Detection of specific microRNA (miRNA) is of great demand due to their essential role in genes regulation, stress response and development of diseases. However, mature miRNAs are small molecules that ...make it difficult to use routine amplification-based methods. Here, we report an approach for detection of miRNA based on a new type of isothermal amplification, namely, multimerization. The proposed technique is simple and versatile, excludes a reverse transcription step, and requires two conventional primers only and no additional stem-loop or fluorogenic probes. Only mature miRNAs can initiate multimerization, thereby, pri- or pre-miRNA are excluded from analysis, ensuring high accuracy of the assay. The approach was approved on miRNA from common wheat Triticum aestivum; the increase of Tae-miRNA159 level for plants affected by Stagonospora nodorum Berk infection was demonstrated. The obtained results allow to perform quantitative analysis, providing determination of specific targets with high reliability (detection limit of about 20 pM).
Display omitted
•Nucleic acids multimerization (MM) allows miRNA detection with high specificity.•Definite size of miRNA determines efficient generation of structures providing MM.•Only mature miRNA initiate MM, while precursors don't involve into reaction.•The proposed technique requires no reverse transcription step.•MM allows accurate quantification of specific miRNA targets.
Full text
Available for:
GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
The use of biocontrol agents based on endophytic bacteria against phloem-feeding insects is limited by a lack of knowledge and understanding of the mechanism of action of the endophyte community that ...makes up the plant microbiome. In this work, the mechanisms of the additive action of endophytic strains
26D and
11VM on the resistance of bread spring wheat against greenbug aphid
was studied. It was shown that
26D secreted lipopeptide surfactin and phytohormones cytokinins, and
11VM produced iturin and auxins into the cultivation medium. Both strains and their lipopeptide-rich fractions showed direct aphicidal activity against greenbug aphid. For the first time, it was shown that
26D and
11VM in the same manner, as well as their lipopeptide-rich fractions, activated the expression of salicylate- and ethylene-dependent
genes, and influenced plant redox metabolism, which led to an increase in plant endurance against aphids. The composition of endophytic strains
26D +
11VM had an additive effect on plant resistance to aphids due to an increase in the number of endophytic bacterial cells, and, as well as due to the synergistic effect of their mixture of lipopeptides - surfactin + iturin, both on the aphid mortality and on the expression of
and
genes. All these factors can be the reason for the observed increase in the growth of plants affected by aphids under the influence of
26D and
11VM, individually and in composition. The study demonstrates the possibility of creating in the future an artificial composition to enhance plant microbiome with endophytic bacteria, which combines growth-promoting and plant immunity stimulating properties against phloem-feeding insects. This direction is one of the most promising approaches to green pesticide discovery in the future.
Full text
Available for:
IZUM, KILJ, NUK, PILJ, PNG, SAZU, UL, UM, UPUK
Cytokinins (CKs) and abscisic acid (ABA) play an important role in the life of both plants and pathogenic fungi. However, the role of CKs and ABA in the regulation of fungal growth, development and ...virulence has not been sufficiently studied. We compared the ability of two virulent isolates (SnB and Sn9MN-3A) and one avirulent isolate (Sn4VD) of the pathogenic fungus
Berk. to synthesize three groups of hormones (CKs, ABA and auxins) and studied the effect of exogenous ABA and zeatin on the growth, sporulation and gene expression of necrotrophic effectors (NEs) and transcription factors (TFs) in them. Various isolates of
synthesized different amounts of CKs, ABA and indoleacetic acid. Using exogenous ABA and zeatin, we proved that the effect of these hormones on the growth and sporulation of
isolates can be opposite, depends on both the genotype of the isolate and on the concentration of the hormone and is carried out through the regulation of carbohydrate metabolism. ABA and zeatin regulated the expression of fungal TF and NE genes, but correlation analysis of these parameters showed that this effect depended on the genotype of the isolate. This study will contribute to our understanding of the role of the hormones ABA and CKs in the biology of the fungal pathogen
.
