Read Erika Greipel´s 2016 (Albitech Ltd.) abstract here!

Erika Greipel Bioengineer M.Sc., researcher Head of laboratory at Albitech Biotechnological Ltd. E-mail: greipel.era@gmail.com Phone: 00 36 20 424 8668

Measurement of intra- and extracellular cytokinin content of algae cultures and application of Scenedesmus sp. cultures for plant growth promotion – resulted the new foliar fertilizer of Albitech Ltd.

Erika Greipel¹, Péter Futó¹, Zsuzsanna Bencsik¹, Attila Bencsik¹, Annamária Kósa², Péter Nyitrai³, Prof. Béla Böddi², József Kutasi¹

¹: Albitech Biotechnological Ltd.
²: Eötvös Loránd Science University Department of Plant Anatomy
³: Eötvös Lóránd Science University Department of Plant Physiology and Molecular Biology

Abstract

Foliar fertilization is a widespread procedure in agriculture for applying inorganic fertilizers or other substances on the foliar surface of plants in order to enhance crop yield. Microalgae suspensions has been used for a long time for the purpose of phytostimulation, owing to their hormone-producing ability. In our work cytokinin content of Scenedesmus sp. BEA D01_12, Chlorella minutissima K2012 41 cultures were investigated. Prolification of algae was stimulated by the addition of Azospirillum brasilense NCAIM 001411 bacterial culture in a low concentration. Scenedesmus sp. cultures were also tested in greenhouse pot trials and small pot field studies. According to the results the algae suspension was effective on fruit, vegetables and cereal crops as well. Cucumber, tomato, apple, sweet pepper and sunflower have shown a significant increase of crop yield. Increased the number of flowers in sweet pepper and tomato, the root weight of sweet pepper, plant height and the number of seeds in maize and the oil content of the seeds in sunflower. In horticulture significant increase in the number of leaves and flowers was observed.

About the author

Born in Budapest (Hungary) and graduated B.Sc. and than M.Sc. degree in Bioengineering at Budapest University of Technology and Economics. She worked on the topic of Heterocypris incongruens immobilization test further development and application of micro-pollutants contaminated testing samples of liquid and solid phase and Isolation of filamentous fungi with cellulase activity from soil samples, perform various enzyme activity assays after shake flask fermentations at the University. She took part a 6 week-long traineeship at the Fermentation plant of Gedeon Richter Plc. (pharmaceutical industry).

Beside M.Sc. courses in 2010 she started to work at Albitech Biotechnological Ltd., she coordinates the inoculum preparation and other manufacturing operations of microalgae-based foliar fertilizer formulation and quality control, moreover she is carrying out research activities with different microalgal and soil bacterial strains. Since 2014 she is PhD. student at Eötvös Lóránd Science University where she study the interactions between

Scenedesmus rubescens green microalga and Azospirillum brasilense soil bacterium.

About the company

Albitech Biotechnological Ltd. is a dinamically developing Hungarian enterprise established in 2007 aiming at microalgae research and development other microalgae based products for agricultural or other purposes.

The fact that algae are suitable for large-scale biomass production could lead to a breakthrough in the biotechnological industry. Our goal is to introduce a new branch of the plant biotechnological industry in Hungary to be utilized in agricultural production by the screening and controlled, industrial-scale culturing of new algal strains capable of producing bioactive compounds. The first fruit of research was the microalgae based Algafix® phytostimulator, a growth-promoting spray-on which was has been authorized in 2010 and produced continuously in the photobioreactor plant at the company.

