Effect of three species of bacteria on damping-off,root rot development,and ectomycorrhizal colonization of lodgepole pine and white spruce seedlings

Interactions between three species of bacteria (Burkholderia cepacia, Pseudomonas chlororaphis and Pseudomonas fluorescens), an ectomycorrhizal fungus (Paxillus involutus), and three root pathogenic fungi (Fusarium moniliforme, Fusarium oxysporum and Rhizoctonia solani) were studied. Burkholderia cepacia significantly reduced the in vitro mycelial growth of P. involutus, whereas, B. cepacia, Ps. chlororaphis, Ps. fluorescens and P. involutus reduced the mycelial growth of F. moniliforme, F. oxysporum and R. solani. Culture filtrates of B. cepacia, Ps. chlororaphis, Ps. fluorescens and P. involutus reduced conidial germination and increased the formation of chlamydospores of F. moniliforme and F. oxysporum. Burkholderia cepacia also reduced the formation of ectomycorrhizal short roots by P. involutus on lodgepole pine (Pinus contorta) and white spruce (Picea glauca) seedlings 2 months after inoculation. However, no significance difference in mycorrhizal short roots was observed after 4 months. The fungicide Anchor (a mixture of carboxine and thiram) significantly reduced root rot severity and increased the survival of lodgepole pine seedlings grown in a growth mix infested with F. moniliforme, F. oxysporum and R. solani. Control of the diseases of white spruce caused by these pathogens was not as successful. Treatment of seeds with either B. cepacia or P. involutus alone significantly increased the survival of seedlings grown in a mix that was inoculated with F. moniliforme and reduced the root rot severity caused by F. moniliforme and F. oxysporum, but not R. solani. Higher seedling survival and lower root rot severity were observed when conifer seeds were concomitantly inoculated with one of the bacterial species, P. involutus and Anchor.     

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.     

The reproductive success of a Quercus petraea × Q. robur F1-hybrid in back-crossing situations

A 56 year old Quercus petraea × Q. robur F1-hybrid was back-crossed to both parental species. Pollen mixes were applied and paternity assigned to offspring based on microsatellite markers. The studied Q. petraea × Q. robur hybrid proved highly fertile and back-crossed well with both Q. robur and Q. petraea with slight but not significant preference for Q. robur. The results do not support the hypothesis about highly unidirectional gene flow between Q. robur and Q. petraea towards Q. robur as the observed back-crossing ability of the hybrid opens a route for nuclear gene flow from Q. robur to Q. petraea. However, Q. petraea × Q. robur hybrids may be rare in nature and the results do not tell us if the (probably more common) reciprocal hybrid also back-crosses easily to Q. petraea.     

Procedure for establishing sheathing mycorrhizas on tree seedlings

A simple procedure is described where seedlings emerging from surface sterilised seeds (of Betula pendula, B. pubescens, Pinus contorta and Picea sitchensis) were grown axenically in a peat/vermiculite mixture which had been previously inoculated, and incubated, with sheating mycorrhizal fungi (including Hebeloma sacchariolens, Paxillus involutus and Amanita muscaria). Mostly, seedlings were grown in split polystyrene tubes (50 mm tall x 13 mm internal diameter) which were arranged $\text{140}\text{150}$ per tray, the trays being covered with transparent propagator lids so forming a unit that could be sealed and sterilised before use with gamma radiation. With the formation of sheating mycorrhizas after about 7 weeks the sterility of the container was broken to enable seedlings to be hardened off prior to transplanting.Roots of Betula pendula seedlings grown in tubes were smaller, as judged by numbers of ‘fragments’, than those of plants which were not ‘restricted’. ‘Tubing’ increased: (a) the proportion of root fragments with mycorrhizas; and (b) the sizes of individual mycorrhizas. The sizes of the ‘tubing’ effects differed with different isolates of A. muscaria, so suggesting, because of the sensitivity of the system, that environmental and genotypic factors must be carefully controlled.     

