Pinus halepensis,Pinus pinaster,Pinus pinea and Pinus sylvestris Essential Oils Chemotypes and Monoterpene Hydrocarbon Enantiomers,before and after Inoculation with the Pinewood Nematode Bursaphelenchus xylophilus

Pinewood nematode (PWN), Bursaphelenchus xylophilus, is the causal agent of pine wilt disease, a serious threat to global forest populations of conifers, especially Pinus spp. A time‐course study of the essential oils (EOs) of 2‐year‐old Pinus halepensis, Pinus pinaster, Pinus pinea and Pinus sylvestris following inoculation with the PWN was performed. The constitutive and nematode inoculation induced EOs components were analyzed at both the wounding or inoculation areas and at the whole plant level. The enantiomeric ratio of optically active main EOs components was also evaluated. External symptoms of infection were observed only in P. pinaster and P. sylvestris 21 and 15 days after inoculation, respectively. The EO composition analysis of uninoculated and unwounded plants revealed the occurrence of chemotypes for P. pinaster, P. halepensis and P. sylvestris, whereas P. pinea showed a homogenous EO composition. When whole plants were evaluated for EO and monoterpene hydrocarbon enantiomeric chemical composition, no relevant qualitative and quantitative differences were found. Instead, EO analysis of inoculated and uninoculated wounded areas revealed an increase of sesquiterpenes and diterpenic compounds, especially in P. pinea and P. halepensis, comparatively to healthy whole plants EOs.     

Screening for PGPR to improve growth of Cistus ladanifer seedlings for reforestation of degraded mediterranean ecosystems

A screening for PGPRs was carried out in the rhizosphere of wild populations of Cistus ladanifer. Two hundred and seventy bacteria were isolated, purified and grouped by morphological criteria. Fifty percent of the isolates were selected and tested for aminocyclopropanecarboxylic acid (ACC) degradation, auxin and siderophore production and phosphate solubilisation. Fifty-eight percent of the isolates showed at least one of the evaluated activities, with phosphate solubilisation and siderophore production being the most abundant traits. After PCR-RAPDs (Randomly amplified polymorphic DNA) analysis, 11 groups appeared with 85% similiarity, revealing the low diversity in the system. One strain of each group was tested in a biological assay, and those that enhanced Cistus growth were identified by 16S rDNA sequencing.Although seven of the 11 assayed strains were phosphate solubilisers and able to produce siderophores, only one was really effective in increasing all biometric parameters in Cistus ladanifer seedlings, the lack of effect of the other six probably being due to the rich substrate used. This suggests that other mechanisms apart from nutrient mobilisation might be involved in growth promotion by this strain. However, the low diversity together with the high redundancy detected by PCR-RAPDs and the predominance of strains able to mobilise nutrients in the rhizosphere of Cistus reveals that the plant selects for bacteria that can help to supply scarce nutrients. This type of plant growth promoting rhizobacteria (PGPR) strains should be succesful in reforestation practices.     

Beneficial native bacteria improve survival and mycorrhization of desert truffle mycorrhizal plants in nursery conditions

Sixty-four native bacterial colonies were isolated from mycorrhizal roots of Helianthemum almeriense colonized by Terfezia claveryi, mycorrhizosphere soil, and peridium of T. claveryi to evaluate their effect on mycorrhizal plant production. Based on the phylogenetic analysis of the 16S rDNA partial sequence, 45 different strains from 17 genera were gathered. The largest genera were Pseudomonas (40.8 % of the isolated strains), Bacillus (12.2 % of isolated strains), and Varivorax (8.2 % of isolated strains). All the bacteria were characterized phenotypically and by their plant growth-promoting rhizobacteria (PGPR) traits (auxin and siderophore production, phosphate solubilization, and ACC deaminase activity). Only bacterial combinations with several PGPR traits or Pseudomonas sp. strain 5, which presents three different PGPR traits, had a positive effect on plant survival and growth. Particularly relevant were the bacterial treatments involving auxin release, which significantly increased the root-shoot ratio and mycorrhizal colonization. Moreover, Pseudomonas mandelii strain 29 was able to considerably increase mycorrhizal colonization but not plant growth, and could be considered as mycorrhiza-helper bacteria. Therefore, the mycorrhizal roots, mycorrhizosphere soil, and peridium of desert truffles are environments enriched in bacteria which may be used to increase the survival and mycorrhization in the desert truffle plant production system at a semi-industrial scale.     

