METHODS FOR IDENTIFYING PLANT PATHOGEN RESISTANCE GENES

摘要

Methods are provided for identifying a plant disease resistance (R) gene for a plant disease of interest. The methods involve using bait sequences to select a subgroup of nucleic acids from a group of nucleic acids that are derived from a mutagenized plant that is susceptible to the plant disease of interest but that was produced by mutagenizing a plant that is resistant to the disease of interest. The bait sequences are designed to hybridize to one or more genes from at least one plant R gene family. The methods further involve sequencing the subgroup of nucleic acids to obtain a collection of nucleic acid sequences, comparing such nucleic acid sequences with corresponding sequences of one or more genes that are derived from a resistant plant, and identifying at least one nucleic acid sequence derived from the mutagenized plant that is not identical in sequence to a corresponding sequence from the resistant plant. Further provided are related methods for identifying a gene associated with a phenotypic change for a trait of interest in plants and other organisms.

主权项

1. A method for identifying a plant resistance (R) gene for a plant disease of interest, the method comprising:
(a) selecting a subgroup of nucleic acids comprising
(i) hybridizing in solution a group of nucleic acids and a set of bait sequences to form a hybridization mixture, wherein the group of nucleic acids are derived from a mutagenized plant that is susceptible to the plant disease of interest and wherein the bait sequences are designed to hybridize to one or more genes from at least one R gene family, and(ii) isolating from the hybridization mixture a subgroup of the nucleic acids that are hybridized to the bait sequences from nucleic acids that are not hybridized to the bait sequences; (b) sequencing the subgroup of nucleic acids to obtain a collection of nucleic acid sequences; (c) comparing the nucleic acid sequences obtained in (b) with corresponding sequences of the one or more genes that are derived from a reference plant that is resistant to the plant disease of interest; and (d) identifying at least one nucleic acid sequence derived from the mutagenized plant that is not identical in sequence to a corresponding sequence from the reference plant, wherein the corresponding sequence comprises a nucleic acid sequence of at least a portion of an R gene for the plant disease of interest.