The Dependence of Quercus robur and Q. petraea Seeding on Cotyledon Potassium, Magnesium, Calcium and Phosphorus during the First Year of Growth

Acorns collected from a range of Quercus robur L. and Q. petraea(Matt.) Liebl. populations were germinated and grown in an unheatedgreenhouse using a standard commercial compost. Total seedlingdry weight did not increase until the end of the growing season,with Q. robur and Q. petraea seedlings showing mean dry weightincreases of 211 per cent and 414 per cent respectively. Oakseedlings appear to depend upon cotyledon reserves of carbohydrate,P, K and Mg during most of the first year of growth, but relyon external sources of Ca. Seedlings become independent of thecotyledons at the end of the first growing season.     

Ectomycorrhizal inoculation of Douglas-fir transplanted container seedlings with commercially produced inoculum

Commercially produced vegetative inocula of Laccaria laccata and Hebeloma crustuliniforme successfully formed ectomycorrhizae with Douglas-fir transplanted container (plug+1) seedlings. After 4.5 months in containers, 83% and 90%, respectively, of short roots were mycorrhizal. L. laccata- or H. crustuliniforme-inoculated seedlings had significantly more mycorrhizal and total short roots than Pisolithus tinctorius-inoculated (4% mycorrhizal root tips) or uninoculated control seedlings. No significant differences were detected in seedling growth at the end of the container phase.After transplantation and growth in nursery beds for 17 months, mean new short root colonization of all seedlings was 80%. H. crustuliniforme persisted as a dominant mycorrhizal fungus on seedlings initially inoculated with this fungus. L. laccata-inoculated seedlings had 40% of their short roots colonized by L. laccata and another 40% by native fungi Rhizopogon and Thelephora spp. All mycorrhizae of control seedlings and those inoculated with P. tinctorius were formed by fungi native to the nusery beds. A significant fungal treatment effect was detected for shoot height only. Control seedlings were significantly taller than L. laccata-inoculated seedlings after transplanting.This article is part of senior author's thesis in partial fulfillment of the Ph.D. degree in the Department of Forest Science at Oregon State University, Corvallis.     

Effect of fertilization on growth and ectomycorrhizal development of container-grown and bare-root nursery conifer seedlings

The effect of three levels of fertilizer on thegrowth of three species of containerized-grownconifer seedlings (Pinus contorta, Picea glauca, and Picea mariana) and twospecies of bare-root conifer seedlings (Pinus sylvestris and Larix sibirica),and on the colonization of these seedlings bysix species of ectomycorrhizal fungi (Hebeloma longicaudum, Laccaria bicolor,Paxillus involutus, Pisolithustinctorius, Rhizopogon vinicolor andSuillus tomentosus), was studied. Thegrowth of the seedlings in both container-grownand bare-root nurseries increased as the levelsof fertilizer increased. For better seedlinggrowth and environmental quality it may be possible to reduce the level of fertilizers in commercial nurseries upto 33% by using selected mycorrhizal fungi.Ectomycorrhizal colonization in all seedlingswas not affected by fertilizer levels. Hebeloma longicaudum, L. bicolor, P.involutus, and P. tinctorius formedwell-developed ectomycorrhizae, whereasectomycorrhizal development by R.vinicolor and S. tomentosus was poor.Native mycorrhizal fungi colonizednon-inoculated control seedlings; however,their colonization was always lower than withinoculated fungi.     

Interactions between Tricholomopsis rutilans and ectomycorrhizal fungi in paired culture and in association with seedlings of lodgepole pine and Sitka‐spruce

Interactions between two isolates (Avondhu and CBS) of Tricholomopsis rutilans and ectomycorrhizal fungi (Hebeloma crustuliniforme, Laccaria laccata and Paxillus involutus) were examined on agar medium in the presence or absence of woodchips. The CBS isolate showed more competitiveness than the Avondhu isolate, when paired with ectomycorrhizal fungi. There was an inhibition of the growth of mycelium of the ectomycorrhizal fungi ranging from overgrowth to avoidance. The ectomycorrhizal fungi exhibited hyphal abnormalities such as discoloration, excessive vacuolation and curling. Formation of mycorrhizas by H. crustuliniforme in Pinus contorta and Picea abies was unaffected by the presence of the CBS isolate, whereas a reduction occurred in the presence of the Avondhu isolate. In non‐mycorrhizal seedlings of both conifers, the Avondhu isolate reduced root and shoot dry mass and number of root laterals and caused a lower number of short roots in lodgepole pine. The significance of these interactions between T. rutilans and ectomycorrhizal fungi in paired culture and during symbiosis is discussed.     

