Phosphorus and root distribution and corn growth as related to long-term tillage systems and fertilizer placement

Soil and fertilizer management during cultivation can affect crop productivity and profitability. Long-term experiments are therefore necessary to determine the dynamics of nutrient and root distribution as related to soil profile, as well as the effects on nutrient uptake and crop growth. An 18-year experiment was conducted at the Federal University of Rio Grande do Sul State (UFRGS), in Eldorado do Sul, Brazil, on Rhodic Paleudult soil. Black oat and vetch were planted in the winter and corn in the summer. The soil management methods were conventional, involving no-tillage and strip tillage techniques and broadcast, row-and strip-applied fertilizer placement (triple superphosphate). Available P (Mehlich-1) and root distribution were determined in soil monoliths during the corn grain filling period. Corn shoot dry matter production and P accumulation during the 2006/2007 growing season were determined and the efficiency of P utilization calculated. Regardless of the degree of soil mobilization, P and roots were accumulated in the fertilized zone with time, mainly in the surface layer (0-10 cm). Root distribution followed P distribution for all tillage systems and fertilizer treatments. Under no-tillage, independent of the fertilizer placement, the corn plants developed more roots than in the other tillage systems. Although soil tillage systems and fertilizer treatments affected P and root distribution throughout the soil profile, as well as P absorption and corn growth, the efficiency of P utilization was not affected.

15N uptake fom manure and fertilizer sources by three consecutive crops under controlled conditions

There are several regions of the world where soil N analysis and/or N budgets are not used to determine how much N to apply, resulting in higher than needed N inputs, especially when manure is used. One such region is the North Central "La Comarca Lagunera", one of the most important dairy production areas of Mexico. We conducted a unique controlled greenhouse study using 15N fertilizer and 15N isotopic-labeled manure that was labeled under local conditions to monitor N cycling and recovery under higher N inputs. The manure-N treatment was applied only once and was incorporated in the soil before planting the first forage crop at an equivalent rate of 30, 60 and 120 Mg ha-1 dry manure. The 15N treatments were equivalent to 120 and 240 kg ha-1 (NH4)2SO4-N for each crop. The total N fertilizer for each N fertilized treatment were 360, and 720 kg ha-1 N. We found very low N recoveries: about 9 % from the manure N inputs, lower than the 22 to 25 % from the fertilizer N inputs. The manure N recovered belowground in soil and roots ranged from 82 to 88 %. The low recoveries of N by the aboveground and low soil inorganic nitrate (NO3-N) and ammonium (NH4-N) content after the third harvested suggested that most of the 15N recovered belowground was in the soil organic form. The losses from manure N inputs ranged from 3 to 11 %, lower than the 34 to 39 % lost from fertilizer N sources. Our study shows that excessive applications of manure or fertilizer N that are traditionally used in this region will not increase the rate of N uptake by aboveground compartment but will increase the potential for N losses to the environment.

Geochronological constraints on post-extinction recovery of the ammonoids and carbon cycle perturbations during the Early Jurassic

This paper presents the first quantitative study of the Early Jurassic recovery of ammonoids after the end-Triassic mass extinction based on detailed U-Pb ID-TIMS (isotope dilution thermal ionization mass spectrometry) geochronology from ash bed zircons placed within a clear phylogenetical and biochronological framework at the subzonal and species level. This study was triggered by the discovery of a rich Peruvian succession of ammonites, deposited concomitantly with an unusually large number of ash beds. Two major phases of rediversification are observed during the Psiloceras spelae and Angulaticeras zones that correspond to positive peaks in the delta C-13(org) curve, providing a possible link between biodiversity and the global carbon cycle. In the case of the post-extinction recovery, the development of the earliest Hettangian ammonites occurs within the genus Psiloceras, which begins with the occurrence of P. spelae and then explodes into worldwide development of smooth psiloceratids of the Psiloceras planorbis group s.l. This rapid biodiversification likely occurred less than 100 ka after the end-Triassic crisis; the genus Psiloceras occupied all the possible ecological niches worldwide, from the Pacific deep waters to the NW European shallow deposits and also in some rare Tethyan occurrences like at Germig in Tibet. This global dispersion allowed the differentiation of the group in several major phyla, the Schlotheimiidae, Discamphiceratinae, Arietitidae and Lytocerataceae, which were the roots of all other Jurassic and Cretaceous ammonites. (C) 2012 Elsevier B.V. All rights reserved.

