Peptide phosphorodiamidate morpholino oligomers plant delivery to reduce pathogens and insect pests
Inventors
Hunter, Wayne B. • Pelz-Stelinski, Kirsten
Assignees
US Department of Agriculture USDA • University of Florida Research Foundation Inc
Publication Number
US-11001842-B2
Publication Date
2021-05-11
Expiration Date
2039-05-15
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Abstract
The current disclosure provides for peptide-conjugated phosphorodiamidate morpholino oligomers (PPMOs) antisense oligonucleotides that target plant pathogens and insect pests by targeting the microbes within the insect pest. Methods of delivering PPMOs to bacteria in plants, in insect carriers, and to the insects via plant feeding are also provided.
Core Innovation
The invention provides peptide-conjugated phosphorodiamidate morpholino oligomers (PPMOs) antisense oligonucleotides that target plant pathogens and insect pests by targeting microbes within insect pests. It includes methods of delivering PPMOs to bacteria in plants, in insect carriers, and to insects via plant feeding.
The problem addressed is the difficulty of targeting plant pathogenic bacteria. Few antibiotics are approved for crops, and concerns about antibiotic resistance have restricted their use. Severe bacterial plant pathogens cause significant crop losses, and current treatment strategies face issues related to cost, registration, and public perception. There is a need for non-antibiotic, safe, and highly specific treatments that can target internal plant pathogens and bacteria harbored by insect pests without harming beneficial microbiota.
The invention offers RNA-targeting compositions and methodologies using PPMOs that inhibit gene expression by binding to complementary RNA sequences, thus interfering with translation. These PPMOs can be delivered to plants and insects via various methods such as root soak, injection, foliar spray, and feeding. PPMOs can induce RNA silencing in bacteria within insect pests and plants, resulting in targeted pathogen and pest control. The compositions can be applied in combination with other agents but focus on the use of PPMOs for effective, specific, and environmentally safer management of plant pathogens and insect pests.
Claims Coverage
The patent contains 7 main inventive features derived from its independent claims, covering oligonucleotide compositions and methods for controlling bacteria and insects using PPMOs.
Oligonucleotide composition with selected sequences
An oligonucleotide comprising at least one phosphorodiamidate morpholino residue, having a sequence selected from a specified group of SEQ ID NOs including SEQ ID NO:2 to SEQ ID NO:49.
Phosphorodiamidate morpholino residue in all oligonucleotide units
Each residue of the oligonucleotide comprises a phosphorodiamidate morpholino residue.
Peptide-conjugated oligonucleotide
The oligonucleotide further comprises a peptide conjugated to the oligonucleotide.
Method of controlling bacteria in insects via feeding on treated food sources
A method involving contacting a food source edible by the insect with a PPMO oligonucleotide having a sequence from a specified group, allowing the insect to feed and ingest the oligonucleotide, thereby inducing RNA silencing in some of the bacteria present in the insect to control the bacteria.
Target bacteria and food sources in the method
The bacteria targeted include Candidatus Liberibacter asiaticus and Candidatus Liberibacter solanacearum; the food source can be plant, bait, artificial diet, or sugar solution; plants can be treated by root soak, injection, or foliar spray; insects include Diaphorina citri or Diaprepes abbreviatus.
Method of controlling plant pathogenic bacteria in plants
A method comprising contacting plants with PPMO oligonucleotides from a specified group by root soak, injection, or foliar spray, allowing distribution of the oligonucleotide through plant tissues, and inducing a detrimental effect on plant pathogenic bacteria present in the plant tissues.
Specific oligonucleotide and plant targets for controlling plant pathogens
The oligonucleotides have specified sequences from a defined group of SEQ ID NOs and the bacteria targeted include Candidatus Liberibacter asiaticus or Candidatus Liberibacter solanacearum; the plants targeted include citrus and potato.
The claims define specific PPMO oligonucleotide sequences, including peptide conjugation, and methods for controlling bacteria in insects via feeding and in plants via application, targeting particular bacterial pathogens and insect pests.
Stated Advantages
PPMOs are effective, specific, and lack non-antisense effects.
PPMOs provide safer, highly specific treatments against internal plant pathogens and bacteria within insect pests.
PPMOs show long-term stability in plant tissues after application.
PPMOs can be delivered using various methods including root soak, foliar spray, injection, and feeding, demonstrating versatile delivery options.
PPMOs reduce bacterial gene expression and viability, leading to control of the target bacteria and increased mortality of insect pests.
Documented Applications
Controlling plant pathogenic bacteria such as Candidatus Liberibacter asiaticus and Candidatus Liberibacter solanacearum in citrus, potato, and other crops.
Controlling insect pests such as the Asian citrus psyllid (Diaphorina citri), Diaprepes abbreviatus, and glassy-winged sharpshooter by delivering PPMOs through plant feeding.
Reducing bacterial endosymbionts in insect pests to increase insect mortality.
Using PPMOs as soil drenches, root soaks, foliar sprays, or injections to deliver control agents systemically in plants and indirectly to feeding insects.
Application in agriculture for managing bacterial diseases and insect pests in fruit crops, potatoes, okra, and other agricultural plants.
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