Grant Information

TRANSCRIPTOME AND SMALL RIBONUCLEIC ACID (RNA) PROFILING STUDIES IN THE ASIAN CITRUS PSYLLID

Sponsoring Institution Agricultural Research Service/USDA
Status ACTIVE
Funding Source USDA INHOUSE
Reporting Frequency Annual
Project Director SHATTERS R G
Accession Number 427147
Project Number 6034-22320-004-016N
Dates 2015-05-01 - 2020-04-30
Recipient Organization Vaccine and Gene Therapy Institute of Florida
9801 SW Discovery Way
Port St. Lucie,FL 34987
Keywords (hlb)
(mirna)
acid
asian
asiaticus
candidatus
citrus
greening
huanglongbing
liberibacter
micro
psyllid
ribonuclic
transcriptome
Research Effort Applied (30%)
Basic (45%)
Developmental (25%)
Classification Parameters
Knowledge AreaSubject of InvestigationField of SciencePercent
211 - Insects, Mites, and Other Arthropods Affecting Plants 910 - Grapefruit 1040 - Molecular biology 40%
215 - Biological Control of Pests Affecting Plants 920 - Orange 1130 - Entomology and acarology 40%
211 - Insects, Mites, and Other Arthropods Affecting Plants 1131 - Wine grapes 1130 - Entomology and acarology 10%
216 - Integrated Pest Management Systems 1130 - Table grapes 1040 - Molecular biology 10%
Goals / Objectives
  1. Matched Proteome and Transcriptome (RNAseq) Analysis of Asian Citrus Psyllid (ACP) Digestive tract and Salivary Glands in Asian citrus psyllid (ACP) with and without the Huanglongbing-Causing Candidatus Liberibacter asiaticus bacterium.
    1. Provide an understanding of genes important in digestive and salivation processes that could be interdiction points for psyllid control.
    2. Differential gene expression as a result of the presence of Candidatus Liberibacter asiaticus (CLas) would provide information about molecular interactions and insect perception .of the bacterium
    3. Could provide huanglongbing (HLB) resistance strategy in 3-4 years.
    4. Implications for control strategies that are applicable across a broad range of insects similar to the psyllid that are major factors effecting global crop production.
  2. Small Ribonucleic acid (RNA) Micro RNA) Analysis of Asian Citrus Psyllids (ACP) with and without the Huanglongbing-Causing Candidatus Liberibacter asiaticus (CLas) Bacterium
    1. Provide an understanding of micro Ribonucleic acid (miRNA) profiles and relationship to important genes within the Asian citrus psyllid.
    2. Showing involvement of micro Ribonucleic acid (miRNAs) in disease development/pathogen transmission would be a significant scientific advancement in our understanding of the huanglongbing disease complex.
    3. Micro Ribonucleic acid therapy could be rapidly commercialized, if their nvolvement in citrus greening is proven.
Methods (unparsed)

I. Matched Proteome and Transcriptome (Ribonucleic acid seq) Analysis of Asian Citrus Psyllid (ACP) Digestive tract and Salivary Glands in Asian citrus psyllid (ACP) with and without the Huanglongbing-Causing Candidatus Liberibacter asiaticus bacterium. 1. Develop 12 Ribonucleic acid (RNAseq) libraries from Polymerase chain reaction (PCR) positive and Polymerase chain reaction (PCR) negative Asian citrus psyllid (ACP) (six from each, three from each of digestive tract and salivary gland libraries). 2. Conduct Illumina Ribonucleic acid (RNAseq) analysis on these libraries (single run 50bp). 3. Provide Bioinformatic support to assemble reads onto a reference gene list developed from other psyllid Ribonucleic acid (RNAseq), Expressed sequence tag (EST) and genomic data. 4. Identify genes and proteins that are differentially regulated in response to the presence of Candidatus Liberibacer asiaticus within the Asian citrus psyllid. 5. Use this information to develop testable Ribonucleic acid interference (RNAi) based gene knockout experiments to study the effect of these knockouts on Candidatus Liberibacter asiaticus (CLas) acquisition by the Candidatus Liberibacter asiaticus. II. Small micro Ribonucleic acid (miRNA) Analysis of Asian Citrus Psyllids (ACP) with and without the Huanglongbing-Causing Candidatus Liberibacter asiaticus (CLas) Bacterium. 1. We have already conducted sequence analysis of two small Ribonucleic acid libraries. Vaccine and Gene Therapy Institute of Florida (VGTI) will provide bioinformatic analysis of these libraries to determine their quality, diversity and proof-of-concept to show that micro Ribonucleic acid (miRNAs) of plant origin can be identified from psyllids as a result of ingestion of these miRNAs. 2. If analysis of existing sequence data looks promising, Vaccine and Gene Therapy Institute of Florida will Conduct Illumina micro Ribonuclic acid sequencing on five other small Ribonucleic acid libraries. 3. Provide Bioinformatic support to assemble reads onto a reference gene list developed from other psyllid Ribonucleic acid seq, Expressed sequence tag (EST) and genomic data. 4. Identify differentially expressed micro Ribonuclic acid (miRNAs) in the psyllid as a result of the presence of Candidater Liberibacter asiaticus and associate these differences with information about the miRNA species origin (i.e. psyllid or citrus) and gene origin (to which gene and metabolic process does the miRNA belong). 5. Use the above information to develop testable hypotheses about the involvement of specific micro Ribonucleic acid (miRNAs) in disease development and psyllid transmission of Candidatus Liberibacter asiaticus (CLas). 6. Develop therapies targeting specific Micro Ribonucleic acid.