Utilization of Boron (10B) derived from fertilizer by sugar cane

The response to B in agricultural systems of sugar cane is still an unexplored issue; B application has however recently been widely publicized and used with a certain degree of frequency. The use of 10B-labeled fertilizers may further contribute to clarify this practice. With the objective of evaluating sugar cane use of B (10B) derived from fertilizer (boric acid), an experiment was conducted under field conditions in the 2005/2006 growing season. The experiment consisted of the installation of microplots (2 x 1.5 m) where 4 kg ha-1 B (boric acid with 85.95 % in 10B atoms) dissolved in water was applied 90 days after planting (May 2005). The solution was applied to the soil on both sides of the plant row at a distance of 20 cm. After harvest (June 2006) the B content and 10B abundance in % atoms in all parts of the sugar cane plants (stalks, dry leaves, tips and roots) were determined. Results showed that the total B accumulated was 471 g ha-1 in the entire plant (35 % in the stalks, 22 % in the dry leaves, 9 % in the tips and 34 % in the roots). The sugar cane plants used on average 14 % of the total accumulated B in the above-ground part (44 g ha-1) and 11 % in the roots (19 g ha-1), totaling 13 % in the entire plant (63 g ha-1). The recovery of 10B-fertilizer by sugar cane plants was low, around 2 % of the total applied amount.

Expression analyses of three members of the AtPHO1 family reveal differential interactions between signaling pathways involved in phosphate deficiency and the responses to auxin, cytokinin, and abscisic acid.

The PHO1 protein is involved in loading inorganic phosphate (Pi) to the root xylem. Ten genes homologous to AtPHO1 are present in the Arabidopsis thaliana (L.) Heyn genome. From this gene family, transcript levels of only AtPHO1, AtPHO1;H1 and AtPHO1;H10 were increased by Pi-deficiency. While the up-regulation of AtPHO1;H1 and AtPHO1;H10 by Pi deficiency followed the same rapid kinetics and was dependent on the PHR1 transcription factor, phosphite only strongly suppressed the expression of AtPHO1;H1 and had a minor effect on AtPHO1;H10. Addition of sucrose was found to increase transcript levels of both AtPHO1 and AtPHO1;H1 in Pi-sufficient or Pi-deficient plants, but to suppress AtPHO1:H10 under the same conditions. Treatments of plants with auxin or cytokinin had contrasting effect depending on the gene and on the Pi status of the plants. Thus, while both hormones down-regulated expression of AtPHO1 independently of the plant Pi status, auxin and cytokinin up-regulated AtPHO1;H1 and AtPHO1;H10 expression in Pi-sufficient plants and down-regulated expression in Pi-deficient plants. Treatments with abscisic acid inhibited AtPHO1 and AtPHO1;H1 expression in both Pi-sufficient and Pi-deficient plants, but increased AtPHO1;H10 expression under the same conditions. The inhibition of expression by abscisic acid of AtPHO1 and AtPHO1;H1, and of the Pi-starvation responsive genes AtPHT1;1 and AtIPS1, was dependant on the ABI1 type 2C protein phosphatase. These results reveal that various levels of cross talk between the signal transduction pathways to Pi, sucrose and phytohormones are involved in the regulation of expression of the three AtPHO1 homologues.

Phloem mobility of Boron in two eucalypt clones

Boron deficiency causes large productivity losses in eucalypt stands in extensive areas of the Brazilian Cerrado region, thus understanding B mobility is a key step in selecting genetic materials that will better withstand B limitation. Thus, in this study B mobility was evaluated in two eucalypt clones (68 and 129), under B sufficiency or B deficiency, after foliar application of the 10B isotope tracer to a single mature leaf. Samples of young tissue, mature leaves and roots were collected 0, 1, 5, 12 and 17 days after 10B application. The 10B:11B isotope ratio was determined by HR-ICP-MS. Samples of leaves and xylem sap were collected for the determination of soluble sugars and polyalcohols by ion chromatography. Boron was translocated within eucalypt. Translocation of foliar-applied 10B to the young tissues, mature leaves and roots was higher in clone 129 than in 68. Seventeen days after 10B application to a single mature leaf, between 14 and 18 % of B in the young tissue was originated from foliar B application. In plants with adequate B supply the element was not translocated out of the labeled leaf.

