Abstract
Candidatus Liberibacter asiaticus ( C Las), the bacteria responsible for citrus greening disease [huanglongbing (HLB)], has become a worldwide threat to citrus ( Citrus sp.) production. HLB has proven difficult to study and treat because of the complex interactions between C Las, the citrus host, and insect vectors. We have selected for single chain fragment variable (scFv) antibodies from a specialized bacteriophage library for binding activity against C Las proteins InvA and TolC. Portions of each protein were chosen as antigens based on predicted binding availability and theorized necessary functions in pathogenicity. Binding affinity for individual scFv-expressing clones was confirmed by phage enzyme-linked immunosorbent assay (ELISA). The scFv sequences were stably transformed under the control of a tandem Cauliflower mosaic virus 35S (CaMV 2x35S) promoter by Agrobacterium tumefacien –mediated transformation into ‘Carrizo’ citrange ( Citrus sinensis × Poncirus trifoliate ), a citrus rootstock cultivar. Replicated plants of single transformations were inoculated by infestation with C Las positive asian citrus psyllid ( Diaphorina citri ), a C Las vector. Inoculation and disease progression was monitored through quantitative real-time polymerase chain reaction. Inoculated transgenic plants showed significantly reduced C Las titer compared with wild types. A subpopulation of transgenic plants displayed no measurable surviving bacteria after 12 months. Interestingly, individual replicated plants from the same transgenic events strongly segregated into two populations by resistance phenotype: a minority that were indistinguishable from wild-type plants and a majority that were highly resistant. Our results are the first step in developing a novel protection strategy for HLB.
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Claims 5
Claim · model-written summary
Individual scFv-expressing clones had confirmed binding affinity.
Verbatim
e0“Binding affinity for individual scFv-expressing clones was confirmed by phage enzyme-linked immunosorbent assay (ELISA).”
e1“binding activity against C Las proteins InvA and TolC”
Claim · model-written summary
ScFv sequences were stably transformed into Carrizo citrange.
- Taxon
- Citrus sinensis × Poncirus trifoliate
Citrus sinensise0 - Material
- ‘Carrizo’e0
- Gene
- scFve0
- Outcome
- stably transformede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The scFv sequences were stably transformed under the control of a tandem Cauliflower mosaic virus 35S (CaMV 2x35S) promoter by Agrobacterium tumefacien –mediated transformation into ‘Carrizo’ citrange ( Citrus sinensis × Poncirus trifoliate ), a citrus rootstock cultivar.”
Claim · model-written summary
Inoculated transgenic citrus plants had significantly reduced C Las titer relative to wild types.
- Treatment
- Inoculated transgenic plantse0
- Interactor
- C Lase0pathogen
- Outcome
- C Las titere0
- Direction
- reducede0
- Comparator
- wild typese0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Inoculated transgenic plants showed significantly reduced C Las titer compared with wild types.”
t0“Citrus”
Claim · model-written summary
Some transgenic citrus plants had no measurable surviving bacteria after 12 months.
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“A subpopulation of transgenic plants displayed no measurable surviving bacteria after 12 months.”
e1“Inoculated transgenic plants showed significantly reduced C Las titer compared with wild types.”
t0“Citrus”
Claim · model-written summary
Replicates of the same citrus transformation events segregated into a wild-type-like minority and a highly resistant majority.
- Interactor
- C Lase1pathogen
- Outcome
- segregated into two populations by resistance phenotypee0
- Qualifier
- a minority that were indistinguishable from wild-type plants and a majority that were highly resistante0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“individual replicated plants from the same transgenic events strongly segregated into two populations by resistance phenotype: a minority that were indistinguishable from wild-type plants and a majority that were highly resistant.”
e1“Inoculated transgenic plants showed significantly reduced C Las titer compared with wild types.”
t0“Citrus”