Ectomycorrhizal inoculation procedures have been developed to improve forestation practice in areas with unfavourable habitat properties. The inoculation of cultivated crops with specially developed inocula of arbuscular-mycorrhizal fungi succeeded in reaching a substantial harvest surplus of plant material, and makes environment-friendly, fertilizer-free farming practices around shallow lakes possible

Bratek, Zoltán

English

Bratek Zoltán

University of Szeged

Volume 46(3-4):187-188, 2002Acta Biologica Szegediensishttp://www.sci.u-szeged.hu/ABS1Dept. of Plant Physiology, Eötvös Loránd University, Budapest, Hungary, 2Research Institute for Soil Science andAgricultural Chemistry, Budapest, HungaryZoltán Bratek1, Ibolya Vörös2, Tünde Takács2, István Parádi1, Szabolcs Rudnóy1, Krisztián Halász1*ABSTRACT                     Ectomycorrhizal inoculation procedures have been developed to improveforestation practice in areas with unfavourable habitat properties. The mycorrhizal inoculationof cultivated crops with specially developed inocula of arbuscular-mycorrhizal fungi succeededin reaching a substantial harvest surplus of plant material, and makes environment-friendly,fertilizer-free farming practices around shallow lakes possible.Acta Biol Szeged 46(3-4):187-188 (2002)KEY WORDSmycorrhizaenvironment-friendly agricultureforestry*Corresponding author. E-mail: bratek@ludens.elte.hu187Advantages of application of mycorrhizated plants in environment-friendly agriculture and forestationsProceedings of the 7th Hungarian Congress on Plant Physiology, 2002S5-06With the usage of appropriate mycorrhizal fungi the plants’stress tolerance can be raised substantially against unfa-vourable environmental effects. Natural forest growth ofdomestic land areas with extreme soil and climate properties,such as salt affected soils, sands, and in mine pits and minespoils is a slow process. The hastening of this process hasbeen made possible by enhanced stress-tolerant/vitalizedmycorrhizal plants has become common practice in manycountries. It is remarkable that some studies were carried outby Hungarian researchers in this field (Bokor 1954; Szántó1994a,b). While growing tree seedlings in nurseries, resis-tance against different diseases improves in cases ofmycorrhizal plants, just as with their growth indicators. Thesurvival of mycorrhizal plants is significantly greater thannon-mycorrhizal ones, therefore mycorrhizal inoculationresults in avoiding unnecessary replacements by workingwith a smaller number of plants.The use of mycorrhiza of cultivated crops (causes) goesalong with the growth in uptake of macro- and micro-elements, making it possible to decrease or even stop theusage of fertilizers. Beside this, the quicker emergence of soillife is made possible, and favourably affects the physicalproperties of the soil on the behalf of plants.Materials and MethodsThe first stage of the work meant the mapping of mycorrhiza-forming fungi with the trees of the territories mentionedbeing developed, as well as the isolation of their sterilecultures, which was followed by the processing of thecultures which had been maintained on solid agar media andaccrued in shake flask cultures (Molina and Palmer 1982;Rudnóy 1999). The molecular identification of the strainswas carried out based on RFLP and sequence analysis of thenuclear ribosomal ITS region (Rudnóy et al. 2000).The greenhouse ectomycorrhizal inoculation wasperformed with the sterile mycelial cultures fragmented in ablender, or with spores from fruit bodies which had beenadded to a pasteurised mixture of peat and vermiculite. Hostplants such as black-locust tree, oak, poplar, and pine treespecies were inoculated in nursery seedbeds. Mycorrhizalcolonization was quantified on one- and two-year oldmycorrhizal seedlings in the nursery. Two years old treeseedlings were outplanted in field soil and the physiologicalparameters (CO2-fixation, chlorophyll content, chlorophyllfluorescence kinetics), the growth and