Boron mobility in eucalyptus clones

Understanding the magnitude of B mobility in eucalyptus may help to select clones that are more efficient for B use and to design new practices of B fertilization. This study consisted of five experiments with three eucalyptus clones (129, 57 and 58) where the response to and mobility of B were evaluated. Results indicated that clone 129 was less sensitive to B deficiency than clones 68 and 57, apparently due to its ability to translocate B previously absorbed via root systems to younger tissues when B in solution became limiting. Translocation also occurred when B was applied as boric acid only once to a single mature leaf, resulting in higher B concentration in roots, stems and younger leaves. The growth of B-deficient plants was also recovere by a single foliar application of B to a mature leaf. This mobility was greater, when foliar-applied B was supplied in complexed (boric acid + manitol) than in non-complexed form (boric acid alone). When the root system of clone 129 was split in two solution compartments, B supplied to one root compartment was translocated to the shoot and back to the roots in the other compartment, improving the B status and growth. Thus, it appears that B is relatively mobile in eucalyptus, especially in clone 129, and its higher mobility could be due to the presence of an organic compound such as manitol, able to complex B.

Effect of abscisic acid on maize root growth. A critical examination

The effect of abscisic acid (ABA) on the growth of maize roots maintained in the dark is investigated in relation to the root varieties and the root age, the mode of application, the concentration used and the duration of both the treatment and the culture. In all the assays, when ABA produces a significant change in root elongation, it shows an inhibitory effect which is enhanced with increasing ABA concentration. The data strongly support the hypothesis that ABA could be one of the growth inhibitors which are formed in or released from the root cap.

Nutrient supply by mass flow and diffusion to maize plants in response to soil aggregate size and water potential

Nutrients are basically transported to the roots by mass flow and diffusion. The aim of this study was to quantify the contribution of these two mechanisms to the acquisition of macronutrients (N, P, K, Ca, Mg, and S) and cationic micronutrients (Fe, Mn, Zn, and Cu) by maize plants as well as xylem exudate volume and composition in response to soil aggregate size and water availability. The experiment was conducted in a greenhouse with samples of an Oxisol, from under two management systems: a region of natural savanna-like vegetation (Cerradão, CER) and continuous maize under conventional management for over 30 years (CCM). The treatments were arranged in a factorial [2 x (1 + 2) x 2] design, with two management systems (CER and CCM), (1 + 2) soil sifted through a 4 mm sieve and two aggregate classes (< 0.5 mm and 0.5 - 4.0 mm) and two soil matric potentials (-40 and -10 kPa). These were evaluated in a randomized block design with four replications. The experiment was conducted for 70 days after sowing. The influence of soil aggregate size and water potential on the nutrient transport mechanisms was highest in soil samples with higher nutrient concentrations in solution, in the CER system; diffusion became more relevant when water availability was higher and in aggregates < 0.5 mm. The volume of xylem exudate collected from maize plants increased with the decrease in aggregate size and the increased availability of soil water in the CER system. The highest Ca and Mg concentrations in the xylem exudate of plants grown on samples from the CER system were related to the high concentrations of these nutrients in the soil solution of this management system.

Response of Epidendrum Ibaguense (orchidaceae) to the application of lime rates to the pot

In the best cultivation methods of orchids, in particular of the genus Phalaenopsis, liming is a common practice. The objective of this study was to evaluate the influence of lime rates (0.0; 1.0; 2.0; 3.0; 4.0; and 5.0 g dm-3 of substrate) applied to the cultivation substrate (xaxim) on the growth of Epidendrum ibaguense seedlings. In a greenhouse, 1-L plastic pots filled with 0.8 dm³ of xaxim were irrigated such that no leachate was lost during the experiment. N, P, K, Ca, Mg, S, Fe, Zn, B, and Mn contents in roots, stems and leaves were measured. Leachate was collected by applying a sufficient water volume to obtain 25 mL from each pot. Fourteen days after lime application of 3 g dm-3, the pH of the collected leachate reached values above 7 and a value of 6.29 with the highest lime rate at the end of the experiment. The lime rate did not influence plant height, probably due to a Zn deficiency at high pH levels and a Ca deficiency in the control. Nevertheless, there was a large increase in leaf production, for number as well as for dry matter mass. There was no statistical difference between treatments in root dry matter production. Maximum dry matter production was obtained at a lime rate of 4.09 g dm-3. Zinc concentrations diminished linearly with increasing lime rates; the concentrations in all treatments were below the levels suggested as adequate in the literature (25-200 mg kg-1). Nutrient concentrations in leaves indicated deficiency of N, S, and B at the highest lime rates (4.0 and 5.0 g dm-3), and of Ca in the treatment without liming.

