Table I. Summary of mutant lines from the LRR-XII library

Mismatches between the spacer and M82 target sequences are underlined. Alleles are combined data from T0 and T1 plants. The genotype of the plants is indicated. Active indicates that Cas9 is actively producing new alleles in those lines. NA, Not applicable; WT, wild type.

LineTarget + PAMTargetLRR-XII Family?T0 and T1 AllelesGenotypeNotes
LRR-1GAAGATATGGTTACTCATGTTGGSolyc07g016010Yes−4 bp deletion/WT, activeM82sp+T1: some weak plants, low seed set, sterility
LRR-2GTCACCACAAAGTGCATCATTGGSolyc06g006040Yes−4/+609 (from Solyc06g006020)M82sp+T0 and T1: high seed weight, low fruit set
GTCACCACAAAGTGCATCATCGGaSolyc06g006020Yes−9/larger deletion
GTCACCACAAAGTGCATCATCGGaSolyc06g006050Yes−1/no amplification (276 upstream)
GTCACCACAAAGTGCATCATCGGaSolyc06g006070Yes−9/−8 (315 upstream)
LRR-3No gRNAN/AN/AN/AM82sp+
LRR-4GGCAATCTCAGCTTCCTAACAGGSolyc10g085110YesWTM82sp+T1: leaf development (1/4)
GAGATGCAACATCCACATCTGGGSolyc03g118330Yes−1/−2,112 bp
GCAGGTGACCAAGTCCATAAGGGSolyc04g014900Yes−1/no amplification (unresolved)
LRR-5GAAAGATCAAAAATTTCAGCAGGSolyc07g018180YesWT, sequence different from the referenceM82sp+
LRR-6GGATTAGTGATTCCTTTCCGGGGSolyc06g006020Yes−2/−3/−7M82sp+Mostly parthenocarpic fruit with small aborted seed; one T1 plant
GGATTAGTGATTCCTTTCCGAGGaSL2.50ch00:17387212No−5/−6/WT
LRR-7GAACTCTCATCATTGTCGGAGGGSolyc03g096190YesWTM82sp+
LRR-8GCTTTGGAATATCTTCACCACGGSolyc03g006100Yes−1/−5M82T1: serrated cotyledons
LRR-9GTAGGGATAGTGTCAACTATGGGSolyc08g075620Yes−7/−1/−3M82sp+Sterile
LRR-10GCTTTAGACTATCTCCATAAAGGSolyc02g068880Yes−2/WT/−∼330M82sp+Sterile
LRR-11GTGTACAAAGGTGCATTGTTTGGSolyc04g014890Yes+1 homozygousM82T1: stunted, bleached plants (1/4)
GTGTACAAAGGTGCATTGTTTGGaSolyc04g014650Yes−2/+1
GTGTACAAAGGCACATTGTTTGGbSolyc04g014900Yes+1/WT
GTGTACAAAGGCACATTGTTTGGbSolyc04g015000YesWT
GTGTACAAAGGCACATTGTTTGGbSL2.50ch04 6829670NoActive
LRR-12GAAAGCAACTTGCTTGGTAACGGSolyc02g068830YesWTM82sp+
LRR-13GCAGGCAAAAAACCGTCCAAGGGSolyc02g068820YesWTM82sp+
LRR-14GTAGGGATAGTGTCAACTATGGGSolyc08g075620YesWTM82T1: no anthocyanin (1/4)
LRR-15GGCTACAGCTTGTTAATAGTCGGSolyc06g005880Yes−7/−87M82sp+
LRR-16GGCCTATTCCGAGTTCATTCGGGSolyc02g031790Yes−4/−5M82sp+
LRR-17GGTCAGATACCTAGTACAATCGGSolyc08g075600Yes−1/−8M82
LRR-18GTCATTTTCATCCTTTCGACCGGSolyc02g072400YesWTM82
LRR-19GGCGGCCAACCATCCACACTTGGSolyc10g085120Yes−1/−3/−4/−5M82sp+Likely tetraploid
LRR-20GAGATTTGAATTTGAGAAGCTGGSolyc02g072380YesWTM82sp+
LRR-21GAGATATTAAACAGTGACGCGGGSolyc04g015980Yes+1/−1/−4M82sp+Mostly parthenocarpic fruit with small aborted seed; one T1 plant
LRR-22GCAGCAGCTTGTTATGACTTTGGSolyc06g006030YesWTM82sp+
LRR-23GGTGAAATTCCTGCCGAGCTGGGSolyc02g070890Yes−2 homozygousM82sp+T1: all are insensitive to flg22 (expected)
GGTGAAATTCCTGCCGAACTAGGbSolyc02g070910YesSomatic active
LRR-24GCAGGCAAAAAACCGTCCAAGGGSolyc02g068820YesWTM82sp+T1: some plants light bleaching, slow-growing plants, low fruit set
GTGAGGTTCCCAATAGAAGGCGGSolyc07g018190YesWT
LRR-25GTGCAGTTAGATAAACCAGGGGGSolyc02g072440Yes−3/−4/−9/−10M82sp+T0 would not self, many parthenocarpic fruit, few seeds from LRR-25 × M82sp+
  • a Unintended targets that were missed during gRNA design.

  • b Off-targets to the gRNA sequence.