The present invention relates to a modified CCA gene which encodes a CCA-adding enzyme, which modified CCA gene leads to resistance against a positive-strand RNA virus having a transfer RNA-like structure (TLS). The invention further relates to plants and seeds comprising the modified genes, methods for making and identifying such plants and use of the gene.

Provided are methods of adding a polymer of non-canonical nucleotides to the 3 end of a ribonucleic acid (RNA). In certain embodiments, the methods comprise combining an RNA, a polynucleotide-3 nucleotidyl transferase, and non-canonical nucleotides, in a reaction mixture under conditions in which the polynucleotide-3 nucleotidyl transferase adds a polymer of the non-canonical nucleotides to the 3 end of the RNA. Such methods may further include analyzing the RNA using a nanopore. According to some embodiments, the methods include identifying the polymer of non-canonical nucleotides added to the 3 end of the RNA, and determining the junction between the 3 end of the RNA and the polymer of non-canonical nucleotides to identify the 3 end of the RNA. Kits that find use, e.g., in practicing the methods of the present disclosure are also provided.

Provided are methods of adding a polymer of non-canonical nucleotides to the 3 end of a ribonucleic acid (RNA). In certain embodiments, the methods comprise combining an RNA, a polynucleotide-3 nucleotidyl transferase, and non-canonical nucleotides, in a reaction mixture under conditions in which the polynucleotide-3 nucleotidyl transferase adds a polymer of the non-canonical nucleotides to the 3 end of the RNA. Such methods may further include analyzing the RNA using a nanopore. According to some embodiments, the methods include identifying the polymer of non-canonical nucleotides added to the 3 end of the RNA, and determining the junction between the 3 end of the RNA and the polymer of non-canonical nucleotides to identify the 3 end of the RNA. Kits that find use, e.g., in practicing the methods of the present disclosure are also provided.

Provided are methods of adding a polymer of non-canonical nucleotides to the 3 end of a ribonucleic acid (RNA). In certain embodiments, the methods comprise combining an RNA, a polynucleotide-3 nucleotidyl transferase, and non-canonical nucleotides, in a reaction mixture under conditions in which the polynucleotide-3 nucleotidyl transferase adds a polymer of the non-canonical nucleotides to the 3 end of the RNA. Such methods may further include analyzing the RNA using a nanopore. According to some embodiments, the methods include identifying the polymer of non-canonical nucleotides added to the 3 end of the RNA, and determining the junction between the 3 end of the RNA and the polymer of non-canonical nucleotides to identify the 3 end of the RNA. Kits that find use, e.g., in practicing the methods of the present disclosure are also provided.

Provided are methods of adding a polymer of non-canonical nucleotides to the 3 end of a ribonucleic acid (RNA). In certain embodiments, the methods comprise combining an RNA, a polynucleotide-3 nucleotidyl transferase, and non-canonical nucleotides, in a reaction mixture under conditions in which the polynucleotide-3 nucleotidyl transferase adds a polymer of the non-canonical nucleotides to the 3 end of the RNA. Such methods may further include analyzing the RNA using a nanopore. According to some embodiments, the methods include identifying the polymer of non-canonical nucleotides added to the 3 end of the RNA, and determining the junction between the 3 end of the RNA and the polymer of non-canonical nucleotides to identify the 3 end of the RNA. Kits that find use, e.g., in practicing the methods of the present disclosure are also provided.

Provided are methods of adding a polymer of non-canonical nucleotides to the 3 end of a ribonucleic acid (RNA). In certain embodiments, the methods comprise combining an RNA, a polynucleotide-3 nucleotidyl transferase, and non-canonical nucleotides, in a reaction mixture under conditions in which the polynucleotide-3 nucleotidyl transferase adds a polymer of the non-canonical nucleotides to the 3 end of the RNA. Such methods may further include analyzing the RNA using a nanopore. According to some embodiments, the methods include identifying the polymer of non-canonical nucleotides added to the 3 end of the RNA, and determining the junction between 3 end of the RNA and the polymer of non-canonical nucleotides to identify 30 end of the RNA. Kits that find use, e.g., in practicing the methods of the present disclosure are also provided.