Endophytic Strain Bacillus subtilis 26D Increases Levels of Phytohormones and Repairs Growth of Potato Plants after Colorado Potato Beetle Damage.

Plant damage caused by defoliating insects has a long-term negative effect on plant growth and productivity. Consequently, the restoration of plant growth after exposure to pathogens or pests is the main indicator of the effectiveness of the implemented defense reactions. A short-term Leptinotarsa decemlineata Say attack on potato tube-grown plantlets (Solanum tuberosum L.) led to a reduction of both the length and mass of the shoots in 9 days. The decrease of the content of phytohormones-indole-3-acetic acid (IAA), abscisic acid (ABA), zeatin and zeatin-riboside-in shoots of damaged potato plants was found. Endophytic strain Bacillus subtilis 26D (Cohn) is capable of secreting up to 83.6 ng/mL IAA and up to 150 ng/mL cytokinins into the culture medium. Inoculation of potato plants with cells of the B. subtilis 26D increases zeatin-riboside content in shoots and the mass of roots of undamaged plants, but does not influence content of IAA and ABA and growth of shoots. The presence of B. subtilis 26D in plant tissues promoted a rapid recovery of the growth rates of shoots, as well as the wet and dry mass of roots of plants after the pest attack, which we associate with the maintenance of a high level of IAA, ABA and cytokinins in their tissues.

Endophytic Bacillus spp. as a Prospective Biological Tool for Control of Viral Diseases and Non-vector Leptinotarsa decemlineata Say. in Solanum tuberosum L.

Viral diseases and their damage causing significant loss to economically important crops have increased by several folds during the last decade. All the conventional approaches are not able to eradicate the viral infection. Therefore, there is a need to look for efficient and eco-friendly viral disease-preventive measures. The genomic material of the majority of deleterious viruses of higher plants is RNA. One of the possible measures to control viruses is the use of ribonucleases (RNases), which can cleave RNA in the viral genome. Based on this, we investigated the RNase activity of endophytic Bacillus spp., which can enrich in 103-105 colony-forming units per gram of wet mass of aboveground part of potato plants. A high level of RNase activity was observed in the culture medium of Bacillus thuringiensis B-6066, Bacillus sp. STL-7, Bacillus sp. TS2, and Bacillus subtilis 26D. B. thuringiensis B-5351 had low RNase activity but high ability to colonize internal plant tissues, Bacillus sp. STL-7 with high RNase activity have relatively low number of cells in internal tissues of plants. B. thuringiensis B-6066, B. subtilis 26D, and Bacillus sp. TS stimulate RNase activity in potato plants for a long time after application. Strains with high ability to colonize internal plant tissues combined with high RNase activity reduced severity of viral diseases symptoms on plants and reduced the incidence of potato viruses M, S, and Y. It is worth noting that Bacillus spp. under investigation reduced the number of Leptinotarsa decemlineata Say. egg clusters and larvae on treated plants and showed antifeedant activity. This results in increase of potato productivity mainly in the fraction of major tubers. B. subtilis 26D and Bacillus sp. TS2 combining endophytic lifestyle, RNase, and antifeedant activity may become the basis for the development of biocontrol agents for plant protection.

Endophytic Strain Bacillus subtilis 26DCryChS Producing Cry1Ia Toxin from Bacillus thuringiensis Promotes Multifaceted Potato Defense against Phytophthora infestans (Mont.) de Bary and Pest Leptinotarsa decemlineata Say.

Novel properties of a previously obtained Bacillus subtilis 26DCryChS strain are described. The B. subtilis 26DCryChS strain is able to produce Cry1Ia δ-endotoxin from B. thuringiensis B-5351 and to exist in internal plant tissues of potato plants in the same manner as the endophytic B. subtilis 26D source strain (487 ± 53 and 420 ± 63 CFU*103/g, respectively). B. subtilis 26DCryChS, as much as the original B. subtilis 26D strain, inhibited mycelium growth of oomycete Phytophthora infestans (Mont.) de Bary and reduced late blight symptoms development on plants by 35% compared with non-treated ones, as well as showed insecticidal activity against Leptinotarsa decemlineata. Production of the fluorescent GFP protein in the B. subtilis 26D genome allowed visualizing the endophytes around damaged sites on beetle intestines. Bacillus strains under investigation induced systemic resistance to P. infestans and L. decemlineata through the activation of the transcription of PR genes in potato plants. Thus, the B. subtilis 26DCryChS strain was able to induce transcription of jasmonate-dependent genes and acquired the ability to promote transcription of a salicylate-dependent gene (PR1) in plants infected with the late blight agent and damaged by Colorado potato beetle larvae. The B. subtilis 26DCryChS strain could be put forward as a modern approach for biocontrol agents design.