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Identification of Candidatus Liberibacter asiaticus effector interactome by proximity labeling coupled with proteomics in citrus

Award Information
Agency: Department of Agriculture
Branch: N/A
Contract: 2023-03957
Agency Tracking Number: 2023-03957
Amount: $650,000.00
Phase: Phase II
Program: SBIR
Solicitation Topic Code: 8.2
Solicitation Number: USDA-NIFA-SBIR-009642
Timeline
Solicitation Year: 2023
Award Year: 2023
Award Start Date (Proposal Award Date): 2023-08-14
Award End Date (Contract End Date): 2025-08-31
Small Business Information
3802 Spectrum BLVD Suite 157
Tampa, FL 33612-9222
United States
DUNS: N/A
HUBZone Owned: No
Woman Owned: No
Socially and Economically Disadvantaged: No
Principal Investigator
 Quinton Allen
 (727) 735-1279
 quinton@soilcea.com
Business Contact
 Yianni Lagos
Phone: (937) 215-8371
Email: yianni@soilcea.com
Research Institution
N/A
Abstract

Huanglongbingdisease("HLB")causedbythebacteriumCandidatusLiberibacterasiaticus
("CLas")haskilledmillionsofcitrustreesandreducedaverageyieldinFloridaby50%costing
billionsinlostrevenue.
CRISPRisthemostpowerfultoolforrapidbreedingofnewHLB-resistantcitrustreesbut
hastobeprecededbyidentifyingspecificgene-editingtargets.Theidentificationoftargetgeneshas
beenchallengingbecauseofpoorunderstandingoftheHLBpathogenesismechanismandtechnical
limitationsofHLB-citrusprotein-interactionscreeningmethods.Apowerfulnewtooltoaddress
theselimitationsisTurboIDanengineeredbiotinligasethatcanlabelitsproximalproteinswhenitis
expressedincells.
TheproposedprojectaddressedthisopportunitybyemployingTurboID-mediatedproximity
labelingtoidentifycitrusproteinsthatinteractwitheffectorproteinsthatarecriticalforHLBCLas
infection. During Phase I research we produced a high-confidence list of CLas Sec-dependent
effector("SDE")-interactingcitrusproteinsbyusingTurboIDcoupledwithmassspectrometry.
InPhaseIIwewillusethediscoveredproteinsasgene-editingtargetsforCRISPRprecision
breedingtodevelopcitrusvarietiesthatdisrupttheeffector-citrusinteractionsleadingtogenetic
resistancetoHLB.
Tothatendwewillperformthreeobjectives.ThefirstisCRISPR-editingofpromising
SDE-interactinggenesandgeneratingnon-transgenicCRISPR-editedcitrusvarieties.Thesecondis
testing CRISPR-edited plants for resistance to HLB through graft inoculation and proteomics
analysis.Thethirdistocommencefieldtrials.
Weanticipatethisprojectwilldevelopnon-GMOHLB-resistantcitrustreesthatwillbesoldto
citrusgrowers.

* Information listed above is at the time of submission. *

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