Genomics and biotechnology of insects of agricultural and medical importance

Group leaders: Profs Mariangela Bonizzoni, Ludvik Gomulski, Giuliamo Gasperi (Contract Professor), Anna Rodolfa Malacrida (Contract Professor)

Collaborators: Francesca Scolari (Postdoc), Paolo Gabrieli (Postdoc), Patrizia Chiari (Technician), Liliana Marcorio (Technician), Alessandro Di Cosimo (PhD student), Grazia Savini (PhD student)

Climate and demographic change, the increase of trade, urbanization and human travel have facilitated the spread of invasive insect species of economic and sanitary importance. These species include the Asian tiger mosquito, a vector of many viruses pathogenic to humans, and species of fruit flies that are important agricultural pests. The risks due to the invasion of these insects are compounded by the lack of vaccines and treatments for the pathogens that they transmit (i.e. Dengue, Chikungunya and Zika viruses) and the emergence of resistance to insecticides, which are currently the most used tool for their control.
The research group deals with genomics, transcriptomics, proteomics and metabolomics of insects of agricultural interest and public health relevance with the aim of developing innovative control strategies that are environmentally sustainable. Additionally, we study their population genetics and evolution in order to understand and prevent their further spread and we use transgenic approaches to study the mechanisms underlying their reproduction in order to improve current genetic control methods.

The insect species currently studied are:

Vectors of disease pathogens:

  • Mosquitoes: Aedes albopictus and Aedes aegypti, vectors of viruses including Zika, Dengue and Chikungunya.
  • Tsetse fly (Glossina) species, vectors of Trypanosomes that cause Human African Trypanosomiasis (HAT or sleeping sickness) and African Animal Trypanosomiasis (AAT or Nagana).

Fruit flies (Diptera, Tephritidae) of African or Asian origin, that are highly invasive and that have recently been introduced into Southern Europe and the Americas where they have produced significant economic damage:

  • Ceratitis capitata, the Mediterranean fruit fly.
  • Bactrocera oleae, the Olive fly.
  • Bactrocera dorsalis sensu stricto, the Oriental fruit fly.
  • Anastrepha fraterculus, the South American fruit fly.

Main Lines of Research

  1. Genome sequencing targeted at the identification of genes related to reproduction, chemoreception, adaptation, immunity and, consequently, host/parasite interactions. These studies are performed within International scientific consortia:
  • Ceratitis capitata (joint project of Baylor College of Medicine Human Genome Sequencing Center, US Department of Agriculture, and the University of Pavia) and the i5K initiative;
  • Aedes albopictus (under EC-FP7 Infrastructure – INFRAVEC coordinated by Imperial College London, and an American-Chinese consortium, coordinated by Anthony James and Xiao-Guang Chen);
  • 5 Glossina species (IGGI Consortium, coordinated by Yale University).

Relevant publications:

 Genome sequence of the tsetse fly (Glossina morsitans): vector of African trypanosomiasis. Science (2014) 344: 380-6.

Genome sequence of the Asian Tiger mosquito, Aedes albopictus, reveals insights into its biology, genetics, and evolution. Proc Natl Acad Sci USA (2015) 112: E5907-15.

A draft genome sequence of an invasive mosquito: an Italian Aedes albopictus. Pathog Glob Health (2015) 109:207-20.

Presence of extensive Wolbachia symbiont insertions discovered in the genome of its host Glossina morsitans morsitans. PLoS Negl Trop Dis. 2014 Apr 24;8(4):e2728.

  1. Origin and spread of the tiger mosquito and fruit flies.

We characterized high resolution molecular markers, such as microsatellites and SNPs, in Ae. albopictus, C. capitata and other fruit flies, and we have applied them to study the genetic links between ancestral and derived populations and to characterize the species invasion processes. We also applied mitochondrial markers, in combination with microsatellites, to study the origin of Ae. albopictus in California and, in collaboration with prof. Torroni, to study the variability of ancestral and derived populations.

Relevant publications:

Relevant genetic differentiation among Brazilian populations of Anastrepha fraterculus (Diptera, Tephritidae). Zookeys. 2015 Nov 26;(540):157-73.

Molecular markers for analyses of intraspecific genetic diversity in the Asian Tiger mosquito, Aedes albopictus. Parasit Vectors. 2015 Mar 28;8:188.

The oriental fruitfly Bactrocera dorsalis s.s. in East Asia: disentangling the different forces promoting the invasion and shaping the genetic make-up of populations. Genetica. 2014 Jun;142(3):201-13.