Effect of endomycorrhizae (VAM) on bamboo seedling growth and biomass productivity

The role of vesicular-arbuscular mycorrhizae (VAM) fungi symbiosis with bamboo seedling was investigated. VAM species Glomus aggregatum, G. fasciculatum and G. mosseae were inoculated individually and in combinations with the bamboo seedlings. The percentage of infection and various growth parameters such as the number of shoots, number of rhizome, internodal distance, leaf length and breadth and total biomass production were studied in myocorrhizal and non-mycorrhizal infected plants. The above findings indicated a significant increase in the growth rate and biomass productivity.     

不同内外生菌根菌接种对米老排苗期的生长效应

9 mycorrhizal fungi, including 4 ectomycorrhizal fungi (ECMF) and 5 arbuscular mycorrhizal fungi (AMF), were used to explore their effect on seedling growth of Mytilaria laosensis. The study shows that M. laosensis is one of the species with both VAM and ECM. Mycorrhizal infection rate of all ECM inoculation methods reached level 3, and that of AMF infection rate was 88%-93%, of which AM91 became the highest. In different periods, different inoculation treatment expressed different effects on seedling growth. At the end of the experiment, the height, ground diameter, underground dry mass and upper ground dry mass of seedling inoculated with PX0801 and 9006 respectively increased by 71%, 45%, 128%, 184%, and 65%, 54%,150%, 208%, which exhibited the best overall effect. Coinoculated seedlings with AM90036, AM3008 and AM91 had advantages over uninoculated ones in all the tested growth indicators, which suggest to significantly promote the growth of M.laosensis. The results obtained provide reference for mycorrhizal fungi application on M. laosensis.     

Changes in water status in seedlings of six species in the Fagaceae after inoculation with Raffaelea quercivora Kubono et Shin-Ito

To clarify the differences in susceptibility of six species in the Fagaceae (Quercus crispula, Quercus serrata, Quercus acutissima, Quercus phillyraeoides, Quercus glauca, and Castanopsis cuspidata var. sieboldii) to Raffaelea quercivora, we inoculated this fungus on seven potted 5-year-old seedlings of each species, observed symptom development, and measured xylem pressure potential (XPP) after inoculation. The first death was observed on the 11th day in Q. crispula and on the 56th day in Q. serrata. The number of dead seedlings of Q. crispula and Q. serrata were five and one, respectively, whereas no mortality was observed in the other four species. The XPP of inoculated seedlings in both Q. crispula and Q. serrata decreased after inoculation. In contrast, the XPP of inoculated seedlings of the other four species remained almost the same as in the control seedlings. These results indicate that R. quercivora is pathogenic to Q. crispula and Q. serrata and that the susceptibility of the six Fagaceae species in our study differed among species.     

Functional response of <Emphasis Type="Italic">Quercus robur</Emphasis> L. to taproot pruning: a 5-year case study

Key message

Quercus robur seedling mass was affected more by planting density than by taproot pruning. Root pruning enhanced stem biomass at the expense of roots in later growth stages. Alteration of biomass allocation due to nursery practices may result in greater susceptibility to injury and death of the seedlings under unfavorable environmental conditions.

Context

Plants adjust their growth and modulate the resource allocation in response to applied treatments and environmental conditions.

Aims

The aim was to examine how taproot pruning in seedlings grown at different densities affected long-term growth of Quercus robur.

