Chitin biosynthesis genes in Diaphorina citri , Asian citrus psyllid

  1. Sherry Miller
  2. Teresa D. Shippy
  3. Blessy Tamayo
  4. Prashant S Hosmani
  5. Mirella Flores-Gonzalez
  6. Lukas A Mueller
  7. Wayne B Hunter
  8. Susan J Brown
  9. Tom D’elia
  10. Surya Saha

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Abstract

The polysaccharide chitin is critical for the formation of many insect structures, including the exoskeleton, and is required for normal development. Here we report the annotation of three genes from the chitin synthesis pathway in the Asian citrus psyllid, Diaphorina citri (Hemiptera: Liviidae), the vector of Huanglongbing (citrus greening disease). Most insects have two chitin synthase (CHS) genes but, like other hemipterans, D. citri has only one. In contrast, D. citri is unusual among insects in having two UDP-N-acetylglucosamine pyrophosphorylase (UAP) genes. One of the D. citri UAP genes is broadly expressed, while the other is expressed predominantly in males. Our work helps pave the way for potential utilization of these genes as pest control targets to reduce the spread of Huanglongbing.

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    This article is a preprint and has not been certified by peer review [what does this mean?].

    Sherry Miller 1Division of Biology, Kansas State University, Manhattan, KS 665062Allen County Community College, Burlingame, KS 66413Find this author on Google ScholarFind this author on PubMedSearch for this author on this siteTeresa D. Shippy 1Division of Biology, Kansas State University, Manhattan, KS 66506Find this author on Google ScholarFind this author on PubMedSearch for this author on this siteORCID record for Teresa D. ShippyBlessy Tamayo 3Indian River State College, Fort Pierce, FL 34981Find this author on Google ScholarFind this author on PubMedSearch for this author on this siteORCID record for Blessy TamayoPrashant S Hosmani 4Boyce Thompson Institute, Ithaca, NY 14853Find this author on Google ScholarFind this author on PubMedSearch for this author on this siteORCID record for Prashant S HosmaniMirella Flores-Gonzalez 4Boyce Thompson Institute, Ithaca, NY 14853Find this author on Google ScholarFind this author on PubMedSearch for this author on this siteORCID record for Mirella Flores-GonzalezLukas A Mueller 4Boyce Thompson Institute, Ithaca, NY 14853Find this author on Google ScholarFind this author on PubMedSearch for this author on this siteORCID record for Lukas A MuellerWayne B Hunter 5USDA-ARS, U.S. Horticultural Research Laboratory, Fort Pierce, FL 34945Find this author on Google ScholarFind this author on PubMedSearch for this author on this siteORCID record for Wayne B HunterSusan J Brown 1Division of Biology, Kansas State University, Manhattan, KS 66506Find this author on Google ScholarFind this author on PubMedSearch for this author on this siteORCID record for Susan J BrownTom D’elia 3Indian River State College, Fort Pierce, FL 34981Find this author on Google ScholarFind this author on PubMedSearch for this author on this siteORCID record for Tom D’eliaSurya Saha 4Boyce Thompson Institute, Ithaca, NY 148536Animal and Comparative Biomedical Sciences, University of Arizona, Tucson, AZ 85721Find this author on Google ScholarFind this author on PubMedSearch for this author on this siteORCID record for Surya SahaFor correspondence: ss2489@cornell.edu

    This work has been peer reviewed in GigaByte (https://doi.org/10.46471/gigabyte.23), which carries out open, named peer-review. These reviews are published under a CC-BY 4.0 license and were as follows:

    **Reviewer 1. Hailin Liu ** Is there sufficient data validation and statistical analyses of data quality?
    No. The validation work is not revealed in the manuscript, such as the qRT-PCR experiment.

    Is the validation suitable for this type of data?
    No. More validation work should be added instead of the RNA-seq data from the public database.

    Is there sufficient information for others to reuse this dataset or integrate it with other data? Yes

    Any Additional Overall Comments to the Author:
    Formatting errors should be corrected, including the tables and the alignment method of words. The introduction and methods seemed to be too simple for readers. More biological meanings should be explained in the manuscript. The basic assessment of the utilized genome should be added.

    Recommendation: Major Revision

    **Reviewer 2. Mary Ann Tuli ** Is the language of sufficient quality?
    Yes. The manuscript reads very well.

    Are all data available and do they match the descriptions in the paper?
    No. Additional Comments :

    1. Line 149. "Multiple alignments of the predicted D. citri proteins and their insect homologs were performed using MUSCLE We need the output of MUSCLE (FASTA).

    2. Line 151. Phylogenetic trees were constructed (figures 1 and 4) using full-length protein sequences in MEGA7or MEGAX. We need the files underlying the phylogenetic tree (newick). Please indicate which version of MEGA was used for each tree.

    3. Line 152. Gene expression levels were obtained from the Citrus greening Expression Network and visualized using Excel and the pheatmap package in R. Please can you provide a file of the raw data used to produce the heatmap (figure 2) and the Expression levels of UAP2 in male and female tissues (figure 5).

    Are the data and metadata consistent with relevant minimum information or reporting standards? Yes. Nomenclature standards have been met.

    All cited INSDC accession numbers are publicly available.

    Is the data acquisition clear, complete and methodologically sound? Yes. The curation workflow used for community annotation is available via protocols.io , nonetheless the manuscript includes a comprehensive summary which is appropriate.

    Is there sufficient detail in the methods and data-processing steps to allow reproduction?
    No. See "Are all data available and do they match the descriptions in the paper?" above. Once the additional files are made available I believe reproduction will be possible.

    Is there sufficient data validation and statistical analyses of data quality? Yes

    Is the validation suitable for this type of data? Yes

    Is there sufficient information for others to reuse this dataset or integrate it with other data? Yes

    Any Additional Overall Comments to the Author:

    1. Line 147. Apollo version should be included in the other D citri manuscripts.

    2. Citation [26] MUSCLE. https://www.ebi.ac.uk/Tools/msa/muscle/.

    • the website suggests users of MUSCLE cite DOI:10.1093/nar/gkz268

    Some of my comments/recommendations are pertinent to the other D. citri manuscripts currently under review.

  2. Version published to 10.1101/2020.09.22.309211v4 on bioRxiv
  3. Version published to 10.1101/2020.09.22.309211v3 on bioRxiv
  4. Version published to 10.1101/2020.09.22.309211v2 on bioRxiv
  5. Version published to 10.1101/2020.09.22.309211v1 on bioRxiv