Sponsoring Institution
Agricultural Research Service/USDA
Status
ACTIVE
Funding Source
USDA INHOUSE
Reporting Frequency
Annual
Project Director
ROGERS E E
Accession Number
435985
Project Number
8044-22000-048-008R
Dates
2018-12-01 - 2022-08-31
Recipient Organization
AGRICULTURAL RESEARCH SERVICE
FT DETRICK,MD 21701
Keywords
asian
asiaticus
bing
candidatus
citrus
greening
hlb
huang
liberibacter
long
phloem
psyllid
transcriptome
Research Effort
Applied (25%)
Basic (50%)
Developmental (25%)
Classification Parameters
| Knowledge Area | Subject of Investigation | Field of Science | Percent |
|---|
| 212 - Pathogens and Nematodes Affecting Plants |
2410 - Cross-commodity research--multiple crops |
1160 - Pathology |
20% |
| 212 - Pathogens and Nematodes Affecting Plants |
5240 - Seeds and other plant propagules |
1160 - Pathology |
20% |
| 212 - Pathogens and Nematodes Affecting Plants |
999 - Citrus, general/other |
1160 - Pathology |
20% |
| 212 - Pathogens and Nematodes Affecting Plants |
1114 - Peach |
1160 - Pathology |
10% |
| 212 - Pathogens and Nematodes Affecting Plants |
1116 - Plum |
1160 - Pathology |
10% |
| 212 - Pathogens and Nematodes Affecting Plants |
1131 - Wine grapes |
1160 - Pathology |
10% |
| 212 - Pathogens and Nematodes Affecting Plants |
1419 - Leguminous vegetables, general/other |
1160 - Pathology |
10% |
Goals / Objectives
This proposal aims to identify genes expressed in citrus phloem that show differential regulation in HLB susceptible and resistant citrus cultivars. CLas is a phloem limited pathogen that manipulates phloem gene regulation to facilitate its life cycle. Phloem tissues are buried in the plant and are extremely difficult to physically separate and study in isolation. The inability to accurately characterize phloem responses to CLas infection remains a significant impediment to understanding CLas-host interactions and to identifying gene targets for engineering resistance.
Methods (unparsed)
Recently, our team developed a new methodology for woody crops that allows gene expression profiling of phloem infected tissues. The method leverages a translating ribosome affinity purification strategy (called TRAP) to isolate and characterize translating mRNAs from phloem specific tissues. The approach is unlike other gene expression profiling methods in that it only samples gene transcripts that are actively being transcribed into proteins and is thus a better representation of active cellular processes than total cellular mRNA. HLB-susceptible sweet orange, and HLB-resistant Poncirus and Carrizo (sweet orange x Poncirus) will be transformed to express the tagged ribosomal proteins under the control of existing phloem-specific promoters. Comparisons of susceptible and resistant phloem cell responses to CLas will identify those genes that are differentially regulated during these host responses. Identified genes will represent unique phloem specific targets for CRISPR knockout or overexpression, permitting the generation of HLB-resistant variants of major citrus cultivars.
Methods
Recently, our team developed a new methodology for woody crops that allows gene expression profiling of phloem infected tissues. The method leverages a translating ribosome affinity purification strategy (called TRAP) to isolate and characterize translating mRNAs from phloem specific tissues. The approach is unlike other gene expression profiling methods in that it only samples gene transcripts that are actively being transcribed into proteins and is thus a better representation of active cellular processes than total cellular mRNA. HLB-susceptible sweet orange, and HLB-resistant Poncirus and Carrizo (sweet orange x Poncirus) will be transformed to express the tagged ribosomal proteins under the control of existing phloem-specific promoters. Comparisons of susceptible and resistant phloem cell responses to CLas will identify those genes that are differentially regulated during these host responses. Identified genes will represent unique phloem specific targets for CRISPR knockout or overexpression, permitting the generation of HLB-resistant variants of major citrus cultivars.