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Home Biotechnologies Increase of plant resistance to diseases, pests and stress with new biostimulants Regoplant, Stimpo and Radostim

Increase of plant resistance to diseases, pests and stress with new biostimulants Regoplant, Stimpo and Radostim

Application of growth regulators Regoplant and Stimpo on cucumbers and sweet peppers in a greenhouse "Agrosvit" Kamenka, Cherkasy region.
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Application of growth regulator Radostim on winter barley in a farm "Yastreb" Velyka Znamenka, Zaporozhye region.
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Application of growth regulator Radostim on wheat in a farm "Zlagoda", Rivnee region.
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In the greenhouse experiments the antipathogenic activity of new polycomponent biostimulants Regoplant, Stimpo and Radostim had been investigated at cultivation of different cultivars of winter wheat, soybean and corn plants on infectious backgrounds.

The best biological efficiency against phytopathogens was obtained at presowing seed treatment and spraying of crops in vegetation period with Regoplant (up to 98%) and Stimpo (up to 89%), less bioprotective efficiency was showed by Radostim (up to 74%) according to control (without treatment with biostimulants).

At 2nd generation of the wheat and chickpea plants (infected by pathogenic micromycetes of Fusarium L. genus without treatment with biostimulants) the increased resistance to pathogenic micromycetes of Fusarium L. genus was found too. Using DOT-blot hybridization method the considerable difference between mRNA of control winter wheat and chickpea seedlings and small regulatory si/miRNA of experimental seedlings (obtained from seeds of the 1st generation of plants infected by pathogenic micromycetes of Fusarium L. genus and treated with biostimulants) was found.

It is proposed that indicated difference connected with partial reprogramming of plant cell genome under the impact of biostimulants – inducers of si/miRNA synthesis with antipathogenic properties.




INTRODUCTION.

Development of economically feasible and environmentally friendly agricultural technologies able to provide stability of agricultural ecosystems, to promote wide use of biocontrol, and to guarantee improvement of quality, is one of the challenges of modern agriculture. Pests (insects, mites, and nematodes), diseases (bacteria, viruses, fungi, nematodes), and weeds cause a significant yield reduction in agricultural production worldwide. According to the Food Agricultural Organization (FAO), the global annual crop losses due to pests, diseases, and weeds reached 20-25% [Stevens and May, 2009].

European corn borer, cutworms, wireworms, grabs cereal flies, aphids, root-knot and leaf weevils, soybean pod borer, spider mites, trips, rape beetles, flea beetles, stink, bugs, and white butterflies belong to the most widespread and dangerous pests that cause significant reduction of yield of important agricultural crops such as corn, wheat, barley, soybean, and rape.

The problem of plant protection against widespread fungi (e.g., Fusarium spp., Fusarium graminearum, Fusarium oxysporum Ciceriae, Cercospora spp., Ascohyta spp., Perronospora spp., Blumeria spp., Pucinia spp., Sclerotinia spp., and Verticillim spp.); bacterial (Pseudomonas spp.), viral (Potyvirus spp.) and phytoparasitic nematodes (such as Heterodera schachtii, Meloidogyne incognita, Globodera rostochiensis, Ditilenchus dipcasi, Rotylenchulus reniformis, Tylenchulus semipenetrans) and other diseases is also economically important [Stevens and May, 2009].

Non-chemical crop protection is important component of sustainable crop production. Development of such compounds was based on achievements in modern microbiology, mycology, biotechnology, soil science, and plant protection.

Long-term research and wide practical application of Stimpo, Radostim, Regoplant and Biolan – the new polycomponent biostimulants developed by the National Enterprise Interdepartmental Science & Technology Center Agrobiotech, Natl. Acad. of Sci. and Min. of Ed. and Sci. of Ukraine, showed that these biostimulants match with economical and environmental demands of modern agriculture. These biostimulants have bioprotective and regulatory effects that are achieved by synergistic action of metabolism products (mixtures of amino acids, carbohydrates, fatty acids, polysaccharides, phytohormones, and microelements) of root fungus-endophyte products of ginseng Panax Ginsed M. as well as of soil streptomycete Streptomyces avermitilis metabolites [Ponomarenko et al., 2010] with phytostimulating, antiparasitic and antipathogenic effect.