Methods

Seedlings, sown as acorns at two planting densities, with or without pruned roots were harvested in the second, fourth, and fifth years of growth. The effect of root pruning on biomass allocation was determined by measuring leaf, stem, and root mass fractions; carbohydrate concentrations in the roots; and C/N ratios. Specific leaf area and root length were also determined to assess morphological adaptations to growth conditions.

Results

Total seedling mass was affected more by planting density than by taproot pruning. After 4 years of growth, root mass fractions were lower and stem mass fractions were greater in seedlings planted at a higher density. Five-year old root-pruned seedlings also had a lower root mass fraction and higher stem mass fractions than unpruned seedlings. Specific root length was not affected by root pruning or planting density.

Conclusion

Decrease of relative root biomass with simultaneous increase of stem biomass may be a long-term consequence of taproot pruning of Q. robur, and the effects may manifest years after the seedling stage.

     

The influence of Desert False Indigo, <Emphasis Type="Italic">Amorpha fruticosa</Emphasis>, and three arbuscular mycorrhizae fungi on the growth of <Emphasis Type="Italic">Populus ussuriensis</Emphasis> seedlings

The influence of Desert False Indigo, Amorpha fruticosa, on the growth of Populus ussuriensis seedlings inoculated with three arbuscular mycorrhizae (AM) fungi (Glomus mosseae, Glomus intraradices, Glomus sinuosa) was studied using the nylon net method. The results showed that all three AM fungi infected P. ussuriensis seedlings and G. intraradices and also G. mosseae infected A. fruticosa. The AM fungi promoted growth of P. ussuriensis and Desert False indigo seedlings. Moreover, under co-cultivation with A. fruticosa, the biomass of P. ussuriensis increased significantly. The concentration of nitrogen in P. ussuriensis grown with A. fruticosa and the concentration of soluble nitrogen in the rhizosphere were also higher than when grown alone. Hypha were found on the two plant seedlings inoculated with G. mosseae and G. intraradices, suggesting that AM fungi may transport nutrients from seedlings of A. fruticosa to the rhizosphere of P. ussuriensis seedlings, which may have promoted the growth of P. ussuriensis. The AM fungi played a critical role on the effect of A. fruticosa on growth of P. ussuriensis.     

Hybridization between Quercus robur and Q. petraea in a mixed oak stand in Denmark

? Hybridization and mating pattern between Quercus robur and Q. petraea was studied in a 5.8 ha mixed forest stand in Jutland, Denmark which comprises in total 135 Quercus robur and 230 Q. petraea trees. Classification of the oak trees into species was performed using canonical discriminant analysis of a range of leaf morphological traits. Adult trees (365) and offspring (582) were genotyped with eight microsatellite markers. Seedlings were sampled in 2003 and acorns were collected in 2004.

? Mating patterns of Q. robur and Q. petraea are expected to be different in the northern range of the distribution area and a larger hybridization rate is expected. It is further expected, that pollination from outside sources will be relatively less in small fragmented forest management systems compared to large scale oak forest. The conclusions should be verified through repeated year to year analysis of the mating pattern.

? Phenological studies revealed that there was no difference in flowering time between species. Data for the adult trees revealed no significant departures from Hardy-Weinberg proportions and there was weak, but significant spatial genetic structure, which supports the idea that the stand is of natural origin. Spatial genetic structure in the first distance class is stronger for Q. petraea. The genetic composition of the offspring was remarkably consistent from year to year. Paternity analysis revealed that, on average, 85% pollination came from fathers within the stand. The direction of the pollen flow varied from year to year. Inter-specific hybridization was high and ranged from 15–17% and from 48–55% for Q. petraea and Q. robur mothers respectively. Paternity analysis revealed that the population was basically outcrossing and only 3.7% of the analysed progeny were the product of selfing. Over the two years of study, approximately 200 trees contributed to the paternity of the next generations.

? The study confirms earlier studies with a greater tendency for Q. robur mothers to produce hybrid seeds than Q. petraea mothers. The rate of hybridization is higher in this Danish stand than in comparable studies elsewhere in Europe. Gene flow from outside sources are relatively low.

