对于单一接种AMF,混合接种的菌根化苗对南方根结线虫病害的抑制作用更好。     

Early events in the life of apple roots: variation in root growth rate is linked to mycorrhizal and nonmycorrhizal fungal colonization

Early events of mycorrhizal and nonmycorrhizal fungal colonization in newly-emerging roots of mature apple (Malus domestica Borkh) trees were characterized to determine the relationship of these events to fine root growth rate and development. New roots were traced on root windows to measure growth and then collected and stained to quantify microscopically the presence of mycorrhizal and nonmycorrhizal fungal structures. Most new roots were colonized by either mycorrhizal or nonmycorrhizal fungi but none less 25 days old were ever internally colonized by both. Compared to nonmycorrhizal colonization, mycorrhizal colonization was associated with faster growing roots and roots that grew for a longer duration, leading to longer roots. While either type of fungi was observed in roots as soon as 3 days after root emergence, intraradical colonization by mycorrhizal fungi was generally faster (peaking at 7 to 15 days) than that by nonmycorrhizal fungi and often occurred more frequently in younger roots. Only 15 to 35% of the roots had no fungal colonization by 30 days after emergence. This study provides the first detailed examination of the early daily events of mycorrhizal and nonmycorrhizal fungal colonization in newly emerging roots under field conditions. We observed marked discrimination of roots between mycorrhizal and nonmycorrhizal fungi and provide evidence that mycorrhizal fungi may select for faster growing roots and possibly influence the duration of root growth by non-nutritional means.     

The survival and development of inoculant ectomycorrhizal fungi on roots of outplanted Eucalyptus globulus Labill

The survival and development of two inoculant ectomycorrhizal fungi (Hebeloma westraliense Bough. Tom. and Mal. and Setchelliogaster sp. nov.) on roots of outplanted Eucalyptus globulus Labill. was examined at two expasture field sites in the south-west of Western Australia. Site 1 was a gravelly yellow duplex soil, and Site 2 was a yellow sandy earth. Plants were grown in steamed or unsteamed soil, in root bags designed as field containers for young growing trees. Three, 6 and 12 months after outplanting, plants were removed from these bags and assessed for dry weights of shoots and ectomycorrhizal colonization of roots.The inoculant ectomycorrhizal fungi (identified on the basis of the colour and morphology of their mycorrhizas) survived on roots of E. globulus for at least 12 months after outplanting at both field sites. At Site 1, however, colonization of new fine roots by the inoculant fungi was low (less than 20% of fine root length). Inoculation had no effect on the growth of E. globulus at this site. In contrast, at Site 2 the inoculant ectomycorrhizal fungi colonized up to 30–50% of new fine root length during the first 6 months after outplanting. There was a corresponding growth response to ectomycorrhizal inoculation at this site, with a close relationship (r²=0.82^**) between plant growth at 12 months and root colonization at 3 months. Plant growth at 12 months was related less closely with root colonization at 6 or 12 months. Root colonization by resident ectomycorrhizal fungi increased with time at both field sites. At Site 2, this increase appeared to be at the expense of colonization by the inoculant fungi, which was reduced to less than 10% of fine root length at 12 months. Steaming the soil had little effect on colonization by the inoculant ectomycorrhizal fungi at either field site, but decreased colonization by the resident ectomycorrhizal fungi.     

Effect of tillage and farming system upon VAM fungus populations and mycorrhizas and nutrient uptake of maize

Low-input agricultural systems that do not rely on fertilizers may be more dependent on vesicular-arbuscular mycorrhizal [VAM] fungi than conventionally managed systems. We studied populations of spores of VAM fungi, mycorrhiza formation and nutrient utilization of maize (Zea mays L.) grown in moldboard plowed, chisel-disked or no-tilled soil under conventional and low-input agricultural systems. Maize shoots and roots were collected at four growth stages. Soils under low-input management had higher VAM fungus spore populations than soils under conventional management. Spore populations and colonization of maize roots by VAM fungi were higher in no-tilled than in moldboard plowed or chisel-disked soil. The inoculum potential of soil collected in the autumn was greater for no-till and chisel-disked soils than for moldboard plowed soils and greater for low-input than conventionally farmed soil. The effects of tillage and farming system on N uptake and utilization varied with growth stage of the maize plants. The effect of farming system on P use efficiency was significant at the vegetative stages only, with higher efficiencies in plants under low-input management. The effect of tillage was consistent through all growth stages, with higher P use efficiencies in plants under moldboard plow and chisel-disk than under no-till. Plants grown in no-tilled soils had the highest shoot P concentrations throughout the experiment. This benefit of enhanced VAM fungus colonization, particularly in the low-input system in the absence of effective weed control and with likely lower soil temperatures, did not translate into enhanced growth and yield.     