Characterization of Plant Growth–Promoting Traits of Bacteria Isolated from Larval Guts of Diamondback Moth <Emphasis Type="Italic">Plutella xylostella</Emphasis> (Lepidoptera: Plutellidae)

Eight bacterial isolates from the larval guts of Diamondback moths (Plutella xylostella) were tested for their plant growth–promoting (PGP) traits and effects on early plant growth. All of the strains tested positive for nitrogen fixation and indole 3-acetic acid (IAA) and salicylic acid production but negative for hydrogen cyanide and pectinase production. In addition, five of the isolates exhibited significant levels of tricalcium phosphate and zinc oxide solubilization; six isolates were able to oxidize sulfur in growth media; and four isolates tested positive for chitinase and β-1,3-glucanase activities. Based on their IAA production, six strains including four that were 1-aminocyclopropane-1-carboxylate (ACC) deaminase positive and two that were ACC deaminase negative were tested for PGP activity on the early growth of canola and tomato seeds under gnotobiotic conditions. Acinetobacter sp. PSGB04 significantly increased root length (41%), seedling vigor, and dry biomass (30%) of the canola test plants, whereas Pseudomonas sp. PRGB06 inhibited the mycelial growth of Botrytis cinerea, Colletotrichum coccodes, C. gleospoiroides, Rhizoctonia solani, and Sclerotia sclerotiorum under in vitro conditions. A significant increase, greater than that of the control, was also noted for growth parameters of the tomato test plants when the seeds were treated with PRGB06. Therefore, the results of the present study suggest that bacteria associated with insect larval guts possess PGP traits and positively influence plant growth. Therefore, insect gut bacteria as effective PGP agents represent an unexplored niche and may broaden the spectrum of beneficial bacteria available for crop production.     

Effect of the Mycorrhizosphere on the Genotypic and Metabolic Diversity of the Bacterial Communities Involved in Mineral Weathering in a Forest Soil

To date, several bacterial species have been described as mineral-weathering agents which improve plant nutrition and growth. However, the possible relationships between mineral-weathering potential, taxonomic identity, and metabolic ability have not been investigated thus far. In this study, we characterized a collection of 61 bacterial strains isolated from Scleroderma citrinum mycorrhizae, the mycorrhizosphere, and the adjacent bulk soil in an oak forest. The ability of bacteria to weather biotite was assessed with a new microplate bioassay that measures the pH and the quantity of iron released from this mineral. We showed that weathering bacteria occurred more frequently in the vicinity of S. citrinum than in the bulk soil. Moreover, the weathering efficacy of the mycorrhizosphere bacterial isolates was significantly greater than that of the bulk soil isolates. All the bacterial isolates were identified by partial 16S rRNA gene sequence analysis as members of the genera Burkholderia, Collimonas, Pseudomonas, and Sphingomonas, and their carbon metabolism was characterized by the BIOLOG method. The most efficient isolates belonged to the genera Burkholderia and Collimonas. Multivariate analysis resulted in identification of three metabolic groups, one of which contained mainly bacterial isolates associated with S. citrinum and exhibiting high mineral-weathering potential. Therefore, our results support the hypothesis that by its carbon metabolism this fungus selects in the bulk soil reservoir a bacterial community with high weathering potential, and they also address the question of functional complementation between mycorrhizal fungi and bacteria in the ectomycorrhizal complex for the promotion of tree nutrition.     