Intraspecific genetic variation in arbuscular mycorrhizal fungi and its consequences for molecular biology, ecology, and development of inoculum

It has been known for some time that different arbuscular mycorrhizal fungal (AMF) taxa confer differences in plant growth. Although genetic variation within AMF species has been given less attention, it could potentially be an ecologically important source of variation. Ongoing studies on variability in AMF genes within Glomus intraradices indicate that at least for some genes, such as the BiP gene, sequence variability can be high, even in coding regions. This suggests that genetic variation within an AMF may not be selectively neutral. This clearly needs to be investigated in more detail for other coding regions of AMF genomes. Similarly, studies on AMF population genetics indicate high genetic variation in AMF populations, and a considerable amount of variation seen in phenotypes in the population can be attributed to genetic differences among the fungi. The existence of high within-species genetic variation could have important consequences for how investigations on AMF gene expression and function are conducted. Furthermore, studies of within-species genetic variability and how it affects variation in plant growth will help to identify at what level of precision ecological studies should be conducted to identify AMF in plant roots in the field. A population genetic approach to studying AMF genetic variability can also be useful for inoculum development. By knowing the amount of genetic variability in an AMF population, the maximum and minimum numbers of spores that will contain a given amount of genetic diversity can be estimated. This could be particularly useful for developing inoculum with high adaptability to different environments.

Growth of seedlings of pigeon pea (Cajanus cajan (l.) millsp), wand riverhemp (Sesbania virgata (cav.) pers.), and lead tree (Leucaena leucocephala (lam.) de wit) in an arsenic-contaminated soil

Phytoremediation strategies utilize plants to decontaminate or immobilize soil pollutants. Among soil pollutants, metalloid As is considered a primary concern as a toxic element to organisms. Arsenic concentrations in the soil result from anthropogenic activities such as: the use of pesticides (herbicides and fungicides); some fertilizers; Au, Pb, Cu and Ni mining; Fe and steel production; coal combustion; and as a bi-product during natural gas extraction. This study evaluated the potential of pigeon pea (Cajanus cajan), wand riverhemp (Sesbania virgata), and lead tree (Leucaena leucocephala) as phytoremediators of soils polluted by As. Soil samples were placed in plastic pots, incubated with different As doses (0; 50; 100 and 200 mg dm-3) and then sown with seeds of the three species. Thirty (pigeon pea) and 90 days after sowing, the plants were evaluated for height, collar diameter and dry matter of young, intermediate and basal leaves, stems and roots. Arsenic concentration was determined in different aged leaves, stems and roots to establish the translocation index (TI) between the plant root system and aerial plant components and the bioconcentration factors (BF). The evaluated species showed distinct characteristics regarding As tolerance, since the lead tree and wand riverhemp were significantly more tolerant than pigeon pea. The high As levels found in wand riverhemp roots suggest the existence of an efficient accumulation and compartmentalization mechanism in order to reduce As translocation to shoot tissues. Pigeon pea is a sensitive species and could serve as a potential bioindicator plant, whereas the other two species have potential for phytoremediation programs in As polluted areas. However, further studies are needed with longer exposure times in actual field conditions to reach definite conclusions on relative phytoremediation potentials.

Study of the regulation of antifungal activity in plant-associated pseudomas fluorescens