The foliar endophytic fungal community composition in Cirsium arvense is affected by mycorrhizal colonization and soil nutrient content

Foliar fungal endophytes are ubiquitous, but understudied symbionts of most plant species; relatively little is known about the factors affecting their occurrence, diversity and abundance. We tested the effects of soil nutrient content and arbuscular mycorrhizal (AM) colonization on the occurrence of foliar endophytic fungi in Cirsium arvense in two field studies. In the first study, we assessed relationships between soil moisture, organic matter, carbon and nitrogen content and plant water, nitrogen and carbon content and AM colonization and the occurrence of foliar endophytic fungal species. In the second study, we manipulated soil nutrient content and AM colonization of potted seedlings and identified differences in endophytic fungal species composition of the leaves and stems. The results reveal that endophytes can occur either more or less frequently, depending on soil nutrient and plant water content and AM colonization. We propose that these patterns were the result of differences in fungal growth responses to nutrient availability in the leaves, which can be affected by resources obtained from the soil or symbiotic fungi in the roots.     

Production of Vesicular-Arbuscular Mycorrhizal Fungus Inoculum in Aeroponic Culture

Bahia grass (Paspalum notatum) and industrial sweet potato (Ipomoea batatas) colonized by Glomus deserticola, G. etunicatum, and G. intraradices were grown in aeroponic cultures. After 12 to 14 weeks, all roots were colonized by the inoculated vesicular-arbuscular mycorrhizal fungi. Abundant vesicles and arbuscules formed in the roots, and profuse sporulation was detected intra-and extraradically. Within each fungal species, industrial sweet potato contained significantly more roots and spores per plant than bahia grass did, although the percent root colonization was similar for both hosts. Mean percent root colonization and sporulation per centimeter of colonized root generally increased with time, although with some treatments colonization declined by week 14. Spore production ranged from 4 spores per cm of colonized root for G. etunicatum to 51 spores per cm for G. intraradices. Infectivity trials with root inocula resulted in a mean of 38, 45, and 28% of bahia grass roots colonized by G. deserticola, G. etunicatum, and G. intraradices, respectively. The germination rate of G. etunicatum spores produced in soil was significantly higher than that produced in aeroponic cultures (64% versus 46%) after a 2-week incubation at 28°C. However, infectivity studies comparing G. etunicatum spores from soil and aeroponic culture indicated no biological differences between the spore sources. Aeroponically produced G. deserticola and G. etunicatum inocula retained their infectivity after cold storage (4°C) in either sterile water or moist vermiculite for at least 4 and 9 months, respectively.     

Colonization potential of in vitro-produced arbuscular mycorrhizal fungus spores compared with a root-segment inoculum from open pot culture

 A reliable inoculum, free from other microorganisms, to produce arbuscular mycorhizal (AM) plants is of the greatest importance when studying the interaction between AM plants and soil microorganisms. We investigated the colonization of leeks from monoxenic in vitro-produced Glomus intraradices spores. The isolated spores were produced using a two-compartment in vitro growth system previously described. A spore suspension was used as inoculum and was compared to the inoculum potential of endomycorrhizal root segments of pot-grown leek (Allium porrum L.) plants. The leeks were grown in a controlled environment and two types of sterilized growth media were tested: calcined montmorillonite clay and a soil mix. Root colonization progressed faster in the soil mix than in the clay. However, in this medium, after an initial delay, root colonization from in vitro-produced spores was essentially the same as that observed with the root-segment inoculum, reaching 44% and 58% respectively, after 16 weeks. Leek roots colonized by the monoxenically-produced spores harbored only the studied AMF fungi while the roots colonized from the root segments were substantially contaminated by other fungi. Accepted: 25 December 1998