Several microalgae strains show phytohormone-like activity. Intensive auxin- and cytokinin-like plant growth hormone production has been observed in green algae species, recently. In agriculture, algae have been used as organic fertilizers or soil conditioners, however, interest has recently grown in their antimicrobial, anti-fungal and plant growth regulating (PGR) properties. The concentration of phytohormones in algae biomass range between 0.3 – 4,0 μg / kg wet weight which approximates that of higher-order plants. Industrial scale culturing of saltwater algae is not economical in Hungary, due to the treatment of large volumes of industrial wastewater with high salt concentrations. Our research confirmed that in optimized conditions, freshwater algae can produce and excrete plant growth promoting hormones, antimicrobial or antifungal substances in higher concentrations than saltwater algae and contain vitamins and trace-elements vital for plant health.

Technology-wise the axenic algal strains are selected and maintained by classic microbiological methods. The identified algal cell lines are preserved and cultured via closed-system sterile photofermentation. According to our new protocol the Scendesmus sp. BEA D01_12 is cultured in air lift photobioreactors. Production of industrial-scale volumes for agricultural use is implemented via scaling up. Laboratory-scale volumes of shaken liquid cultures are produced which go through culturing in several 2-10 l volume photobioreactors to reach 30 and 200 litre photobioreactors. Adequate light intensity is provided by high-power light sources to ensure proper growth rate. The production capacity has quadrupled in the past 4 years. A new technology developed in the last few years makes it possible to achieve a five-fold increase in the alga cell count.

In Albitech Ltd. there are currently six people working in our laboratories: Dr. József Kutasi PhD., co-owner and head of research, Attila Bencsik, co-owner and head of informatics and technology, two bioengineers: Erika Greipel MSc. (PhD-student at Eötvös Lórand Sicence University), Péter Futó MSc., an administrator, a laboratory and a fermentation assistant. Our young researchers have participated in both national and international conferences with oral and poster presentations. In the framework of granted research projects, our company has hosted bioengineering undergraduates from Budapest Technical University for traineeship and thesis work.

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¹ Research Department, Agricultural Research and Development Station of Braila, Sos. Vizirului, km. 9, Braila, Romania.

² Agrochemical Laboratory, BRAICOOP Agricultural Cooperative, No. 38, Imparatul Traian Street, Braila, Romania

³ Agriculture Department, “Dunarea de Jos” University of Galati, Engineering and Agronomy Faculty of Braila, No. 29, Calea Calarasilor, Braila, Romania

Abstract

Experience with bio-fertilizers included two sub-experiences,with maize and sunflower. Placing of experiences was performed by
the method of Latin,rectangle in three repetitions, on Braila Plain, Romania, in 2014 year. The biological products used were Bactofil and
Algafix, in four doses (V2 – Bactofil 285g / ha + Algafix 1.5 l / ha; V3 – Bactofil 500g / ha + Algafix 2 l / ha; V4 – Bactofil 571g / ha +
Algafix 2.5 l / ha; V5 – Bactofil 642,8g / ha + Algafix 3 l / ha). The control was variant without fertilizers. Biometric measurements were
performed throughout the growing period and at harvest were determined elements of productivity and average production to determine
optimal doses for increased production in terms of quantity and quality.

Keywords: bio-fertilizers, doses, corn, sunflower, yield.

INTRODUCTION

The importance of biological fertilization increases continuously to practice sustainable agriculture and decrease environmental pollution. The specific consumption of nutrients for the species studied is very big: corn consumed 27.5 kg N, P2O5 and 16.5 kg K2O 12.5kg and sunflower consumed 36.5kg N, P2O5 and 50.0 kg 17.5kg K2O per ton of main and secondary crop (Davidescu, 1999). In case of using organic products, these doses decreased significantly (Trifan, 2015). Maize and sunflower are very important in world agriculture, but requires knowledge about growing areas of hybrids and varieties, about fertilization and the sowing and harmonization of ecological factors with rational use of technological measures specific to each culture, while also improving the physic, chemical and biological parameters of soil and implementation of incentive income for farmers (Rinchita, 2008).