Microsatellite markers from the ‘South American fruit fly’ Anastrepha fraterculus: a valuable tool for population genetic analysis and SIT applications. BMC Genet. 2014;15 Suppl 2:S13.

Genetic analysis of invasive Aedes albopictus populations in Los Angeles County, California and its potential public health impact. PLoS One (2013) 8:e68586.

A new threat looming over the Mediterranean basin: emergence of viral diseases transmitted by Aedes albopictus mosquitoes. PLoS Negl Trop Dis. 2012;6(9):e1836.

The utility of microsatellite DNA markers for the evaluation of area-wide integrated pest management using SIT for the fruit fly, Bactrocera dorsalis (Hendel), control programs in Thailand. Genetica. 2011 Jan;139(1):129-40.

Uncovering the tracks of a recent and rapid invasion: the case of the fruit fly pest Bactrocera invadens (Diptera: Tephritidae) in Africa. Mol Ecol. 2009 Dec;18(23):4798-810.

Isolation and characterization of microsatellite markers in the newly discovered invasive fruit fly pest in Africa, Bactrocera invadens (Diptera: Tephritidae). Mol Ecol Resour. 2008 Nov;8(6):1509-11.

Globalization and fruitfly invasion and expansion: the medfly paradigm. Genetica. 2007 Sep;131(1):1-9.

 

  1. Analyses of reproductive processes using functional genomics, proteomics and metabolomics techniques. RNA-Seq, microarray, proteomics and RNA interference approaches are applied to identify genes and proteins that are related to sexual maturity and mating.

Relevant publications:

The Spermatophore in Glossina morsitans morsitans: Insights into Male Contributions to Reproduction. Sci Rep. 2016 Feb 5;6:20334.

How functional genomics will impact fruit fly pest control: the example of the Mediterranean fruit fly, Ceratitis capitata. BMC Genet. 2014;15 Suppl 2:S11.

Transcriptome profiling of sexual maturation and mating in the Mediterranean fruit fly, Ceratitis capitata. PLoS One. 2012;7(1):e30857.

Transcriptional profiles of mating-responsive genes from testes and male accessory glands of the Mediterranean fruit fly, Ceratitis capitata. PLoS One. 2012;7(10):e46812.

Gene discovery in an invasive tephritid model pest species, the Mediterranean fruit fly, Ceratitis capitata. BMC Genomics. 2008 May 23;9:243.

 

  1. Analysis of reproductive behaviour using molecular and transgenic (GMOs) approaches. Molecular and transgenic markers are used to analyse mating dynamics, sperm competition and sperm use in different species in the laboratory and in nature. These studies are fundamental for the implementation of environmentally friendly control methods such as the Sterile Insect Technique (SIT).

Relevant publications:

Polyandry in the medfly – shifts in paternity mediated by sperm stratification and mixing. BMC Genet. 2014;15 Suppl 2:S10.

Towards mosquito sterile insect technique programmes: exploring genetic, molecular, mechanical and behavioural methods of sex separation in mosquitoes. Acta Trop. 2014 Apr;132 Suppl:S178-87.

Polyandry is a common event in wild populations of the Tsetse fly Glossina fuscipes fuscipes and may impact population reduction measures. PLoS Negl Trop Dis. 2011 Jun;5(6):e1190.

Safe and fit genetically modified insects for pest control: from lab to field applications. Genetica. 2011 Jan;139(1):41-52.

Sperm storage and use in polyandrous females of the globally invasive fruitfly, Ceratitis capitata. J Insect Physiol. 2010 Nov;56(11):1542-51.

Site-specific recombination for the modification of transgenic strains of the Mediterranean fruit fly Ceratitis capitata. Proc Natl Acad Sci U S A (2009) 106:18171.

Fluorescent sperm marking to improve the fight against the pest insect Ceratitis capitata (Wiedemann; Diptera: Tephritidae). N Biotechnol. 2008 Jun;25(1):76-84.

 

  1. Analysis of chemoreception. The genes and the encoded proteins involved in insect-host and insect-insect interactions are being characterised. Specifically we are studying the genes and their encoded proteins that are involved in the perception of: 1) odours emitted by plant hosts, that are involved in attraction/repulsion of insect pests; 2) odours emitted from a vertebrate host, that are involved in the attraction of blood-sucking insects; 3) odours/pheromones involved in mating and oviposition processes. An integrated approach is used involving functional genomics, proteomics, structural biology, electrophysiology coupled to gas chromatography and mass spectrometry. This line of research has significant implications for the development of repellents and attractants.

Relevant publications:

Sniffing out chemosensory genes from the Mediterranean fruit fly, Ceratitis capitata. PLoS One (2014) 9:e85523.

