PR-proteins with ribonuclease activity and plant resistance against pathogenic fungi

Филипенко, Е. А.; Кочетов, А. В.; Kanayama, Yoshiharu; Malinovsky, V. I.; Shumny, V. K.

doi:10.1134/s2079059713060026

Cited by 30 publications

(17 citation statements)

References 71 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Another class of proteins involved in plant defense is ribonuclease (RNAse), a protein involved in the processing and degradation of RNA. Although its mechanisms are not known, RNAse participates in complex plant defense responses to pathogens, primarily against pathogenic fungi [ 44 , 45 ].…”

Section: Discussionmentioning

confidence: 99%

In vitro anthelmintic effects of Spigelia anthelmia protein fractions against Haemonchus contortus

et al. 2017

View full text Add to dashboard Cite

Gastrointestinal nematodes are a significant concern for animal health and well-being, and anthelmintic treatment is mainly performed through the use of chemical products. However, bioactive compounds produced by plants have shown promise for development as novel anthelmintics. The aim of this study is to assess the anthelmintic activity of protein fractions from Spigelia anthelmia on the gastrointestinal nematode Haemonchus contortus. Plant parts were separated into leaves, stems and roots, washed with distilled water, freeze-dried and ground into a fine powder. Protein extraction was performed with sodium phosphate buffer (75 mM, pH 7.0). The extract was fractionated using ammonium sulfate (0–90%) and extensively dialyzed. The resulting fractions were named LPF (leaf protein fraction), SPF (stem protein fraction) and RPF (root protein fraction), and the protein contents and activities of the fractions were analyzed. H. contortus egg hatching (EHA), larval exsheathment inhibition (LEIA) and larval migration inhibition (LMIA) assays were performed. Proteomic analysis was conducted, and high-performance liquid chromatography (HPLC) chromatographic profiles of the fractions were established to identify proteins and possible secondary metabolites. S. anthelmia fractions inhibited H. contortus egg hatching, with LPF having the most potent effects (EC50 0.17 mg mL-1). During LEIA, SPF presented greater efficiency than the other fractions (EC50 0.25 mg mL-1). According to LMIA, the fractions from roots, stems and leaves also reduced the number of larvae, with EC50 values of 0.11, 0.14 and 0.21 mg mL-1, respectively. Protein analysis indicated the presence of plant defense proteins in the S. anthelmia fractions, including protease, protease inhibitor, chitinase and others. Conversely, secondary metabolites were absent in the S. anthemia fractions. These results suggest that S. anthelmia proteins are promising for the control of the gastrointestinal nematode H. contortus.

show abstract

Section: Discussionmentioning

confidence: 99%

In vitro anthelmintic effects of Spigelia anthelmia protein fractions against Haemonchus contortus

et al. 2017

View full text Add to dashboard Cite

show abstract

“…The expression of different genes in the PR-4 family is induced by very different stimuli, including pathogen invasion, elicitors, tissue wounding, methyl jasmonate, abscisic acid, ethylene, ozone, drought, salinity, cold, UV-light, and heat shock (for a review, see [4]). Some (but not all) PR-4 proteins exhibited both RNase activity and fungicidal properties in the in vitro experiments.…”

Section: Discussionmentioning

confidence: 99%

“…Ribonucleases belong to the apoplast dynamic proteome and their synthesis increases in response to various stimuli. In particular, some extracellular S-like RNases and pathogenesis-related protein 4 (PR-4) possessing ribonuclease activity are elevated both locally and systemically after wounding or pathogen invasion [4]. It has been assumed that extracellular ribonucleases participate in defense against viruses with RNA genomes.…”

Section: Introductionmentioning

confidence: 99%

Expression of an extracellular ribonuclease gene increases resistance to Cucumber mosaic virus in tobacco

et al. 2016

View full text Add to dashboard Cite

BackgroundThe apoplast plays an important role in plant defense against pathogens. Some extracellular PR-4 proteins possess ribonuclease activity and may directly inhibit the growth of pathogenic fungi. It is likely that extracellular RNases can also protect plants against some viruses with RNA genomes. However, many plant RNases are multifunctional and the direct link between their ribonucleolytic activity and antiviral defense still needs to be clarified. In this study, we evaluated the resistance of Nicotiana tabacum plants expressing a non-plant single-strand-specific extracellular RNase against Cucumber mosaic virus.ResultsSevere mosaic symptoms and shrinkage were observed in the control non-transgenic plants 10 days after inoculation with Cucumber mosaic virus (CMV), whereas such disease symptoms were suppressed in the transgenic plants expressing the RNase gene. In a Western blot analysis, viral proliferation was observed in the uninoculated upper leaves of control plants, whereas virus levels were very low in those of transgenic plants. These results suggest that resistance against CMV was increased by the expression of the heterologous RNase gene.ConclusionWe have previously shown that tobacco plants expressing heterologous RNases are characterized by high resistance to Tobacco mosaic virus. In this study, we demonstrated that elevated levels of extracellular RNase activity resulted in increased resistance to a virus with a different genome organization and life cycle. Thus, we conclude that the pathogen-induced expression of plant apoplastic RNases may increase non-specific resistance against viruses with RNA genomes.