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Abstract

Foliar fertilization is a widespread procedure in agriculture for applying inorganic fertilizers or other substances on the foliar surface of plants in order to enhance crop yield. The efficiency of foliar fertilization can be 8-20% better than solid fertilizers. Certain axenic, high-concentration algal cultures have a growth promoting effect on field cultivations, due to nitrogen-fixation and production of growth-promoting hormone-like substances. These concentrated suspension of microalgae and/or cyanobacteria – or its extract – efficiently enhance growth of plants by increasing plant height and leaf surface, improving germination capacity of seeds, or even increasing the clorophyll content of leaves, or the starch, oil and protein concentration of the fruit.

Our product, Algafix® is a liquid culture of microalgae, containing phytohormones, macro- and microelements. For home gardens, greenhouses, agricultural and horticultural use for mono- and dycotyledon cultivars. It contains Scenedesmus rubescens/obtusiusculus, a freshwater green microalga isolated from the Lake Balaton, Hungary. Between it’s plant hormone producing capacity it produses different lipids and molecules with antioxidant capacity which can delay the scenescence of leaves. About agricultural experiments it can be used for cereals, sunflower, corn, vegetables, fruit-trees, grape and also for indoor plants. According to the patent HU 1200251 Scenedesmus strains are cultivated together with the (plant growth promotion rhizobacteriae) Azospirillum brasilense NCAIM (P) 001411(National Collection of Agricultural and Industrial Microorganisms )reached surprisingly higher algae cell number and more effective plant stimulating properties. These bacteria are able to produce auxins, in the first line indole-3-acetic acid whitch can stimulate the proliferation and hormone production of algae. In this way cultivated microalga culture has 10⁸cell/ml cell concentration, stimulate the germination, has antimicrobial and antifungal activity, furthermore in case of field plants stimulate the stem elongation and blossoming and increase the yield.

The cytokinine and gibberellin-like plant hormon production of axenic cultures of Scenedesmus sp. BEA(Banco Espanol de Algas) D01_12(Algafix®,  Patent HU 1200251) Chlorella minutissima K 2012 (Albitech collection) and Botryococcus braunii green  alga strains were investigated. In case of S. rubescens/obtusiusculus we found kinetin production an order of magnitude higher than in case of the other two strains. Trans-zeatin-riboside whitch is one of the most effective natural cytokinin was found in evapored extracts of B. braunii, C. minutissima and in S. obtusiusculus cultures as well. B. braunii cultures haven’t investigated in this field previously. It seems that the S. obtusiusculus, the strain of Algafix® foliar fertilizer is really suitable to use for regulating plants. These results can serve as a good basis for the development of algae-based foliar fertilizer products and for a better understanding of their mode of action.

Erika  Greipel, Zsuzsanna Bencsik, Péter Futó, József Kutasi:

Preparated  Cladophora  cellulose,  as a  substrate  of  cellulase fermentation

Abstract

Some filamentous algae collected from the sea are used for paper production. Moreover, it can serve as a raw material of cellulose production as a more efficient substrate than plant cellulose.  Moreover algae are currently exlioited by the food industry for a range of ingradients. The filamentous freshwater green algae Cladophora contents the cellwall cellulose mostly in the crystalline form. In contrast to the plant cell wall cellulose in this case the Iα whitch is more accessible to the enzymes is present in 63% while the Iβ form in 37%. Plants have reverse ratio making it difficult to process the fermentation plant cellulose. Cladophora cellulose forms gel structures at cellulose concentrations as low as 0.2% (w/v), whereas the lower threshold for the commercially available analog is around 1.5% (w/v) solids contents (Mihranyan 2011). It’s powder  could be a useful alternative to commercially available dispersible cellulose grades when very low quantities of stabilizing agent are required.

With the extracted cellulose from Chladophora the cellulase enzyme induction in soil bacteria were investigated for facilitating and to enhance the production of enzymes. The bacterial digestion was performed on dried and milled, but untreated, Cladophora fibers as well. The success of the experiment can be explained by that the algae cell wall doesn’t contain lignin, which would inhibite the access of cellulase enzymes to the untreated material.

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