三株固氮类芽孢杆菌的特点及其对中国青菜的产量和土壤酶活性的影响

【目的】本研究分析三株固氮菌PGPR性状特征及其对中国青菜产量和土壤酶活的影响。【方法】氮(N)-修复(固氮)细菌被认为是一种能够促进植物生长和增产的施氮方式。在本研究中,我们用无氮培养基分离出了30株根际固氮细菌:11株来自小麦根际,16株来自中国青菜根际和3株来自莲花根际。基于16S r DNA序列分析,对小麦、中国青菜和莲花等植物根际中属于类芽孢杆菌属的主要固氮细菌进行研究。【结果】本研究从这30株固氮菌中筛选出三株属于类芽孢杆菌属(Paenibacillus)的细菌,分别命名为P-4、W-7和L-3,它们的固氮酶活性不但高于对照组(圆褐固氮菌),而且可以有效抑制两种或三种植物病原菌的生长,即核盘菌(Sclerotinia sclerotiorum)、玉蜀黍赤霉(Gibberella zeae)和棉花黄萎病菌(Verticillium dahliae)。菌株W-7还具有溶解难溶磷的能力,中国青菜在接种菌株W-7和L-3后,其鲜重显著增加,同时改变了田间土壤蔗糖酶、磷酸酶和过氧化氢酶的活性;而接种了菌株P-4对植物的生长和酶活性没有显著的影响。【结论】土壤蔗糖酶、磷酸酶和过氧化氢酶活性与中国青菜的生物量呈正相关。同时,菌株W-7和L-3具有促进植物产量和提高土壤质量的良好潜力。     

Diversity of endophytic bacteria in ginseng and their potential for plant growth promotion

Endophytic bacteria have been found in virtually every plant studied, where they colonize the internal tissues of their host plant and can form a range of different beneficial relationships. The diversity of bacterial endophytes associated with ginseng plants of varying age levels in Korea was investigated. Fifty-one colonies were isolated from the interior of ginseng stems. Although a mixed composition of endophyte communities was recovered from ginseng based on the results of 16S rDNA analysis, bacteria of the genus Bacillus and Staphylococcus dominated in 1-year-old and 4-year-old plants, respectively. Phylogenetic analysis revealed four clusters: Firmicutes, Actinobacteria, α-Proteobacteria, and γ-Proteobacteria, with Firmicutes being predominant. To evaluate the plant growth promoting activities, 18 representative isolates were selected. Amplification of nifH gene confirmed the presence of diazotrophy in only two isolates. Half of the isolates solubilized mineral phosphate. Except four, all the other endophytic isolates produced significant amounts of indole acetic acid in nutrient broth. Iron sequestering siderophore production was detected in seven isolates. Isolates E-I-3 (Bacillus megaterium), E-I-4 (Micrococcus luteus), E-I-8 (B. cereus), and E-I-20 (Lysinibacillus fusiformis) were positive for most of the plant growth promoting traits, indicating their role in growth promotion of ginseng.     

Characterization of Bacteria Associated with Pinewood Nematode Bursaphelenchus xylophilus

Pine wilt disease (PWD) is a complex disease integrating three major agents: the pathogenic agent, the pinewood nematode Bursaphelenchus xylophilus; the insect-vector Monochamus spp.; and the host pine tree, Pinus sp. Since the early 80''s, the notion that another pathogenic agent, namely bacteria, may play a role in PWD has been gaining traction, however the role of bacteria in PWD is still unknown. The present work supports the possibility that some B. xylophilus-associated bacteria may play a significant role in the development of this disease. This is inferred as a consequence of: (i) the phenotypic characterization of a collection of 35 isolates of B. xylophilus-associated bacteria, in different tests broadly used to test plant pathogenic and plant growth promoting bacteria, and (ii) greenhouse experiments that infer the pathogenicity of these bacteria in maritime pine, Pinus pinaster. The results illustrate the presence of a heterogeneous microbial community associated with B. xylophilus and the traits exhibited by at least, some of these bacteria, appear to be related to PWD symptoms. The inoculation of four specific B. xylophilus-associated bacteria isolates in P. pinaster seedlings resulted in the development of some PWD symptoms suggesting that these bacteria likely play an active role with B. xylophilus in PWD.     

