Coinoculation of bioinoculants improve <Emphasis Type="Italic">Acacia auriculiformis</Emphasis> seedling growth and quality in a tropical Alfisol soil

We conducted a study to find out if arbuscular mycorrhizal (AM) fungi (Acaulospora scrobiculata, Scutellospora calospora) and phosphate solubilizing bacteria (PSB, Paenibacillus polymyxa) inoculation either individually or in combinations can improve Acacia auriculiformis seedling growth, uptake of nutrients and quality in a phosphorus deficient tropical Alfisol. The seedlings were assessed for various growth and nutrient uptake parameters after 60 days of treatment. Inoculation with P. polymyxa stimulated mycorrhizal formation. Seedling height, stem girth, taproot length, number of leaves and leaf area, plant dry matter production, nodulation, and nodular dry weight were significantly higher for seedlings that were either dual inoculated or triple inoculated compared to individual inoculation of AM fungi or PSB, and uninoculated seedlings. Dual and triple application of AM fungi and PSB also significantly improved the nutrient contents of shoots and roots and nutrient uptake efficiencies. The calculated seedling quality indexes of the AM fungi and PSB inoculated seedling were 25–208% higher than uninoculated seedlings. These findings show that A. auriculiformis seedlings when dual inoculated or triple inoculated performed better than seedlings inoculated with the microbes individually and compared with uninoculated control seedlings. We conclude that bioinoculation is important for the production of high-quality A. auriculiformis seedlings in tree nurseries for planting in nutrient deficient soils.     

Growth, nitrogen fixation and mineral acquisition of Alnus sieboldiana after inoculation of Frankia together with Gigaspora margarita and Pseudomonas putida

The role of tetrapartite associations among Frankia, Gigaspora margarita (an arbuscular mycorrhizal fungus), Pseudomonas putida (rhizobacterium), and Alnus sieboldiana in growth, nitrogen fixation, and mineral acquisition of A. sieboldiana was investigated. Seedlings of A. sieboldiana were inoculated with Frankia isolated from root nodules of alder, followed by inoculation of G. margarita and P. putida, and were grown for 5 months in a greenhouse. The seedlings inoculated with Frankia and G. margarita together produced the highest biomass of shoots and root nodules. Nitrogen-fixation activity, measured by acetylene reduction assay, was observed when Frankia was inoculated. The activity, on a per-nodule gram basis, decreased after G. margarita inoculation, but on a per-plant basis there was no significant difference in the activity among inoculation treatments. The mineral content in the seedlings changed after inoculation with Frankia, but not after inoculation with P. putida and/or G. margarita. The results showed a synergistic interaction among Frankia, the mycorrhizal fungus, and the rhizobacterium on the growth of A. sieboldiana.     

Improving drought tolerance of <Emphasis Type="Italic">Casuarina equisetifolia</Emphasis> seedlings by arbuscular mycorrhizas under glasshouse conditions

Effects of mycorrhizal association on improving tolerance of host plant under stress environments have received attentions in recent years. In this paper, six isolates of AMF (arbuscular mycorrhizal fungus) were inoculated to Casuarina equisetifolia seedlings under glasshouse conditions to investigate the effects of AMF on growth and drought tolerance of host plants. All the six isolates which belong to Glomus showed high mycorrhizal colonization (88.5–96.0%) with C. equisetifolia seedlings. Seedlings were subjected to drought stress without watering for 7 days and survival of the seedlings inoculated with Glomus caledonium Gc90068, G. versiforme Gv9004 and G. caledonium Gc90036 increased by 36.6, 23.3 and 16.6%, respectively compared with uninoculated seedlings. Limited influence of AMF on seedling height growth was found, but the effects of AMF on total biomass increment were very significant; the increment ranged from 25.7 to 118.9% compared with uninoculated treatment, and it was noted that AMF exerted more influences on root biomass than shoot biomass. Based on the changes in physiological and biochemical parameters among different treatments caused by drought stress, it was concluded that AM (arbuscular mycorrhizal) associations improve the drought tolerance of C. equisetifolia seedlings by means of some physiological and biochemical responses, such as lowering permeability of plasma membrane and MDA (malondialdehyde) contents, enhancing concentrations of P nutrition, soluble sugar, soluble protein and activities of POX (Peroxidase) of C. equisetifolia seedlings.     

