1 Biostimulators IN MODERN AGRICULTURE Ornamental and special plants E D I T O R : Aleksandra Łukaszewska W a r s a w
2 1 Biostimulators IN MODERN AGRICULTURE Ornamental and special plants EDITOR: Aleksandra Lukaszewska Warsaw 2008
3 2 The series of monographs under a common name BIOSTIMULATORS IN MODERN AGRICULTURE contains a review of recent research related to this subject and consists of the following parts: GENERAL ASPECTS FIELD CROPS SOLANACEOUS CROPS VEGETABLE CROPS FRUIT CROPS ORNAMENTAL AND SPECIAL PLANTS EDITORIAL BOARD: Andrzej Sadowski, Department of Pomology, Warsaw University of Life Sciences (WULS) chairman Zbigniew T. D¹browski, Department of Applied Entomology, WULS Helena Gawroñska, Laboratory of Basic Natural Sciences in Horticulture, WULS Aleksandra ukaszewska, Department of Ornamental Plants, WULS Adam S³owiñski, Arysta LifeScience Poland PRODUCTION EDITORS: Aleksandra ukaszewska Warsaw University of Life Sciences (WULS) Anna Karbowniczek, Arysta LifeScience Poland Ada Krzeczkowska, Wieœ Jutra Halina Skrobacka, Wieœ Jutra REVIEWERS: Aleksandra ukaszewska, Warsaw University of Life Sciences (WULS) Karol Chyliñski, Warsaw University of Life Sciences (WULS) Halina Laskowska, University of Life Sciences in Lublin Monika Latkowska, Warsaw University of Life Sciences (WULS) Leonard Indeka, Warsaw University of Life Sciences (WULS) Bo ena Matysiak, Research Institute of Pomology and Floriculture, Skierniewice Irena Olszewska-Kaczyñska, Warsaw University of Life Sciences (WULS) Andrzej Pacholczak, Warsaw University of Life Sciences (WULS) Ewa Skutnik, Warsaw University of Life Sciences (WULS) Piotr Urbañski, Poznan University of Life Sciences Katarzyna Wróblewska, Wroc³aw University of Environmental and Life Sciences This edition was supported by Arysta LifeScience Cover: Plantpress ISBN Published by the Editorial House Wieœ Jutra, Limited Janowskiego Warszawa phone: (0 22) Printed by Ryko Copies 300, publishing sheets: 6.0
4 3 CONTENTS PREFACE... 5 THE EFFECT OF BIOPREPARATIONS ON EARLY STAGE OF GROWTH OF SELECTED GRASS SPECIES USED FOR RECLAMATION OF MUNICIPAL WASTE DUMPS... 7 Daria G¹bka, Karol Wolski EFFECT OF ASAHI SL BIOSTIMULATOR ON ORNAMENTAL AMARANTH (AMARANTHUS SPP.) PLANTS EXPOSED TO SALINITY IN GROWING MEDIUM...15 Mariola Wrochna, Barbara ata, Bo enna Borkowska, Helena Gawroñska THE EFFECTS OF BIOSTIMULATORS ASAHI SL AND SIAPTON 10L ON THE GROWTH OF BERGENIA CORDIFOLIA ((HAW.) STERNB.) `ROTBLUM` AND HOSTA (TRATT.) `SUM AND SUBSTANCE` AND `MINUTEMAN`...33 Justyna Krajewska, Monika J. Latkowska EFFECT OF ASAHI SL ON THE INITIAL DEVELOPMENT OF WILLOW CUTTINGS AT VARIED SOIL MOISTURE Gra yna Harasimowicz-Hermann, Krzysztof Czy THE EFFECT OF ATONIC PLANT GROWTH STIMULATOR ON PHYSIOLOGICAL INDICATORS OF THE BASKET WILLOW (SALIX VIMINALIS L.) CULTIVATED IN ANTHROPOGENIC SOIL...47 Jacek Wróbel, Anna WoŸniak INFLUENCE OF ETEPHON ON FLOWERING OF EASY POT FREESIA CULTIVATED IN SPRING-SUMMER SEASON...56 Piotr urawik POLISH SUMMARIES... 63
5 4 PREFACE The high yield potential of modern cultivars is often restrained by various environmental stresses both of biotic and abiotic nature, affecting the crop status. The present approach in pro-ecological plant protection from such biotic stresses as weeds, diseases and pests emphasises enhancement of naturally occurring compounds, organisms or plant defence mechanisms. These compounds should fill the gap resulting from the regulatory decisions of national authorities in many countries, leading to restrictions in use of a number of synthetic pesticides. Extensive research carried out in the last two decades has shown that some natural products may be efficiently used in enhancing the plant s endogenous resistance or tolerance to the biotic and abiotic stresses. A group of such active products is presently classified as biostimulators. When reduction of the chemical input is expected, the use of biostimulators becomes a particularly promising option. Biostimulators are defined as compounds of biological origin and should act by increasing natural capabilities of plants to cope with stresses. Biostimulators do not act neither as nutrients nor affect directly the stress factors making them less harmful for plants. The efficacy of biostimulators is not limited to reducing effects of biotic and abiotic stresses. They stimulate growth and development of plants under unfavourable soil and climatic conditions. Although the effects of biostimulators are not so spectacular and not always stable over the years due to interaction with other used chemicals and/or environmental factors the interest of farmers in using biostimulators is successively increasing over time. According to the national legislation, biostimulators are related to the category of plant protection