Shatterproof genes and mutations

Abstract

The present disclosure provides shatterproof (SHP) genes and plants and/or plant cells bearing one or more mutations in a shatterproof gene; as well as methods of making and using such plants. In some embodiments the plant or plant cell is resistant to preharvest dehiscence.

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         47 . A plant or part thereof comprising at least one mutation in at least three, at least four, at least five, at least six, at least seven, or eight nucleic acid sequences encoding SHATTERPROOF (SHP) genes. 
     
     
         48 . The plant or part thereof of  claim 47 , wherein the nucleic acid sequences have at least 90% sequence identity, at least 95% sequence identity, at least 98% sequence identity, or at least 99% sequence identity to nucleic acid sequences selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, and SEQ ID NO: 8. 
     
     
         49 . The plant or part thereof of  claim 47 , wherein the nucleic acid sequences are selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, and SEQ ID NO: 8. 
     
     
         50 . The plant or part thereof of  claim 47 , wherein the mutation is a frameshift mutation. 
     
     
         51 . The plant or part thereof of  claim 50 , wherein the frameshift mutation results in one or more nucleotide insertions or deletions as compared to the corresponding endogenous gene without the frameshift mutation. 
     
     
         52 . The plant or part thereof of  claim 50 , wherein the frameshift mutation results in a premature stop codon. 
     
     
         53 . The plant or part thereof of  claim 50 , wherein the mutation reduces or eliminates the expression or activity of the SHP gene and/or SHP polypeptide. 
     
     
         54 . The plant or part thereof of  claim 47 , wherein the plant exhibits reduced susceptibility to preharvest dehiscence. 
     
     
         55 . The plant of  claim 47 , wherein the plant is selected from the group consisting of  Brassica napus, Brassica rapa, Brassica oleracea, Brassica juncea, Brassica species, Raphanus sativus, Pisum sativum, Phaseolus vulgaris, Lens culinaris, Glycine max , and Fabaceae species. 
     
     
         56 . A method of producing the plant of  claim 47 , comprising the steps of:
 a) introducing mutations into plant cells, wherein the mutations are at least one mutation in at least three, at least four, at least five, at least six, at least seven, or eight nucleic acid sequences encoding SHP genes;   b) selecting plant cells containing the mutations; and   c) regenerating a plant having the mutations;   wherein the plant exhibits reduced susceptibility to preharvest dehiscence.   
     
     
         57 . The method of  claim 56 , wherein the mutations are introduced using one or more vectors, wherein the vectors comprise gene editing components selected from the group consisting of a nuclease, an RNA-guided DNA endonuclease, a CRISPR/Cas9 system, a TALEN, a zinc finger, and a meganuclease designed to target a nucleic acid sequence encoding a SHP gene. 
     
     
         58 . The method of  claim 56 , wherein the mutations are introduced using a GRON system designed to target a nucleic acid sequence encoding a SHP gene. 
     
     
         59 . The method of  claim 58 , wherein the GRON system comprises one or more modifications selected from the group consisting of a Cy3 group, 3PS group, and a 2′O-methyl group. 
     
     
         60 . The method of  claim 56 , wherein the nucleic acid sequences have at least 90% sequence identity, at least 95% sequence identity, at least 98% sequence identity, or at least 99% sequence identity to nucleic acid sequences selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, and SEQ ID NO: 8. 
     
     
         61 . The method of  claim 56 , wherein the nucleic acid sequences are selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, and SEQ ID NO: 8. 
     
     
         62 . The method of  claim 56 , wherein the mutation is selected from the group consisting of a frameshift mutation, a frameshift mutation resulting in one or more nucleotide insertions or deletions as compared to the corresponding endogenous gene without the frameshift mutation, and a frameshift mutation resulting in a premature stop codon, and wherein the mutation reduces or eliminates expression of the SHP gene and/or SHP polypeptide. 
     
     
         63 . The method of  claim 56 , wherein the plant is selected from the group consisting of  Brassica napus, Brassica rapa, Brassica oleracea, Brassica juncea, Brassica species, Raphanus sativus, Pisum sativum, Phaseolus vulgaris, Lens culinaris, Glycine max , and Fabaceae species. 
     
     
         64 . An F1 plant, wherein the F1 plant has the plant of  claim 47  as a parent. 
     
     
         65 . A method of making plant seeds, the method comprising crossing the plant of  claim 47  with another plant and harvesting seed therefrom. 
     
     
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         67 . A plant produced by growing the seed of  claim 65 , wherein the plant has all the physiological and morphological characteristics of a plant comprising at least one mutation in at least three, at least four, at least five, at least six, at least seven, or eight nucleic acid sequences encoding SHATTERPROOF (SHP) genes.