Effect of Frankia inoculation on the growth of Alnus sieboldiana on unsterilized soil

The effect of inoculation with Frankia, a N-fixing actinomycete, on the growth of Alnus sieboldiana seedlings was studied on unsterilized soil from a nursery and an alder stand (forest of Alnus firma). The seedlings of A. sieboldiana were inoculated with Frankia before or after a 2-month culture on sterilized vermiculite, during which they nodulated, and transplanted to unsterilized soil from the nursery and the alder stand. The control seedlings were also cultured on sterilized vermiculite for about 2 months and transplanted to unsterilized soil without Frankia inoculation. The seedling growth, nodulation and N-fixing activity were measured 3, 10 and 16 weeks after the transplantation. Growth and nodule biomass of the seedlings inoculated with Frankia and those grown on the alder soil were better than those without inoculation with Frankia and grown on the nursery soil, respectively. The seedlings inoculated before spontaneous nodulation grew better than those inoculated at the transplantation. Nitrogen-fixing activity measured by acetylene reduction assay at 16 weeks after the transplantation was higher in the seedlings grown on the soil from the nursery than on the soil from the alder stand.     

Effect of the arbuscular mycorrhizae <Emphasis Type="Italic">Glomus fasciculatum</Emphasis> and <Emphasis Type="Italic">G. macrocarpum</Emphasis> on the growth and nutrient content of <Emphasis Type="Italic">Cassia siamea</Emphasis> in a semi-arid Indian wasteland soil

The effect of two arbuscular mycorrhizal (AM) fungi, Glomus fasciculatum and G. macrocarpum on shoot and root dry weights and nutrient content of Cassia siamea in a semi-arid wasteland soil was evaluated. Under nursery conditions, mycorrhizal inoculation improved growth of seedlings. Root and shoot dry weights were higher in mycorrhizal than non-mycorrhizal plants. The concentration of P, K, Cu, Zn and Na was significantly higher in AM inoculated seedlings than in non-inoculated seedlings. Mycorrhization led to decrease in alkalinity of the rhizosphere soil from pH 8.5 to 7.4. Under nursery conditions, the degree of mycorrhizal dependency increased with age of C. siamea seedling. On transplantation to the field, the survival rate of mycorrhizal seedlings (75%–90%) was higher than that of non-mycorrhizal seedlings (40%). Besides better survival rate, AM inoculation improved the growth performance of seedlings in terms of height and stem diameter. Among the two AM fungi used, the efficiency of Glomus macrocarpum was higher than that of G. fasciculatum under both nursery and field conditions.This revised version was published online in May 2005 with corrected page numbers.     

Inoculation of fumigated nursery soil increases nodulation and yield of bare-root red alder (Alnus rubra Bong.)

To determine if inoculation increases nodulation and yield of bare-root red alder (Alnus rubra Bong.), fumigated nursery plots were treated with inoculum and ammonium sulfate (28 kg N ha^–1) in a factorial experiment. Inoculum was alder soil with 100 infective units of Frankia g^–1. Seedlings were evaluated for nodulation at age 10 wk and when lifted, at age 9 mo. Inoculation produced earlier and more extensive nodulation and increased seedling root collar diameter, height, and dry weight. Fertilization decreased seedling height, but did not decrease nodulation. No interaction of fertilization with inoculation was found. Inoculated unfertilized plots had the highest yield of packable seedlings (257 m^–2), and uninoculated fertilized seedlings had the lowest yield (126 m^–2).     