products. Therefore they must comply with all rules for registration and hence prior to formal approval for use they must be tested for safety to humans and the environment. The dynamic increase of research projects on biostimulators and of farmers interest in their use in agriculture and horticulture production provoked an idea of the international conference on Biostimulators in Modern Agriculture. It was organized by the Laboratory of Basic Sciences in Horticulture, at the Faculty of Horticulture and Landscape Architecture at the Warsaw University of Life Sciences. The conference has attracted a large group of scientists and graduate students from universities and research institutions involved in basic and applied research
6 in agriculture as well from the industry. About three hundred sixty participants included also representatives of farmers and distributors of agricultural supplies. The extensive and creative discussions during the conference and interest in conference materials as well as suggestions from participants indicated the urgent need for dissemination of the state of knowledge on biostimulators. This inspired the organizers of the Conference to co-ordinate preparing reviews on recent scientific achievements in the field of biostimulators, including the practical aspects of their application on various crops. Following suggestions appearing at the Conference, the organisers invited scientists having experience and achievements in work on biostimulators to prepare relevant reviews related to particular products and crops. Based on the submitted manuscripts the Editorial Board decided to publish a series of monographs entitled: BIOSTIMULATORS IN MODERN AGRICULTU- RE comprising the following six volumes: General Aspects, Field Crops, Solanaceous Crops, Vegetable Crops, Fruit Crops and Ornamental and Special Plants. The Editors hope that this publication would fill the gap in knowledge on the mechanisms of action of various biostimulators and on the conditions for their high efficacy. We are very grateful to the authors who willingly agreed to contribute to these books. 5 EDITORS
8 7 THE EFFECT OF BIOPREPARATIONS ON EARLY STAGE OF GROWTH OF SELECTED GRASS SPECIES USED FOR RECLAMATION OF MUNICIPAL WASTE DUMPS Daria G¹bka, Karol Wolski Wroclaw University of Environmental and Life Sciences, Wroclaw, Poland INTRODUCTION Recently, we have been observing the effects of the years lasting human activity which resulted in environmental pollution exemplified, among others, by devastated and degraded areas which have to be subjected to land reclamation processes. Restoring the value of soils by special biological treatments has become a challenge to contemporary scientists. Crucial for the successful soil reclamation is an appropriate selection of plants able to thrive in the extremely unfavorable habitat conditions since they often have to face pollution of all environmental components, namely water and air pollution, as well as soil contamination. Grasses have proved to be very advantageous plants regarding biological reclamation of soil as they have low nutrition requirements, high resistance to drought and frost as well as tolerance of salinity and heavy metals. They also posses an ability to incorporate toxic substances into their biomass and a considerable immunity to plant diseases and pathogens. Grasses do successfully fulfill three basic functions of reclamation process: antierosion, soil-building and decoration. Yet, in spite of all the features mentioned above, the environment is sometimes polluted to such a degree that additional introduction of the so-called biopreparations, also known as natural biological stimulators, seems to be fully justified. Biopreparations consist solely of natural substances like free amino acids in their biologically active form, oligoproteins, soluble humates, plant extracts, growth factors, selected microorganisms, valuable nutrients, probiotic and enzymatic factors, essential for many biological functions. Because of their rich composition biopreparations are of high quality and have a wide spectrum of action. Biopreparations act on the root system increasing its capacity of nutrient absorption from soil or any substrate, they possess chelating properties for microelements which become easily available for plants thus decreasing a risk of various forms of chlorosis, they stimulate and nourish week and stressed plants. They fulfill a useful function of carriers in relation to mineral elements and fertilizers, and increase plants resistance to climate and environmental stresses. The objective of this study was to evaluate the effect of the applied biopreparations on early growth of grass species destined for reclamation of municipal waste dump slopes at Swojec in Wroclaw.
