The Genome of the Yellow Mealworm, Tenebrio molitor: It’s Bigger Than You Think

Author:

Oppert Brenda1ORCID,Dossey Aaron T.23ORCID,Chu Fu-Chyun2,Šatović-Vukšić Eva4ORCID,Plohl Miroslav4ORCID,Smith Timothy P. L.5ORCID,Koren Sergey6,Olmstead Morgan L.1ORCID,Leierer Dewey7,Ragan Gail1ORCID,Johnston J. Spencer8

Affiliation:

1. USDA Agricultural Research Service, Center for Grain and Animal Health Research, 1515 College Ave., Manhattan, KS 66502, USA

2. All Things Bugs LLC, 755 Research Parkway, Suite 465, Oklahoma City, OK 73130, USA

3. Invertebrate Studies Institute, 2211 Snapper Ln, Midwest City, OK 73130, USA

4. Division of Molecular Biology, Ruđer Bošković Institute, Bijenička 54, 10000 Zagreb, Croatia

5. USDA ARS U.S. Meat Animal Research Center, Clay Center, NE 68933, USA

6. Genome Informatics Section, Computational and Statistical Genomics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20894, USA

7. Department of Molecular Biology and Biophysics, Kansas State University, Manhattan, KS 66506, USA

8. Department of Entomology, Texas A&M University, College Station, TX 77843, USA

Abstract

Background: Insects are a sustainable source of protein for human food and animal feed. We present a genome assembly, CRISPR gene editing, and life stage-specific transcriptomes for the yellow mealworm, Tenebrio molitor, one of the most intensively farmed insects worldwide. Methods: Long and short reads and long-range data were obtained from a T. molitor male pupa. Sequencing transcripts from 12 T. molitor life stages resulted in 279 million reads for gene prediction and genetic engineering. A unique plasmid delivery system containing guide RNAs targeting the eye color gene vermilion flanking the muscle actin gene promoter and EGFP marker was used in CRISPR/Cas9 transformation. Results: The assembly is approximately 53% of the genome size of 756.8 ± 9.6 Mb, measured using flow cytometry. Assembly was complicated by a satellitome of at least 11 highly conserved satDNAs occupying 28% of the genome. The injection of the plasmid into embryos resulted in knock-out of Tm vermilion and knock-in of EGFP. Conclusions: The genome of T. molitor is longer than current assemblies (including ours) due to a substantial amount (26.5%) of only one highly abundant satellite DNA sequence. Genetic sequences and transformation tools for an insect important to the food and feed industries will promote the sustainable utilization of mealworms and other farmed insects.

Funder

Defense Advanced Research Projects Agency

Cooperative Research and Development Agreement

Publisher

MDPI AG

Subject

Genetics (clinical),Genetics

Reference56 articles.

1. Dossey, A.T., Morales-Ramos, J.A., and Guadalupe Rojas, M. (2016). Insects as Sustainable Food Ingredients: Production, Processing and Food Applications, Academic Press.

2. Transcriptome analysis of life stages of the house cricket, Acheta domesticus, to improve insect crop production;Oppert;Sci. Rep.,2020

3. Dossey, A.T., Oppert, B., Chu, F.-C., Lorenzen, M.D., Scheffler, B., Simpson, S., Koren, S., Johnston, J.S., Kataoka, K., and Ide, K. (2023). Genome and genetic engineering of the house cricket (Acheta domesticus): A resource for sustainable agriculture. Biomolecules, 13.

4. Dossey, A.T., Morales-Ramos, J.A., and Guadalupe Rojas, M. (2016). Insects as Sustainable Food Ingredients: Production, Processing and Food Applications, Academic Press.

5. Dossey, A.T., Morales-Ramos, J.A., and Guadalupe Rojas, M. (2016). Insects as Sustainable Food Ingredients: Production, Processing and Food Applications, Academic Press.

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