Field survival of containerized red and jack pine seedlings inoculated with mycelial slurries of ectomycorrhizal fungi

Red pine and jack pine seedlings growing in styroblocks were inoculated 8 wk after sowing with mycelium/agar slurries of 3 mycorrhizal fungi (Laccaria bicolor, Scleroderma citrinum, and an unidentified basidiomycete), and one suspected mycorrhizal fungus (Cantharellula umbonata). Seedlings inoculated with L. bicolor developed mycorrhizae earlier and in greater numbers than the other inoculation treatments, with red pine out-performing jack pine in both respects. At 34 wk following sowing, seedlings were outplanted on a cleared xeric site in Baraga Co., in Michigan's Upper Peninsula. Seedlings inoculated with C. umbonata failed to form mycorrhizae and were not outplanted. Inoculation treatments did not affect shoot or root weight at outplanting. Red pine inoculated with L. bicolor averaged 21% and 19% greater survival compared with control seedlings after one and two years in the field, respectively. Other inoculation treatments failed to increase seedling survival for either tree species. Jack pine demonstrated higher overall survival than did red pine for both years in all corresponding inoculation treatments.     

Influence of Microbial Inoculations on Cedrus deodara in Relation to Survival,Growth Promotion and Nutrient Uptake of Seedlings and General Soil Microflora

Abstract

Selected microorganisms (Laccaria laccata, Trichoderma viride, Bacillus subtilis, Pseudomonas corrugata and Azotobacter chroococcum) were used as inoculants to study their influence on seed germination, seedling survival and subsequent growth in Cedrus deodara. The microbial inoculations positively affected these parameters over control, in the following order: Azotobacter chroococcum Pseudomonas corrugata Bacillus subtilis Trichoderma viride Laccaria laccata.The major cause of seedling mortality was found to be (1) wilting of emerged seedlings by Fusarium oxysporum, and (2) cutworm attack. Bacterial inoculations were found to be superior in suppressing the proliferation of Fusariumsp., thereby enhancing seedling survival. One of the bacterial inoculants, B. subtilis, resulted in 76% seed germination, compared to 54% in control. Though inoculation with L. laccataresulted in maximum mycorrhizal infection (up to 72%) compared to48%in control seedlings, it afforded least protection against Fusariumsp., as well as minimal growth promotion. On the other hand, inoculation with T. viridesignificantly controlled wilting of seedlings and improved plant growth. Microbial inoculations were found to influence the soil microflora, nutrient status of the rhizosphere soil and that of different parts of seedlings. Enrichment of carbon was recorded in the rhizosphere soil. Enhanced nitrogen, phosphorus and potassium content of various plant parts indicated a positive influence also on nutrient uptake. The present study has implications for large scale forestry programs.     

Effects of Acidic Precipitation and Ectomycorrhizal Inoculation on Growth,Mineral Nutrition,and Xylem Water Potential of Juvenile Loblolly Pine and White Oak

Abstract

Individual and interactive effects of simulated acidic rainfall and mycorrhizal inoculation on growth and nutrient and water relations of loblolly pine (Pinus taedaL.) and white oak (Quercus albaL.) grown in a loam soil were examined. Seedlings of each species inoculated with basidiospores of the ectomycorrhizal fungus Pisolithus tinctorius(Pers.) Coker and Couch, a known my-cobiont of both loblolly pine and white oak, and uninoculated control seedlings received two simulated rains per week of either pH 3.6, 4.2, or 4.8 for 26 weeks. Higher acidity rainfall reduced the growth but increased mycorrhizal colonization of loblolly pine, while both loblolly pine and white oak exposed to these rains exhibited greater foliar injury. Inoculation with P. tinctoriusincreased growth and reduced foliar injury of both species. Foliar concentrations of P, S, and Cu in loblolly pine and white oak, Ca in loblolly pine, and Fe and Zn in white oak decreased with increasing rain acidity while the Al concentration of both species increased. Higher rainfall acidity also reduced soil pH and Ca and Mg concentrations while increasing soil AI. Foliage of inoculated seedlings of both species had higher N and P concentrations and lower Al concentrations than control seedlings. Following the final rain applications, a drought cycle was simulated by withholding irrigation for two weeks during which seedling xylem pressure potential and soil water potential were measured. One day after cessation of irrigation, xylem pressure potential of loblolly pine that had received pH 3.6 rains was lower than that of other treatments. Thereafter, xylem pressure potential and soil water potential of the inoculated treatment decreased below those of the control treatment in both species. These results suggest that acid deposition is detrimental to juvenile loblolly pine and white oak, but the magnitude of this effect is less than the positive response to ectomycorrhizal inoculation.     