In our molecular-genetic experiments, we have showed that positive effects of the above-mentioned biostimulants were revealed in quantitative and qualitative changes in gene expression as a consequence of plant cell genome reprogramming with biostimulants [Tsygankova et al., 2010; Tsygankova et al., 2011].

We have found also [Tsygankova, Ponomarenko et al., 2012; Tsygankova, Stefanovska et al., 2012] that these biostimulants significantly enhanced plant resistance to different pathogens due to stimulation of the synthesis of cellular small regulatory si/miRNA that participate in RNAi (RNA interference) process. This process called posttranscriptional gene silencing (PTGS) was found in plants, animals, and fungi [Filipowicz et al., 2005; Bakhetia et al, 2005; Katiyar-Agarwal, 2006]. Small regulatory si/miRNA with 22-24 nt size plays a leading role in silencing: together with site-specific endo- and exonucleases of RNA-induced silencing complex (RISC), it blocks (silences) translation of variable cell mRNA with imperfect structure as well as mRNA of pathogens and parasites, or enzymatically cleaves these target mRNA molecules causing its degradation [Vaucheret, 2006; Voinnet, 2008; Zhang, 2007].

The purpose of our work was verifying the possibility of increasing of the plant resistance to pathogens and parasites by above mentioned biostimulants and determination of genetic mechanisms of these biostimulants’ post-action (i.e. effect of inheritance of increased with biostimulants plant resistance to pathogenic and parasitic organisms).

MATERIALS AND METHODS

In the greenhouse experiments we compared efficiency of biostimulants in combination with standard insecticides in controlling ground beetle Zabrus tenebrioides, turnip moth Scotia segetum and Chloropidae spp., and wheat nematode Anguina tritici. The experiments on comparative efficiency of biostimulants and standard insecticides were conducted in pots of 25 cm x 25 cm size; each experiment was replicated four times. Soil samples were contaminated separately by nematode, ground beetle, and turnip moth. We used winter wheat of Dalnytska cultivar, soybean of Arcadia Odeska cultivar, and hybrid corn of Kobza МV cultivar. Seeds were sown in a box after treatment with biostimulants. 50 plants were studied in the experiments on determination of the amount of damaged plantlets (in pcs/m2) and biological efficiency (in %).

The effect of biostimulants on stability, productivity, and quality of obtained seeds, as well as on wheat, soybean and corn plants’ resistance to infections was determined. The experiments with winter wheat, soybean and corn rot and mildew diseases caused by pathogens, such as Mucor spp., Rhizopus spp., Aspergillus spp., Penicillium spp., and Trichothecium roseum, were conducted on the artificial infectious backgrounds. Efficiency against pathogens was studied at low level of spores, i.e., 0.1 g of spores per 1 kg of seeds, and at high level of spores, i. e., 1 g of spores per 1 kg of seeds.

In laboratory experiments we studied also a post-effect of biostimulants on the second generation of winter wheat plants of 2 cultivars: Lastivka and Princess Olga and chickpea Cicer arietinum L plants of 2 cultivars: Rosanna and Triumph. These plants were not treated with biostimulants; however, they were obtained from seeds of the first generation of winter wheat and chickpea plants that were infected by pathogenic micromycetes Fusarium graminearum and Fusarium oxysporum f. ciceris and were not treated with biostimulants (control plants) and treated with biostimulants (experimental plants). In our experiments we used control and experimental seeds (obtained from the first generation of control and experimental plants) that were sprouted in Petri-dishes on a filter paper wetted by distilled water (without adding biostimulants) on an infected background (at the presence of pathogenic micromycetes Fusarium graminearum and Fusarium oxysporum f. ciceris). Specificity of biostimulants post-effect was determined according to: 1) integral indices of growth and development of control and experimental 7-day old seedlings; 2) molecular-genetic indices: the difference in the level of homology between the basic constituents of plant immune system – si/miRNA isolated from experimental 7-day old seedlings to mRNA of control seedlings. Small regulatory si/miRNA was isolated from experimental seedlings by our elaborated method [Tsygankova, Andrusevych, 2011].

