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System and methods for the biocontrol of plant pathogens

A technology for plant pathogens, control systems, applied in the fields of botanical equipment and methods, biochemical equipment and methods, plant products, etc., capable of solving problems such as inability to effectively solve one or more problems or challenges, lack of understanding, etc.

Inactive Publication Date: 2019-11-01
PEBBLE LABS USA INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0014] Due to a lack of understanding, attempts to meet these longstanding needs may not effectively address one or more of the problems or challenges identified here

Method used

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  • System and methods for the biocontrol of plant pathogens
  • System and methods for the biocontrol of plant pathogens
  • System and methods for the biocontrol of plant pathogens

Examples

Experimental program
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Embodiment 1

[0168] Example 1: Demonstrates the construction of a bacterial plasmid encoding a hairpin (hp) RNA construct encoding a target green Snapback RNA for Chromochrome Fluorescent Protein (GFP).

[0169] As demonstrated by the present inventors, using from New England BioLabs Inc. The HiFiDNA Assembly Cloning Kit constructs hpRNA encoding plasmid vectors. Here, as an illustrative example, seven hpRNA vectors were constructed to target green fluorescent protein (GFP) encoding mRNA expressed from pJL-TRBO-G (Lindbo 2007).

[0170] The maps of these plasmids are shown in the figure below: 1=7: pAD-WRKY-GHY1 ( figure 1 ), pAD-ADH-GHY2 ( figure 2 ), pOXB-WRKY-GHY3 ( image 3 ), pOXB-ADH-GHY4 ( Figure 4 ), pAD-WRKY-GHY5 ( Figure 5 ), pAD-ADH-GHY6 ( Figure 6 ). An hpRNA designed using the TMV motor protein: pAD-WRKY-GHY7 ( Figure 7 ) was constructed to target kinesin and GFP from PJL-TRBO-G. Bold bases are overlapping sequences. Bases in italics are insertion sequence...

Embodiment 2

[0173] Example 2: Demonstration of a bacterial plasmid encoding an inserted intron between the sense and antisense strands of the GFP gene target build.

[0174] An intron of a putative WRKY-type DNA binding protein from Arabidopsis thaliana is 287 bp in length and was cloned from plasmid pJawohB-RNAi (Accession No. AF404854) (Table 9: SEQ ID NO. 26).

[0175] The present inventors generated another intron from Zea mays alcohol dehydrogenase intron Adhl with a length of 566 bp and cloned it from plasmid pMCG161 (Accession No. AY572837) (Table 9: SEQ ID NO. 27-29).

Embodiment 3

[0176] Example 3: Demonstration of construction and sequence listing of exemplary hpRNAs.

[0177] In this example, exemplary hpRNAs comprising SEQ ID NO. 30-36 listed in Table 10 below were constructed.

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Abstract

The inventive technology relates to novel systems, methods, and strategies for the control of plant pathogens and herbivores. The inventive technology may specifically include novel compositions, systems, and methods configured to deliver inhibitory RNA molecules to a pathogen-infected plant. In a preferred embodiment, the invention may include genetically engineered endophytic bacteria configuredto deliver one or more inhibitory RNA molecules to a pathogen-infected plant. One preferred embodiment may include a novel trans-kingdom delivery of hairpin RNA targeting viral encoded proteins resulting in the reduction of viral protein accumulation levels.

Description

[0001] This application takes priority from U.S. Provisional Application No. 62 / 430,671, filed December 6, 2016. The entire specification and drawings of the above-mentioned application are hereby incorporated by reference in their entirety. [0002] sequence listing [0003] This application contains a Sequence Listing, which has been filed in ASCII electronic format and is hereby incorporated by reference in its entirety. technical field [0004] The field of the invention relates generally to plant molecular biology and plant biotechnology. More specifically, it relates to structures and methods for inhibiting the expression of or down-regulating a target gene. The present technology also relates to biological control of plant pathogens, pests and / or herbivores. In particular, the present invention may relate to new techniques, systems and methods for the biological control of disease-spreading plant pathogens through the delivery of inhibitory RNA molecules by genetica...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N63/02C12N15/10A01N63/60
CPCA01H3/00A01H17/00C12N15/113C12N15/111C12N15/1131C12N2310/14A01N63/60C12N15/8218C12N15/8283C12N2310/531
Inventor 理查德·塞瑞凯伦·尹佩德罗·科斯塔·努内斯
Owner PEBBLE LABS USA INC
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