Sponsoring Institution
Agricultural Research Service/USDA
Status
ACTIVE
Funding Source
USDA COOPERATIVE AGREEMENT
Reporting Frequency
Annual
Project Director
HECK M L
Accession Number
431131
Project Number
8062-22410-006-10A
Dates
2016-02-01 - 2018-12-31
Recipient Organization
COLUMBIA UNIVERSITY
630 WEST 168TH STREET
NEW YORK,NY 10032
Keywords
asiaticus
candidatus
citrus
clas
hlb
igem
liberibacter
psyllid
Research Effort
Applied (25%)
Basic (75%)
Developmental (0%)
Classification Parameters
| Knowledge Area | Subject of Investigation | Field of Science | Percent |
|---|
| 215 - Biological Control of Pests Affecting Plants |
2410 - Cross-commodity research--multiple crops |
1000 - Biochemistry and biophysics |
100% |
Goals / Objectives
The overall goal of our project is to develop technologies that block Asian citrus psyllid (ACP) transmission of Candidatus Liberibacter asiaticus (CLas) and to develop a simple yeast biosensor that will enable the identification of CLasharboring insects in the field.
Methods (unparsed)
This project will utilize a panel of isofemale ACP lines which have been characterized for their difference in CLas transmission efficiency. Analysis of differences between natural populations of ACP varying in their ability to acquire or transmit the citrus greening pathogen may lead to new strategies for the management of HLB. We will determine the genetic basis of CLas transmission by Diaphorina citri, the Asian citrus psyllid (ACP), including how interactions between the insect and its bacterial endosymbionts contribute to the CLas transmission process. Candidatus Profftella armatura is an endosymbiont specific to the ACP which produces high amounts of a polyketide toxin, diaphorin, hypothesized to play a key role in enabling CLas transmission by the insect vector. Functional analysis of diaphorin, including identification and characterization of its protein binding partners, will be performed. Citrus tristeza virus (CTV) gene silencing technology will be used to knock down production of ACP proteins in insects feeding on nontransgenic CTV-silenced plants. Proteomic data revealing specific ACP peptides found only in insects harboring CLas will be exploited to develop a yeast biosensor to identify individual ACP harboring CLas that can easily be used by growers. Our project will have an education component: we will involve Cornell University undergraduate researchers in the International Genetically Engineered Machine (iGEM) program in the development of the CLas biosensor using synthetic biology.
Methods
This project will utilize a panel of isofemale ACP lines which have been characterized for their difference in CLas transmission efficiency. Analysis of differences between natural populations of ACP varying in their ability to acquire or transmit the citrus greening pathogen may lead to new strategies for the management of HLB. We will determine the genetic basis of CLas transmission by Diaphorina citri, the Asian citrus psyllid (ACP), including how interactions between the insect and its bacterial endosymbionts contribute to the CLas transmission process. Candidatus Profftella armatura is an endosymbiont specific to the ACP which produces high amounts of a polyketide toxin, diaphorin, hypothesized to play a key role in enabling CLas transmission by the insect vector. Functional analysis of diaphorin, including identification and characterization of its protein binding partners, will be performed. Citrus tristeza virus (CTV) gene silencing technology will be used to knock down production of ACP proteins in insects feeding on nontransgenic CTV-silenced plants. Proteomic data revealing specific ACP peptides found only in insects harboring CLas will be exploited to develop a yeast biosensor to identify individual ACP harboring CLas that can easily be used by growers. Our project will have an education component: we will involve Cornell University undergraduate researchers in the International Genetically Engineered Machine (iGEM) program in the development of the CLas biosensor using synthetic biology.