9 8 MATERIALS AND METHODS The experiment was established on 19 th April 2007 at the northern slope of municipal waste dump at Swojec in Wroclaw (Fig. 1), which is located in the eastern part of the city, in the river Widawa basin. This dumping ground has been utilized since 1973 at the former clay pit, on the area of permeable bedding, without any protection against pollution of ground waters with effluents from waste dump. In 1991 exploitation of this municipal dumping ground ended and the stage of dump technical reclamation was completed [Account 2005]. Climate conditions (air temperature and rainfall) on the area of municipal dumping ground in 2007 and in the years are shown in table 1. THE EXPERIMENTS Doœwiadczenia THE MUNICIPAL WASTE DUMPS Sk³adowisko odpadów komunalnych FIGURE 1. LOCATION PLAN MUNICIPAL DUMPING GROUND AT SWOJEC IN WROCLAW WITH EXPERIMENTAL PLOTS SOURCE: OWN STUDY. Rysunek 1. Sk³adowisko odpadów komunalnych Swojec we Wroc³awiu z lokalizacj¹ za³o onych doœwiadczeñ ród³o: badania w³asne.
10 9 TABLE 1. WEATHER CONDITIONS DATA FROM WEATHER STATION AT SWOJEC IN WROCLAW Tabela 1. Dane meteorologiczne ze stacji Wroc³aw Swojec MONTH Miesi¹c 0 AVERAGE DAILY TEMPERATURE [ C] IN PARTICULA R MONTH IN Œrednia dzienna temperatura [ C] z miesi¹ca w 2007 r. A VERAGE DAILY TEMPERATURE [ 0 C] INTHE YEARS Œrednia dzienna temperatura [ C] za lata T OTAL MONTHLY PRECIPITATION [ mm] IN 2007 M iesiêczna suma opadów [ m m] w 2007 r. TOTAL PRECIPITATION [ mm] IN THE YEARS Suma opadów [mm] w latach SOURCE: OWN STUDY. ród³o: badania w³asne. IV V VI VII VIII IX , , Agriculturally used areas can be found in the vicinity of the dumping ground (arable land, meadows and pastures). The south-west part of waste dump, however, adheres to the area of stores and warehouses. Soil grain size analysis proved the occurrence of silty soil (sand silt) featuring advantageous protective properties (sorption of contaminants) [Account 2002]. The whole object of investigation was divided into two experiments with different sowing rates (1 or 2 caryopsis/cm 2 ). The experiments were established according to a split-plot design with two factors (1) particular grass species and cultivars, (2) biopreparations applied (Greenflorid, GreenBio) to be compared to an untreated control (Tab.2). TABLE 2. EXPERIMENTAL SCHEME MUNICIPAL WASTE DUMP AT SWOJEC IN WROCLAW Tabela 2. Schemat doœwiadczeñ na sk³adowisku odpadów komunalnych Swojec we Wroc³awiu SPECIES Gatunek Festuca arundinacea Lolium perenne Festuca rubra Poa pratensis Festuca ovina CULTIVAR Odmiana `ASTERIX` `INKA` `PINIA` `ADIO` `NIMBA` `ALICJA` `NANDU` `NONI` `WITRA` SOURCE: OWN STUDY. ród³o: badania w³asne. I EXPERIMENT 1 CARYOPSIS/ cm 2 doœwiadczenie 1 ziarniak/c m I 2 CONTROL APPLIED (UNTREATED) BIOPREPARATION Obiekt kontrolny Biopromotor II EXPERIMENT 2 CARYOPSES/ cm 2 I doœwiadczenie 2 ziarniaki/c m I 2 CONTROL (UNTREATED) Obiekt kontrolny APPLIED BIOPREPARATION Biopromotor
11 10 Grass species and cultivars were selected to meet the requirements of sowing on difficult habitat areas (land reclamation species) and biopreparations introduced were meant to support plant development, as well as to increase grass resistance to unfavorable environmental conditions. The experiments involved different sowing rates and various rates of biopreparation according to the species sown. To assess efficiency of biopreparations on seedling emergence for two sowing rates (unit/m 2 ) results of observations were subjected to analysis after 4 and 6 weeks (biopreparations versus untreated control). RESULTS AND DISCUSSION Observations of the early growth of grass on experimental plots (biopreparations vs untreated control) showed that biopreparations fulfilled their function which was to contribute to a better growth and development of plants and had beneficial effects on their vegetation in the unfavorable environmental conditions. Natural humic substances, as components of soil organic matter and included in biopreparations, play a vital role in soil fertility and environmental quality and are beneficial for plant growth. They increase fertilizer efficiency or reduce soil compaction. Our work indicated that plant stimulation provided by increased micro-elements iron and zinc availability, in the presence of humic substances, was the most important factor. Biopreparations facilitate