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. 2019 Apr 17:10:483.
doi: 10.3389/fpls.2019.00483. eCollection 2019.

RNAi-Based Biocontrol of Wheat Nematodes Using Natural Poly-Component Biostimulants

Konstantin B Blyuss  1 , Farzad Fatehi  1 , Victoria A Tsygankova  2 , Liudmyla O Biliavska  3 , Galyna O Iutynska  3 , Alla I Yemets  4 , Yaroslav B Blume  5
Affiliations

RNAi-Based Biocontrol of Wheat Nematodes Using Natural Poly-Component Biostimulants

Konstantin B Blyuss et al. Front Plant Sci. .

Abstract

With the growing global demands on sustainable food production, one of the biggest challenges to agriculture is associated with crop losses due to parasitic nematodes. While chemical pesticides have been quite successful in crop protection and mitigation of damage from parasites, their potential harm to humans and environment, as well as the emergence of nematode resistance, have necessitated the development of viable alternatives to chemical pesticides. One of the most promising and targeted approaches to biocontrol of parasitic nematodes in crops is that of RNA interference (RNAi). In this study we explore the possibility of using biostimulants obtained from metabolites of soil streptomycetes to protect wheat (Triticum aestivum L.) against the cereal cyst nematode Heterodera avenae by means of inducing RNAi in wheat plants. Theoretical models of uptake of organic compounds by plants, and within-plant RNAi dynamics, have provided us with useful insights regarding the choice of routes for delivery of RNAi-inducing biostimulants into plants. We then conducted in planta experiments with several streptomycete-derived biostimulants, which have demonstrated the efficiency of these biostimulants at improving plant growth and development, as well as in providing resistance against the cereal cyst nematode. Using dot blot hybridization we demonstrate that biostimulants trigger a significant increase of the production in plant cells of si/miRNA complementary with plant and nematode mRNA. Wheat germ cell-free experiments show that these si/miRNAs are indeed very effective at silencing the translation of nematode mRNA having complementary sequences, thus reducing the level of nematode infestation and improving plant resistance to nematodes. Thus, we conclude that natural biostimulants produced from metabolites of soil streptomycetes provide an effective tool for biocontrol of wheat nematode.

Keywords: RNA interference; cereal cyst nematode (Heterodera avenae); crop protection against nematodes; streptomycete-derived biostimulants; wheat (Triticum aestivum L.).

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Figures

Figure 1
Figure 1
Plant hormone composition of biostimulants.
Figure 2
Figure 2
(A) Effects of biostimulants on viability of plants grown on non-invasive or invasive background in perlite-filled cuvettes from seeds treated in planta by biostimulants, as compared to a viability of control plants grown in perlite-filled cuvettes without biostimulants. (B) Reduction in infestation with H. avenae larvae of plants grown in perlite-filled cuvettes from seeds treated in planta by biostimulants, as compared to control plants grown in perlite-filled cuvettes in a nematode-infested background without biostimulants. The error bars in plot (B) are not visible, since they are almost 3 orders of magnitude smaller than the reported values.
Figure 3
Figure 3
Effects of biostimulants on increasing the synthesis of si/miRNA in nematode-infested plants grown using biostimulants in planta that are complementary with mRNA from control plants (A), and with nematode mRNA (B), scaled by the amount of si/miRNA isolated from nematode-infested plants grown under the same conditions but without the use of biostimulants. The error bars are not visible, since they are three orders of magnitude smaller than the reported values.
Figure 4
Figure 4
Effects of biostimulants on silencing activity of si/miRNA isolated from nematode-infested plants grown using biostimulants in planta, i.e., a percentage of inhibition of protein synthesis on the template of nematode mRNA in a wheat germ cell-free system, measured per 1μg of protein. Horizontal black lines denote the respective levels of silencing activity achieved by si/miRNA isolated from nematode-infested plants grown without the use of biostimulants.
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