Isolation and Characterization of Beneficial Bacteria Associated with Citrus Roots in Florida

Cultivable diversity of bacteria associated with citrus was investigated as part of a larger study to understand the roles of beneficial bacteria and utilize them to increase the productive capacity and sustainability of agro-ecosystems. Citrus roots from Huanglongbing (HLB) diseased symptomatic and asymptomatic citrus were used in this study. A total of 227 and 125 morphologically distinct colonies were isolated and characterized from HLB asymptomatic and symptomatic trees, respectively. We observed that the frequency of bacterial isolates possessing various plant beneficial properties was significantly higher in the asymptomatic samples. A total of 39 bacterial isolates showing a minimum of five beneficial traits related to mineral nutrition [phosphate (P) solubilization, siderophore production, nitrogen (N) fixation], development [indole acetic acid (IAA) synthesis], health [production of antibiotic and lytic enzymes (chitinase)], induction of systemic resistance [salicylic acid (SA) production], stress relief [production of 1-amino-cyclopropane-1-carboxylate deaminase] and production of quorum sensing [N-acyl homoserine lactones] signals were characterized. A bioassay using ethidium monoazide (EMA)-qPCR was developed to select bacteria antagonistic to Candidatus Liberibacter asiaticus. Using the modified EMA-qPCR assay, we found six bacterial isolates showing maximum similarity to Paenibacillus validus, Lysinibacillus fusiformis, Bacillus licheniformis, Pseudomonas putida, Microbacterium oleivorans, and Serratia plymutica could significantly reduce the population of viable Ca. L. asiaticus in HLB symptomatic leaf samples. In conclusion, we have isolated and characterized multiple beneficial bacterial strains from citrus roots which have the potential to enhance plant growth and suppress diseases.     

Potential role of rhizobia to enhance chickpea-growth and yield in low fertility-soils of Tunisia

Plant growth-promoting rhizobacteria are bacteria that improve plant growth and reduce plant pathogen damages. In this study, 100 nodule bacteria were isolated from chickpea, screened for their plant growth-promoting (PGP) traits and then characterised by PCR-RFLP of 16 S rDNA. Results showed that most of the slow-growing isolates fixed nitrogen but those exhibiting fast-growth did not. Fourteen isolates solubilized inorganic phosphorus, 16 strains produced siderophores, and 17 strains produced indole acetic acid. Co-culture experiments identified three strains having an inhibitory effect against Fusarium oxysporum, the primary pathogenic fungus for chickpea in Tunisia. Rhizobia with PGP traits were assigned to Mesorhizobium ciceri, Mesorhizobium mediterraneum, Sinorhizobium meliloti and Agrobacterium tumefaciens. We noted that PGP activities were differentially distributed between M. ciceri and M. mediterraneum. The region of Mateur in northern Tunisia, with clay–silty soil, was the origin of 53% of PGP isolates. Interestingly, we found that S. meliloti and A. tumefaciens strains did not behave as parasitic nodule-bacteria but as PGP rhizobacteria useful for chickpea nutrition and health. In fact, S. meliloti strains could solubilize phosphorus, produce siderophore and auxin. The A. tumefaciens strains could perform the previous PGP traits and inhibit pathogen growth also. Finally, one candidate strain of M. ciceri (LL10)—selected for its highest symbiotic nitrogen fixation and phosphorus solubilization—was used for field experiment. The LL10 inoculation increased grain yield more than three-fold. These finding showed the potential role of rhizobia to be used as biofertilizers and biopesticides, representing low-cost and environment-friendly inputs for sustainable agriculture.