uptake of trace metals like iron and zinc and, at the same time, provide simple and less expensive methods of plant nutrition eliminating or reducing the risk of environmental contamination. Investigation by Clapp et all  confirmed such an action of biopreparations and proved the improvement of grass growth by humic substances. In Agrostis stolonifera peat extract addition caused the increase in root and shoot dry mass from 46 to 65%, with the more pronounced increase being observed in roots. Soil organic matter is also a source of nitrogen and other nutrients. Kerek et al.  proved that nitrogen can be found in soil mobile fraction. Nitrogen content in soil mobile fraction increases as the sward age increases. The mentioned preparations affect soil microbiological activity and therefore, the root zone. Due to that fact the quality of grass becomes significantly ameliorated. Soil microorganisms make use of biopreparations and consequently, the visual quality of grass is considerably improved after application of biopreparations and this improvement is not a result of a nutrients source. Nevertheless, these observations do not exclude the possibility that biopreparations can have additional ability to improve the plant tolerance to excessive drought and moisture [Mueller 2005]. Soil physico-chemical properties affect a behavior and reproduction of Listeria and Yersimia bacteria. Such factors as high content of humic acids and fulvic acids in humus, as well as high temperature of soil stimulate bacteria reproduction in soils. High content of humus, predominance of humates and low temperature of soil inhibit bacteria development, which was proved by Sidorenko et all . The research carried out by Zhang revealed an effect of temperature on activity of enzymes in two cultivars of Cynodon dactylon. Catalase and peroxidase activity decreased in response to cold acclimation in both cultivars. The cold acclimation triggered
12 accumulation of sugars and proline in grass and affected structure of carbohydrates and protein, but this effect was cultivar-dependant [Zhang et al. 2006]. According to Duo Li An et al.  the addition of EDTA significantly increased heavy metal accumulation in grasses The concentrations of heavy metals in Lolium perenne and Festuca arundinacea increased with the increasing EDTA supply. The EDTAinduced increase in the accumulation of heavy metals in turfgrass was plant and metalspecific. Lolium perenne had a relatively high ability to accumulate Cr, Ni, and Zn. Additionally EDTA caused increase in the chlorophyll and proline contents and improved caryopsis germination [Duo Li An et al. 2005]. Amino acids and peptides constitute an essential part of biomolecules which form biostimulants. Cambri  carried out his research using Aminoplast preparation which, among others, diminishes yield losses resulting from stress conditions due to abiotic factors (salinity, extreme temperature, drought), especially in the crucial stages of plant growth, e.g. during flowering, which consumes a considerable amount of energy, therefore, the important processes taking place in plants can be easily disturbed. Aminoplast applied to plant roots in the conditions of soil salinity completely prevented inhibition of plant growth resulting from a high salt concentration. There are other reports confirming normalization of such plant physiological parameters as contents of the following elements: K, Na, Cu, Zn and Fe, as well as stomatal conductance, CO 2 absorption or transpiration which in untreated plants usually undergo negative alterations. Application of particular amino acids does not ensure balanced improvement of gene expression and only synergetic amino acids activity brings about a satisfactory result. Investigation involving Aminoplast, which is amino acids solution, proved that activity of this preparation is due to stimulation of plant defensive reactions [Cambri 2008]. Growth regulators are organic substances which, applied in low amounts, can stimulate, inhibit or modify in other ways physiological processes taking place in plants. Investigation by Starczewski et al.  suggest that an appropriate selection of grass cultivars, as well as introduction of growth regulators can markedly decrease the costs of plant cultivation and labor input without lowering the quality of lawn grass. In