Fig. 2

Target sites of the gRNA in the OsNAC2d gene and recombining diagram of the pYLCRISPR/Cas9-OsNAC2d-T12 vector A: the position and nucleotide sequences of two gRNA targets in the OsNAC2d gene locus; B: the recombining diagram of two gRNA cassettes and pYLCRISPR/Cas9-MT vector."

Table 1

Nucleotide sequence and corresponding primer sequences used in this study"

Fig. 1

Relative expression patterns of OsNAC2d gene in rice A: the differential expression level of OsNAC2d gene in the root, stem, leaf, anther, and grain of wild type ZH11; B: the temporal expression of OsNAC2d in leaves and roots of rice incubated in the nutrient solution including 15% PEG-6000, and samples were collected at 0, 0.5, 3, 6,12, and 24 hours after treatment, respectively. Different letters above the columns are significantly different at the 0.05 probability level."

Fig. 3

Sequencing detection of two target sequences of pYLCRISPR/Cas9-OsNAC2d-T12 recombinant vector"

Fig. 4

Identification of the positive transgenic plants -: the negative control is an equal volume of water instead of the template; +: the positive control is an equal volume of the positive plasmid as a template; 1-11: transgenic lines; M: 2 kb DNA ladder."

Fig. 5

Sequence alignment of OsNAC2d between the osnac2d mutants and its wild type The blue letters are the target genome sequence, the yellow highlighted letters denote PAM, dashes strikethrough indicate the deleted bases, and insertion nucleotides are shown in red lowercase letters. -: deletion; +: insertion; WT: wild type (ZH11)."

Fig. 6

Phenotype of osnac2d mutants and its wild type ZH11 incubated in the nutrient solution including 15% PEG-6000 A and D denote wild type ZH11, B and E denote osnac2d-6 mutant, C and F denote osnac2d-4-2 mutant. The control group is the seedlings incubated in the complete nutrient solution. The duration of PEG-6000 treatment is 14 days."

Fig. 7

Growth status of osnac2d and wild type ZH11 incubated in the nutrient solution including 15% PEG-6000 A: the length of root; B: the length of seedling; C: the length of whole plant. The control group is the seedlings incubated in the complete nutrient solution. The duration of PEG-6000 treatment is 14 d. Different lowercase letters above the columns are significantly different at the 0.05 probability level."

Fig. 8

Biomass of osnac2d and wild type ZH11 incubated in the nutrient solution including 15% PEG-6000 A: the root biomass; B: the above-ground biomass; C: the total biomass of whole plant. The control group is the seedlings incubated in the complete nutrient solution. The duration of PEG-6000 treatment is 14 days. Different lowercase letters above the columns are significantly different at the 0.05 probability level."

Fig. 9

Relative expression levels of OsNAC2d gene in the osnac2d-6, osnac2d-4-2, and wild type ZH11 incubated in the nutrient solution including 15% PEG-6000 The control group is the seedlings incubated in the complete nutrient solution. Different letters above the columns are significantly different at the 0.05 probability level."

Table 2

Agronomic traits of different osnac2d mutants and its corresponding wild type grown in paddy field"