Effect of inoculation with ecto- and arbuscular mycorrhizae and Rhizobium on the growth and nitrogen fixation by black locust,Robinia pseudoacacia

Experiments were made to determine the effect of inoculation withRhizobium, the ectomycorrhizal fungus Hebelomamesophasem and the arbuscular mycorrhizal fungus Glomuscaledonium on the growth and nitrogen fixation of black locust(Robinia pseudoacacia Linn.) seedlings grown invitro and in pot culture. The results showed that both mycorrhizalfungi and Rhizobium stimulated the growth and nitrogenfixing ability of inoculated seedlings. Inoculation with all three microbestogether produced the most beneficial effects on nitrogen fixation, mycorrhizaldevelopment and seedling growth.     

Source of mycorrhizal inoculum influences growth of Faidherbia albida seedlings

Poor land use management and practice inhibit the growth and establishment of tree seedlings in dryland areas.We assessed arbuscular mycorrhizal fungi(AM)status of Faidherbia albida(Del.)A.Chev.trees grown on different land uses.We quantified the growth and nutrient uptake of F.albida seedlings inoculated with AM from different sources.These efforts were based on soil and fine root samples from the rhizosphere soils of F.albida trees.AM root colonization was determined using the gridline intersect method.Spores were extracted by the wet sieving and decanting method and identified to genus level.The seedling experiment had a completely randomized onefactorial design with four treatments and five replications.Faidherbida albida seedlings were grown in a greenhouse.All in situ F.albida trees were colonized by AM fungi.AM root colonization of F.albida trees was significantly higher(P<0.0086)in area exclosures than on lands used for grazing or cultivation.Spore abundance was significantly higher(P<0.0014)in area exclosures followed by cultivated land and grazing land.Glomus was the dominant genus in all land-uses.AM-inoculated F.albida seedlings grew better(P<0.05)than non-inoculated controls.Seedlings inoculated with AM from area exclosure had significantly(P<0.05)higher growth and nutrient uptake than those inoculated with AM from grazing and cultivated land.This emphasizes the importance of the native soil AM potential for better establishment of seedlings to achieve optimum plant growth improvement and assist in rehabilitation of degraded arid lands.     

Peat carrier increases inoculation success with Frankia on red alder (Alnus rubra Bong.) in fumigated nursery beds

Inoculation trials were set up in fumigated nursery beds for red alder (Alnus rubra Bong.) bare-root seedling production. Frankia inoculum was applied either as nodule homogenate or as pure culture (strain ArI5). The plots were laid out in 4 blocks of 8 treatments consisting of: control, nodule suspension, and three levels each of cell suspension and cells applied with a peat carrier. Numbers, height, and percentage nodulation on the seedlings were determined at mid-season. Numbers, size, dry weights, and degree of nodulation were determined at lifting. The peat inoculum treatment produced larger seedlings than the other treatments, both at mid-season and at lifting. The other treatments had little effect on growth. Size differences paralleled differences in degree of nodulation. Differences in percentage of seedlings nodulated were most pronounced at midseason, indicating that inoculation confered primarily an early-season advantage.     

Inoculation of ectomycorrhizal fungi contributes to the survival of tree seedlings in a copper mine tailing

To advance our understanding of the effects of inoculation with ectomycorrhizal fungi (EMF) on seedling colonization in mine wastelands, we conducted a field experiment in a copper tailing. Six-month-old seedlings of Japanese red pine (Pinus densiflora) and oak (Quercus variabilis) separately inoculated with three EMF species (Pisolithus sp., Cenococcum geophilum, Laccaria laccata) were transplanted to the copper tailing. The survival rates of tree seedlings were monitored monthly, and growth (biomass and height), contents of nutrients and heavy metals (K, P, Ca, Mg, Cu, Zn), and mycorrhizal infection rates of seedlings were determined 6 months after planting. Oak seedlings exhibited higher survival rates than pine seedlings after 6 months of growth on the tailing. EMF inoculations of pine seedlings significantly enhanced their survival, growth, and nutrient uptake. In contrast, EMF inoculations of oak seedlings improved growth only in terms of biomass. Additionally, EMF inoculation caused pine seedlings to accumulate more Cu and Zn in roots compared to non-inoculated seedlings, whereas inoculation inhibited the accumulation of heavy metals in shoots. However, similar results were not observed in oak seedlings. Observations of roots indicated that the rates of mycorrhizal infection of both tree species had dramatically declined at harvest time. In conclusion, ectomycorrhizal symbioses can improve the survival and performance of pine seedlings in mine tailings. The present study provided direct evidence of the importance of EMF inoculation of seedlings to the reforestation of mine wastelands.     