the experiment at Swojec the most important reactions resulting from biopreparations application appeared 6 weeks after sowing, while in the experiment with two caryopses per one area unit (cm 2 ) biopreparation activity was recorded after 4 weeks. Generally speaking, it takes biopreparations about 4 weeks to reveal their activity. The first assessment of seedling emergence in experiment 1 did not reveal any activity of biopromotors since for all grass cultivars the number of units per 1 m 2 was higher for the control. Yet in experiment 2 several grass cultivars showed higher values on the biopromotor treated plots as compared to the control one: Festuca arundinacea `Asterix`, Festuca rubra `Adio`, Festuca ovina `Noni` and `Witra`. The most evident results could be observed in both experiments 6 weeks after sowing, because grass seedlings emergence was definitely higher on the objects with biopromotors in comparison to the control treatment. For the following grass cultivars the higher emergence values (unit/m 2 ) were recorded: Festuca arundinacea `Asterix`, Lolium perenne `Pinia`, Festuca rubra `Adio`, 11
13 12 TABLE 3. SEEDLING EMERGENCE OF SELECTED GRASS SPECIES 4 AND 6 WEEKS AFTER SOWING FOR TWO SOWING RATEST ( 1 OR 2 CARYOPSES/ c m 2 ) 2 T abela 3. Wschody traw po 4 6 tygodniach od za³o enia doœwiadczenia dla ró nych norm wysiewu (1 lub 2 ziarniaki/cm ) SPECIFICATION Wyszczególnienie SPECIES Gatunek Festuca arundinacea Lolium perenne Lolium perenne Festuca Festuca Poa Poa Festuca Festuca rubra rubra pratensis pratensis AVERAGE Œrednia ovina ovina L SD a = 0.05 AFTE R 4 WEEK S N IR a = po 4 tygodniac h CULTIVAR odmiana `ASTERIX` `INKA` `PINIA` `ADIO` `NIMBA` `ALICJA` `NANDU` `NONI` `WITRA` L LSD a = 0.05 AFTE R 6 WEEK S N IR a = po 6 tygodniac h SOURCE: OWN STUDY. ród³o: b adani a w³asne. CONTROL ( UNTREATED ) Obiekt kontrolny 1 CARYOPSIS/ c m ziarniak/cm APPLIED BIOPREPARATION Biopromotor SOWING RATE Norma wysiewu CONTROL ( UNTREATED ) Obiekt kontrolny 2 CARYOPSES/ c m ziarniaki/cm APPLIED BIOPREPARATION Biopromotor CONTROL ( UNTREATED ) Obiekt kontrolny AVERAGE Œrednia APPLIED BIOPREPARATION Biopromotor WEEK Tydzieñ , , , i
14 Festuca ovina `Witra`, while in experiment 2 higher values characterized : Festuca arundinacea `Asterix`, Lolium perenne `Inka`, `Pinia`, Festuca rubra `Adio`. Inhibition of grass growth under the influence of biopreparation in experiment 1 was recorded for the following cultivars: Lolium perenne `Inka`, Festuca rubra `Nimba`, Poa pratensis `Alicja`, `Nandu`, Festuca ovina `Noni`, and in experiment 2 for Festuca rubra `Nimba`, Poa pratensis `Alicja`, `Nandu`, Festuca ovina `Noni`, `Witra`. Increased sowing rate accompanied by introduction of a biopreparation resulted in growth inhibition in some grass species and their cultivars (Tab. 3). The analysis of the weather conditions in 2007 allows to notice that high temperature in July affected grass seedlings emergence as 6 weeks after sowing a smaller number of units/m was determined than after 4 weeks on the object treated with biopromotor (Festuca arundinacea `Asterix`), which can be explained by the effect of air temperature on the biopreparation activity. Not only high but also low temperature affects the activity of microorganisms, components of biopromotors, either stimulating or inhibiting their activity. In July, i.e. 6 weeks after grass sowing, the highest amount of rainfall was recorded at the experimental site. However, April, when sowing took place, featured the lowest amount of rainfall (2,7 mm), considerably less than the average value (30,5 mm) recorded for the years The latter parameter could affect the early stage of grass growth and result in reduced growth of some grass species, among others Poa pratensis. Biopreparation activity depends both on grass species and cultivar, as well as on a sowing rate and weather conditions. The factors mentioned above can affect biopreparation activity in either positive or negative way. CONCLUSIONS 1. The effect of biopreparations becomes apparent about 4-6 weeks after their application, when their activation in soil is established. 2. Biopreparation activity depends on grass species and cultivar, as well as on a sowing rate and weather conditions. 