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Hexokinase as a versatile molecular genetic marker for Microsporidia

Published online by Cambridge University Press:  15 October 2018

Yuri S. Tokarev Open the ORCID record for Yuri S. Tokarev [Opens in a new window] ,
Sergei A. Timofeev ,
Julia M. Malysh Open the ORCID record for Julia M. Malysh [Opens in a new window] ,
Alexander A. Tsarev ,
Anastasia N. Ignatieva ,
Oksana G. Tomilova  and
Vyacheslav V. Dolgikh
Yuri S. Tokarev
Affiliation:
All-Russian Institute of Plant Protection, Podbelskogo, 3,196608 St. Petersburg, Pushkin, Russia
Sergei A. Timofeev
Affiliation:
All-Russian Institute of Plant Protection, Podbelskogo, 3,196608 St. Petersburg, Pushkin, Russia
Julia M. Malysh*
Affiliation:
All-Russian Institute of Plant Protection, Podbelskogo, 3,196608 St. Petersburg, Pushkin, Russia
Alexander A. Tsarev
Affiliation:
All-Russian Institute of Plant Protection, Podbelskogo, 3,196608 St. Petersburg, Pushkin, Russia
Anastasia N. Ignatieva
Affiliation:
All-Russian Institute of Plant Protection, Podbelskogo, 3,196608 St. Petersburg, Pushkin, Russia
Oksana G. Tomilova
Affiliation:
Institute of Animal Systematics and Ecology SB RAS, Frunze, 11, 630091 Novosibirsk, Russia
Vyacheslav V. Dolgikh
Affiliation:
All-Russian Institute of Plant Protection, Podbelskogo, 3,196608 St. Petersburg, Pushkin, Russia
*
Author for correspondence: Julia M. Malysh, E-mail: [email protected]
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Abstract

Hexokinase (HK) is a core glycolytic enzyme of Microsporidia which regulates host cell metabolic processes. The goal of the present study was to test for the utility of HK for molecular phylogenetics, species identification and molecular detection of microsporidia in infected insects. HK sequence-based reconstructions were essentially similar to those based upon largest subunit RNA polymerase (RPB1) gene sequences, as well as previously published rRNA gene and genome-based trees. Comparing HK sequences allowed clear differentiation of closely related taxa, such as Nosema bombycis and Nosema pyrausta. In Nosema ceranae, unique SNPs were found for an isolate from wild colonies of the Burzyan dark honey bee as compared with the isolates from domesticated European honey bee. Similarly, in Encephalitozoon cuniculi, HK was as effective as RPB1 for discrimination of isolates belonging to different ITS genotypes. Amplification using species-specific primers flanking short fragments at the 3′-end of HK gene showed the presence of infection in insect tissues infected with N. pyrausta, Nosema ceranae and Paranosema (Antonospora) locustae. For the latter parasite species, HK expression was also demonstrated at early stages of infection using total mRNA extracts of locust larvae. These results indicate the suitability of HK as a novel tool for molecular genetic studies of Microsporidia.

Keywords

Gene expressionhexokinaseMicrosporidiamolecular detectionmolecular phylogenyspecies identification
Type
Research Article
Information
Parasitology , Volume 146 , Issue 4 , April 2019 , pp. 472 - 478
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Copyright
Copyright © Cambridge University Press 2018 

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