RésuméEn agriculture d'énormes pertes sont causées par des champignons telluriques pathogènes tels que Thielaviopsis, Fusarium, Gaeumannomyces et Rhizoctonia ou encore l'oomycète Pythium. Certaines bactéries dites bénéfiques, comme Pseudomonas fluorescens, ont la capacité de protéger les plantes de ces pathogènes par la colonisation de leur racines, par la production de métabolites secondaires possédants des propriétés antifongiques et par l'induction des mécanismes de défenses de la plante colonisée. P. fluorescens CHAO, une bactérie biocontrôle isolée d'un champ de tabac à Payerne, a la faculté de produire un large spectre de métabolites antifongiques, en particulier le 2,4- diacétylphloroglucinol (DAPG), la pyolutéorine (PLT), le cyanure d'hydrogène (HCN), la pyrrolnitrine (PRN) ainsi que des chélateurs de fer.La plante, par sécrétion racinaire, produit des rhizodéposites, source de carbone et d'azote, qui profitent aux populations bactériennes vivant dans la rhizosphere. De plus, certains stresses biotiques et abiotiques modifient cette sécrétion racinaire, en terme quantitatif et qualitatif. De leur côté, les bactéries bénéfiques, améliorent, de façon direct et/ou indirect, la croissance de la plante hôte. De nombreux facteurs biotiques et abiotiques sont connus pour réguler la production de métabolites secondaires chez les bactéries. Des études récentes ont démontré l'importance de la communication entre la plante et les bactéries bénéfiques afin que s'établisse une interaction profitant à chacun des deux partis. Il est ainsi vraisemblable que les populations bactériennes associées aux racines soient capables d'intégrer ces signaux et d'adapter spécifiquement leur comportement en conséquence.La première partie de ce travail de thèse a été la mise au point d'outils basés sur la cytométrie permettant de mesurer l'activité antifongique de cellules bactériennes individuelles dans un environnent naturel, les racines des plantes. Nous avons démontré, grâce à un double marquage aux protéines autofluorescentes GFP et mCherry, que les niveaux d'expression des gènes impliqués dans la biosynthèse des substances antifongiques DAPG, PLT, PRN et HCN ne sont pas les mêmes dans des milieux de cultures liquides que sur les racines de céréales. Par exemple, l'expression de pltA (impliqué dans la biosynthèse du PLT) est quasiment abolie sur les racines de blé mais atteint un niveau relativement haut in vitro. De plus cette étude a mis en avant l'influence du génotype céréalien sur l'expression du gène phlA qui est impliqué dans la biosynthèse du DAPG.Une seconde étude a révélé la communication existant entre une céréale (orge) infectée par le pathogène tellurique Pythium ultimum et P. fluorescens CHAO. Un système de partage des racines nous a permis de séparer physiquement le pathogène et la bactérie bénéfique sur la plante. Cette méthode a donné la possibilité d'évaluer l'effet systémique, causé par l'attaque du pathogène, de la plante sur la bactérie biocontrôle. En effet, l'infection par le phytopathogène modifie la concentration de certains composés phénoliques dans les exsudats racinaires stimulant ainsi l'expression de phi A chez P.fluorescens CHAO.Une troisième partie de ce travail focalise sur l'effet des amibes qui sont des micro-prédateurs présents dans la rhizosphere. Leur présence diminue l'expression des gènes impliqués dans la biosynthèse du DAPG, PLT, PRN et HCN chez P.fluorescens CHAO, ceci en culture liquide et sur des racines d'orge. De plus, des molécules provenant du surnageant d'amibes, influencent l'expression des gènes requis pour la biosynthèse de ces antifongiques. Ces résultats illustrent que les amibes et les bactéries de la rhizosphere ont développé des stratégies pour se reconnaître et adapter leur comportement.La dernière section de ce travail est consacrée à l'acide indole-acétique (LA.A), une phytohormone connue pour son effet stimulateur sur phlA. Une étude moléculaire détaillée nous a démontré que cet effet de l'IAA est notamment modulé par une pompe à efflux (FusPl) et de son régulateur transcriptionnel (MarRl). De plus, les gènes fusPl et marRl sont régulés par d'autres composés phénoliques tels que le salicylate (un signal végétal) et l'acide fusarique (une phytotoxine du pathogène Fusarium).En résumé, ce travail de thèse illustre la complexité des interactions entre les eucaryotes et procaryotes de la rhizosphère. La reconnaissance mutuelle et l'instauration d'un dialogue moléculaire entre une plante hôte et ses bactéries bénéfiques associées? sont indispensables à la survie des deux protagonistes et semblent être hautement spécifiques.SummaryIn agriculture important crop losses result from the attack of soil-borne phytopathogenic fungi, including Thielaviopsis, Fusarium, Gaeumannomyces and Rhizoctonia, as well as from the oomycete Pythium. Certain beneficial microorganisms of the rhizosphere, in particular Pseudomonas fluorescens, have the ability to protect plants against phytopathogens by the intense colonisation of roots, by the production of antifungal exoproducts, and by induction of plant host defences. P. fluorescens strain CHAO, isolated from a tobacco field near Payerne, produces a large array of antifungal exoproducts, including 2,4-diacetylphloroglucinol (DAPG), pyoluteorin (PLT), hydrogen cyanide (HCN), pyrrolnitrin (PRN) and iron chelators. Plants produce rhizodeposites via root secretion and these represent a relevant source of carbon and nitrogen for rhizosphere microorganisms. Various biotic and abiotic stresses influence the quantity and the quality of released exudates. One the other hand, beneficial bacteria directly or indirectly promote plant growth. Biotic and abiotic factors regulate exoproduct production in biocontrol microorganisms. Recent studies have highlighted the importance of communication in establishing a fine-tuned mutualist interaction between plants and their associated beneficial bacteria. Bacteria may be able to integrate rhizosphere signals and adapt subsequently their behaviour.In a first part of the thesis, we developed a new method to monitor directly antifungal activity of individual bacterial cells in a natural environment, i.e. on roots of crop plants. We were able to demonstrate, via a dual-labelling system involving green and red fluorescent proteins (GFP, mCherry) and FACS-based flow cytometry, that expression levels of biosynthetic genes for the antifungal compounds DAPG, PLT, PRN, and HCN are highly different in liquid culture and on roots of cereals. For instance, expression of pltA (involved in PLT biosynthesis) was nearly abolished on wheat roots whereas it attained a relatively high level under in vitro conditions. In addition, we established the importance of the cereal genotype in the expression of phi A (involved in DAPG biosynthesis) in P. fluorescens CHAO.A second part of this work highlighted the systemic communication that exists between biocontrol pseudomonads and plants following attack by a root pathogen. A split-root system, allowing physical separation between the soil-borne oomycete pathogen Phytium ultimum and P. fluorescens CHAO on barley roots, was set up. Root infection by the pathogen triggered a modification of the concentration of certain phenolic root exudates in the healthy root part, resulting in an induction ofphlA expression in P. fluorescens CHAO.Amoebas are micro-predators of the rhizosphere that feed notably on bacteria. In the third part of the thesis, co-habitation of Acanthamoeba castellanii with P. fluorescens CHAO in culture media and on barley roots was found to significantly reduce bacterial expression of genes involved in the biosynthesis of DAPG, PLT, HCN and PRN. Interestingly, molecular cues present in supernatant of A. castelanii induced the expression of these antifungal genes. These findings illustrate the strategies of mutual recognition developed by amoeba and rhizosphere bacteria triggering responses that allow specific adaptations of their behaviour.The last section of the work focuses on indole-3-acetic acid (IAA), a phytohormone that stimulates the expression of phi A. A detailed molecular study revealed that the IAA-mediated effect on phi A is notably modulated by an efflux pump (FusPl) and its transcriptional regulator (MarRl). Remarkably, transcription of fusPl and marRl was strongly upregulated in presence of other phenolic compounds such as salicylate (a plant signal) and fusaric acid (a phytotoxin of the pathogenic fungus Fusarium).To sum up, this work illustrates the great complexity of interactions between eukaryotes and prokaryotes taking place in the rhizosphere niche. The mutual recognition and the establishment of a molecular cross-talk between the host plant and its associated beneficial bacteria are essential for the survival of the two partners and these interactions appear to be highly specific.