3. In spite of unfavorable ambient conditions biopreparations applied in this investigation positively affected growth and development of the following grass species and cultivars: Festuca arundinacea `Asterix`, Lolium perenne `Pinia`, Festuca rubra `Adio`. 4. In grass species and cultivars such as: Lolium perenne `Inka`, Festuca rubra `Nimba`, Poa pratensis `Nandu`, Festuca ovina `Noni`, biopreparations inhibited plant growth, especially at increased sowing rate. REFERENCES Account with monitoring research among of the municipal waste dumps Swojec in Wroclaw in year. 2005: Arcadis Ekokonrem, Wroclaw. Account with monitoring research number 5 among of the municipal waste dumps Swojec in Wroclaw. 2002: Arcadis Ekokonrem. Wroclaw, Department of Environment protection and Landscape Development in Wroclaw. 13
15 14 Cambri D. 2008: Amino Acids: the scientific basis of the biostimulation. [In:] Materia³y konferencyjne abstrakty. SGGW, Warszawa. Clapp C.E., Chen Y., Hayes M., Cline V., Palazzo A., Molina J., White D., Baker J.M. 2002: Humic substances for enhancing plant growth. International Humic Substances Society Conference. p University of Minnesota, USA. Duo Li An., Gao YuBao, Zhao ShuLan 2005: Heavy metal accumulation and ecological responses of turfgrass to rubbish compost with EDTA addition. J. Integ. Plant Biol., 47(9): Kerek M., Rhae A., Drijber R., Gaussoin E. 2003: Labile soil organic matter as a potential nitrogen source in golf greens. Soil Biol. Biochem., col. 35, No. 12, s Mueller S.R., Kussow W.R. 2005: Biostymulant influences on turfgrass microbial communities and creeping bentgrass putting green quality. Hort. Sci., 40(6): Report with monitoring research among of the municipal waste dumps Swojec in Wroclaw in year. 2005: Arcadis Ekokonrem, Wroclaw. Report with monitoring research number 5 among of the municipal waste dumps Swojec in Wroclaw. 2002: Arcadis Ekokonrem, Wroclaw. Sidorenko M., Buzoleva L.S., Kostenkov N.M. 2006: The effect of soil properties on the preservation and reproduction of Listeria and Yersinia. Biology Soil Science Institute DVO RAN. Vladivostok, Russia. Pochvovedenie, (2): Starczewski K., Affek-Starczewska A., Jankowski K. 2008: Ryegrass turf grasses regrowth rate, depending on the applied phytohormones. [In:] Materia³y konferencyjne abstrakty. SGGW, Warszawa. Zhang X.Z., Ervin E. H., LaBranche A.J. 2006: Metabolic defense responses of seeded bermudagrass during acclimation to freezing stress. Crop Sci., 46(6):
16 15 EFFECT OF ASAHI SL BIOSTIMULATOR ON ORNAMENTAL AMARANTH (AMARANTHUS SPP.) PLANTS EXPOSED TO SALINITY IN GROWING MEDIUM Mariola Wrochna 1, Barbara ata 1, Bo enna Borkowska 2, Helena Gawroñska 1 1 Warsaw University of Life Sciences, Warsaw, Poland 2 Institute of Pomology and Floricutlure, Skierniewice, Poland INTRODUCTION Plants cultivation in urban areas meets several difficulties, which mostly result from anthropopressure leading to acid rain, accumulation of heavy metals, soil degradation and accumulation of particulate matter in air [Zimny 2004, Gawroñski, Gawroñska 2007, Gawroñski et al. 2007]. In northern hemisphere there is also a problem of salinity due to application of sodium chloride for deicing roads [Wrochna et al. 2005, Wrochna 2007]. Soil salinity causes reduction of biomass accumulation and adverse changes in physiological and biochemical processes of plants. It was stated that salinity lead to the decrease of water potential in leaves (y w ), loss of turgor, drop of relative water content (RWC) [Morant-Manceau et al. 2004, Kacperska 2005] and decrease of photosynthesis rate, which results both from stomata closure and from toxic effects of Na + and Cl - ions on mesophyll cells and chloroplasts [Ueda et al. 2002]. In salinity environment, both the chlorophyll a fluorescence [Maxwell, Johnson 2000, Wrochna et al. 2007] and level of reactive oxygen species (ROS) increase [Bartosz 1995, Foyer, Noctor 2000], the enzyme activity of antioxidizing system changes [Munns 2002, Parida, Das 2005, Wrochna 2007] and accumulation of osmoprotectants increases [Chen, Murata 2002, GwóŸdŸ 2004, Wrochna 2007]. These adverse changes eliminate from cultivation a wide range of plant species sensitive to salinity and drought. Plants, which are able to impel a various protective mechanisms minimizing salt stress effects, have the possibility to grow in sites with increased salt concentration. Therefore, in addition to selection and breeding toward increased