RESULTS AND DISCUSSION

We tested bioprotective properties of new biostimulants at cultivation of winter wheat, soybean, and corn on the infected backgrounds. The results were compared with the effect of modern pesticides produced by leading agrochemical companies such as Alpha-Cypermethrin insecticide, Yunta Quattro insecto-fungicide, Lamardor, Selest Top, Imidacloprid, Terios and Microplant micronutrients. Use of biostimulants in combination with chemical pesticides caused increase of plant resistance against different diseases caused by microbial pathogens. Biostimulants reduced phytotoxicity of chemical protectants and stimulated immune reactions of plants. As a result, commercial grain yield and seed material quality improved. The bioprotective effect of biostimulants was found sufficiently high at their use on crops infected by nematodes, ground beetle, turnip moth, and chloropid flies hat (Table 1). biostimulants effect did not exceed an effect achieved at the use of insecto-fungicides, e.g., Yunta Quattro and Selest Top. However, the level of efficiency shown by Regoplant against wheat nematode, ground beetle, and turnip moth, and by all the biostimulants studied against chloropid flies was just set as high taking into account their economic effect and environmental safety.

Regoplant and Stimpo showed also antipathogenic activity against causative agents of winter wheat (Odeska semidwarf cultivar) rot and mildew (Table 2). However, we do not consider the use of these biostimulants as alternative to chemical pesticides reliable, especially on the highly infected background. On the low infected backgrounds, the biostimulants application is quite possible, taking into account the level of their potential efficiency.

We studied also Regoplant and Stimpo bioprotective effect on soybean and corn plants infected by dangerous pathogens of soybean and causative agents of rot and mildew of corn (Table 2 and Table 3). We found that biostimulants has a positive effect on growth and development of soybean and corn seeds. They reduced the infection impact on seed development and commodity grain quality.

Our laboratory experiments demonstrated the inheritance of wheat and chickpea plants resistance to pathogenic organisms. We found that plants of the 2nd generation which were not treated with biostimulants, maintain high viability which is close to the results obtained on the 1st generation of plants treated with biostimulants on infected background [Tsygankova, 2012]. The molecular-genetic analysis by the DOT-blot method hybridization si/miRNA with mRNA populations [Tsygankova, 2010; Maniatis, 1982] showed high level of homology between immuno-protective small regulatory si/miRNA and mRNA of experimental plants and lower level of homology in respect to mRNA of control plants (Table 4). We called this effect ‘quasi-heterosis’. It was found that Regoplant, Stimpo and Biolan strongly increased growth rate of heterosis plants as well as resistance to pathogenic organisms. We concluded that principal mechanism of these biostimulants in plant cells includes almost twofold increasing of the synthesis (abundance) of small regulatory si/miRNA, which has antipathogenic properties.

CONCLUSIONS

In the greenhouse experiments the bioprotective activity of new РGRs: Regoplant, Stimpo and Radostim had been investigated at cultivation of different cultivars of winter wheat, soya and corn plants on infected backgrounds. It was found that presowing treatment of plants and spraying of crops in vegetation period with biostimulants Regoplant, Stimpo and Radostim increased plant viability and resistance to different phytopathogens. In the laboratory experiments was shown that the seedlings of the 2nd generation of winter wheat and chickpea plants (that were grown on infected background without treatment with biostimulants), obtained from the seed of the first generation of plants (that were grown on infected background and were treated with biostimulants: Regoplant, Stimpo and Biolan), show high viability and resistance to pathogenic micromycetes of Fusarium L. genus due to integral indexes of germination and growth comparatively with control seedlings. The molecular-genetic indexes -% homology si/miRNA of experimental plants to mRNA of control plants testify that at embryogenesis during forming of plant seeds there is observed reprogramming genome of plant seed embryos through the way of “switching on” genes of the antipathogenic si/miRNA which synthesis is induced under biostimulants’ action.

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Contact Details

Ponomarenko Sergiy P.
Organization: State Enterprise ISTC Agrobiotech of National Academy of Sciences of Ukraine and Ministry of Education & Science of Ukraine
Adress: 50, Kharkivske shose, Kyiv, 02160 Ukraine.
+380-44-559-66-17, 558-07-10.
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