show abstract

“…The complex of bacterial extracellular enzymes provides the pool of low-molecular weight nutrients which can be absorbed by plants. Being a cytotoxic, RNases can contribute to anti-pathogen defense (Deshpande and Shankar, 2002;Filipenko et al, 2013). Finally, 2¢-3¢-cGMP which is formed during RNA hydrolysis by guanyl-preferring RNases can act as signal molecule which has an influence on proliferation and programmed cell death (Kessler and Steinberg, 1973;Sokurenko et al, 2015;Van Damme et al, 2014).…”

Section: Resultsmentioning

confidence: 99%

Phylogenetic distribution of extracellular guanyl-preferring ribonucleases renews taxonomic status of two <i>Bacillus</i> strains

Ulyanova

Mahmud

Dudkina

et al. 2016

J. Gen. Appl. Microbiol.

View full text Add to dashboard Cite

† The GenBank accession number for the 16S rRNA gene sequence of Bacillus altitudinis B-388 is JX129389, for the rpoB gene sequence of Bacillus altitudinis B-388 is JX129391, for the 16S rRNA gene sequence of Bacillus pumilus 7P is JX129390 and for the rpoB gene sequence of Bacillus pumilus 7P is JX129392 correspondingly.None of the authors of this manuscript has any financial or personal relationship with other people or organizations that could inappropriately influence their work. IntroductionBarnase, an extracellular alkaline guanyl-preferring ribonuclease first isolated from Bacillus amyloliquefaciens (Hartley and Rogerson, 1972), was shown to possess a wide range of biological activities and is of special interest for research and practical applications in biotechnology and medicine (Kempe et al., 2014;Spång et al., 2012;Sreenivasan et al., 2013;Ulyanova et al., 2011). Ribonucleases similar to barnase (UniProtKB/Swiss-Prot accession number P00648) were purified from B. intermedius 7P (P00649), B. circulans BCF-256 (P35078), B. pumilus KMM-62 (P48068), B. coagulans BCF-247 (P37203), and B. thuringiensis var. subtoxicus B-388 (Q9R5D7, GenBank acc No. AAB28984.1). B. intermedius RNase referred to as binase is as well studied as barnase. Its antitumor and antiviral properties are documented (Mironova et al., 2013;Mitkevich et al., 2014;Shah Mahmud and Ilinskaya, 2013;Ulyanova et al., 2011) while the strain itself is an undescribed taxon which is The potential of microbial ribonucleases as promising antitumor and antiviral agents, determines today's directions of their study. One direction is connected with biodiversity of RNases. We have analyzed completed and drafted Bacillus genomes deposited in GenBank for the presence of coding regions similar to the gene of an extracellular guanyl-preferring RNase of Bacillus amyloliquefaciens (barnase). Orthologues of the barnase gene were detected in 9 species out of 83. All of these belong to "B. subtilis" group within the genus. B. subtilis itself, as well as some other species within this group, lack such types of RNases. RNases similar to barnase were also found in species of "B. cereus" group as a part of plasmid-encoded S-layer toxins. It was also found that taxonomic states of culture collection strains, which were initially described based on a limited set of phenotypic characteristics, can be misleading and need to be confirmed. Using several approaches such as matrix-assisted laser desorption ionizationtime of flight mass spectrometry (MALDI-TOF MS), sequencing of genes for 16S ribosomal RNA and RNA polymerase subunit beta followed by reconstruction of phylogenetic trees, we have re-identified two RNase-secreting Bacillus strains: B. thuringiensis B-388 which should be assigned as B. altitudinis B388 and B. intermedius 7P which should be renamed as B. pumilus 7P. Therefore, small secreted guanyl-preferring RNases are the feature of "B. subtilis" group only, which is characterized by distinctive lifestyle and adaptation strategies to environment.

show abstract

PR-proteins with ribonuclease activity and plant resistance against pathogenic fungi

Cited by 30 publications

References 71 publications

In vitro anthelmintic effects of Spigelia anthelmia protein fractions against Haemonchus contortus

In vitro anthelmintic effects of Spigelia anthelmia protein fractions against Haemonchus contortus

Expression of an extracellular ribonuclease gene increases resistance to Cucumber mosaic virus in tobacco

Phylogenetic distribution of extracellular guanyl-preferring ribonucleases renews taxonomic status of two <i>Bacillus</i> strains

Contact Info

Product

Resources

About