salinity tolerance also the attempts to use biostimulators are made, which by definition should improve plant protective mechanisms. It is believed that biostimulators, including Asahi SL, have a protective impact on plants thus, it seems important to study mechanisms underlying their mode of action. Results showed an increase of biomass accumulation by plants treated with biostimulators, which can be the effect of better development of both the above ground part and roots [Djanaguiraman et al. 2004, Krajewska, Latkowska 2008, Wróbel, WoŸniak 2008], and more effective water and ions uptake [Zhao, Oostherhuis, 1997, Maciejewski et al. 2008]. Biostimulators increase activity of antioxidizing system enzymes [Djanaguiraman et al. 2004], accelerate transport of electrons in photosynthesis process and flow of assimilates in a plant [Chitu et al.1998]. All the men
17 16 tioned above changes due to biostymulators application might have a protective impact on processes disturbed by salinity. The study objective was the assessment of whether and to what extent Asahi SL stimulator has a protective effects on plants growing under increased salt concentration in growing medium. MATERIALS AND METHODS The study object were plants of 3 varieties of ornamental amaranth: Amaranthus paniculatus L. `Copper Mountain` and `Monarch` and Amaranthus caudatus L. `Pony Tail`. Experiments were carried out at the Laboratory of Basic Research of Horticulture, Faculty of Horticulture and Landscape Architecture, Warsaw University of Life Sciences (WUSL), except for: (1) measurements of chlorophyll fluorescence a conducted in Physiology Department of the Institute of Pomology and Floriculture in Skierniewice, (2) determination of ions concentration in plant material carried out in Center of Analysis of WULS. Two experiments were carried out under controlled conditions in Faculty greenhouses. In both experiments a uniform four-week-old seedlings of amaranth were transplanted either to substrate or hydroponics culture and agricultural practices were applied as needed. EFFECTS OF ASAHI SL BIOSTIMULATOR ON BIOMASS PRODUCTION AND ACCUMULATION OF SELECTED IONS In this experiment, seedlings of `Copper Mountain` and `Pony Tail` were planted in pots (O16 cm), filled with 1 dm -3 of peat and sand mixture (3:1 v/v). The substrate was prepared to have ph 6.5 (using 12 kg of agricultural chalk per 1 m 3 ), and the content of mineral elements respectively to the level of: N-310 mg. dm -3, P-100 mg. dm -3, K-400 mg. dm -3, Mg 110 mg. dm -3 (MIS 3 p. A). Microelements were added in the form of MIS 3. Salinity of this substrate reached 2 g NaCl. dm -3 (unit used in agriculture). Tenweek-old plants were treated with deicing salt (98% NaCl) in concentrations of 100 and 240 mm, and half of plants were sprayed with 0.1% solution of Asahi SL. After 14 days plants were harvested and fresh weight of above ground part was recorded. Immediately after harvesting, the plants were oven dried for 24 h at 105 o C and then for 72 h at 75 o C. After drying and weighing the plant material was grinded in a grinder (W 1). Sub-samples (1 g) of powdered material were dry mineralized in muffle stove (Nobertherm) in temperature of 450 o C, dissolved in 0.5 mole HCI. The concentration of Na +, K +, Ca 2+, Mg 2+ was determined by flame atomic absorption spectrometry FAAS and P 3+ by the method of ICP-AES. Level of Cl ions was determined by Moohr`s titration method after extraction with water from 1 g of dry, powdered plant material and total nitrogen with Kieydahl method.
18 EFFECT OF ASAHI SL BIOSTIMULATOR ON SELECTED PHYSIOLOGICAL AND BIOCHEMICAL PROCESSES In this experiment, seedlings of `Copper Mountain`, `Monarch` and `Pony Tail` were placed in hydroponics, where the plants grew for subsequent 6 weeks in Hoagland`s nutrients solution, in which salt concentration was 3 g NaCl dm -3. After that time, half of plants was sprayed with 0.1% of Asahi SL preparation and 17 or 50 mm of NaCl was added to the medium. After 16 days of treatment, the plants were harvested, dried and weighed as in experiment 1 st. Data for the following processes and parameters were collected during the experiment: efficiency of photosynthetic apparatus based on: chlorophyll content (Chlorophyll Content Meter CCM-200, Opti-Science, USA) and chlorophyll a fluorescence i.e.: Yield, F