     

Ectomycorrhizal fungal communities of Dipterocarpus alatus seedlings introduced by soil inocula from a natural forest and a plantation

We attempted to introduce ectomycorrhizal (ECM) fungi onto seedlings of Dipterocarpus alatus (Dipterocarpaceae) by soil inocula collected from tree stands of D. alatus. Top soils collected from beneath the trees of D. alatus in a dry evergreen forest and a 15-year-old plantation were inoculated onto germinating seeds of D. alatus. After 7 months of seedling cultivation, ECM fungal communities in the seedlings were investigated based on the sequences of ITS rDNA. The ECM fungi detected were divided into 19 phylotypes by molecular analysis. Most of the phylotypes were identified as ECM fungal taxa, i.e., Clavulina, Laccaria, Lactarius, Tomentella, Pyronemataceae, and Tricholomataceae. Accordingly, we can confirm that soil inoculation is a simple method to induce ECM formation with diverse fungi in pot cultured seedlings, which would be useful for introducing diverse ECM fungi to dipterocarp plantations.     

Inoculation with the ectomycorrhizal fungi <Emphasis Type="Italic">Pisolithus arhizus</Emphasis> and <Emphasis Type="Italic">Scleroderma</Emphasis> sp. improves early growth of <Emphasis Type="Italic">Shorea pinanga</Emphasis> nursery seedlings

Trees of the family Dipterocarpaceae are the dominant trees in Southeast Asian tropical forests where they play an important ecological role and are also important commercially. An experiment was conducted to determine the effect of ectomycorrhizal fungi on the growth of dipterocarp species in peat soils. Seedlings of Shorea pinanga were inoculated with spores of two ectomycorrhizal fungi, Pisolithus arhizus and Scleroderma sp. were grown in pots containing sterilized peat soil for 7 months. The percentage of ectomycorrhizal colonization on S. pinanga exceeded 86%. Colonization of S. pinanga roots by ectomycorrhizal fungi resulted in increased shoot height, stem diameter, number of leaves, and shoot fresh and dry weight. Survival rates of S. pinanga were greater for inoculated seedlings than control seedlings. These results suggest that inoculation of ectomycorrhizal fungi can improve the early growth of S. pinanga grown in tropical forests and that this technique will accelerate the rehabilitation of degraded dipterocarp forests.     

Effects of endophytic and ectomycorrhizal basidiomycetes on Quercus virginiana seedling growth and nutrient absorption

Quercus virginiana (Fagaceae), native to the southeastern United States, has recently been introduced to the Yangtze Delta (China) and is often planted in landscaping and in coastal shelter forests. Here, we performed pot inoculation assays to evaluate the effects of two types of basidiomycetous root fungal symbionts, across four orders, on the growth and nutrient absorption of Q. virginiana seedlings. The results indicated that two isolates showed plant growth-promoting activities 2 months after inoculation. In particular, the ectomycorrhizal Sistotrema sp. (Cantharellales) had a pronounced effect on root development and morphology. With regard to nutrient absorption, the total nitrogen content in aboveground tissues was improved in inoculated seedlings, although most differences were not significant. Meanwhile, the total phosphorus contents in seedlings were either positively or negatively affected, probably depending on the symbiont type. Interestingly, the total potassium content in stems and leaves was significantly enhanced in all treatment groups. In addition, indole-3-acetic acid and abscisic acid production were determined in Sistotrema sp. and Atractiella rhizophila (Pucciniomycotina) by high-performance liquid chromatography tandem mass spectrometry. Our findings suggest that both endophytic and ectomycorrhizal fungi have beneficial effects on Q. virginiana growth, and highlight the potential for the development of mycobiome-based seedling raising techniques.     