     

Biocontrol Potential of Soybean Bacterial Endophytes Against Charcoal Rot Fungus, <Emphasis Type="Italic">Rhizoctonia bataticola</Emphasis>

A total of 137 bacterial isolates from surface sterilized root, stem, and nodule tissues of soybean were screened for their antifungal activity against major phytopathogens like Rhizoctonia bataticola, Macrophomina phaseolina, Fusarium udam, and Sclerotium rolfsii. Nine bacterial endophytes suppressed the pathogens under in vitro plate assay. These were characterized biochemically and identified at the genus level based on their partial sequence analysis of 16S rDNA. Eight of the isolates belonged to Bacillus and one to Paenibacillus. The phylogenetic relationship among the selected isolates was studied and phylogenetic trees were generated. The selected isolates were screened for biocontrol traits like production of hydrogen cyanide (HCN), siderophore, hydrolytic enzymes, antibiotics, and plant growth promoting traits like indole 3-acetic acid production, phosphate solubilization, and nitrogen fixation. A modified assessment scheme was used to select the most efficient biocontrol isolates Paenibacillus sp. HKA-15 (HKA-15) and Bacillus sp. HKA-121 (HKA-121) as potential candidates for charcoal rot biocontrol as well as soybean plant growth promotion.     

Effect of inoculation with putative plant growth-promoting rhizobacteria isolated from Pinus spp. on Pinus pinea growth, mycorrhization and rhizosphere microbial communities

Aims: In this study, 10 putative plant growth-promoting rhizobacteria (PGPR) were assayed for their ability to improve Pinus pinea growth and mycorrhization. Methods and Results: After an inoculation assay, except for two, all strains stimulated plant growth. All bacteria altered rhizosphere microbial communities as revealed by phospholipid fatty acid analysis; associating plant growth promotion with a decrease in biological diversity. Three strains were tested for their ability to enhance pine mycorrhization with wild fungi species. Only strain BB1 increased the total number of mycorrhizal root tips. Mycorrhizas present in the roots of each treatment were identified by ribosomal RNA sequencing and denaturing gradient gel electrophoresis analysis, detecting specificity between mycorrhizal species colonizing the roots and the inoculated PGPR. Conclusions: In conclusion, BB1 appears to be a good candidate to be developed into a biofertilizer directed to enhance pine growth and mycorrhization, which should result in a better establishment rate for plants used in reforestation. Significance and Impact of the Study: This study shows the potential of PGPR to improve fitness of forest tree specie. Moreover, the specificity between the bacteria inoculated and the mycorrhiza that the plant selects involve a potential biotechnological use in production of value-added fungi.     

Characterization of α-pinene synthase gene in <Emphasis Type="Italic">Pinus pinaster</Emphasis> and <Emphasis Type="Italic">P. pinea</Emphasis> in vitro cultures and differential gene expression following <Emphasis Type="Italic">Bursaphelenchus xylophilus</Emphasis> inoculation

Pinus pinaster and P. pinea are two important pine species in Portugal. These two pine species show different susceptibility to Bursaphelenchus xylophilus, the nematode causing pine wilt disease, as well as a diverse volatile composition. To clarify the role of terpenes in plant–nematode interactions, the α-pinene synthase gene expression was studied, using P. pinaster and P. pinea in vitro axenic shoot cultures. Identification and isolation of α-pinene synthase genes from both pine species was performed, together with functional characterization of the genes, revealing that the translated amino acid sequences between both species shared 97.3 % pairwise identity. Heterologous expression of full and truncated sequences, devoid of the 48 amino acids of the transit peptide, proved the functionality of both, with the production of α-pinene as the major final product. Relative quantification of protein activity showed a twofold increase of α-pinene production at 4 °C in comparison to assays performed at 21 and 37 °C. Both MnCl₂ and KCl were required for substrate conversion. Furthermore, the variation in gene expression was studied by RT-PCR, using both axenic in vitro shoot pine cultures and co-cultures with B. xylophilus. In P. pinaster there was no difference between co-cultures and control cultures, while in P. pinea α-pinene synthase gene was upregulated in the co-cultures, with a peak of expression at 24 hpi (h post inoculation).     