v /F m, Fv/Fo, Fo/Fm, qp, qn, and NPQ (MINI-PAM, Walz, Germany), Relative Water Content, membrane integrity (conductometrically, MultiLevel 1, WTW, Germany), level of superoxide anion-radical (O 2 - ) by spectrometry (UV/VIS Spectrometer Lambda Bio 10, Perkin Elmer), activity of ascorbate peroxidase (APX) [Nakano, Asada 1987], catalase (CAT) [Beers 1952], glutation reductase (GR) [Foyer, Halliwel 1976] by spectrometry (UV/ VIS Spectrometer Lambda Bio 10, Perkin Elmer). Level of O 2 -, and activity of APX, CAT and GR were measured at wave lengths of 580, 240, 290 and 340 nm respectively. In both experiments the controls were plants without NaCl and not treated with Asahi SL. Results were subjected to statistical analysis using the two- and three-factorial analyses of variance ANOVA of Statgraphics 4.1 Plus. Significance of differences between the treatments was assessed with t-student test at a The data present means from five (experiment 1) or four (experiment 2) biological replications (single plant) and each measurement was repeated twice on separate sub-sample. RESULTS Plant exposure to salinity in growing medium resulted in adverse changes manifested by strong wilting followed by drying of edges of leaf blades particularly in older leaves. Besides, the visual symptoms of salt impact, also a significant adverse effects of increased salinity on all studied physiological and biochemical processes were recorded. Application of Asahi SL biostimulators, in general, decreased negative effects of salt stress (Photo 1). 17
19 18 A B Asahi SL +ASAHI SL Asahi SL +ASAHI SL PHOTO 1. EFFECT OF ASAHI SL ON A. PANICULATUS `OPPER MOUNTAIN` PLANTS GROWING FOR 7 DAYS IN PRESENCE OF 100 (A) And 240 (B) mm of NaCl Fotografia 1. Wp³yw Asahi SL na roœliny A. paniculatus `Copper Mountain` rosn¹cych przez 7 dni w obecnoœci 100 (A) i 240 (B) mm NaCl EFFECT OF ASAHI SL BIOSTIMULATOR ON BIOMASS PRODUCTION AND SELECTED IONS ACCUMULATION Increase of salinity reduced accumulation of both fresh weight and dry matter while application of Asahi SL biostimulator, in most cases, increased it. Fresh weight increased mostly in Asahi SL treated plants of `Pony Tail` growing without salt (33% of untreated). However, in plants of `Copper Mountain` it was lower than in control (by ~5%) (Fig. 1). Accumulation of dry matter also increased after Asahi SL application, but in some treatments with Asahi SL it was also slightly lower (Fig. 2). Salinity of growing medium caused multiple increase in the amount of accumulated toxic Na + (even over 100 fold) and Cl - ions and changes in concentration of macroelements. Reaction of tested genotypes of amaranths on spraying with Asahi SL depended on the cultivar. Thus, in plants of `Pony Tail` decrease of Na + and Cl - accumulation was noted along with increase of accumulation of other tested ions, particularly in the highest applied salt concentration (Tab. 1). That resulted in smaller changes in proportion of K + /Na + than it was in case of the plants growing in salinity environment but not treated with Asahi SL (from to 0.74 in control and from to 0.97 after treatment with Asahi SL). Similar changes were recorded in proportion of sum of K + +Ca 2+ +Mg 2+ /Na + (Tab. 1). However, in plants of `Copper Mountain` application of Asahi SL led to increased accumulation of Na + and Cl - and greater reduction of proportions of K + /Na + from to 0.90 and of K + +Ca 2+ +Mg 2+ /Na + from to 0.76 (Tab. 1). Application of Asahi SL caused significant increase in calcium accumulation in plants of both varieties (Tab. 1).
20 19 TABLE 1. EFFECT OF ASAHI SL ON ION ACCUMULATION IN ABOVE GROUND PLANT PARTS OF ARE MEAN OF 6 REPLICATIONS (3 PLANTS WITH TWO MEASUREMENTS IN EACH) Tabela 1.Wp³yw Asahi SL na akumulacjê jonów w czêœciach nadziemnych roœlin szar³atu ozdobnego. Dane dwa pomiary z ka dej) CULTIVAR Odmiana COPPER MOUNTAIN PONY TAIl SALINITY Stê enie soli. [ mm NaCl ] CONTROL Kontrola A SAHI SL N-TOTAL % D.M. N-ca³kowity % s.m. ORNAMENTAL przedstawiaj¹ AMARANTH PLANTS. œrednie z 6 powtórzeñ (3 DATA roœliny Na Cl Ca K Mg P K:N a (K+Ca+Mg):N a m g g - 1 D.M. m g g - 1 s.m CONTROL Kontrola L SD a = 0,05 FOR ASAHI S L N IR a = 0,05 dla Asahi S L po L SD a = 0,05 FOR SALINIT Y N IR a = 0,05 dla stê enia sol i L SD a = 0,05 FOR CULTIVA R N IR a = 0,05 dla odmian y SOURCE:OWN STUDY. ród³o: badania w³asne