Inoculation of Acacia holosericea with ectomycorrhizal fungi in a semiarid site in Senegal: growth response and influences on the mycorrhizal soil infectivity after 2 years plantation

The main objectives of this study were to determine the influence of several strains of ectomycorrhizal fungi on the growth of Acacia holosericea in a dry tropical environment and their short-term effect on indicators of ecosystem soil conditions such as biological soil properties (microbial biomass, mycorrhizal soil infectivity). Three fungal strains have been tested: Pisolithus albus IR100, P. albus COI024 and Scleroderma dictyosporum IR109. Ectomycorrhizal inoculation has significantly enhanced the growth (shoot and root biomass) of A. holosericea seedlings after 4 months culture in glasshouse conditions in a disinfected soil. In field conditions, the transplanting shock was less important for the ectomycorrhized trees. After 2 years, the inoculated trees had a better growth than that recorded in the control treatment. Ectomycorrhizal inoculation has significantly stimulated height, leave and wood biomass, root biomass of A. holosericea trees. Moreover, this fungal inoculation has significantly modified the leaf nutrient contents for P, N and phenols. Microbial biomass and mycorrhizal soil infectivity were also larger in the inoculated plots.Ectomycorrhizal inoculation could be of great relevance to improve the reafforestation process of degraded areas with legumes fast growing trees. This biological practice could also ameliorate soil characteristics such as microbial biomass or mycorrhizal soil infectivity.     

Changes in nitrogen level and mycorrhizal structure of Scots pine seedlings inoculated with Thelephora terrestris

? We investigated the influence of Thelephora terrestris, an ectomycorrhizal fungus, on the concentration of nitrogen in needles of Pinus sylvestris L. seedlings and their mycorrhizal structure within four seasons of growth.

? Seedlings were grown in four treatments: (I) inoculated + 0.030 g N, (II) inoculated + 0.045 g N, (III non-inoculated + 0.030 g N, (IV) non-inoculated + 0.045 g N under laboratory conditions and later planted in post-agricultural land.

? For inoculated treatments, statistically significant differences in N concentration of needles were observed for two- and four-year-old seedlings. The number of ectomycorrhizae and concentration of N in needles were negatively correlated. After four seasons of growth the greatest numbers of ectomycorrhizae were present on seedlings from treatment II, and the smallest on seedlings from treatment IV.

? Two-year-old seedlings had been colonized by at least six different fungal taxa, i.e. Cenococcum geophilum, Hebeloma crustuliniforme, Rhizopogon sp., Suillus bovinus, Thelephora terrestris and a non-identified one. Treatments I and II were dominated by the ectomycorrhizae of T. terrestris and their number was significantly greater than in treatments III or TV. On four-year-old seedlings ectomycorrhizae of T. terrestris were dominated in all treatments, while ectomycorrhizae of Hebeloma sp. were not found.

     

Growth response, phosphorus content and root colonization of Polylepis australis Bitt. seedlings inoculated with different soil types

Polylepis forests are one of the most endangered high mountain ecosystems of South America and reforestation with native Polylepis species has been highly recommended. Greenhouse bioassays were set up to determine the influence of three different soils on growth and phosphorous nutrition of Polylepis australis seedlings. Soils were collected from a grassland, a rare mature forest and a forest degraded due to repeated fires. We identified the arbuscular mycorrhizal fungi (AMF) present in the three soils and after 12 months we harvested the seedlings to evaluate root and shoot biomass, plant P content and root colonization by native AMF and dark septate endophytes (DSE). The soil inocula contained 26 AMF morphospecies. Grassland inoculum showed the highest AMF richness, and mature forest showed a different AMF community assembly from grassland and degraded forest inocula. Root biomass and root colonization were highest in seedlings inoculated with mature forest soil, meanwhile shoot biomass and plant P content were similar between all treatments. AMF colonization correlated negatively with DSE and root biomass was negatively correlated with DSE colonization, thus these fungal symbionts could be competing for resources. Our results indicate that AMF inoculum from the mature forest stand has the potential to improve P. australis performance, probably due to the dominance of Glomeraceae and Acaulosporaceae families. However, other soil microorganisms could be together with AMF in the natural inocula, affecting the growth response of P. australis seedlings. Future studies evaluating the effect of these inocula under field conditions should be carried out.