Methods to evaluate traits
The present disclosure provides methods for testing agronomic performance of transgenic traits and genome edits in plants and for accelerated selection of such plants. Methods evaluated include testing constructs for transgenic trait performance (T) using isolines. The methods comprise crossing events into different genetic backgrounds. Methods also comprise combining isoline data with F2:3 bulk data and developing breeding values across different genetic backgrounds for a higher level of confidence in selecting events.
1 . A method for accelerated selection of a genome edited plant comprising at least five site-specific genome edits introduced via CRISPR/Cas, wherein said site-specific genome edits comprise targeted mutagenesis and do not comprise stable integration of exogenous polynucleotides, the method comprising:
(a) crossing a plant line or a population of plants that is homozygous positive for the genome edits with a diverse panel of plant lines that do not contain said genome edits;
(b) crossing an isogenic null of (a) with the same diverse panel of plant lines of (a) that do not contain said genome edits, wherein the isogenic null of (a) is homozygous negative for the genome edits;
(c) collecting segregating offspring of said crosses of (a) and (b) to produce a bulk pool of genome edits positive and genome edits negative hybrids;
(d) phenotyping or genotyping the bulk pool of hybrids that are positive for the genome edits and the hybrids that are negative for the genome edits;
(e) assigning a breeding value to each genome edited plant or a subset of the genome edits in the genome edited hybrids based on the phenotyping or the genotyping; and
(f) selecting the genome edits based on the breeding value.
2 . The method of claim 1 , further comprising growing the positive and negative hybrids in adjacent yield plots having similar environmental conditions.
3 . The method of claim 2 , further comprising comparing agronomic characteristics of said positive and negative plant.
4 . The method of claim 3 , wherein the agronomic characteristics are selected from the group consisting of emergence, early vigor, growth, flowering time, flowering duration, height, maturity, and yield.
5 . The method of claim 1 , wherein whole genome molecular markers are used to characterize the hybrid plants.
6 . The method of claim 1 , wherein whole genome marker by genome edits is used to predict a win/loss and genome edit breeding value of said genome edits.
7 . The method of claim 3 , further comprising setting a criterion for acceptable agronomic phenotype.
8 . The method of claim 1 , wherein the breeding value is assigned based on both phenotyping and genotyping.