Gliocladium virens in an alginate prill ineffective as a biological control of Fusarium root disease in container-grown Douglas-fir

An alginate prill formulation of Gliocladium virens (GL-21) was added as a top-dressings (54 g per m²) or incorporated into medium (1.2 kg per m³) used to grow Douglas-fir seedlings in styrofoam containers. Seedlings in the top-dress treatment were similar to control seedlings; infection and colonization by naturally-occurring Fusarium was unaffected by treatment. Incorporated G. virens reduced seedling growth and increased occurrence and colonization intensity of Fusarium. In a laboratory experiment, inoculating Douglas-fir seedlings with G. virens (10% w/w) prior to inoculation with Fusarium increased survival time when compared to concurrent inoculations of fungi.University of Idaho, Idaho Forest, Wildlife and Range Experiment Station Contribution No. 742.     

Litter of an alien tree, <Emphasis Type="Italic">Casuarina equisetifolia</Emphasis>, inhibits seed germination and initial growth of a native tree on the Ogasawara Islands (subtropical oceanic islands)

We investigated the effects of the accumulation of litter of an alien tree, Casuarina equisetifolia, on the initial establishment of Schima mertensiana, a tree native to the Ogasawara (Bonin) Islands, in the northwestern Pacific Ocean, in field and greenhouse experiments. The field experiment compared emergence of seedlings in forests dominated by C. equisetifolia with those in native forests, with and without litter removal. The greenhouse experiment compared seedling germination and biomass among 12 treatments that included two soil types (collected from the C. equisetifolia and native forests), two litter types (C. equisetifolia and native litter), and three amounts of litter (5, 15, or 45 g/pot). Significantly fewer S. mertensiana seedlings emerged at sites dominated by C. equisetifolia than at sites of native species in the field experiment. Litter removal tended to increase seedling emergence. Fewer and smaller S. mertensiana seedlings germinated with greater litter cover regardless of soil type or litter type in the greenhouse experiment. Our results demonstrate that C. equisetifolia litter suppresses germination and initial growth of S. mertensiana, and that the main cause of the suppression is the greater amount of litter accumulated under C. equisetifolia forest floor rather than factors such as the nature of the soil or litter between native and alien species.     

Comparison of procedures to evaluate the pathogenicity of Ceratocystisfimbriata sensu lato isolates from Eucalyptus in South Africa

Ceratocystis fimbriata sensu lato (s.l.) is an important pathogen of Eucalyptus. Pathogenicity of isolates has typically been evaluated by inoculating seedlings under greenhouse conditions. It is, however, not clear how accurately this reflects pathogenicity under field conditions. In this study, five techniques to potentially screen C. fimbriata isolates for their relative pathogenicity to Eucalyptus were compared. These included: in vitro growth comparisons on artificial media; inoculations on apples; inoculation on Eucalyptus seedlings in a greenhouse; inoculations on Eucalyptus bolts freshly cut from stems of young trees; and field inoculations on young trees. Eight isolates of C. fimbriata s.l. collected from various areas in South Africa were used. There was considerable variation in growth in culture and aggressiveness of the eight isolates. Field inoculations on young trees were best correlated with inoculations of bolts (r = 0.76). Lower correlation coefficients were obtained with seedlings (r = 0.59), apple inoculations (r = 0.56), and in vitro colony growth (r = 0.42). Inoculation of bolts provides a rapid and reliable method to screen isolates of C. fimbriata s.l. for pathogenicity to Eucalyptus.     