Root bacteria from nematicidal plants and their biocontrol potential against trichodorid nematodes in potato

Trichodorid nematodes (Nematoda: Trichodoridae) are vectors of tobacco rattle virus (TRV), one of the causal agents of spraing disease in potato. Root bacteria from nematicidal plants and their control potential against Trichodoridae were the focus of this study. Bacteria isolated from the roots of 12 nematicidal plants and potato were characterized for their production of hydrolytic enzymes, hydrogen cyanide, phenol oxidation ability and antifungal activity towards the potato pathogen Rhizoctonia solani. Based on these functional traits, bacteria isolates were selected and tested in greenhouse conditions on potato (cv. Saturna) for their effect on plant growth, and screened for nematicidal activity against Paratrichodorus pachydermus and Trichodorus primitivus in naturally infested soil. Sixteen bacteria isolates out of 44 reduced nematode densities by 50–100%. Nine selected isolated were further tested by bacterizing potato tubers (cv. King Edward) which were planted in a trichodorid and TRV-infested soil. Four bacterial isolates consistently reduced nematode densities (by 56.7–74.4%) with no visual negative effect on plant growth. These isolates were tentatively identified, partly by fatty acid methyl ester (FAME) analysis as: Stenotrophomonas maltophilia, Bacillus mycoides, Pseudomonas sp., and one unidentified bacterium. The isolates originated from potato, Plantago major, Thymus vulgaris and Asparagus officinalis, respectively. Two Pseudomonas isolates obtained from Zinnia elegans and selected for their strong nematicidal activity in soil screening tests, did not reduce the nematode population when tested on potato. It is concluded that plants releasing nematicidal compounds may harbour nematode-antagonistic bacteria as well.     

Isolation and Characterization of Nonrhizobial Plant Growth Promoting Bacteria from Nodules of <Emphasis Type="Italic">Kudzu</Emphasis> (<Emphasis Type="Italic">Pueraria thunbergiana</Emphasis>) and Their Effect on Wheat Seedling Growth

The leguminous vine Kudzu (Pueraria thunbergiana) is an introduction into the N. W. Himalayan region of India. Despite its value as a fodder and cover crop, little is known about the nature of the nodule microflora. In an attempt to study the nodule bacteria, we isolated and characterized three nonrhizobial plant growth promoting bacteria from surface sterilized nodules of Kudzu. Based on the sequencing of the 16 S r RNA gene, the isolates were designated as Bacillus thuringiensis KR-1, Enterobacter asburiae KR-3, and Serratia marcescens KR-4. Crystalline bodies were detected in the isolate KR-1, confirming its identity as B. thuringiensis. Under in vitro conditions, all three isolates were found to produce indole acetic acid. Other plant growth promotion attributes such as P solubilization, hydrogen cyanide production, and ammonia production varied among the isolates. All of the three isolates promoted growth and positively influenced nutrient uptake parameters of wheat seedlings.     

Introduction of some new endophytic bacteria from Bacillus and Streptomyces genera as successful biocontrol agents against Sclerotium rolfsii

Endophytic bacteria live inside plant tissues without causing disease and not only promote plant growth but can also protect plants against plant pathogens. During 2010–2011 crop years, some endophytic bacteria were collected and are then biochemically and molecularly identified (16srRNA) from bean farms of East Azarbaijan, Iran. Among these bacteria isolates, four isolates from Bacillus genera and four isolates from Streptomyces genera were selected for evaluation of their ability for biocontrol of Sclerotium rolfsii in laboratory and glasshouse conditions. Except one isolate named Streptomyces parvus, the rest of isolates could significantly inhibit mycelial growth in dual culture on PDA medium. All seven selected isolates showed significant inhibition in disease treatments in glasshouse experiments. Biological traits, such as length, wet and dry weight of roots and stems in endophytic bacterial treatment showed no differences with healthy control.     

