Influencia de diferentes medios de cultivo y niveles de pH en el crecimiento in vitro de 6 cepas del género Suillus

Se analizó el comportamiento in vitro de 6 cepas de las especies Suillus luteus, S, granulatus y S. bellinii, asociados a plantaciones de Pinus radiata. Se evaluó las variables crecimiento radial (CR), velocidad media de crecimiento(VMC) y biomasa (B) creciendo en los medios PDA (Papa Dextrosa Agar), MNM (Medio Melin-Norkrans) y BAF (Biotina Aneurina ácido Fólico) y en 5 niveles de pH (4,8, 5,3; 5,8; 6,3; y 6,8). A nivel de cepas se analizó las interacciones presentes entre los factores medio de cultivo y pH. Se observó comportamientos variables de cada cepa para las variables CR y VMC frente a los factores evaluados. Para la variable B, se observan promedios significativos para los medios de cultivo PDA y/o BAF, dependiendo de la cepa estudiada, con valores significativos de pH que variaron para cada cepa. Comparaciones entre valores del CR y de B, muestran alta correlación entre ellos para cepas de S. granulatus, baja para cepas de S. luteus y mediana a nula para cepas de S. bellinii.

Ectomycorrhizal Inoculation Enhances the Salt Tolerance of Quercus mongolica Seedlings

Salt stress harms the growth and development of plants, and the degree of soil salinization in North China is becoming increasingly severe. Ectomycorrhiza (ECM) is a symbiotic system formed by fungi and plants that can improve the growth and salt tolerance of plants. No studies to date have examined the salt tolerance of Quercus mongolica, a typical ectomycorrhizal tree species of temperate forests in the northern hemisphere. Here, we inoculated Q. mongolica with two ectomycorrhizal fungi (Gomphidius viscidus; Suillus luteus) under NaCl stress to characterize the effects of ECM. The results showed that the symbiotic relationship of Q. mongolica with G. viscidus was more stable than that with S. luteus. The cross-sectional area of roots increased after inoculation with the two types of ectomycorrhizal fungi. Compared with the control group, plant height, soluble sugar content, and soluble protein content of leaves were 1.62, 2.41, and 2.04 times higher in the G. viscidus group, respectively. Chlorophyll (Chl) content, stomatal conductance (Gs), and intracellular CO2 concentration (Ci) were significantly higher in Q. mongolica inoculated with ectomycorrhizal fungi than in the control, but differences in the net photosynthetic rate (Pn), transpiration rate (Tr), and photosystem II maximum photochemical efficiency (Fv/Fm) were lower. The relative conductivity of Q. mongolica inoculated with the two ectomycorrhizal fungi was consistently lower than that of non-mycorrhizal seedlings, with the effect of G. viscidus more pronounced than that of S. luteus. The malondialdehyde (MDA) content showed a similar pattern. Peroxidase (POD) and catylase (CAT) levels in mycorrhizal seedlings were generally higher than those of non-mycorrhizal seedlings under normal conditions, and were significantly higher than those of non-mycorrhizal seedlings on the 36th and 48th day after salt treatment, respectively. Overall, the results indicated that the salt tolerance of Q. mongolica seedlings was improved by ectomycorrhizal inoculation.

Promoting the Growth of Pinus sylvestris var. mongolica Seedlings and Improving Rhizosphere Fungal Community Structure through Interaction between Trichoderma and Ectomycorrhizal Fungi

In this study, pot experiments were conducted on the seedlings of Pinus sylvestris var. mongolica to study the influence of Trichoderma (Trichoderma harzianum E15) and Ectomycorrhizal fungi (Suillus luteus N94) on the growth of these seedlings. In particular, the effects of these fungi on the fungal community structure in the rhizosphere soil of the seedlings were investigated. Inoculation with Trichoderma harzianum E15 and Suillus luteus N94 significantly (P < 0.05) promoted the growth of the Pinus sylvestris seedlings. The non-metric multidimensional scaling (NMDS) results indicated a significant difference (P < 0.05) between the fungal community structures in the rhizosphere soil of the annual and biennial seedlings. In the rhizosphere soil of annual seedlings, the main fungi were Ascomycota, Basidiomycota, Zygomycota. Ascomycota, Basidiomycota, Mortierellomycota, and p-unclassified-k-Fungi were the main fungi in the rhizosphere soil of biennial seedlings. The dominant genus in the rhizosphere soil and a key factor promoting the growth of the annual and the biennial seedlings was Trichoderma, Suillus, respectively. Both of them were negatively correlated with the relative abundance of microbial flora in the symbiotic environment. Trichoderma had a significant promoting effect on the conversion of total phosphorus, total nitrogen, ammonium nitrogen, nitrate nitrogen, and the organic matter in the rhizosphere soil of the seedlings, while Suillus significantly promoted the conversion of organic matter and total phosphorus.