Pathogenitätstests mit Bursaphelenchus mucronatus an Kiefern und Fichtensämlingen in Deutschland

Pathogenicity tests with Bursaphelenchus mucronatus on pine and spruce seedlings in Germany Inoculation tests on Pinus sylvestris seedlings with a German, a Siberian and a French isolate of Bursaphelenchus mucronatus and on Picea abies seedlings using only the German isolate have clearly shown, under German field conditions, that the nematode can cause wilt symptoms on the apex of about 60% of the inoculated P. sylvestris plants. The nematodes in all these tests mostly remained near the inoculation site. However, further spread of the nematodes and wilt symptoms occured when a climate chamber, at 25°C, was used or when the French isolate was used outdoors. At 25°C, 10% of the inoculated plants with at least 1000 nematodes per seedling died. Seedlings' apices wilted if more than 50 nematodes per trunk were present, whereas smaller numbers of nematodes could cause partial wilting. Nevertheless, a few pine seedlings did tolerate high nematode-population densities, up to 966 per plant. No further progress of wilt of plants with symptoms was observed the following year. Inoculations of P. abies with a German isolate resulted in population development near the site of inoculation. Spruces largely tolerated nematode infestations without any development of wilt symptoms.     

Selection of efficient VA mycorrhizal fungi for Casuarina equisetifolia—second screening

An investigation was carried out to screen and select efficient vesicular arbuscular mycorrhizal (VAM) fungi for inoculating the forest tree species, Casuarina equisetifolia. The seedlings were inoculated with 10 different VAM fungi, obtained from various sources. Inoculated seedlings generally had greater plant height, stem girth, biomass and P content than uninoculated plants. They also had more mycorrhizal root colonization and spore numbers in root zone soil. C. equisetifolia seedlings responded best (in biomass) to inoculation with Glomus mosseae (Nicolson and Gerdemann) Gerdemann and Trappe, closely followed by Acaulospora laevis Gerdemann and Trappe and G. fasciculatum (Thaxter Sensu Gerdemann) Gerdemann and Trappe; all the three being statistically on par with each other.     

Nursery Sterilization and Inoculation Regimes for Alder Production

Nursery experiments using common alder (Alnus glutinosa) toexamine the interaction of soil sterilization and inoculationwith Frankia are described. Seedling growth was stimulated whensoil was sterilized with methyl bromide or dazomet at currentrecommended rates, but nodulation was reduced. Inoculation withcultured Frankia or a crushed nodule suspension significantlyimproved nodulation, especially if the inoculant was appliedbefore seed germination. However, seedling growth was significantlylarger using cultured Frankia than the crushed nodule inoculant.If the latter is used to inoculate alder seedlings, a reductionin the recommended rate of dazomet application to c. 285 kgha^–1 is likely to encourage nodulation, though seedlinggrowth will be reduced slightly.     

Growth of Nursery-grown Bamboo Inoculated with Arbuscular Mycorrhizal Fungi and Plant Growth Promoting Rhizobacteria in two Tropical Soil Typeswith and without Fertilizer Application

A nursery experiment was conducted to assess the effect of bioinoculants (Glomus aggregatum, Bacillus polymixa, Azospirillum brasilense) on seedling growth promotion of bamboo (Dendrocalamus strictus (Roxb.) Nees.) in two soil types (alfisol, vertisol) with or without fertilizer application. Bamboo seedlings were grown in the presence or absence of bioinoculants either individually or in all combinations for 180 days in field soil under tropical nursery conditions. Shoot, rhizome and root length, dry masses, nutrient concentrations and arbuscular mycorrhizal (AM) colonized root lengths were determined at harvest. Under the experimental condition tested combined inoculation of AM fungi, PSB and A. brasilense resulted in maximum growth response both under fertilized and unfertilized conditions in both soil types. Fertilizer application enhanced the efficiencies of N, P and K uptake, whereas reduced their usage efficiencies. Though soil type did not affect microbial inoculation response, fertilizer application significantly affected plant response to microbial inoculation.