Isolation and characterization of non-Frankia actinobacteria from root nodules of Alnus glutinosa, Casuarina glauca and Elaeagnus angustifolia

Actinobacterial isolates randomly obtained on nitrogen-free BAP medium from surface sterilized root nodules of Alnus glutinosa, Casuarina glauca and Elaeagnus angustifolia sampled from fields were reported. They were assigned on the basis of partial 16S rRNA sequences to Micromonospora, Nocardia and Streptomyces genera. The isolates have been screened for hydrolytic activities, indole acetic acid (IAA) and siderophores production, phosphate solubilization and antagonistic activities. Results suggest putative traits as plant growth promoting bacteria proprieties of the isolates that occur in unique association in root nodules of the three analysed actinorhizal host species.     

Dynamics,Diversity and Function of Endophytic Siderophore-Producing Bacteria in Rice

Siderophore production confers to bacteria competitive advantages to colonize plant tissues and to exclude other microorganisms from the same ecological niche. This work shows that the community of endophytic siderophore-producing bacteria (SPB) associated to Oryza sativa cultivated in Uruguayan soils is dynamic and diverse. These bacteria were present in grains, roots, and leaves, and their density fluctuated between log₁₀ 3.44 and log₁₀ 5.52 cfu g⁻¹ fresh weight (fw) during the plant growth. Less than 10% of the heterotrophic bacteria produced siderophores in roots and leaves of young plants, but most of the heterotrophic bacteria were siderophore-producers in mature plants. According to their amplified restriction DNA ribosomal analysis (ARDRA) pattern, 54 of the 109 endophytic SPB isolated from different plant tissues or growth stages from replicate plots, were unique. Bacteria belonging to the genera Sphingomonas, Pseudomonas, Burkholderia, and Enterobacter alternated during plant growth, but the genus Pantoea was predominant in roots at tillering and in leaves at subsequent stages. Pantoea ananatis was the SPB permanently associated to any of the plant tissues, but the genetic diversity within this species—revealed by BOX-PCR fingerprinting- showed that different strains were randomly distributed along time and plant tissue, suggesting that a common trait of the species P. ananatis determined the interaction with the rice plant. Several isolates were stronger IAA producers than Azospirillum brasilense or Herbaspirillum seropedicae. In vitro inhibition assays showed that SPB of the genus Burkholderia were good antagonists of pathogenic fungi and that only one SPB isolate of the genus Pseudomonas was able to inhibit A. brasilense and H. seropedicae. These results denoted that SPB were selected into the rice plant. P. ananatis was the permanent and dominant associated species which was unable to inhibit two of the relevant plant growth-promoting bacteria.     

Ectomycorrhizal fungi of Pinus pinea L. in northeastern Spain

 Although Pinus pinea L. is an important forest species in the Mediterranean region, few reports exist on its ectomycorrhizal associates. Sixty isolates, obtained from fungal sporocarps collected in mixed forests of P. pinea in Catalonia (northeastern Spain), were tested for ectomycorrhiza formation on containerized P. pinea seedlings when applied as mycelial inoculum produced in peat-vermiculite. A total of 17 isolates, in 8 genera (Amanita, Hebeloma, Laccaria, Lactarius, Pisolithus, Rhizopogon, Scleroderma and Suillus), formed ectomycorrhizas and the percentages of mycorrhizal short roots varied among isolates and species from 13% to 89%. Some of these fungi are cited for the first time in association with P. pinea. The results indicate further fungal candidates for controlled inoculation of P. pinea seedlings in the nursery. Accepted: 29 October 1998