survival characteristicsof them were measured continuously.We isolated monosporic arbuscular-mycorrhizal fungistrains from a number of soil types (brown forest soil andcarbonated field soil) in the Balaton drainage basin. We usedperennial ryegrass (Lolium perenne) and ribwort plantain(Plantago lanceolata) as snare plants in sterile soil. Weaugmented the cultures further under semisterile laboratoryconditions, using alfalfa (Medicago sativa) and red clover(Trifolium pratense). The large scale augmentation of thecultures was performed in the greenhouse with the help ofSudanese grass (Sorghum sudanense). Those inoculumswhich were gained in such a way, which were comprised ofthe roots of the host plant, the soil, and those mycorrhizalparticles present in both, were used for field mycorrhization,where the augmented strains, or their mixtures, so calledcoctail inoculums were used. The growth of AMF-colonization and the physiological parameters (CO2-fixation,chlorophyll content, stoma conductance) of crops (winterwheat, corn, alfalfa) used in the field experiments wereexamined continuously.Mycorrhizal colonization was quantified by the methodsof Phillips and Hayman (1970) and Trouvelot et al. (1986).Dry matters were determined after drying at 80oC untilconstant weight. Total percentage plant phosphorus, nitrogenand kalium contents (% of d. wt) were measured by an ICPspectrometer (Thermo Jarrell Ash ICAP 61E) after digestaccording to Buzás (1988). Chlorophyll a+b concentration(Porra et al. 1989) and chlorophyll fluorescence kinetics (Fv/Fm) by a PAM (Pulse Amplitude Modulation) ChlorophyllFluorometer (Heinz Walz GmbH, Effeltrich) were measured(Schreiber 1986). 14CO2 fixation capacities of excised leaveswere determined using a closed illuminated glass chamberwith saturating light intensity (1000 µmol m-2 s-1) and with1880.1 % CO2 concentration (Láng et al. 1985). Radioactivity ofthe leaf samples was determined by a liquid scintillationapparatus (Beckman LS 5000 TD).Results and DiscussionLists of fruit body forming mycorrhizal fungi species livingin domestic saliferous and sandy forests as well as theyounger forests of mining sites was drawn up, and thetypization and identification (where possible) of theuncovered, cleaned and washed ectomycorrhizal roots wasperformed (Halász et al. 2000; Rudnóy et al. 2000; Barna etal. 2001; Berecz 2001). The isolation of several hundredectomycorrhizal fungal strains succeeded in the areasmentioned, and the identification of the strains was rein-forced by molecular methods (Rudnóy et al. 1999-2000;Kovács et al. 2001), and their maintenance was solved bycompiling a collection. Methods for the economical large-scale production of mycelium-based inoculum as well as fortheir mycorrhizal inoculation procedures were succesfullydeveloped. The mycorrhizal synthesis resulted in many casesof heavy mycorrhizal colonization without the use ofmycorrhization helping bacteria (MHBs). The growth ofmycorrhizal trees usually surpassed substantially that of non-mycorrhizal ones followed by increased values of CO2-fixation. The high falling out rate was detected only in thecase of non-mycorrhizal white poplars growing undergreenhouse conditions. In order to decrease contamination,increase mycorrhizal colonization and select of plant cloneswith higher mycorrhizal colonization further studies arerequired.Among AM-fungi strains isolated from plants used assnares, those ones were selected which had high indicatorvalues for mycorrhization and growth of ploughland crops.During the small parcel field experiments, directedmycorrhizal inoculation provided extraordinary results in theamount of biomass and the growth somewhat in alfalfa butespecially in corn. We took physiological characteristics aswell as biomass values into consideration when comparinginocula. These values of the artificially mycorrhizated plantsvaried greatly compared to those experienced with controlplants, but in most experimental versions surpassed thecontroll. Based