Abstract
The phytopathogenic fungus Venturia inaequalis causes scab of apple. Once this fungus penetrates the plant surface, it forms a specialized body called a stroma between the inner cuticle surface and the epidermal cell wall. A novel Venturia inaequalis 5704 (Cin3) and three expressed sequence tags (ESTs); 38, 6987, and 4010 are strongly up-regulated in the early stages of infection. The CIN3 and three ESTs using two vectors pMAL-c2 and pET 21 were expressed in Escherichia coli. Recombinant proteins expression, solubility and yields were analyzed. 38, 5704 (Cin3) and 6987 re- combinant proteins were expressed in soluble form and while 4010 was expressed in inclusion bodies. Re- solution on native-PAGE, the recombinant proteins; 38, 5704 (Cin3), 6987 were shown to be present in dimmer, tetramer and polymer. A method was de- veloped, consisting of induction of expression at va- rious temperatures, and using enriched broth with 4% glycerol together with slow shaking, led to a decrease in concentration of nascent polypeptide and production of soluble recombinant proteins of; 38, 5704 (Cin3), 6987 and 4010. Resolution on native- PAGE, the recombinant proteins were shown to be present as monomer.
Highlights
Ventura inaequalis (Cooke) Wint. causes black spot or scab of apple (Maluspumila (Mill.) Henry) [1]
The phytopathogenic fungus Venturia inaequalis causes scab of apple. Once this fungus penetrates the plant surface, it forms a specialized body called a stroma between the inner cuticle surface and the epidermal cell wall
Once the cuticle has been breached, the fungal hyphae grow between the inner cuticle surface and the epidermal cell wall
Summary
Ventura inaequalis (Cooke) Wint. causes black spot or scab of apple (Maluspumila (Mill.) Henry) [1]. Once the cuticle has been breached, the fungal hyphae grow between the inner cuticle surface and the epidermal cell wall At this time the hyphae differentiate, producing a specialized body called a stroma that is made up of single or multiple layers of pseudoparenchyma, a laterally dividing cell-type which is distinct from the usual tubular form of hyphal growth [1,2]. Differentiation of sub-cuticular hyphae and stromata can be simulated by growing V. inaequalis in vitro on cellophane discs [2]. Full-length versions of these genes were found in an expressed sequence tag library from apple leaves infected with V. inaequalis, and quantitative real-time PCR (qRT-PCR) indicated that these genes were up-regulated over 1000-fold during the early stages of infection compared with in vitro growth [2]. Three ESTs: 38(7006), 6987 and 4010 were selected since qRT-PCR indicated that these genes were up-regulated during the early stages of infection compared with in vitro growth (Gene accession number EB135294)
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