Identification of pheromone components and their binding affinity to the odorant binding protein CcapOBP83a-2 of the Mediterranean fruit fly, Ceratitis capitata. Insect Biochem Mol Biol (2014) 48:51-62.

 

  1. biochemical approaches to study saliva of the tiger mosquito.
    The mosquito females require a blood meal for egg development and, when they bite an individual infected with a virus, they can acquire the pathogen. The mosquito saliva contains pharmacologically important proteins, with vasodilatory function, anticoagulants and anti-haemostatic activities that facilitate blood acquisition. The saliva proteins are also known to modulate and enhance viral transmission. In this project, we aim primarily to characterize the components of the saliva of Ae. albopictus and then we will study the biochemical and molecular functions of a subset of these proteins.

 

  1. Transcriptomics of resistance to insecticides and development of new bio-insecticides. RNA-seq approaches are used to identify the mechanisms of resistance to insecticides, primarily pyrethroids. Additionally, we are attempting to identify new molecules with bio-insecticide activities produced by soil bacteria.

Relevant publications:

RNA-seq analyses of changes in the Anopheles gambiae transcriptome associated with resistance to pyrethroids in Kenya: identification of candidate-resistance genes and candidate-resistance SNPs. Parasit Vectors (2015) 8:474.

Comparative transcriptome analyses of deltamethrin-resistant and -susceptible Anopheles gambiae mosquitoes from Kenya by RNA-Seq. PLoS One (2012) 7:e44607.

 

Collaborations

National:

  • University of Pavia: Andrea Mattevi, Federico Forneris, Antonio Torroni, Alessandra Albertini, Luigi Casella, Enrico Monzani, Eugenio Regazzini, Federico Bassetti, Riccardo Bellazzi, Giovanni Maga, Federico Focher, Daniele Merli, Barbara Mannucci, Antonella Lisa.
  • University of Perugia: Andrea Crisanti, Philippos Papathanos
  • University Federico II: Francesco Pennacchio, Giuseppe Saccone
  • Fondazione Edmund Mach
  • Istituto Zooprofilattico Sperimentale della Lombardia e dell’Emilia Romagna, delle Venezie, del Piemonte, Liguria e Valle d’Aosta
  • Istituto Superiore di Sanità
  • Centro Agricoltura Ambiente

International:

  • FAO/IAEA (Vienna, Austria)
  • Yale School of Public Health, & Dept. of Ecology and Evolutionary Biology, New Haven, USA
  • Institut Pasteur
  • Imperial College, London
  • European Bioinformatics Institute, Hinxton, UK
  • Baylor College of Medicine, Houston, TX, USA
  • USDA, US Department of Agriculture
  • ICIPE, International Centre of Insect Physiology and Ecology
  • University of California, Irvine
  • Johns Hopkins University
  • Vanderbilt University, Nashville, Tennessee, USA
  • Southern Medical University, Guangzhou, China
  • Royal Museum for Central Africa, Tervuren, Belgium
  • Bioiberica, S.A., Barcelona, Spain
  • University of Goettingen, Germany
  • University of Giessen, Germany
  • Slovak Academy of Sciences, Bratislava, Slovakia
  • Institute of Entomology a Ceské Budejovice, Czech Republic
  • CNRS di Gif-sur-Yvette, Francia

 

Active Research Grants 

  • Biotechnology and Biological Sciences Research Council (BBSRC) (PI Nikolai Windbichler, Malacrida & Scolari collaborators)
  • Bioiberica S.A., Barcelona, Research Contract (01/07/2015-current) (PI Scolari)
  • ERC Consolidator NIRV_HOST_INT (2016-2021) (PI Bonizzoni)
  • FAO/IAEA research CRP project # 17896 (2013-18) (PI Scolari)
  • FAO/IAEA Program of the United Nations project #17630 (2013-18) (PI Malacrida)
  • FAO/IAEA research CRP project #19049R0 (2015-2020)(PI Gomulski, Gasperi)
  • “Fight the mosquito bite” (Universitiamo Crowd-funding initiative, University of Pavia) (2015-present) (PI Gasperi, Malacrida)
  • Fondazione Bussolera Branca (2015-17) (PI Gasperi, Albertini)
  • NIH R21Al109263-01 (2014-17) (PI Malacrida)
  • USDA – University of Pavia – Baylor College “Consortium for genome sequencing of the Medfly, Ceratitis capitata” (2010 – present) (PI Gasperi)
  • Vectorbase-NIH Project “Comparative Genomics of 5 Glossina species” (PI Malacrida)