Molecular characterization of cyclophilin A-like protein from Piriformospora indica for its potential role to abiotic stress tolerance in E. coli

Abstract

BackgroundCyclophilins (CyP), conserved in all genera, are known to have regulatory responses of various cellular processes including stress tolerance. Interestingly, CyP has a crucial role as peptidyl-prolyl cis–trans isomerases (PPIases). Our earlier in silico based approach resulted into the identification of cyclophilin family from rice, Arabidopsis and yeast. In our recent report, we discovered a new OsCYP-25 from rice. Here, we identified a novel cyclophylin A-like protein (PiCyP) from Piriformospora indica which is responsible for abiotic stress tolerance in E. coli.ResultsCyclophylin A-like protein (CyPA) (accession number GQ214003) was selected from cDNA library. The genomic organization CyPA revealed a 1304 bp of CyPA in P. indica genome, showing 10 exons and 9 introns. Further, CyPA was evident in PCR with gDNA and cDNA and Southern blot analysis. The phylogenetic examination of CyPA of P. indica showed that it is closed to human cyclophilin. The uniqueness of PiCyPA protein was apparent in western blot study. Kinetics of purified PiCyPA protein for its PPIas activity was determined via first order rate constant (0.104 s-1) in the presence of 1 μg of PiCyPA, with increasing PiCyPA concentration, in the presence of cyclosporin A (CsA) and the inhibition constant (4.435 nM) of CsA for inhibition of PiCyPA. The differential response of E. coli harbouring pET28a-PiCypA was observed for their different degree of tolerance to different abiotic stresses as compared to empty pET28a vector.ConclusionsOverexpression of PiCyPA protein E. coli cells confer enhanced tolerance to wide range of abiotic stresses. Thus, this study provides the significance of PiCypA as a molecular chaperone which advanced cellular stress responses of E. coli cells under adverse conditions, and it, furthermore, confirms the mounting the sustainability of E. coli for exploitation in recombinant proteins production. Additionally, the PiCyPA gene cooperates substantial functions in cellular network of stress tolerance mechanism, essentially required for various developmental stages, and might be a potential paramount candidate for crop improvement and its sustainable production under adverse conditions.

The endophytic fungus Piriformospora indica enhances Arabidopsis thaliana growth and modulates Na + /K + homeostasis under salt stress conditions

“…The increase in chlorophyll content, stimulated by P. indica, under salt stress explains the common observation that P. indica colonized plants exhibit greater fresh and dry weight [55]. Previous studies showed that P. indica colonization triggers several stress-tolerance mechanisms in host plants [41,42,56], including an increase in the levels of detoxifying and antioxidant-forming enzymes, in addition to several indirect mechanisms [57,58]. For instance, P. indica induces leaf dehydroascorbate reductase (DHAR) activity [41] and increases ascorbic acid content in colonized barley roots under salt stress conditions [56].…”

Influence of the root endophyte Piriformospora indica on the plant water relations, gas exchange and growth of Chenopodium quinoa at limited water availability

“…Of these, Piriformospora indica , a plant‐root‐colonizing basidiomycete fungus, is analogous to arbuscular mycorrhizal fungi (AMF) in many morphological, functional and growth promotional aspects (Rai & Varma, ). Recently, P. indica has been confirmed to be a potent new candidate symbiont for a broad spectrum of mono‐ and dicot plants, conveying several benefits to the host plants including growth promotion along with better resistance to various biotic and abiotic stresses (Ansari, Gill, & Tuteja, ; Franken, ; Oelmüller et al., ; Trivedi et al, ,b; Varma et al., ). P. indica inoculation was found to improve tremendously the growth and biomass of diverse hosts including Oryza sativa, Triticum sativum, Zea mays, Setaria italica, Sorghum vulgare (Achatz et al., ; Baltruschat et al., ; Varma, Rai, & Sahay, ; Varma, Verma, Sudah, & Franken, ), Arabidopsis thaliana (Peškan‐Berghöfer et al., ; Shahollari, Varma, & Oelmüller, ; Sherameti et al., ), Hordeum vulgare (Achatz et al., ; Baltruschat et al., ; Waller et al., , ), Nicotiana attenuata (Barazani, Dahl, & Baldwin, ), Brassica chinensis (Sun et al., ) and Lycopersicon esculentum (Sarma et al., ; Trivedi et al, ,b).…”

The peptidyl-prolyl cis-trans isomerase activity of the wheat cyclophilin, TaCypA-1, is essential for inducing thermotolerance in Escherichia coli

“…Few of the cyclophilins have been implicated in abiotic stress tolerance of plants [9] , [10] , [17] but the role of wheat cyclophilin, TaCypA-1, is still a matter of conjecture. Our previous studies demonstrated that TaCypA-1 is an enzymatically active protein and can catalyse cis-trans isomerisation of peptidyl-prolyl bond in vitro [11] .…”

“…Expression of cyclophilins in plants is modulated by different abiotic stresses such as heat-, cold-, drought- and salt stress [4] , [5] , [6] , [7] , [8] , [9] , suggesting a role of these proteins in stress adaptation. The role of cyclophilins in abiotic stress tolerance is further supported by recent studies which demonstrated that ectopic expression of cyclophilin genes from pigeon pea ( CcCyp ) [9] , rice ( OsCyp2 ) [4] , [5] and a xerophytic fungus Piriformospora indica ( PiCypA ) [10] in the transgenic plants resulted in enhanced tolerance to multiple abiotic stress conditions. Wheat is one of the most important grain crops, and our recent studies demonstrated that one of its cyclophilin, TaCypA-1, possesses PPIase activity [11] .…”