Full text
Available for:
IZUM, KILJ, NUK, PILJ, PNG, SAZU, UL, UM, UPUK
26D is a plant growth-promoting endophytic bacteria capable of inducing systemic resistance through the priming mechanism, which includes plant genome reprogramming and the phenomenon of RNA ...interference (RNAi) and microRNA (miRNAs). The phloem-feeding insect bird cherry-oat aphid
L. is a serious pest that causes significant damage to crops throughout the world. However, the function of plant miRNAs in the response to aphid infestation remains unclear. The results of this work showed that
26D stimulated aphid resistance in wheat plants, inducing the expression of genes of hormonal signaling pathways
,
,
,
,
,
and
. In addition,
26D activated the RNAi mechanism and regulated the expression of nine conserved miRNAs through activation of the ethylene, salicylic acid (SA), and abscisic acid (ABA) signaling pathways, which was demonstrated by using treatments with phytohormones. Treatment of plants with SA, ethylene, and ABA acted in a similar manner to
26D on induction of the expression of the
,
and
,
genes, as well as the expression of nine conserved miRNAs. Different patterns of miRNA expression were found in aphid-infested plants and in plants treated with
26D or SA, ethylene, and ABA and infested by aphids, suggesting that miRNAs play multiple roles in the plant response to phloem-feeding insects, associated with effects on hormonal signaling pathways, redox metabolism, and the synthesis of secondary metabolites. Our study provides new data to further elucidate the fine mechanisms of bacterial-induced priming. However, further extensive work is needed to fully unravel these mechanisms.
Endophytic plant-growth-promoting microorganisms can protect plants against pathogens, but they have rarely been investigated as potential biocontrol agents and triggers of induced systemic ...resistance (ISR), regulated by phytohormones, against viruses. We studied the role of endophytic strains
26D and
Ttl2, which secrete ribonucleases and phytohormones, in the induction of tomato plant resistance against potato virus X and potato virus Y in a greenhouse condition. The endophytes reduced the accumulation of viruses in plants, increased the activity of plant ribonucleases and recovered the fruit yield of infected tomato plants. Both the 26D and Ttl2 strains induced ISR by activating the transcription of genes related to salicylate- and jasmonate-dependent responses. The 26D and Ttl2 strains increased the content of cytokinins and decreased the level of indolacetic acid in plants infected with PVX or PVY. PVY led to an increase of the abscisic acid (ABA) content in tomato plants, and PVX had the opposite effect. Both strains reduced the ABA content in plants infected with PVY and induced ABA accumulation in plants infected with PVX, which led to an increase in the resistance of plants. This is the first report of the protection of tomato plants against viral diseases by foliar application of endophytes.
Full text
Available for:
IZUM, KILJ, NUK, PILJ, PNG, SAZU, UL, UM, UPUK
Ethylene, salicylic acid (SA), and jasmonic acid are the key phytohormones involved in plant immunity, and other plant hormones have been demonstrated to interact with them. The classic phytohormone ...cytokinins are important participants of plant defense signaling. Crosstalk between ethylene and cytokinins has not been sufficiently studied as an aspect of plant immunity and is addressed in the present research. We compared expression of the genes responsible for hormonal metabolism and signaling in wheat cultivars differing in resistance to
in response to their infection with fungal isolates, whose virulence depends on the presence of the necrotrophic effector SnTox3. Furthermore, we studied the action of the exogenous cytokinins, ethephon (2-chloroethylphosphonic acid, ethylene-releasing agent) and 1-methylcyclopropene (1-MCP, inhibitor of ethylene action) on infected plants. Wheat susceptibility was shown to develop due to suppression of reactive oxygen species production and decreased content of active cytokinins brought about by SnTox3-mediated activation of the ethylene signaling pathway. SnTox3 decreased cytokinin content most quickly by its activated glucosylation in an ethylene-dependent manner and, furthermore, by oxidative degradation and inhibition of biosynthesis in ethylene-dependent and ethylene-independent manners. Exogenous zeatin application enhanced wheat resistance against
through inhibition of the ethylene signaling pathway and upregulation of SA-dependent genes. Thus, ethylene inhibited triggering of SA-dependent resistance mechanism, at least in part, by suppression of the cytokinin signaling pathway.
Full text
Available for:
IZUM, KILJ, NUK, PILJ, PNG, SAZU, UL, UM, UPUK
Display omitted
•When plants and pests interact, their microbiomes interact too.•Endophytic strain Bacillus subtilis 26D decreases the survival of L. decemlineata.•B. subtilis 26D inhibits the growth ...of L. decemlineata microsymbionts.