Growth promotion of pineapple 'vitória' by humic acids and burkholderia spp. during acclimatization

In vitro propagation of pineapple produces uniform and disease-free plantlets, but requires a long period of acclimatization before transplanting to the field. Quicker adaptation to the ex vitro environment and growth acceleration of pineapple plantlets are prerequisites for the production of a greater amount of vigorous, well-rooted planting material. The combination of humic acids and endophytic bacteria could be a useful technological approach to reduce the critical period of acclimatization. The aim of this study was to evaluate the initial performance of tissue-cultured pineapple variety Vitória in response to application of humic acids isolated from vermicompost and plant growth-promoting bacteria (Burkholderia spp.) during greenhouse acclimatization. The basal leaf axils were treated with humic acids while roots were immersed in bacterial medium. Humic acids and bacteria application improved shoot growth (14 and 102 %, respectively), compared with the control; the effect of the combined treatment was most pronounced (147 %). Likewise, humic acids increased root growth by 50 %, bacteria by 81 % and the combined treatment by 105 %. Inoculation was found to significantly increase the accumulation of N (115 %), P (112 %) and K (69 %) in pineapple leaves. Pineapple growth was influenced by inoculation with Burkholderia spp., and further improved in combination with humic acids, resulting in higher shoot and root biomass as well as nutrient contents (N 132 %, P 131 %, K 80 %) than in uninoculated plantlets. The stability and increased consistency of the host plant response to bacterization in the presence of humic substances indicate a promising biotechnological tool to improve growth and adaptation of pineapple plantlets to the ex vitro environment.