on the results so far, in the case of certain hostplant species mycorrhizal inoculation makes application offertilizers unnecessary or greatly reducable, which –especially in the vicinity of surface water courses - couldbecome an important factor in environment-friendly farmingpractise.AcknowledgmentsWe wish to express our thanks to the Silvanus Group Ltd. andthe Silvanus Nursery Inc. as well as to the NEFAG Inc. forthe support they gave to our research.ReferencesBarna T, Jakucs E, Bratek Z, Szántó M (2001) Az erdôsítések eredmé-nyességét fokozó közös mikorrhiza kutatások az ESZTK-ERTI-ELTErészvételével. In Erdészeti Tudományos Intézet Kiadványai,Gondolatok az erdôvédelemrôl az ezredfordulón. pp. 169-178.Berecz B  (2001) „Bauxit meddôk és bányagödrök mikorrhizagombái”.Thesis, ELTE.Bokor R (1954) A mykorrhiza gombákkal történô talajoltások újagrokémiai eljárása. Erd Kut Bp 4:27-45.Halász K, Bratek Z, Jakucs E, Albert L, Csiha I. Ökológiailag stabilszikfásítás. In Kutatói Nap 1998-1999. eds., Horváth B, AlföldiErdôkért Egyesület, pp. 91-93.Halász Krisztián (2000) Szikes talajú tölgyesek ektomikorrhiza-gombáinaktáptalaj-szelekciója és ITS-régiójuk szekvenciaanalízise. Thesis, ELTEKovács G, Rudnóy Sz, Vágvölgyi Cs, Lásztity D, Rácz I, Bratek Z (2001)Intraspecfic invariability of the ITS region of rDNA of Terfeziaterfezioides in Europe. Folia Microbiol 46:423-426Láng F, Sárvári É, Szigeti Z.(1985) Apparatus and method for rapiddetermination of photosynthetic CO2 fixation of leaves. BiochemPhysiol Pflanzen 180:333-336.Molina R, JG Palmer (1982) Isolation, Maintenance, and Pure CultureManipulation of Ectomycorrhizal Fungi. In: Schenk, N. C., ed.,Methods and Principles of Mycorrhizal Research. St Paul, MN: AmerPhytopat Soc 115-129.Phillips JM, Hayman DS. Improved procedures for clearing roots andstaining parasitic and vesicular-arbuscular mycorrhizal fungi for rapidassessment of infection. – Trans. Brit. Mycol. Soc. 55: 158-160, 1970.Porra RJ, Thompson WA, Kriedemann PE. Determination of accurateextinction coefficients and simultaneous equations for assayingchlorophylls a and b extracted with four different solvents: verificationof the concentration of chlorophyll standards by atomic absorptionspectroscopy. Biochim Biophys Acta 975:384-394, 1989.Rudnóy Sz, Bratek Z, Albert L, Barna T, Lásztity D (2000) Kiskunságifehérnyárasok mikorrhiza viszonyainak feltárása. In Kutatói Nap 1998-1999. Szerk.: Horváth B, Alföldi Erdôkért Egyesület, pp. 94-97.Rudnóy Sz, Bratek Z, Halász K, Rácz I, Lásztity D (1999-2000) A fehérnyárral (Populus alba L.) ektomikorrhizás kapcsolatban álló kétgombafaj táptalaj szelekciója és molekuláris biológiai vizsgálata. BotKözl 86-87:121-128Rudnóy Sz (1999) Ektomikorrhizás gombatörzsek táptalaj-optimalizálásaés molekuláris biológiai vizsgálata. Thesis, ELTE.Schreiber U. Detection of rapid induction kinetics with a new type of highfrequency chlorophyll fluorometer. Photosynth Res 9:261-272, 1986.Szántó M (1994a) Mikorrhizált erdei- és feketefenyô (Pinus sylvestris L.,Pinus nigra Arn.) csemeték összehasonlító vizsgálata 1. Növekedés-vizsgálat, a magasság alakulása, hossznövekedés. Mikol Közl 34:64-74.Szántó M (1994b) Mikorrhizált erdei- és feketefenyô (Pinus sylvestris L.,Pinus nigra Arn.) csemeték összehasonlító vizsgálata 2. A növényektömegviszonyai. Mikol Közl 34:42-47.Trouvelot A, Kough JL, Gianinazzi-Pearson V. Mesure du taux demycorhization VA d’un systéme radiculare. Recherche de méthodesd’estimation ayant une signification fonctionelle. In: Mycorrhizae:physiology and genetics-Les mycorhizes: physiologie et génétique. -Proceedings of the 1st ESM/1er SEM, Dijon, 1-5 July 1985-INRA,Paris: 217-221,1986.

Advantages of application of mycorrhizated plants in environmentfriendly agriculture and forestations

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Advantages of application of mycorrhizated plants in environmentfriendly agriculture and forestations

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