Plants are exposed to pervasive attack by diverse attackers, such as pathogens and pests. But plants have their own endophytic microflora as well as the attacking insects. These microbiomes contact face to face in the nature. It has been found that the endophytic strain Bacillus subtilis 26D increases mortality of Colorado potato beetles, disturbing the development of insect microsymbionts Enterobacter ssp. and Acinetobacter ssp.
Full text
Available for:
GEOZS, IJS, IMTLJ, KILJ, KISLJ, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UL, UM, UPCLJ, UPUK, ZRSKP
One of the main insect potato pests is the Colorado potato beetle (
Leptinotarsa decemlineata
Say). It contains some microbial associates which can affect diverse interactions between host plant and ...the herbivorous insect. Previously, the most common
L. decemlineata
microsymbiont isolated from anterior and posterior parts of beetle gut was defined as
Enterobacter
BC-8. The role of
Enterobacter
BC-8 in manipulating plant defenses was investigated using antibiotics-treated beetles and model system simulating beetles attacks (wounding plants treated with
Enterobacter
BC-8 suspension). We demonstrated that the symbiotic bacteria suppressed plant defenses such as hydrogen peroxide and phenolic compounds accumulation and activity of peroxidases and trypsin inhibitors. It is worth noting that the influence of the insect symbionts on potato plants stimulated salicylate-sensitive genes and the marker of salicylate signaling pathway. Transcription activities of jasmonate-sensitive genes which encode some defense proteins against herbivores, were suppressed. So,
Enterobacter
BC-8 plays the role in salicylate/jasmonate crosstalks manipulating to suppress plant defense mechanisms.
Full text
Available for:
EMUNI, FIS, FZAB, GEOZS, GIS, IJS, IMTLJ, KILJ, KISLJ, MFDPS, NLZOH, NUK, OBVAL, OILJ, PNG, SAZU, SBCE, SBJE, SBMB, SBNM, UKNU, UL, UM, UPUK, VKSCE, ZAGLJ
Plant viruses are globally responsible for the significant crop losses of economically important plants. All common approaches are not able to eradicate viral infection. Many non-conventional ...strategies are currently used to control viral infection, but unfortunately, they are not always effective. Therefore, it is necessary to search for efficient and eco-friendly measures to prevent viral diseases. Since the genomic material of 90% higher plant viruses consists of single-stranded RNA, the best way to target the viral genome is to use ribonucleases (RNase), which can be effective against any viral disease of plants. Here, we show the importance of the search for endophytes with protease and RNase activity combined with the capacity to prime antiviral plant defense responses for their protection against viruses. This review discusses the possible mechanisms used to suppress a viral attack as well as the use of local endophytic bacteria for antiviral control in crops.
The aim of the study was to access the extended occlusion of the intercostal arteries by a stent graft in the development of postoperative spinal cord injury during aortic arch surgery using the ...frozen elephant trunk technique.
A total of 37 consecutive patients underwent total aortic arch surgery using the frozen elephant trunk technique between March 2012 and July 2017. The mean age of the patients was 54.7 ± 10.5 years. Type A and Type B aortic dissections were the indications for surgery. Moderate hypothermia and antegrade cerebral perfusion via the innominate artery were utilized. The mean diameter of the implanted stent graft was 27.7 ± 2 mm (range 24-30 mm).
No permanent spinal cord injuries occurred. The distal edge of the stent graft was in the T7-T12 range. Its lower edge was implanted at the T9-T12 level in 25 (67.6%) cases. Preoperatively, the mean number of intercostal arteries was 10 ± 1 on the left side and 10 ± 2 on the right side (P = 0.59). Postoperatively, the mean number of open segmental arteries was 3 ± 2 on the left and 4 ± 1 on the right (P = 0.003).
The frozen elephant trunk procedure is associated with the occlusion of most (two-thirds) of the intercostal arteries. Maintenance of adequate blood flow in the subclavian and iliac arteries is an integral prerequisite for a favourable outcome. The level of the deployment of the distal edge of the stent graft does not play a defining role.
PDF