Ectomycorrhizal Fungi Associated with Pinus densiflora Seedlings under Flooding Stress

Flooding is an environmental stress for plants that not only limits aeration and nutrient acquisition, but also disturbs underground plant-associated fungal communities. Despite frequent flooding, red pine (Pinus densiflora) seedlings thrive in streamside environments. However, whether the compatible ectomycorrhizal fungi (EMF) of red pine are affected by natural flooding is unclear. As EMF are vital symbionts for the development of many trees and allow them to overcome various environmental stresses, in this study, the EMF species associated with red pine seedlings in a streamside environment in Korea were investigated after flooding. The EMF species in 47 seedlings collected from the streamside site were identified by observing their different morphotypes using internal transcribed spacer sequence analysis, and a total of 10 EMF species were identified. The EMF species diversity was lower than that in samples collected from a nearby forest analyzed as a control. The dominant EMF species of streamside seedlings included Amphinema spp., Rhizopogon luteolus, Suillus luteus, and Thelephora terrestris. This study could serve as a basis for investigating the mechanisms by which advantageous EMF aid plant development under flooding stress.

Proteomic Insight into the Symbiotic Relationship of Pinus massoniana Lamb and Suillus luteus towards Developing Al-Stress Resistance

Global warming significantly impacts forest range areas by increasing soil acidification or aluminum toxicity. Aluminum (Al) toxicity retards plant growth by inhibiting the root development process, hindering water uptake, and limiting the bioavailability of other essential micronutrients. Pinus massoniana (masson pine), globally recognized as a reforestation plant, is resistant to stress conditions including biotic and abiotic stresses. This resistance is linked to the symbiotic relationship with diverse ectomycorrhizal fungal species. In the present study, we investigated the genetic regulators as expressed proteins, conferring a symbiotic relationship between Al-stress resistance and Suillus luteus in masson pine. Multi-treatment trials resulted in the identification of 12 core Al-stress responsive proteins conserved between Al stress conditions with or without S. luteus inoculation. These proteins are involved in chaperonin CPN60-2, protein refolding and ATP-binding, Cu-Zn-superoxide dismutase precursor, oxidation-reduction process, and metal ion binding, phosphoglycerate kinase 1, glycolytic process, and metabolic process. Furthermore, 198 Al responsive proteins were identified specifically under S. luteus-inoculation and are involved in gene regulation, metabolic process, oxidation-reduction process, hydrolase activity, and peptide activity. Chlorophyll a-b binding protein, endoglucanase, putative spermidine synthase, NADH dehydrogenase, and glutathione-S-transferase were found with a significant positive expression under a combined Al and S. luteus treatment, further supported by the up-regulation of their corresponding genes. This study provides a theoretical foundation for exploiting the regulatory role of ectomycorrhizal inoculation and associated genetic changes in resistance against Al stress in masson pine.

Ectomycorrhizal Fungal Assemblages of Nursery-Grown Scots Pine are Influenced by Age of the Seedlings

Scots pine (Pinus sylvestris L.) is the most widely distributed pine species in Europe and is relevant in terms of planted areas and harvest yields. Therefore, each year the demand for planting stock of Scots pine is exceedingly high, and large quantities of seedlings are produced annually throughout Europe to carry out reforestation and afforestation programs. Abundant and diverse ectomycorrhizal (ECM) symbiosis is critical for the success of seedlings once planted in the field. To improve our knowledge of ECM fungi that inhabit bare-root nursery stock of Scots pine and understand factors that influence their diversity, we studied the assemblages of ECM fungi present across 23 bare-root forest nurseries in Poland. Nursery stock samples were characterized by a high level of ECM colonization (nearly 100%), and a total of 29 ECM fungal taxa were found on 1- and 2-year-old seedlings. The diversity of the ECM community depended substantially on the nursery and age of the seedlings, and species richness varied from 3–10 taxa on 1-year-old seedlings and 6–13 taxa on 2-year-old seedlings. The ECM fungal communities that developed on the studied nursery stock were characterized by the prevalence of Ascomycota over Basidiomycota members on 1-year-old seedlings. All ecological indices (diversity, dominance, and evenness) were significantly affected by age of the seedlings, most likely because dominant ECM morphotypes on 1-year-old seedlings (Wilcoxina mikolae) were replaced by other dominant ones (e.g., Suillus luteus, Rhizopogon roseolus, Thelephora terrestris, Hebeloma crustuliniforme), mostly from Basidiomycota, on 2-year-old seedlings. Across all nurseries, negative correlations were identified for diversity metrics and soil N or C, indicating that mineral and organic fertilization contributes to the differences in the ECM fungal communities in nurseries. We discuss the ecological and practical implications of the composition and diversity of ECM fungi occurring on bare-root planting stock of Scots pine.